 Let's start this session, the second day of this webinar on TF4-resistant banana varieties. Thank you very much for your participation and welcome again to another session of our TF4 Global Network webinar series, which aims to provide updated news on TF4. My name is Victor Prada and I am the secretary of the World Banana Forum, a multistakeholder platform of us simply to address the main challenges of the banana industry. Our platform is hosted in FAO headquarters accurately in the Trade and Markets division and I would like to acknowledge the support provided so far to make these webinar available by the World Banana Forum, TF4 Global Network members, TF4 Task Force, our colleagues in FAO, in the Supreme Office in Panama, because without them this wouldn't be possible. These ongoing series of dialogues on a topic that is urgent, that is complex, as multidimensional, is also an opportunity to strengthen the relations of different stakeholders with different methodologies and varieties for the benefit of millions of rural families who depend on banana production to survive. So today, as I mentioned before, our webinar is on TF4 resistant varieties. Very quick, I would like to remind you the housekeeping rules. Kindly keep your microphone always muted or while you are not speaking of course. If you would like to intervene, we would like to ask you to raise your hand or write in the chat box and we will give you the floor as soon as possible. As is customary, the webinar will be recorded and this recording will be available in both the TF4 Global Network website and the FAO YouTube channel. Please use the question and answer section that you will find close to the chat box, to the chat bottom, to formulate your questions because if you formulate your questions in the chat box, it is difficult for panelists or even for us in the secretariat in FAO to address those questions and those questions get diluted in a vast amount of messages received in the chat box. So please use the question and answer section to formulate your questions for panelists. Then important for our interpreters already providing simultaneous interpretation in Spanish and English in the interpretation bottom is that you try to speak clear and slowly. If you feel that your sound quality is not sufficient, we can wait and fix those problems. If it's possible, do not use the built-in computer microphone or a conference called microphone. It will be very useful if you use a headset. Try to connect of course if it's possible with a wire instead of Wi-Fi and then I think that's it. It would be important if you can use your video, you can switch on your camera while you are speaking because that makes the work of the interpreters easier and of course without a mask if the distancing rules in your place allow you to do it. So that being said, I would like to give the floor of the screen to Pascal Liu, Senior Economist as you know in the Trade and Markets Division and Tim Lida of the Responsible Global Value Chain Team. Pascal, you have the screen. Thank you. Thank you very much Victor and good morning, good afternoon, good evening ladies and gentlemen. On behalf of FAO, welcome to the second day of the webinar on TR4-Resistant Banana Varities, organized by the World Banana Forum and FAO Subregional Office for Central America. I will quickly repeat the background information that I presented yesterday because not all participants today were already present in the session of yesterday. So just a little bit of background for the newcomers today. The World Banana Forum is a multi-stakeholder platform created in 2009 and it includes all stakeholder groups of the global banana sector from producer organizations, exporters, importers, supermarket chains, governments, research institutes, development agencies, unions of workers, consumer associations, environmental associations and other civil society organizations. The forum operates through three working groups, each of one corresponding to one of the three dimensions of sustainability, economic, social and environmental, and it also operates through task forces. Now the Fuselium-Wild TR4 is a great concern for the WBF and its members. It's been spreading from Asia and the Pacific to other regions, going westward, and then reached the Americas in 2019 with the first reported case in Colombia and was also reported in Peru last April. As a result, from the spread over decades of the disease westward, the steering committee of the World Banana Forum created in 2013 the Task Force on TR4, which gathers experts from research institutes, national plant protection organizations, companies, development agencies, producer organizations and non-governmental organizations. Moreover, in 2020, two years ago, the World Banana Forum and AFO launched the global network on TR4, which plays the role of a global hub for dissemination of knowledge and good practices on TR4 and how to combat it. Also, it plays the role of early warning system for producers and exporters. It also disseminates extension materials and training materials. It also plays a role in coordinating global efforts to ensure that these efforts are made in synergy and to reduce the application of efforts. The global network on TR4 has hundreds of participants and it reaches out to over 2,500 people involved in the sector. It can count on the experts from FAO's Plant Protection Division and also experts from the Regional and Subregional Office of FAO and the WF Secretariat and those also the experts of the Task Force on TR4. So why is the TR4 so challenging for the World Banana Industry? The fact is that once it is in the soil of a banana farm, TR4 is almost impossible to eliminate and it can stay there in the soil for decades. No conventional management method is able to get rid of it and also the spores of TR4 spread very easily. For example, they can be spread by floodings, by wind, by the movement of people, animals or materials and equipment. So it requires heavy and costly biosafety measures to be contained and to protect the farms and this is already very costly for medium and large scale farms, but for small producers it's even more of a challenge due to their lack of resources. So developing varieties of bananas that are resistant to TR4 or at least tolerant of TR4 looks like one of the most sustainable approach to addressing this challenge. So today we will hear about work streams in this area. We already heard yesterday about a few examples of development of varieties. So today we will hear more examples and Victor will introduce the agenda for today. So Victor, the floor is back to you. Thank you. Thank you very much Pascal. Also thanks to all of you for your participation yesterday. Now as you can see in your screen, the agenda for the second day. As we did yesterday, we divided the session today in three blocks, in three sections. The first one is on conventional breeding and resistant varieties to TR4 provided by CATAS. The second one will be on gene editing and TR4 resistant in Cavendish provided by James Dale and Dina Tripati and also by Eli Kajat. And the third block will be provided by Miguel Dita on obstacles and opportunities for banana cultivar diversification. So then as his customary and really appreciated Raisa Jauger from our office in Panama in Mesoamerica will provide the closing remarks. So I'd like to remind you that this webinar is the first one on varieties out of a series of webinars on TR4 organized by the TR4 Global Network. Then it's important for you to know that we try today to have as many relevant specialists as possible, taking into account the length that a webinar can have. But of course, this is not totally comprehensive. There are many other panelists and specialists or entities working on TR4 varieties and they could also be here. So this is the first one of a series that we would like to replicate in the near future. So these entities would be, for example, the International Atomic Energy Agency in the United Nations or people in China, in Wendong, people from Emberapa, which received some information from India. Therefore, we really appreciate if you have information out of entities working on new varieties and you think that that could be useful for us and the TR4 Global Network, we really appreciate if you can send us an email or communicate with us and inform on these entities or scientists and then we will include them as panelists in future locations. So very quick reminder for panelists, please stick to the time allocation agreed. If you need more time, as we will have all the webinars in the future, we will have the opportunity to continue the discussion. But it will be good if we can stick to the time allocation there for everybody, every panelist will have the chance to speak. Then I would like to start with this section on breeding TR4-resistant cultivars and applications in South China. This information will be provided by Professor Yuhana Liu and Professor Shouxin Wei that will share a presentation on this resistant germ-plus materials developed by mixed techniques conventional and mutagenesis by the Chinese Academy of Tropical Agricultural Sciences, Katas. Professor Liu is a researcher and professor from the Chinese Academy of Tropical Agricultural Sciences. Her research focuses on banana important characters, regulation regulation and genetic improvement. She is responsible for more than 15 national and provincial level scientific research projects. She has invented and registered more than 15 patents. As one of the important breeders, her work developed a new technique of radiation mutagenesis and directional breeding and developed the thongre number one, banana viarity, with high resistance to prosarion TR4. Together with her, Professor Wei is a researcher and professor from the Chinese Academy of Tropical Agricultural Sciences, Katas again, and the head of the banana germ-plus resource nursery in Hainan province. His research is mainly focused on banana germ-plus resources and cultivation technology. He formulated several agricultural industry standards in the country, received several hours and led the development of the Boa Dao and Ba Bedou viaries. Consider one of the best alternatives for prosarium TR4 in infected areas in China. So Dr. Liu, the screen is yours. I will stop sharing now because I understood that you would like to share your screen. So please press the bottom to share your screen. Okay, perfect. We can see it really well. The screen is yours. Ladies and gentlemen, nice to meet you in this webinar. My name is Ju Huan Liu. My colleague Professor Xu Xinwei is sitting side by side with me. We come from Katas. We are members of China Agriculture Research System. It's my great honor of having this opportunity to talk something about breeding TR4, resistant carnivores and advocating in South China. The outline includes five aspects. First, in the background, the threat of banana-fusarium viaries are quickly spread of TR4, lack applicable resistant carnivores, no matching cultivation techniques. In order to jointly tackle these problems, the government concentrates the national scientific research forces to fund China Agriculture Research System, Katas, headquartered at Katas. This system is under the leadership of chief of the chief scientists. 13 scientists including germplans collection and the evaluation, breeding techniques, variety improvements, and cultivation etc. are responsible for resistant carnivores breeding and the techniques development. Other land integrative testing stations which are scattered in main banana producing regions are responsible for promotion and application of resistant carnivores with corresponding cultivation techniques. The purpose is to support the sustainable and the highest development of banana industry. With the efforts of Katas, several TR4 resistant carnivores are bred and selected. The one is Zhenrui, number one, which is bred by radiation mutagenesis and then Baoduo, which is selected from the lateral mutation. Rui Ke, number two, and Nan Tianfeng are also selected from the lateral mutation. Fanzha, number one, is bred by hybridization. Next, I take Zhenrui, number one as an example to help reintroduce its breeding process and take Baoduo as an example to carefully introduce the planting techniques and application to breeding Zhenrui, number one. Nowadays, lateral mutation is the main way to breed new varieties. The essence of lateral mutation is somatic mutation. However, this method exists two short comments of time consuming and low efficiency. Radiation is another way to breed new varieties. This method also exists short comments of time consuming and low efficiency. After 12 years investigation, we developed a new technique for radiation mutagenesis and the direction of breeding, improving the efficiency of somatic mutation. First, the breeding process. The embryonic kelly and the vintages shoots are regenerated from the immature male flower of Baxi, which is a blonde cavendish. These materials were irradiated by a cobalt-60 after recovery culture. These materials were infected by a faulty alpha-crude toxin and then directed screening. Hundreds of young plants this method improved beneficial mutation rate for four percent. Two, TR4 selection of young plants in liquid culture. We designed a liquid culture device for tissue-herd culture plantarites and construct a symbiotic system for seedling and the president's force realized directed the screening of TR4 by using GFP as a marker. It is intuitively committed nearer to controllability and higher efficiency. Three, higher TR4 incidence field screening of young plants induced by radiation. After planting in the higher TR4 incidence field and screening, a total of 44 beneficial mutations have been self-cultivated. One of the thoughts is January number one. Three, characteristics. January number one showed higher resistance to TR4. Under the same conditions, the rate of disease for January number one is only four percent, while the CK reached 93.3 percent. Even so, symptoms are later onset. Later symptoms, stronger recovery ability, later obstruction of vascular. January number one showed higher fruit quality for its beautiful shape. Dark green color smoothed fruit surface with no spots. The nutrient composition is the same as Bashi. January number one has obtained the carnival authorization on June 18, 2021, granted by the Ministry of Agriculture and the Royal Affairs of the People's Republic of China for integrated planting techniques. Because resistant carnivals showed special characteristics, such as sensitive to low or higher temperature or damage. It is very important to develop planting techniques suitable for them. We develop a five-year integrated control system. This system includes sire-paceton detection, sire regulation, resistant carnivores, biophotoleather, low or less tinnates. In this system, the resistant carnivores are the core. The sire-paceton detection is the instruction. Sire regulation is the foundation. Biophotoleather and low or less tinnates are supplements when sire-paceton detection. This detection can understand the relationship between the paceton spores and disease rates. For example, if each gram of sire contains paceton spores, at least then 1,500, the disease rate will be less than 5%. And we will give the plant a suggestion that this sire is suitable for planting any carnivores. If the spot number between 2,000 and 500 to 3,000 and 500, the disease rate will be 10 to 20%. And we suggest the planters use resistant carnivores with sire regulation. If the spot number are more than 5,000, the disease rate will be more than 50%. And we suggest the plant or farmers not that this sire is not suitable for any carnivores to sire regulation. This technique includes sire disinfection, pH value regulation, and the increased organic matter. This is the acrophotoleather. The application of acrophotoleather can regulate the pH value. Sire regulation can increase microbial diversity, improve ecological environments, create a comfortable environment for banana growth. 3. Biophotoleather will isolate rather square microorganisms such as lactococcus, bacillus, striped mesis, pseudomonas, and then fermented. After fermentation with organic matters, the biophotoleather are produced. 4. No-or-less tinnage. This technique is trial-based to avoid crossing friction and root damage. 5. Application of the integrated control system. This system was successfully applicable in South China. This is in Guangdong. This is in Guangxi. This is in Yunnan. This is in Hainan. 2. Technical training. 213 training courses and onset meetings were conducted. And 14,000 100 land-to-land enterprises, planters, and farmers were trained with this system. 3. Achievements. A total of land key testing stations and 45 fixed monitoring points have been established. The rate of disease was controlled, at least 10%. Harmless and manageable control of TR4 was realized. The application area reached 30,000 hectares. Summary. Under the leadership of the government and health, five human integrated control systems were developed. Scientists trained this system to interpret these planters and farmers. Thus, therefore, shortened the distance from selection to market demand. This is the KS family. It's a big family. Professor Jiang Fei Xie is the chief scientist of KS. Professor Zhi Qiangqing, Professor Shuxing Wei, and Professor Yi Xianxie are scientists. It's me. All of the family members are participated in the breeding of junior number one and the application of a border. Thank all the family members for cooperation and contribution. Thank all the Blenheim-Fahren secretariat. Thank FAO. Thank you for your listening. Thank you very much, Dr. Liu. How do you appreciate it? Is then your presentation finished, isn't it? Did you finish? Yes. Can you hear me, Dr. Liu? Can you hear me? Yes. Yes. We have maybe considering the time, because it's very late for you, we were planning to have the question and answer section afterwards, but maybe we saw that there's a question in the chat box that might be good for you. I would like to ask Mateus, if you can at least formulate one of the questions. Mateus, can you please open the microphone? Yes, Mr. Thank you. Thank you very much, Dr. Liu. And there is many questions actually in the Q&A section. I will select one of them. There's a lot of interest regarding the spores, because you mentioned the quantity of spores, so there is one question asking regarding the amount of the proportion of plamidal spores and other spores in the soil, and other questions relevant asking about how the toxins are obtained, if you can answer very quick these two or you can merge in order to answer. Thank you. The microphone is closed. First, we caught the TR4 for their own words, bacterial, and then sheking, and it is extracted from the bacteria, liquid bacteria. Okay. Concentrated. Okay. Dr. Liu, thank you very much for your support and for your reply to this question. Maybe we can address those in the chat box, or maybe in the future by email. But we really appreciate your presentation. It was extremely insightful. And then we really appreciate your participation in this webinar. Okay, then let's continue then with the next panelist. Thank you again. Thank you. Okay. So, let me then share my screen. Now we go to Gene, Edited, and TR4, the systems in Cavendish. I guess that you will be able to see my screen now. Dr. James Dale, when I introduce you, would you like to share your screen? Can you please let me know? Yes, I will. Okay, then I now need to share mine. Wait a second. Okay. Then our distinguished professor James Dale is, as you know, because most of you know him. He's a banana, he's part of the banana research program leader in the Center for Tropical Crops and Biocomorities at the Queensland University of Technology. He received several awards, Dr. Dale holds a PhD in plant sciences by the University of Sydney, and has been involved in biotechnology research for more than 30 years, with a specific focus in development of disease resistant and genetically modified bananas among several other topics. His team has recently developed an efficient protocol to generate non-GM, gene-edited Cavendish plants, and his experimental station is conducting experiments with resistant bananas to Fusarium Tia IV for more than five years with interesting results. We are pleased to pass the floor now, to pass the screen to Dr. James Dale, that will present the advancements and his research on genetically modified and gene-edited Cavendish bananas for resistant to Tia IV. Dr. Dale, thank you very much. Please, we cannot see your screen yet. Yeah, I'm just going to it now. Fantastic. Yeah, but it's not coming up. Isn't that annoying? Otherwise, we are ready to jump in and put it for you, but this is coming. Here it is. Fantastic. Thank you. Okay. Well, thank you very much, Victor, and good morning, good afternoon, good evening to everybody around the world listening. I'm going to rattle along because I'm covering two quite large projects, one on genetically modified and the other one on gene-edited Cavendish for Tia IV resistance. So when we think about what the reality of the future of export bananas is, and I'm looking slightly longer termed here, the two major strategies are either going to be through conventional breeding, which will come up with a banana calvary, which is different to Cavendish, but obviously with disease resistance as high yielding, transportability, extra taste and texture. The other opportunity is to genetically modify or gene-edited Cavendish and other similar bananas, but developing those with multiple disease resistances. And we're obviously in that second strategy. We've been working with genetic modification for quite a long time now. We identified, or nearly 20 years ago, a gene called RGA2, which is a resistance gene from a wild banana. And we transferred this gene by genetic modification by agro-bacterium transformation into Cavendish and began small-scale field trials in 2012, and they ran for three years. And essentially, what we did is we took this RGA2 gene, which is an NBSLRI gene that occurs in miserocuminata subspecies malachensis, and we transferred that into Cavendish's grand name. I should say that this gene, RGA2, occurs in virtually every banana, but in the vast majority is not expressed. So we took a number of lines to our field trial site at Darwin-Frit Farm. This is a commercial plantation area. It's been severely affected by TR4 for more than 10 years. And so we were planting on a commercial site, which had already been heavily invested, and we collected the complete history of every plant in the trial and confirmed all infections by PCR and many also by sequencing. After three years, and we have also had four controls in there, GCTCV2 and 8, Williams and a grand name Cavendish and DPM25. This is the result after three years and five cycles. You remember yesterday, most people were talking about two cycles when they reported their results. We took this one out to five cycles, and we had four lines, RGA2, RGA2, RGA2, 3, 2, 4 and 2, 5 all had high levels of resistance, one of which RGA2, 3 had very, very low and no infection. So that was pretty exciting. So one of the things we also did was look at the accumulated infection rate over that three year period. And as you'll see, the red lines, the controls, particularly 2, 1, 8, Williams, DPM25 and grand name, you'll see over those five cycles that the incidence continues to go up quite dramatically over those five cycles, whereas the transgenics either tracked right along the bottom there, that's RGA2, 3, and the other ones were less than 15% infection. So it was pretty exciting. And we published that quite a few years ago. So most people are aware of that result. However, we then took those plants to a much larger field trial and with the idea of taking the best lines through to deregulation. So this is a field trial site. This is the actual field trial, as you see in that top, it's a large field trial. Again, it's in the northern territory of Australia, Darwin Fruit Farm. So this time what we did was to mimic a commercial plantation. We wanted to really see how these plants performed under a commercial plantation situation. It was a complete randomized block design. And we took four of the RGA2 grand name lines that we identified in phase one. We collected yield cycle time, long-term histrific plant. And we're just finalizing that manuscript. Okay, so this is after three years. This is the infection rates. So the grand name control after three years was around about 58%. We now have renamed these lines QCAB 2, 3, 4, 5, etc. The best one QCAB 4 after three years had no infection. And Williams was up to about 68%. Another QCAB 3, very good, around about 6%. This is after three years. So this graph actually shows six cycles. So this is from the plant crop right through to the fifth return. And you'll see at the top there, Williams, which is a titico to control. The grand name is a cell line control. So that's exactly the same cell line we use for transformation. And the two best lines QCAB 4 and QCAB 3, tracking along the bottom, it was only in return four did we see one plant of QCAB 4 with symptoms of TR4. And then another plant in return five, so very, very low levels of infection after those six cycles. So essentially after six crops, QCAB 4 had only two harvests out of 300 that were lost to TR4 infection. But very importantly, we've been collecting this data. The bunch weight of QCAB 4 is equivalent to the grand name control. And that's again over six cycles and the grand name control obviously were those ones that were not infected. The cycle time of QCAB 4 was significantly shorter than the grand name control. So essentially, we were getting better yields or equivalent yields with QCAB 4 as we were with grand name. And I should reiterate that QCAB 4 was transformed with a single banana gene, RGA 2, which as I said previously occurs naturally virtually all domesticated bananas. So essentially, we've developed a Cavendish banana with close to immunity to TR4. And this has no impact on yield. We're now in the process of putting together all the data to deregulate that QCAB 4 in Australia and we'll be looking to expand the geographical distribution of field trials as soon as that can be organised. So that's gone away very well. And that really means that we have a safety net of a Cavendish banana which is resistant to TR4 that will yield essentially as good as the grand name that that is grown around the world at the present time. However, we've also commenced our gene editing program of Cavendish and also of other bananas. And one of the important things I think I should stress is that with genetic modification, this is the transgenics and cisgenics. Essentially, this involves the addition of new DNA into the plant through transformation that almost always includes either new or foreign DNA. And that's always been some of the concerns of some consumers and regulators. Gene editing on the other hand, and I'm going to particularly talk about non-GN gene editing. So this is really the precise targeted modification of a plant's DNA and can include deletions, substitutions and through the addition of entire genes. So for non-GN editing, this is really important, there must be no new foreign DNA added to those lines. So that's very important. So very early, we had to firstly work out whether it was possible to edit Cavendish bananas. And we did this. We use the technology, the CRISPR technology, CRISPR-Cas9 technology as our editing tool. And what we've done is we've targeted the phyto-ND saturase gene. And the reason we've done that is when you completely knock out that phyto-ND saturase gene, you end up with a banana that is completely bleached. It's an albino. So that gives you a very, very good visual marker of the success of that editing. And so we were able to do that. This was the original experiments were through stable transformation. So these were GM. But we were able to demonstrate that a high proportion of the transgenic lines contained that edited sequence. And of course, there are three copies where Cavendish is a triploid, we were getting more than 60% of what we call triallelic knockouts where all of the three alleles had been knocked out. So that's a very, very efficient way of knocking out CRISPR-Cas9. And we published that CRISPR-Cas9 works very, very well in bananas. Okay, the next stage, and this is where we're moving to now and making this a practical application, is we firstly had to develop a platform for regenerating Cavendish bananas that had been edited, but very importantly do not contain any foreign or new DNA. And that can be adapted to many different banana cultivars. So that was stage one. I'll go through the results of each of these as we go along. And then we needed to identify the Cavendish genes that we were going to target to generate resistance. Now, and this includes both up regulation and down regulation. And it can provide new traits in addition to TR4 resistance, we can look at other things as well. And then using edits that are acceptable in different countries. And that's quite a complex regulatory situation, which I won't talk about today. Okay. So firstly, developing that non-GM gene editing platform in Cavendish. So we had to firstly get a system in place, a protocol in place where we could do the edits and regenerate plants. We had to confirm that those edits had been done. And we do that by saying a sequencing, so amplifying across the target region, and then confirming by PCR, the absence of integrated DNA. And then what we do now is sequence the entire genome, and then we can then go in and be absolutely certain that there is no foreign DNA being added to those edited plants. So to develop this platform, again, we use the PDS gene, the phyto-reduce-acturase gene, because it's such a convenient visual marker. So we start primarily with embryogenic cell suspensions, even though we have looked at generating through meristems, et cetera, but primarily from embryogenic cell suspensions. From those, we can make viable protocols. We can actually make protoplasts that are very, very high efficiency now, up to 10 to 15 million protoplasts per cell of settled cell volume. So a very, very efficient way of making protoplasts directly from those embryogenic cells. And the advantage of protoplasts, of course, is they're single cells. So it's possible to go in and edit just one of those cells and regenerate from there. So we've also been able to develop a very efficient way of transfecting those protoplasts. In this example, we're using GFP. And so we can take those embryogenic cell-derived protoplasts and transfect those with a plasmid-expressing GFP. And we're getting very high levels of transfection. So the really important part, of course, is to be able to regenerate a whole plant. And so we've now gone through that. And in this instance, we're giving the example of using protoplasts, taking a protoplast single cell and then being able to demonstrate that going through various stages of multiplication to micro-cali formation, finally forming those somatic embryos. And that takes around about four months from initiation. And once we get to that stage, we can take those right through. So we now have a very efficient system of regenerating cavities plants from a single protoplast. So that's fabulous. Where we are now, and this is the example you see up there on the top left. This is an example of editing banana cells and knocking out PDS. And so you'll see little white plants there. So those are the ones that have been knocked out. Some of these haven't been knocked out because we don't get 100% editing as we go through these. And because there's no selectable marker, we get escapes coming through as well. We can then determine by PCR across the site which we targeted that we're getting those edits in place. And we can see exactly what the edits have been. But most importantly, then we go to PCR to determine whether any of the plasma DNA, because we're using plasma, any of that plasma DNA has been integrated into the banana genome. So we do that firstly by PCR. And this is the result here. This is one of the albinos where we've knocked out PDS. This is one of the ones where the placement had actually been integrated. And you'll see that that's picked up by various pairs of primers that we're using there. We then go and sequence the entire genome and look for any of the sequences derived from the placement. So you'll see here three different lines. Now here's wild type, sorry, wild type at the top there. And two different lines which contain none of that plasma DNA. Here's a stable integrated transgenic line. So that's a difference. And you'll be able to pick that up very, very quickly. So the reality now is that we've got non-GM gene added to Cavendish bananas. These ones look quite there, obviously pretty useless from a practical perspective, but they demonstrate that we can now do that at a very, very efficient rate. So the next part is to identify the Cavendish genes that we're going to use for editing. And we go through various stages from there. We have access to some fabulous resources. One of the major resources we've used is our QCAB-4 line compared to Cavendish Grand Names. So these two Cavendish Grand Names and QCAB-4 are essentially isolines. They're exactly the same genotype differing by one gene and that is the RGA2 gene. And the difference is this one, Cavendish Grand Name, is susceptible to TR4 and QCAB-4 is highly resistant. And so we've been able to go in there and say, okay, what's been up-regulated and what's been down-regulated in the comparison of those two. We've also got another set of very, very good genotypes, Malachensis 852, which is resistant and Malachensis 848, which is susceptible. And we've done the same thing. So what we've got there is we've identified a panel of 24 candidate genes. Some of these genes would be the sorts of genes that you would expect to identify. Some of them are completely novel and you would not expect or would have previously recognised as targets for TR4 resistance. So we've identified a panel of 24 candidate genes. We've now we're about just over halfway through editing Cavendish cells for those different genes. And they include genes that we're up-regulating and genes that we're down-regulating and genes that we're knocking out. And so that's been going on since 2021 and obviously will continue into 2023. And now when the process of regenerating those early ones and regeneration is coming along very well and they will be challenged in the screen house and finally going to progressing the promising lines to the field. So screen house will start to the screen house screening will start to occur at Darwin Fruit Farm mid this year, mid 2022. And the early promising lines we believe will be moving into the field in Darwin in probably late 2022. So not very far away at all. And based on our previous field trials we'll know within around about 12 to 18 months whether any of those lines compared to the controls have promising levels of resistance. But of course we'll do what we have done previously and that's we take them through at least three years and at least five years of cycles to be certain. Okay so where are we? We've developed an efficient technology to generate non-gm edited Cavendish bananas. So we've identified numerous Cavendish genes that potentially could increase to our full resistance in Cavendish. And I should say we're increasing the number of genes as well. We haven't stopped looking. And we've commenced that editing. We're actually quite a long way through that now and plants it's coming through. So importantly gene editing allows a continuous improvement and currently accepted banana cultivars. That's that's very important. So we can go and edit these these Cavendish for our for resistance. But at some later stage we can go back and edit for some other trait. But for instance once we are a handle on on the genes involved in Black Cigar Toga resistance that would be certainly what we were doing. And essentially Cavendish and Tia for resistance is just the start. And with with plant genomics and banana genomics moving at such an incredibly rapid rapid pace. This is going to provide us with the opportunity to identify many more genes and traits and that can be disease resistances, food quality, nutrition and plant architecture. So things are looking pretty pretty exciting from a banana gene editing perspective. So I'm going to I'm going to finish there. Obviously the sorts of work we do take a large team of people and on the left hand side that's the group that is currently involved in the program. And we're getting very good funding support from Fresh Del Monte, Horticulture and Innovation Australia, and also a company called the Australian Banana Research Company. So thank you very much. Excellent Dr. Del, highly appreciated. Okay I am mindful of the time so I'm going to share my screen right away. Wait a second. Yeah we go. Then it was a very interesting presentation Dr. Del. I really appreciate your participation in spite of the hour. And then here we go. Okay Mateos can you confirm if you can see my screen? Now yes. Okay perfect. So our next panelist is Dr. Elina Tripatis. She's the director of the Eastern Africa Hub and leader of the biotechnology program at the International Institute of Tropical Agriculture ETA. She holds a PhD in plant molecular biology and she has experience in the genetic improvement of important staple food drops. Dr. Tripatis leads the transgenic and gene editing research at ETA based at Biosciences for East and Central Africa. Her primary research focuses on genetic improvement of banana and plantain, cassava and jam for disease and pest resistance. Dr. Tripatis will present the CRI SPR but the CAS9 technique in the development of the aphorosystem diabetes. Dr. Tripatis you have the screen. Can you hear me? Yeah thank you very much Dr. Victor for your kind introduction. So if you stop sharing your screen that's so that I can share my screen. Yeah absolutely please go ahead please. We've allocated 15 minutes for your presentation. It will be good if we can stop in yeah 15 minutes. Yeah I'll finish in 15 minutes yeah. Thank you I'll appreciate it. I apologize for this and for your information we'll have to cut a little bit the Q&A sessions right after because we are a bit late with the agenda but I think we'll manage to solve that later. Apologies Dr. Tripatis the screen is yours. Thank you. Yeah thank you very much and hi everyone. So I'm going to talk about the CRISPR-Cas9 technique in the development of the aphorosystem bananas and James has already touched on it so you know it's my task is a little bit easier now. Okay so as he explained genome editing is the technique used to precisely and efficiently making specific changes to the DNA of a cell or organism. So it's like the cell the endogenous DNA of the cell itself is not the foreign DNA and what happens in this technique is that there are several several enzymes which can like make a cut at the DNA at the specific site and is a double stranded cut and after that the repair happens using the the cells machine rate cells and the repair can be the non-homologous and joining repairing which can create some some mutation or it can be the homologue directed repair and and then that's where actually the insertion can happen. So genome editing can be used to either to add to remove or to alter the DNA in the in the genome. This is basically not new nature has been editing the genome for a very long time creating variation but in 20th century actually mutations were accelerated using the chemicals and the radiations and later on the technologies were developed for like more precise targeted mutations and then so like as you can see like you know starting from 1895 when that's when the radiation discovery was happened and in 1927 there was the first plant mutated using the radiations. So but then like later on the CRISPR has rapidly became the most popular genome editing approach for several reasons and then in in 2020 actually the CRISPR cast technology won the Nobel Nobel Prize and this is still the technology is still very much evolving and that's when the new breeding pools are are coming up. So when we talk about the gene editing actually not all type of gene editing are considered as the non-GMO. So that's why it's very very very important to understand like what type of gene editing and it needs to be distinguished in terms of like how it's going so so that to avoid the regulatory because the regulatory approaches are different for different types. So as I explained that these these side directed nucleases they make the cut and then the repair can be the non-homologous and joining or the HR type. So if it is a non-homologous and joining then there is actually no donor template is basically the cut and then the repair happens and during the repair there are the mutations. So these are the targeted but random mutation and this can lead to the gene silencing, gene knockout or a change in the the function of of the gene and it's called SDN1. But then in the the homologous directed repairing the HR type it can be like a small change where there is a small repair happens you have to provide the donor donor DNA and so it allows the introduction of the mutation at the target site but is a small mutation and it's called SDN2 type and then there is SDN3 type where there is a donor template is like a complete gene insertion or or the or the replacement and it allows actually the introduction you can actually insert the full gene there and this gene can be actually from the same species so it can be like cisgenus or transgenic it can be foreign gene as well. So SDN1 and SDN2 types are very similar to the mutation obtained through chemical mutagenesis or irradiation or spontaneous natural mutation and in many of the countries like these are the these are the type of the gene editing which are not regulated as GMOs. So there are a lot of benefits of gene editing as I said like you know these are the two ladies who got the Nobel Prize in 2020 and so there are a lot of technologies for gene editing is not only the CRISPR but then the CRISPR is now actually the most common if you see like almost like 68% of the products which are in the pipeline they are using the CRISPR technology and you can use this technology starting from the functional genomics for the like developing the biotech stress resistance, abiotic stress resistance, herbicide tolerance, also enhancing the nutrition value and then the yield improvement. So this is not only in theory this is now in the reality. So non-browning mushroom it was approved in US and Canada then there is high olexuabine oil which is the first gene edited product which came on the US market. In the gene edited like blight resistant rice was also approved by USDA and Colombian regulators as non-GMO and then recently last year Japan approved the first gene edited tomato rich in agaba which is actually which can fight the high blood pressure and this is actually now ready for commercialization. So in the in banana at IATA for the crop improvement actually we use all different types of tools. So starting from the conventional breeding so we have a very strong conventional breeding program and in the conventional breeding actually it can take a little bit longer but it's a low cost and there is no regulatory issues involved if you are using the conventional breeding but you know the crossing the two parents you need a parent which has the resistant like if you're talking about the disease resistant a resistance gene but if that trait is not present in the germplasm actually you cannot use the conventional breeding then you can you have to look for the alternate procedures and that can be the genetic modification and in the genetic modification actually you introduce the foreign gene it can be from from plant species from bacteria from viruses from wherever so so this is like in terms of the time it it takes a little bit lesser than than the conventional breeding so it's a medium time is a high cost and mainly the high cost is because of the biosafety regulation and it requires and and then you know each country has its own biosafety regulatory guideline and you have to follow that so that's the biggest hurdle with this technique and then there is now this gene editing in gene editing actually you can precisely manipulate the gene the genome of the banana itself so suppose if you want to develop the the resistant varieties then you know you can work on the genes which are responsible for making those varieties susceptible and is comparatively shorter in time is less costly in comparison to genetic modification and there are if if you are going through SDN1 type there is no regulatory issue similar to GMO you know that all the products are regulated but they are not regular these so SDN1 type are not regulated as GMOs and then but then the regulations depend upon the country policies not not all countries has actually the clear guideline in place so depending upon the urgency depending upon what is available we can choose like from which of these three technologies we want to apply we are not saying if we are doing gene editing we are leaving the the genetic modification or conventional breeding out so we are actually using all the three together so at IIT our our genetic improvement program of banana and now I will only focus on the disease resistance actually it starts from the selection of the resistant varieties so if we can select so this is the short term then we have the conventional breeding program we have the genetic modification program so we are working on banana zenithomonas world for using genetic engineering the nematode resistance and also the control of the banana bunti top virus for gene editing we are focusing on banana streak virus which we have now the proof of concept we also have developed recently the proof of concept for banana zenithomonas world resistance and using gene editing and recently we started working on fusarium world resistance and the black sticker toka resistance using gene editing so at IIT we started this program for genome editing in 2015 so this was the first time approval in Kenya from the national biosafety authority for gene editing of banana we were trying to actually develop the varieties for the virus resistance which is the banana streak virus actually we are trying to inactivate the endogenous integrated virus for the streak virus so in 2018 actually we developed the proof of concept for knocking out of the endogenous banana streak virus we demonstrated that in plantain but in order to actually go for any trait as James explained you have to have a tool or the system in place so we also developed the system initially for plantain plantain has a genome and the b genome it is also triploid so we developed the system using the cell suspension so we were delivering the guide RNA into the cell suspension and so we use again the same marker which James talked about 520 saturates because this is a very good visual marker you can get if you knock out the gene make it non-functional and you get the complete albino plants if all the all the alleles are knocked out so and that's what we have developed the system but later on actually we expanded this and we have we have actually validated the system for all various genomic group of banana so like you know triple A's the cavendish cross-michael matukya varieties like you know different type of varieties then in 2020 actually we started working for editing for the bacterial disease and and the fusarium wilt disease so now we have proof of concept for the bacterial disease and we are still working for for fusarium wilt and our plan is that by the end of this year we should be able to start the field trial at least with the bacterial disease in in Kenya so our approach is very much similar to what James is doing so you know we are trying to identify the targets mainly in in the based on the information from the originators of bananas like the wild type wild type bananas and then we transfer that information into the susceptible cultivated varieties so either we knock out the susceptibility genes once we identify the susceptibility genes we knock out we are actually testing tons of susceptibility genes we are also trying to knock out individually or also sometimes are knocking out multiple susceptible genes together and then the second approach is like we are also trying to overexpress the defense genes like if you we know the the R genes but you know maybe the R genes are present but they are not expressed in enough quantity to develop to develop the resistance so we are overexpressing those defense genes into the susceptible varieties we are working very closely with the our breeding program so IATA has this project on accelerated breeding of better bananas so we and there are as IAT is working with a lot of partners on that project and so that information for the TR4 which is because that project was working on TR4 for for a few years so the information which is available from that project is actually is now fed in into my program and we are actually validating the disease resistance genes which are identified under those those program and and and then the second approach as I explained we are also developing the the TR4 resistance this is like by knocking down and the candidate susceptibility or susceptible genes using CRISPR-Cas and then this is the product actually which will be will be the non-GMO because we are trying to deliver that in a DNA-free fashion then the up regulation we are also trying to do the up regulation of some of these R genes but also some of the antimicrobial protein there we are we are either doing it with single or multiple and then we are using the CRISPR activation tool where we modulate the transcriptional activation domain and we have some good result there by showing we can actually enhance the expression from two to tenfold but we haven't started the glass houses screening yet so we have only reached up to the level of the enhancing expression but then these products if the product will come out from here this product will be actually similar to the GM and the important thing what we are doing is like we are trying to integrate the gene additive CRISPR-Cas based gene editing into the into our breeding program so we are actually working very closely with the breeders improving the parents so that you know the gene editing can become the integral part of the of the breeding and in that way you know the okay so the whatever is even though if the foreign gene are getting integrated we can segregate out later on during the in the program and then another aim here is that you know we are also trying to do the stacking of the multiple traits you know so the TR4 but then in addition to TR4 it can be for the nutrition enhancement for the resistance to the other diseases as well like BXW because Cavendish is also very susceptible to the xanthomonas well disease so when we when we talk about the the gene additive product basically there there is a biosafety concern there so one is the unwanted genetic changes in the plants due to the off-target mutations so off-target mutations can be minimized because by if we start from the improving the guide RNA at the starting and also the delivery method if you use the DNA free approaches you can minimize but then in the end when you have the product you can do the whole genome sequencing to rule out any any off-target mutation there the second the second concern is the transient integration so to avoid that we use the DNA free genome editing and in the DNA free actually we are using the similar approach as as James was explaining using the protoplast and then also we are trying to develop some other other approaches apart from the CRISPR cast to develop the DNA free genome edited approaches or you can also if there is we are using the parents so you can back cross and then segregate the transient out and just just retain with the with the mutation so that's the another approach we can use it so just briefly giving you some overview of the legislation for genome editing so like the first time in 2015 Argentina developed the first regulation in the world for the gene edited product and like if the gene edited crops if there is no foreign gene integration in them they are not regulated as GMOs so if you look at this map all the countries which are highlighted in green these are the countries where the gene edited crops with no foreign gene integration are not regulated as GMOs in Africa Nigeria has approved the guideline last year and so they are the first African country where the they have clear guideline for the gene edited product but then there are several other countries which are which are actually developing the guidelines and Kenya is actually quite advanced and only the the countries which are in red these are the countries where the genome edited crops are actually treated similar to the to the GMOs so this is my final slide key take home messages is that the application of gene editing tool has potential to develop the TR4 resistant banana and CRISPR-Cas has rapidly become the most popular gene edited approach due to its simplicity is efficiency is a specificity and easy to adopt the gene edited crops with no foreign gene integration are not regulated as GMOs in several countries and at IIT we have shown the application of CRISPR-Cas based gene edited for developing the disease resistant bananas particularly for the virus and the bacterial resistance we have already the proof of concept so in the end I would like to acknowledge the my team the bioinformatics team the breeding group and also the partners like from the University of Queensland, Bayer, Wagon and University and the financial support from USCID CIGIR research program and Bayer thank you very much. Dr. Tripati thank you very much for the excellent presentation I'd like now to pass the floor to Dr. Eni Kalyat I wonder if you would like to share your screen Dr. should I do it for you? No I can share my screen no problem as you consider appropriate okay then I will provide a brief introduction for you why you share your screen Dr. Eni Kalyat is the scientific director of Raham Meristem and retired professor of the Hebrew University of Jerusalem he holds a PhD in Biokinistry and has more than 30 years of experience with plant research Dr. Kalyat holds several patents on in two biographies and technologies and has developed a method for development of T4 resistant banana biographies based on in vitro mutagenesis Raham Meristem is an agro biotech company specialized in propagation and reading of bananas and plantings by tissue cultures Dr. Kalyat will present the role of in vitro mutagenesis in T4 resistant in Cavendish bananas. Dr. Kalyat thank you very much for your participation you have the screen thank you very much thank you very much Victor and thank you very much to the organizers for inviting me to present our work so I will start by saying that the background of our work was on a variety called GAL we are using also now a second variety that will be shown in this presentation as well and GAL is a very well known variety for many banana growers throughout the world anywhere from Australia to Costa Rica and the the Latin America everywhere it's really our best variety and our clients are very happy with this variety so this was used as a background okay one second yeah so I would like to start my presentation with showing the different levels of validation that we did to prove that the varieties that we are producing are really resistant varieties so the levels of validation start with a molecular proof of induced mutagenesis so we show molecularly that the induction was performed the next step was to show in an ex vitro state where we took the plants to a greenhouse inoculation experiment at the University of Wageningen at the group of Professor Kema and Dr. Fernando Garcia was the person that to perform this screening experiment the next level of validation was a comparison between the wild type and the mutated variety and that was done on a whole genome sequencing and I will show some of this evidence finally we did a field trial in three places in the Philippines in very highly contaminated areas that the areas that previous to this experiment really all the plants died from fusarium wilt the final step is in preparation we are taking the plants or the clones that we have selected to Laguajira in Colombia and we have the approval and we'd like to send our appreciation to Ica and Agro-Savya that allowed us to perform this experiment in Laguajira is the area that was detected that's contaminated in Colombia so the plants are already in Colombia they are being propagated now and they are tested by Ica for other diseases possibly viruses and so on and so in the second quarter of 2022 this year they will be planted in the field but as I said they're already being tested in the Philippines for more than 100 weeks of testing and I will show the results so being a commercial company we also were very careful to show that all the yield components are not affected by the mutations and also the fruit quality is not it does not have a downside to it so all of the parameters of yield and fruit quality were measured in all of the experiments and I will show how the experiment was designed so we all know the very common mutations in Cavendish starting from the dwarf mutation over here and this is a little bit more rare but a lot of times we see pigmentation on the leaves we see this mutant which is a I call it the narrow long and narrow leaf compared to the to the wild type you see the Masada mutation which is really a very strange mutant because the number of chromosomes is 34 instead of 33 and also you can see that the arrangement of the stomach is different it's really not like the wild type which is in straight lines like all the monocled plants but here it's really dispersed all over the leaf so taking that into account we started with the our elite clone which is the gal clone and we performed an in vitro mutagenesis which I will show how we did it and we did we did it in the tissue culture and we used the many many generations of tissue culture and we also used TDZ which is a strong cytokine in order to activate cell division very very quickly and we did it on a very small on very small Mary stems in order to activate the mutations in the in the in the very highest part of the Mary stem if you want the stem cells of the Mary stem okay so we in the final step of the tissue culture we divided the population into three parts this is each cluster that was developed in the last step of the tissue culture was divided into three parts and we call it siblings so one set of siblings was sent for the greenhouse inoculation and selection in Bachmingen the second set and the third set were sent to two different parts of two different fields in order to perform at the same time that we are finishing up the work we looked at different phenotypes especially agro agronomical phenotypes like the components of yield and the the quality of the fruit finally we will see a little bit what we did with the whole genome analysis and here we compared the wild type to the mutants okay so the way we perform the mutagenesis is by activation of retro transposable elements these are elements that exist in the genome in very large parts especially in bananas bananas are known to have high levels of retro transposable elements and we activated the transcription of this retro trans transposable element and the amplification of these elements in the genome by simply demethylation of DNA and I will show the compound that we used in vitro to demethylate the DNA or in essence what we did is we inhibited the enzyme the metal transferase that is responsible for methylation in mitosis during cell division so these are three different types of transposons but the this is class two transposons that we did not bother to use and class electron transposons that we also did not bother to use so we used the class one retro transposable elements that undergo transcription translation reintegration into the genome and by reintegrating into the genome they cause mutations so this is a sudden block using this compound AZA which is a compound that does exactly what I described it inhibits the enzyme that is responsible for methylation of DNA so using this compound in the tissue culture you can see before using the compound this is the sudden block and we used as a probe a small part from a retro element so you can see that the retro elements exist in the genome but you can see that after the induction with the demethylation of the sorry of demethylation of the meristems you can see that new bands appeared in the genome and these new bands were analyzed by us and these new bands are mutations that appear in the in the chromosomes either in existing parts of the part of the chromosomes where had in the past small amounts of retro transposable elements or new areas in the chromosomes where landing pads are now newly being created in the in the chromosomes and this is the structure of a retro transposable element which is very similar to retro viruses that exist in many species of plants and animals okay so in essence what we did is we used our technology to create a library of mutants in tissue culture and we created this library which we can show that each individual plant is somewhat different from his siblings by having these retro transposable elements being amplified either at the different levels or in different positions in the chromosomes so this is clear and very easy to see because we get different patterns when we use our DNA hybrid so when we use the southern blocks we can see different patterns and we choose the pattern that is best for us to conform the resistance which was pre-studied so here is an experiment that was done by Fernando which I mentioned before at the Wageningen University to demonstrate that the screening method is really very reliable and very effective so you can see that this is one of our Cavendish developed cultivars it's called Adi it's a dwarf Cavendish it's a dwarf cultivar and you can see that if you infected with TR4 you can see the infection very clearly same with Granane and other Cavendish and the same with Gross Michel which also is not tolerant it's very susceptible to TR4 now if we use if we inoculate the plants or Fernando inoculated the plants with race one that you can see here that the Cavendish varieties were white they did not get infected while the Gross Michel which is susceptible to gross to TR4 is infected so that this gave us the direction that we really were looking for that really this system that was developed at the University of Wageningen can be used very effectively in our experimental system and we let them do perform the selection so in the first batch of selection we gave on our gull material we saw that from approximately a little bit less than 10 000 plants we saw about the 8.4 percent of the plants which is 415 plants where what we call or what Fernando would call asymptomatic in other words they did not show the symptoms that I showed in the previous slide so here is more you can see the the entire plants in the in the greenhouse and you can see that most of the plants showed symptoms and the symptoms were internally and also you can see it on the leaves externally and but these plants had no symptoms and these were the ones that were selected and repropagated in a lab in in the Netherlands and then the plants were transported to the Philippines so following the the the experiments that were done in the Netherlands and also the the propagation of each one of the clones we performed a field experiment which we took from each clone we propagated approximately 120 plants and dispersed them in a field that was previously as I said before heavily contaminated with TR4 actually we did it first in two areas and now it is in three areas all the same clones giving us exactly the same result and you can see that the level of resistance is very high but I want to mention here that we really were very careful not to use plants that perhaps will have tolerance to the disease so even 99.5 percent of the plants that are resistant we did not like we liked only clones that had over now it's over a hundred weeks in the field okay so it's like more than two years or approximately two years with zero infection absolutely zero infection from each one of the clones if we had one plant that was infected the entire clone was discarded so here you can see the the wild types or the control plants that all were infected and this is the the plants that stayed in the field with no infection you can see the bunches are normal this is a normal gull we made all the measurements which you will see in my next slide all the measurements show that all of the other parameters that are important to the growers are exactly exactly like gull it's a perfect it's a perfect variety okay we also in the next step we chose we made sure to take only clones that were surrounded by controls that all died so the red dots represent plants that died from TR4 and the 8794 it's one example of a clone that survived despite the fact that the approximately two and a half meters from it there were or several plants that died completely died and were infected by the disease by the way many people came to the field trials and saw our field trials from everywhere in the world many of the banana growers in from Central America and the South America came to see these field trials in their own eyes okay so looking at the parameters of the other parameters of compared to to the to the gull so it is really exactly like the gull there is no difference in any of the parameters and here i'm not going to go into details we have we don't have enough time but it is exactly like a gull there is no difference whatsoever no significant difference anyways so looking at the whole genome sequence very interestingly we found one locus where there was a new landing pad that is exactly in the center of a cluster of our genes so our explanation is and of course we did not prove it as yet but our explanation is that with so many with so many sequences of this transposable element with so many fragments of these elements really we changed the conformation of the of the of the gene of the chromatin that position and by changing the conformation of the chromatin we allowed transcription factors or all kinds of other elements to transcribe genes that perhaps in the past were silenced so this is one example we have other examples and actually we're doing a lot of bioinformatics now on the comparison of the resistant and the wild type yeah i'm sorry to interrupt you we use already 20 minutes and i see you still have four slides i would really appreciate for the shake of you know allowing the other panelists to to also provide the presentations if you can perhaps i will need another two minutes okay perfect thank you very much okay so this is case two on a variety that is called Valerie doing exactly the same experiment this is a Valerie that using this technology we made it shorter because Valerie is a very tall mutant i mean Cavendish and here again at the University of Wachningen this time we put two plants in a pot and we only chose pot that one plant died and one plant stayed alive and was not infected was completely or asymptomatic as we call it and this will go on all of these plants will go again as i said to La Guajira both from both experiments we have the authorization from IKA and agrosavia and the government of Colombia to use this field in order to validate our results i just want to thank my group and also our people in our laboratory both laboratories in Colombia and in the Philippines thank you very much thank you very interesting information provided i'd like now to pass the screen to Mateus Lima to ask two questions one per panelist we we are a bit late we will see we have some time available after the last panelist so Mateus please you have the screen thank you Victor and thank you for our panelist for the very interesting presentations we select one question for each panelist as mentioned by Victor first to Dr. Dale Dr. Dale there is a question asking what is the gene editing efficiency the proportion of positive transformation using CRISPR-Cas you mentioned that not all plants are transformed so there is this question asking if there is a proportion Ellie happy for you to answer my question okay efficiency what we're finding and so this is quite important is that now that we're doing practical editing for for particularly knocking out or down regulating susceptibility genes and if we get 20 regenerants remember there's no selection in process here 20 regenerants about a third of them are edited and of that third more than half of those appear to be completely DNA free so it's a it's a real it's a relatively efficient protocol thank you very much Dr. Dale the other question now we have to Dr. Ellie Kayaf is how it can be ensured that plants tend to be tested in different countries such as the one you mentioned that will be tested in Colombia are free of diseases such as viruses and other important diseases that can pose as dangers to the production of the country okay so we we are working under supervision in Colombia of course of ICA which is the phytosanitary authorities of Colombia and we are doing the same thing we will do in Ecuador so we already have the agreement of the of the country of Ecuador to so they will the plants will be quarantined and they will be tested for a bunch of top virus and the other viruses banana brach mosaic virus and the other diseases as well and as far as we know fungi do not overcome the tissue culture process that was that was proven in Australia and in Costa Rica and by us and by many groups so there will be really it's almost impossible that there will be any TR4 but the plants are all tested for that okay so they are all diagnosed for all these diseases thank you very much sorry please Victor no please thank you very much Dr. Dale go Dr. Kayaf for the excellent presentations now I'm mindful of the time so I would like to continue with the next block the next section on obstacles and opportunities for banana cultivar diversification we have Miguel I'm going to switch to Spanish maybe now for interpreters to know to change the the channel and and also for um English speakers now we'll move to Spanish so you need to switch your channel into English in order to receive the English version uh don Miguel Dita tenemos la tenemos exactamente 41 minutos quizás raisa puede proveer las conclusiones finales en cinco minutos tenemos entonces 30 35 minutos un poquito menos de lo que habíamos calculado hay hemos pensado que la presentación fuera 20 minutos digo esto porque podemos seguir con esa presentación de 20 minutos pero la sesión de preguntas y respuestas tendrá que reducirse un poquito entonces en tiene un doctorado en fitopatología por la universidad federal de visosa y tiene más de 20 años de experiencia liderando proyectos de investigación en todo el mundo su amplia experiencia radica en el desarrollo de herramientas de diagnóstico el diseño de planes de contingencia para respuestas rápidas y coordinadas a plagas y enfermedades emergentes además como la integración de enfoques positivos para la naturaleza de la salud de las plantas en balanos el señor dita presentará obstáculos y oportunidades para diversificación de de plantas y cultivar es don miguel creo que estás con nosotros si gracias victor me puede dar por favor permiso para compartir mi pantalla ahora está full bienvenido muchas gracias ok muchísimas gracias victor por la invitación también a todos los colegas que que que se han conectado a la parte del mundo que han saludado muchos saludos a todos que estén bien también gracias a los panelistas que que han hecho un brillante trabajo de realidad yo me lo he disfrutado muchísimo el desafío que tenemos un poco nosotros es intentar hacer un resumen un poco y una visión poco crítica de personas desafíos obstáculos oportunidades para la diversificación de variedades y para empezar yo creo que es importante entender cómo está hoy la cadena de de bananos en términos de exportación si estamos hablando de diversificar portafolio de variedades de banana a través de los canales de exportación tenemos que entender que esto hace parte de una cadena que es multiactores multi intereses y que es bastante compleja si y que en el día de hoy o sea es realmente orientada hacia cabines si las variedades que se ven en el mercado residentes rasguardo que a veces son del tipo cabines pues esa diversificación de esos tipos de cabines pues no tendría mayor mayor problema porque porque pero si no lo son si tendríamos problema tendría mayor problema digo porque realmente cuando cuando vemos el mundo hoy jida y va cambiando pero hace muchos años más esa sentada que cada vez dice mantiene los cinco continentes o sea que es es una desafío enorme que tendemos si no tenemos variedades que son del tipo cabines habiendo dicho esto si habiendo dicho eso yo creo que podríamos entonces entrar a discutir básicamente estos cinco puntos la parte de desarrollo de variedades que ya ha sido tocado por muchos de las cualidades que presentaron la parte de propiedad intelectual la parte de ese proceso de importación de evaluación en campo luego de esa que ya se tenga las variedades a multiplicación a larga escala y de distribución que no son siempre procesos juntos lógicamente nosotros tenemos que hacer a lo largo de de esta presentación un poco de consideraciones de que nuevas variedades se implican para los sistemas de producción y mercado y nosotros como si ya ya tenemos también un poquito que cuál puede ser nuestro granito de arena si bueno empezando inicialmente por desarrollo de cultivar yo creo que que hay cinco puntos importantes no se trata apenas de buscar esa bala de plata que no vaya a salvar como fue el cabines para los michel si hay diferentes herramientas necesidades regulaciones ya se fueron discutidos hasta ahora lo que hay en mercado vamos a discutir más en detalles son variedades que son tolerantes o parcialmente resistentes y eso hace falta un poco que se entienda mejor que es que implica si porque ahí me lleva al tercer punto que es este que está aquí que que negocio cuando se planta un variedades que no son totalmente resistentes pues cambia cambia un poco aquí ayer ya se discutió un poco el doctor Adolfo de de fía lo mencionó las oportunidades que hay para otras variedades y aquí para nosotros también es muy importante que tengamos o sea asegurar que aquellos pequeños productores sobre todo pequeños pues no se quedan atrás que tienen también acceso a estas nuevas variedades que van a ser desarrolladas que ya están desarrolladas porque están en proceso si aquí realmente tenemos que ser clar es la única alternativa que hay cuando sabemos hasta lo que fue presentado aquí comercialmente disponible es son estos estos clones o suma clones y civil 2018 que son tolerantes o parcialmente resistentes aquí hay tres preguntas que están en el interior de su pantalla no lo voy a no voy a discutir porque se han discutido en otras presentaciones pero básicamente lo que nos preocupó con poco creo que a todos es esta cuestión de otras enfermedades y otra cuestión de nuevas razas sobre todo la cuestión de el sistema producción de mono cultivo intensivo pero regresando a las variedades que son parcialmente resistentes tolerantes como se quieran llamar yo creo que la presentación a ser fueron muy claras el doctor altus fue muy claro en esto que es una resistencia que cuantitativa si que ella si en determinado momento con minores niveles pero se se enferma así el doctor altus también mencionó no son inmuni inmuni quiere decir para que el colega que pregunto que que no es el mismo caso de cabines para raza uno si estamos acostumbrado a cabines para raza uno lo que estas variedades tienes si un nivel de resistencia es la única alternativa que tenemos pero pero tienen ese esa característica y aquí muy importante que yo creo que es el punto número dos que que se necesitan programas de entrenamiento fuerte para que los productores entiendan cómo se maneja esas enfermedades y ahí o sea esas esas variedades son progles que probablemente algunos que va a llegar al mercado tiene ese tipo de resistencia también cómo se manejan si que cuáles son las cosas que cambian y aquí las experiencias en mosambique y las experiencias en filipinas que se fueron presentada ayer son super super importantes tenemos que entender también que que hay un riesgo si para la diseminación de patón ya sea a cortas o a largas distancias a través también de material de siembra y eso lo voy a tocar un poquito después cuando toque nuestros temas esto se habla un poquito ayer probablemente lo que viene en términos de de diversificación de variedades un portafolio de cabines o casi cabines que está llegando pues se está trabajando fuerte en eso y básicamente tenemos esos desafíos que que que yo mencioné anteriormente sobre que están allá bajo el lit también lo mencionado como si la tocan negra etc y aquí un poco lo que está pasando con los límites máximo de residuos y las otras pladas que están saliendo con cambio climático sobre todo plagas incertes pladas de racinos que no podemos olvidarnos de esto y un tema que lo voy a tocar rápidamente que el tema de cambio climático en relación a sequía o estrés hídrico y es algo que nos está afectando mucho más y ojalá que dentro de todo este este proceso también estas cuestiones de cambio climático de de de agua sobre todo sean consideradas me gustó mucho también la observación que su doctor ador fallero sobre la diversificación de la oportunidad que hay para otras variedad si hay hay oportunidades para otras variedades y ojalá que poco a poco sabemos que hay una presión muy fuerte para para resolver el problema de carmenis pero que estas otras variedades que eventualmente también son resistentes o y como disculpen también son susceptible de raza cuato y cal sean también de alguna manera atendidas porque hay nicho para esa variedad de muchos productores diferentes países que también las usan y son súper importantes para el comercio local y también para para asegurar alimentaria que es mucho más importante sí básicamente si resumimos esa parte hay muchas variedades de banana otras a que puesto tres de los michel y orito que están esperando también una luz al final del túnel para vencer esa batalla con tras raza cuatro tópica el segundo tema que quería abordar el tema de propiedad individual es un tema un poco más delicado no es una especialidad nuestra pero aquí quería llamar la atención para estos tres puntos principales ese proceso de negociación inicial y he puesto aquí lenguaje lenguaje que no es solamente la cuestión de idioma que uno habla es lenguaje que se usa para negociación su proceso a veces no está muy claro y se necesita en terminado momento un soporte legal de sobre todo inclusive en las leyes de recursos médicos de países a veces eso no está claro en segundo punto las regulaciones que tienen las organizaciones nacionales de tradición filosanitaria las capacidades que tienen instalada en los países que son asimétricas también y luego la habilidad que se cuentan entre esos procesos para atender las necesidades o a veces deseos del de ser todo productivo básicamente como ya dicho el principio de ese acceso de estos nuevos clones nuevas variedades patente y todo de los pequeños productos es algo que necesita ser discutido de desde un inicio si esto me lleva al tercer punto supongamos que tenemos la variedad que se llama se va a importar un país y hay un ejemplo interesante y el doctor elín mencionó sobre lo que se ha avanzado en colombia pero aquí me parece que hay tres puntos cuando miramos de manera global y a veces yo me siempre un poco más en américa latino y caribe porque es donde yo estoy y aquí yo he listado cinco puntos puesto uno el proceso de cuarentena post entrada si los roles institucionales cuál es el rol por ejemplo de la organización nacional de sanitaria la ONPF cuál es el rol de la instituto de investigación si qué facilidades hay por ejemplo para aclimatar a esas plantas en los países y para cumplir con los requisitos de cuarentena número tres qué capacidades existen para ser un diagnóstico o un indexado multi multi patógenos ya se tocó un poquito el tema diferentes virus por ejemplo hay capacidades para el vtb para banana bunch of virus hay capacidades por ejemplo para el virus de la vaca esos tipos de cosas hay un poco que realizarlas y adelantándolas para cuando llegue el tiempo a cada país de llegar a esta fase número tres la otra parte que le hemos puesto que la parte de auditorías control de calidad o sea cuántas plantas se mostraría vamos a discutir rápidamente y al final cuando llega esa liberación de se cultivar para los avalaciones en campo también ese proceso a veces no está muy claro en algunos países aquí por ejemplo y es una preocupación que tenemos en la región y a la gente lo menciona es importante cuando se hace el proceso de importación y por ejemplo el banana bunch of virus yo puesto que como big big 3 virus para que la gente entiende que esto realmente es una gran amenaza por lo menos para América Latina y caribe porque pues porque todavía nosotros somos región libre la enfermedad incrementado bastante hay artículos recibes de los colegas de la ita y mostrando esto nuestra región todavía libre pero nuestra región se está en vector que es pentadomia nibromelos sólo venimos repitiendo yo creo a partir de la mitad del año pasado porque realmente un riesgo que no debemos no debemos odiar aquí es importante también yo creo que aquí es un poquito de granito arena que podemos poner nosotros que es que hay experiencia para la introducción y movimientos seguros de la plasma hay guía que se puede adaptar a estar el international transit center en bélgica que tiene muchísima experiencia más de 110 países más de 30 usuario enviando material de siempre o material de germoplasma por nuestra manera de manera segura entonces quizás ahí se podría un poco aprovechar de esa experiencia en el tc y ver cómo cómo cómo se integra todo este proceso sí el otro punto que no solamente en áfrica o sea el esculpen en en en Europa nosotros también a América Latín y caribe ahora con esta alianza biologist internacional sea en marzo se debe inaugurar el edificio se llama semillas del futuro yo creo que que tenga más interés no podemos conversar pero hay una oportunidad grande también para que dentro de esta experiencia que tiene sea que si el también tiene una mitad de fitosanidad de germoplasm así poder también aprovechar todas las experiencias todas las capacidades para que tengamos por lo menos en América Latín y caribe esas capacidades o aprovechar de la experiencia que ya existe en el otro punto voy a tomar dos preguntas desde la sesión de ayer que fue que fueron recurrentes a hoy también un poco y le agradezco a rafael y a luis por hacerlas ayer y tienen que ver que cómo reaccionan estas variedades bajo la cultura de bajos insomnos en periodos de sequías prolongados cuál es la respuesta a otras enfermedades etcétera eso me lleva al cuarto punto que es súper importante que el punto de evaluación en campo estamos hablando aquí de campo si aquí tres puntos fundamentales los ensayos multisitios en diferentes sitios si lo que se resolvió en filipinas va a funcionar en peru o lo que funcionó en peru por ejemplo en piura va a funcionar en en en por ejemplo en selva central o en colombia etcétera es importante esos sitios esos ensayos multisitios porque el condición es ambiental son diferentes vamos a ver pero sobre todo también porque hay dos escenarios aquí si lo probamos en un índice en áreas afectadas por hace cuatro tropicales y también en áreas que son libres para que para estudiar los características morpheronómicas de producción etcétera hay un punto importante que el toerí también mencionó que es la parte de tener protocolos que sean confiable de repetibles y que se pueden comparar también si y aquí el ayer fue discutir un poco también sobre si se usa inoculo artificial si es mucho si se usa sobre el núcleo natural si se usan dos y si se usó natural como garantizar que esas plantas que sobrevivir estaban realmente en contacto con una cantidad suficiente que esos otros aspectos discutir del inoculo si es importante y sobre todo en la parte de evaluación en aquellos países que la enfermedad todavía bajo está bajo control oficial inclusive en aquellos que no también pero la cuestión de bioseguridad aquí lo que quiero llamar atención en la parte de evaluación es que de este de este triángulo famoso de epidemiología de enfermedad donde tenemos que tener una planta verdad susceptible del ambiente y el patógeno nosotros en este seminario están midando apenas una esquina de ese triángulo que es la planta si tenemos que entender que cuando hablamos de esos ensayos multisitios estamos hablando de ensayos patógenos probablemente que pueden tener diferente comportamiento dependiendo de suelo del ambiente etcétera y por eso que es importante hacerlo aquí algo que también podríamos aprender lo que pasamos en el pasado si nosotros en bayores international isía que la alianza trabajaba con redes y tenemos acciones en los tres continentes en estos tres continentes con tres redes pero en el pasado también hubo un program internacional para evaluación de variedades que se pueda mejorar que se puede revisar quizás es aprender de lo que se hizo también el pasado quien sabe tener programas regionales para la evaluación y distribución después de esa variada de resistentes a tener aquí para como una idea de verdad luego hablando de evaluación para que los colegas que no conocen el año pasado publicamos esta guía con algunas elementos estamos hablando de una guía verdad no es que queramos decir que esto no tienen que hacer los países no es una guía para que personas tengan una idea de que variables pueden ser utilizados y no solamente para la fusaria también está para siga toca y algo que nos preocupa mucho vuelvo a decir lo que es la cuestión de la cuestión de de la secra o de la secuilla esto me lleva entonces ya al punto número cinco que es la multiplicación a larga escala ya compres una variada digamos o tolerante o resistentes o ya que está lista para multiplicar y estoy yendo un poquito delante porque esa etapa sabe que pasarles aquí hay tres puntos importantes que me gustaría analizar el primero certificación y certificación de material libre de enfermedades versus las capacidades nacionales vamos a discutir el segundo es poquito del control de calidad y aquí voy y después quizás a la parte de decir lo voy a explicar en una información y lo que existe desde méxico argentina por ejemplo lo conozco y esto es el grupo también en otros continentes también que es un intercambio informal de material de siempre básicamente todo esto tiene que ir integrado a los temas o los programas nacionales de producción de semilla en el promedio es limpia que muchos países sabemos que no lo tiene donde que ahora es que cuando está variando y ella tiene que entrar en el final esos canales de distribución pasan por estos procesos aquí voy a poner un ejemplo por ejemplo sabemos que el cultivo de tejido debe ser que es el material de siempre la fuente que debe ser utilizada y voy a poner después también e incentivada si y cada país tiene sus políticas sus capacidades etcétera pero a veces hemos visto que las metodologías por ejemplo para para para selección para para monestrear o para hacer aquel genote o aquella diagnóstico a larga escala para multiplicar no no están disponibles a veces tenemos situación que ya si están en los viveros y aquí me gustaría enfatizar nuevamente que hace unos minutos el material de cultivo de tejido se ha oído plantas para muchos países america tiene que ir a ese término debe ser recomendado e incentivado pero recordemos que esto no es una garantía per se de que está libre enfermedades porque depende mucho de cómo ese proceso de aclimatización ocurre y además de eso porque después que sale de los viveros también tiene otros procesos como de transporte como intermediarios como de manejo y eso no siempre está muy digamos alineado o claro en los países sabemos que la expectativa que tenemos es esta que está mi izquierda y tu planta material de siempre ha certificado libre de plagas enfermedades pero en realidad que tenemos hasta ahora por lo menos es es muy diferente es esta otra que tenemos en la derecha y aquí podemos ver un ejemplo de que cómo sucede esto en campo por diferentes por diferentes motivos y pregunto un poco aquí es que cuando eso va a resistir que o tolerante este un campo y no muestra síntomas y se sepa que hay una variante diferente en la región y se ha de decir por ejemplo si qué pasa con esos hijos y si no van a entrar entonces en ese canal de distribución informal de material de siempre al final llegamos a lo que llamó a lo que llegamos a lo que llamó de la cadena de valor de verdad se lo ponen a la parte izquierda de su pantalla saliendo nosotros estamos discutiendo un poco y esta parte aquí de sitio específico mercado etcétera y aquí un flujo que a mí siempre funciona por ejemplo los colegas que están y mutación que están directamente aquí otros colegas también están porque ya pasan por todo esto pero estamos aquí cuando esa variedad llega ya se empieza a consolidar y eso hemos visto en otros países que ya estamos diciendo una variedades pues hay muchísimas otras preguntas que hacemos a lo largo de la cadena de valores ojalá que a muchas de ellas sean respondidas para llegar a lo que en teoría soñamos que son sistemas de producción presidente en todos este proceso señores para para ser convenios para ser integración para hacer colaboración porque una variedad de fin del cabo es una variedad pero tiene que enfrentarse a todo este proceso sin hablar ya lo que mencioné en principio que es la cadena ya de mercado nosotros en la alianza viva es el que nace una silla tiene cgiar tenemos ese alcance digamos como global tenemos está en todos esos lugares donde ven esos círculos más oscuros y estamos dispuestos como siempre hemos trabajado a crear fuentes a ayudar a tener proyectos a fortalecer capacidades no solo a la técnica libre pero también facilitar ese intercambio que tenemos en otros países para desarrollar resoluciones basadas en investigación en datos y los sólidos para ayudar a que este este proceso de diversificación de variades realmente se hace un éxito y que nuestro sistema de producción llegan a ser realmente más sostenibles yo creo que como una reflexión final yo creo que todo el mundo estamos de acuerdo de que necesitamos más de una no un solo tipo de de banana yo creo que eso ya ya no sabemos pero también yo creo que tenemos que llegar a entender que el desarrollo de esa variada de esa variedad o de esas variedades es apenas uno de los desafíos que tenemos adelante es una pregunta que me gustaría dejar para para la audiencia es un poco así estamos listos para para realmente enfrentar este desafío con eso me gustaría agradecer realmente todas las instituciones que partísima con nosotros o con antes etc. lo que uno presenta aquí realmente es el fruto de muchas interacciones de muchos años de muchos esfuerzos muchas veces anónimos y me gustaría realmente agradecer a todos los que colaboran con nosotros o que no colaboran pero que están trabajando en pro de que realmente este problema sacó de tropical y los otros que también que también viene en la cadena de Guamano se han resuelto de la mejor manera posible muchísimas gracias por su atención Victor nuevamente las colegas Victor nuevamente gracias a bachman e-mail si por una cuestión de tiempo no tenemos posibilidad de poner todas las preguntas por favor siéntanse libre de contactarme por esa bien muchísimas gracias perfecto Miguel como siempre una una presentación excelente con unos contenidos excelentes voy a dar la palabra ahora al señor Mateos Lima para que haga una pregunta y abrimos la rueda de preguntas quizás durante cinco minutos adelante Mateos estimado por Miguel muchísimas gracias por la presentación muy completa y una visión muy holística del tema una pregunta que recibimos es cuál es el rol de las ONPF's en el movimiento de germoplasmas de banano con relación a lo que hablaron de riesgo de introducción de enfermedades de cuarentena y etc. Muchas gracias Miguel. Gracias Mateos, si yo creo que hay colegas de las ONPF's aquí comentados es una pregunta donde me parece que cambia un poco de país para país pero no nos olvidemos que para una plaza valentinaria por menos para América Tínica y para el control oficial quien tiene digamos la última palabra en todos los movimientos esto es la ONPF's si entonces desde la parte de bilanzas de diagnóstico está bajo hasta donde yo entiendo para el mandato de las ONPF's y hay que seguir gratuitamente las regulaciones que estas organizaciones tienen. Si, yo he quedado el falemanto de la mano, he donado el falemante, por favor. Si, buenos días. A mí me gustaría hacer un comentario sobre la propiedad de las variedades que se están produciendo tanto en Honduras como Israel, Australia y otros países. Yo veo que la mayor parte de los participantes en este foro son representantes de gobiernos y obviamente estas variedades todas van a ser patentadas y todas van a tener pago de regalías. O sea que lo más posible es que no van a estar a disposición de pequeños productores a no ser que algún gobierno o algún ente internacional decida hacerlo. Probablemente las únicas que van a estar disponibles para pequeños productores son las que haga el IITTA en África. Si es que el IITTA lo decide así. Pero en el caso nuestro, en fía nosotros no podemos y yo estoy seguro de que las empresas en Inglaterra, en Israel y en Australia todo van a querer recuperar los costos de producción de nuevas variedades. El costo nuestro hacer una variedad es de cerca de tres millones de dólares. Ese es nuestro costo básico y eso después de tener casi cincuenta años de experiencia y mucha tecnología local y bajos costos de mano de obra en Honduras. O sea que me imagino que nuestra parte va a ser más alto. O sea que siempre no crean que cuando ya hay una variedad sólo hacer pedirlo. No, no se la va a entregar. Se van a tener que patentar, hacer convenios y va a tener todo un costo. Muchas gracias. Muy bien. Domingo, no sé si qué es reaccionar a eso. Si yo tengo alguna pregunta para ti. No, yo creo que Alfonso Adolfo fue muy claro en eso. Por eso yo lo puse en toque porque hay que agarrar eso la institución, los gobiernos o las agencias que realmente van a conseguir un puesto. Tienen que estar muy muy claras en lo que mencionó Adolfo, que esto tiene un costo. Estas variedades tienen. Sí, es bueno que se vaya discutiendo en su principio para que sepan que esto no va a estar probablemente como un accionado Adolfo gratis. Eso tienen pavos reales y hay que estar preparado para eso, para no tener sorpresas al final. Yo como ves, y esto es una pregunta directamente, como ves el futuro de la financiación de nuevas variedades sobre la continuación con esta investigación, teniendo en cuenta estos temas tan importantes de la propiedad intelectual y que, bueno, pago de regadías, patentes, etc. Gracias, Victor. Es una pregunta exactamente dedicada, verdad? Yo como lo veo y nosotros pensábamos, hay que hay iniciativas que ya empezaron. Por ejemplo, Adolfo mencionó muy bien ayer, y ya se están trabajando con expresas, con sorcios privados. Yo creo que hay espacio todavía para tener con sorcios donde se ponga recursos, donde se garantice que esas variedades van a estar excesivas de pequeños productores. Entonces, ya sean asociaciones, ya sean gobiernos. Pero si se crea un consorcio, digamos, donde que no empiezan a ser, quizás, pero donde se junten las fuerzas. Pero, al final de cuentas, estamos hablando que Banano emplea millones de trabajadores, y que esas empresas que exportan y todo, al final de cuentas, Banano son activos que requieren muchísimas manas de obra. Y sí que es importante para proceder para estabilidad social en muchos países. Entonces, hay que tener una exclusión bastante seria con esto, inclusive con las personas que entienden procesos legales. Yo no soy especialista en la propiedad intelectual. Yo lo he puesto porque me parece que es algo que hay que ir discutiendo, porque poco, en breve, ojalá que más tarde, más temprano que tarde, tengamos variedades disponibles y alguien que me sentarse a negociar. Pero yo creo que hay posibilidades para tener consorcios para que estas variedades estén disponibles. Pero, lógicamente, hay que discutir caso a caso, quién está aquí dispuesto a hacerlo de esa manera. Hay que entender también, y hay una cosa que eso les pongo un poco a, por ejemplo, a Adolfo o Ali también. Por ejemplo, vamos a poner con un productor OX que sí le pagó a Fia, por ejemplo, le distribuido su material de siembra porque tiene derecho a productores pequeños que le venden ahí, ¿sí? Entonces, ¿qué pasa con eso? Por eso me refería un poquito el control de calidad. ¿Qué pasa con esos hijos? Esas plantas, una vez que tienen territorio, eso es un tipo poco complicado. Y yo creo que hay que respetar la propiedad intelectual, que respetar el trabajo, la obligación que se hizo, y hay que empezar a discutir y negociar de una manera que no se afecte realmente lo que Adolfo mencionó. Las inversiones que se han hecho en todo este proceso hay que ser realistas en todo este proceso y intentar encontrar mecanismos donde que sean ganar-ganar, win-win situations, ¿sí? Entonces, es delicado, pero hay que empezar a discutir. Perfecto, muchas gracias. Sí, la verdad es que el tema de pequeños productores y productores de media escala es un motivo de preocupación en estos asuntos. Hay otra pregunta también para el señor Dita. ¿Cómo el cambio climático puede afectar la dispersión de fusaríos en bananas? ¿Hay alguna organización en la institución que está investigando sobre el efecto y diferentes factores ambientales en la productividad de bananas? Sí, gracias. Es una pregunta excelente. Sí, nosotros hemos hecho algo, sí, hay cosas muy interesantes sobre factores de prediposición a fusaríos. Por ejemplo, ahí estamos escribiendo unos productores en Brasil que están lloviendo muchísimo, que han visto la noticia, inundación y todo esto. Pero, más allá de la dispersión, en ese momento hay una aceleración de fusaríos en muchos casos por condiciones de predisposición, por ejemplo, encharcamiento. Hay que dividir dos cosas. Una cuestión es dispersión, ¿verdad? Y, pues, a través del agua, de los canales, etc. La otra parte, de factores de predisposición. ¿Cómo esa planta está más predispuesta o menos apta para sobrevivir o para enfrentar, digamos, al fusario? Y aquí hemos visto que podría periód prolongados de seca, por después periód excesivos de lluvia, dispara de enfermedad. Y depende mucho, también, de la condición en suero, que sea un dinogluo, etc. Se necesita un mayor estudio, básicamente, y nosotros siempre decimos que estos estudios tienen que ser sitio específico. Lo que se aplica a uno es un lugar, no se aplica al otro. Por ejemplo, en Perú tenemos suelos a muchos lugares alcalinos, a muchos lugares suelos más ácidos, y es un multifactorial que hay que estudiar. Ahora, si tiene una variedad, digamos, que es totalmente inune, podría, o sea, totalmente resistente, y eso ayuda mucho más, como es el caso, que hemos tenido con Caines y Raza. Muy bien. Muchas gracias. Estoy consciente del tiempo. No sé si Raísa o Mateos tienen alguna buena pregunta más de las 40 preguntas, casi que tenemos en la sección de preguntas y respuestas que, como decimos, responderemos de forma bilateral después, y daremos, incluso, meterlas en el informe y ver si nuestros panelistas pueden responderlas. Pero si tienen alguna pregunta más, antes de pasar a las conclusiones, Mateos y Raísa. No, gracias, Víctor. Hay muchas preguntas que yo creo que la idea también es pues en el informe que hagamos y que vamos a compartir con todos poder, pidiéndole obviamente el apoyo a los panelistas dar respuesta a las preguntas que hoy están en el chat. Absolutamente. No sé si antes de que pase más, porque estoy viendo que nos quedan todavía 9 minutos, antes de que pasemos a las conclusiones finales, no sé si alguno de los panelistas presentes quería comentar con respecto a este último tema, que quizás es también muy importante, no solamente la identificación de variedades, sino cómo luego están esas variedades disponibles para los pequeños productores medianos, y viendo que Don Adolfo está presente en la llamada, no sé si Don Adolfo querría, por ejemplo, dar su opinión al respecto, cómo hace esas variedades disponibles para el público en general. Adelante. Bueno, para el público en general, no sé, pero si le digo que para los socios nuestros, lo que se ha pensado es, tenemos tres empresas que están patrocinando el programa. El programa, como les digo, cuesta aproximadamente un millón de dólares al año. Los híbridos se le han mandado ya las tres empresas, Australia, Guatemala, y aquí en Honduras, Andorra. Ahora, por lo que hemos hablado, y todavía no tenemos un reglamento bien escrito, es que cada empresa tiene derecho a producir esos híbridos en sus plantaciones, sin restricción. Ahora, si ellos quieren tener, por ejemplo, en el caso de cualquiera de los tres, quieren tener productores asociados, porque las tres empresas son grandes y las tres mercadean banan. Si ellos quieren tener productores asociados, produciendo la misma variedad, ellos le pueden dar el material genético y le van a cobrar por caja de banano producida, porque van a tener ahorros, especialmente inicialmente va a ser el gran ahorro en el control de Cigatoca. Control de Cigatoca en América Central puede costar entre mil y dos mil dólares por hectárea. Y si usted se va a ahorrar de eso el 80%, pues es una barbaridad de dinero. Y bien puede pagar 50 centavos de dolar por caja o 60 centavos de dolar por caja para sufragar los gasos que la empresa incurrió en el desarrollo del híbrido. Y también como FIA es parte de ese arreglo, pues de ese pago, también una parte vendría para FIA el que se reinvertería en el programa y continuoríamos con el mejoramiento en ético de otros materiales. Esta es una idea que se ha mencionado. Ahora, la otra es que es el asunto de las patentes. Esto hoy en día, esto habría que patentarlo en todos los países, en uno por uno, y eso es muy caro. Eso es muy caro. FIA tiene un poco de experiencia patentando variedades en Estados Unidos, en México, solo creo que esos dos países. Y en realidad lo hicimos hace varios años solo para aprender cómo se patentaban variedades de banana y plátano, porque en una época no se podía. Ahora sí se puede. Ahora, ¿qué quiere decir? Que yo tengo una patente de un banano en Estados Unidos y en Estados Unidos no se produce. Bueno, tal vez Hawaii Puerto Rico. Eso quiere decir que si usted lo produce ilegalmente en cualquier otro país y lo envían a Estados Unidos y yo lo detecto, el banano se decomisa, se multa y se destruye. Esa es la protección que me daría a la patente en los países donde lo podamos patentar en Estados Unidos y Europa en los mercados de exportación. Pero la idea es que el productor que acoja esta variedad en la que se ha producido le pague un canon a cualquiera de las empresas asociadas con ellos para resarcir los gastos que se han invertido. Muchas gracias. Como le digo, hasta ahora ni ningún gobierno, perdón, Víctor, ningún gobierno ha expresado interés en unirse a este proyecto. Todo lo que nosotros vemos es que todos siguen interesados en contención, prevención, regulaciones y demás. Pero ninguno ha dicho, yo quiero participar en el desarrollo de nuevas variedades. Aquí está mi aporte. Eso no lo hemos visto nosotros hasta ahora. Gracias. Gracias. Adelante, Don Miguel. Gracias, Víctor, gracias, Adolfo. Yo creo que nada más discusión. Hay una pregunta cortita para eso que usted acabó de mencionar, porque se prevé que paguen quizás algo por un tiempo, no para realmente pertenecer a eso, solamente, o sea, el libre, ¿no? Pero ahí, yo sé que están todavía trabajando en ese proceso, pero habría eventualmente un tiempo, por ejemplo, un pedido de la aporte, después serían libres o eso depende de las leyes de cada país, yo creo, ¿no? Eso no se ha decidido todavía, pero yo eso lo que sí veo es, pues se habla mucho de, yo hice, mencionó relativamente poco en estos días sobre la diversidad de las otras musacias, y veo que los esfuerzos que están dirigidos a mejoramiento genético en este momento, los principales son para acá en de eso. A mí me gustaría aquí, personalmente, tener un poco más de financiamiento adicional solo para Platanas, o solo para otras variedades diferentes, de tipos, tipo prata, con alto contenido de taroteno, etcétera, una lista que les mantiene ayer, y para eso no tenemos financiamiento. Los fondos que tenemos ahora son prácticamente, yo diría el 75% es para Cavendish, y el otro 25% es para Grossmischel y Cavendish, puros o convenados, eso es todo lo que tenemos, pero los demás materiales tenemos las madres listas, pero lo estamos haciendo absolutamente nada por falta de pongo. Vale, solo para completar, para aprovechar lo que tenían ahí, pero yo creo simplemente ya un poco saliendo a la parte de esa red de variedades, yo sé que es el principal objetivo de este seminario, pero yo creo que es mi deber un poquito también de decir que todavía es mucho más, es mucho, pero mucho, es muy, muy importante todavía mantener países libres de enfermedad, y continuar con programas de contención, en lo que una variedad realmente está disponible para los países. O sea, estamos hablando de variedades, yo digo para el público que está importantísimos, están discutiendo esto, yo estoy súper contento de que veo que hay avances, de verdad, y todo eso, pero todavía así del quita, una y con la otra, y no vas a tener problemas, en teoría pasaría con Grossmischel, siendo resubstituido por Kameleis, en mi experiencia lo que hemos visto todavía, pero es que no, o sea, muy importante discutir la parte de variedades, pero todavía seguir importante con cuestiones de prevención y contención y todo esto que tienen los países. Yo estoy de acuerdo un poco con Adolfo, que hay que trabajar en paralelo, hay que pensar que es un día puede llegar y ya tener, por ejemplo, adelantado, porque desarrollar variedades cuesta, pero tampoco olvidarnos quizás de la biodiversidad en la fin que la prevención, no haciendo claves, al menos para Mérida y Baty Nicarin. Absolutely. I have a say in English, I wonder if any of the panelists we have before James Day, Eli Kallat, Frederick Barkley, Althuss, do you join? I wonder, or Mr. Sorensen, I wonder if they have any input or any suggestion on this aspect of the intellectual property of new varieties. I can speak, this is Eli Kallat. Please go ahead. Yeah, so I think like every other technology, it's really, it's important to protect the rights by patent or breeder's rights. I think that we are doing, especially us that work for commercial companies, it's part of our tasks, to protect the rights of the company. And I don't see any, why should anybody be inhibited from, because the growers will benefit from it, as well as we do. And so it's only normal that we issue patents and actually on this TR4 resistance that we have developed, we have patents worldwide, probably in more than 20 countries. Anywhere where bananas are being produced or imported, we issue the protection method. And our patent is on the method itself. So it's a utility patent. It's not just a breeder's rights or a plant patent, but it's a utility patent. And it's important that the breeders and the people that spend a big part of their lives will benefit from their work, unless they get funded from international organizations or something like that. Perfect, excellent. I'd like not to give the screen to the next one, the next who raises hand, Dr. or Mr. Sorensen, please, you have the screen. Thank you very much. I just wanted to comment on the issue of varieties and how they could be available and how that could turn into a sustainable system, I would say, for bananas. I think there's a lot to learn from other sectors that are working with plants in general, and where a system which has been mainly fueled by the concept of plant breeders rights has turned into an industry that is continually delivering innovations in plants because there's a system of rewarding the breeder for his efforts by paying a price for the material, which will then continue a breeding process. And after a number of years, this system is a best guarantee for a sustainable continuous delivery of novel innovations, also for bananas. So I don't think banana industry has to reinvent the wheel. There are many plant sectors where this is working very well that we can copy once we get out of this monoculture and into a diversity of banana varieties in the market. Excellent, thank you very much. Dr. Maikri, yes, please. Thank you for giving me the talk. For any way, I think that any activities of banana breeding or impoverished activities must be found. And the difficulty is to reflect about the way we want to choose to fund these activities. And clearly, we have to be remembered by Ellie. We can't put patent on technologies. We can have plant-reader rights. But maybe regarding the particularity of this plant, which can be easily, vegetatively propagated, I'm not absolutely convinced that plant-reader rights can be recognized and established for most of the countries around the world. So I think that probably we have to think about the system of partnership with common partnership with many stakeholders from the industry, banana industry, or for research, and so on, which will finance the research and then which will have the priority to the research, giving them a commercial advantage in terms of competitivity to reach some markets. Because it will be extremely difficult to make recognizable the plant-reader rights in most parts of the world. It was just a commentary. Thank you very much. OK, I'm mindful of the time. We exceeded the time in five minutes. We have interpretation available. So for the sake of time and also your agenda, of course, I would like now to pass the screen to this is Raiza Jauger, a cultural officer in the office in Mesoamerica to provide the closing remarks. Raiza, you have the screen. Thank you. That's the word. Go ahead. Yes, Victor, thank you very much. And really, thank you very much to all the panelists and to the colleagues who have joined us in these two days of the webinar. Como bien dijiste al inicio, o sea el tema de las variedades, tendremos otros momentos para discusión y llevar a cabo esta agenda que venimos realizando con el Forum Mundial Bananero en el fortalecimiento de las capacidades de la región. Han sido dos días de webinario con temas que hoy ocupan una de las prioridades dentro de la agroindustria bananera. Se han tratado temas de gran interés con respecto a todo lo que puede ser la tecnología para la edición de genes. Y por otra parte, también todo lo que abordaron con temas para la introducción de germoplasma en las diferentes regiones. Es importante también ver hoy hacia dónde se enmarca las nuevas investigaciones en las políticas de variedades y de manera general cabe señalar la importancia de tener siempre un movimiento de germoplasma seguro, evitando, por supuesto, los riesgos de entrada e ingresos de otras probables enfermedades cuarentanarias presentes o no en diferentes regiones de a nivel global. Y la necesidad, por supuesto, de programas de entrenamiento, de capacitación para los productores con el fin de conocer cómo manejar estas nuevas variedades tolerantes o parcialmente resistentes a la raza cuatro tropical. Una vez más, se resalta la importancia de la vinculación de la agenda científica con las agendas también que llevan tanto diferentes instituciones del sector público como el sector privado para tener una mejor estrategia en el manejo de la raza cuatro tropical. Hay una gran reflexión sobre la diversificación de clones para consumos diferentes a Cabin Dish y, por supuesto, todo lo relacionado con la disposición a todos los sectores, a todo público y todo lo que tiene que enfrentar también la cadena comercial. Se resalta, una vez más, la necesidad de fortalecer la institucionalidad y los roles de los diferentes sectores y organismos tanto del sector público y privado para asegurar un movimiento de gelmoplasma con bioseguridad. Para cerrar, aunque estemos en un seminario fundamentalmente este webinario para el tema de variedades, sí queremos siempre continuar con el llamado para los países que no tienen presente la raza cuatro tropical y continuar todos los esfuerzos en cuanto a la prevención y, además, también en los que están presentes y a nivel global en la contención de la enfermedad, de manera que nos podemos preparar aún mejor para enfrentar la raza cuatro tropical y otras enfermedades cuarentenarias también que son de gran importancia para la industria del banano. Agradecemos a todos los colegas que se han unido con nosotros estos dos días de webinario. Estaremos compartiendo también a través del Forum Mundial de Banano todo un resumen de estas sesiones de webinario. Y, por supuesto, Víctor, como hemos pensado, continuaremos con todos estos esfuerzos en capacitación, a no solo para la región de América Latín y Caribe, sino a nivel global también. Gracias, Víctor. Ahora se me escucha. Muchas gracias, Raíza. Excellente cierre. Y repetir lo que ha dicho, seguimos trabajando, seguimos informando desde FAO y, sobre todo, desde la plataforma de la red global de R4T. Continuaremos, sobre todo, con este tema de variedades que creo necesitamos más ponencias y más discusión. Y eso será, seguramente, en los próximos dos, tres meses organizaremos el próximo webinario. Entonces, hago también un llamado para que, si tienen panelistas que ofrecer, que nos indiquen para poder que, de general, está agenda y tener discusiones interesantes al respecto. Muchas gracias por estar con nosotros estos dos días y me despido con un abrazo y esperando verles pronto y seguir trabajando con ustedes. Bueno, gracias. Gracias, gracias a todos.