 I'm Bouddhika. Can you hear me? Yes we can. Hi everyone. I'm Bouddhika from the Centre for Crop Health at the University of Southern Queensland. Today I'm going to share some of my PhD research which was focused on characterization of pyranophora terraces and mapping of virus genes. A net blotch in Bali is caused by pyranophora terraces and will cost you to net blotch can exceed 60 million Australian dollars. So net blotch can appear as two forms. As you can see in this figure, two figures, net form net blotch and square form net blotch. So net form net blotch is caused by pyranophora terrace form terrace and square form net blotch is caused by pyranophora terrace form maculata. So these two forms are also called PTT and PTM respectively and the hybridization between the two forms is rare. However, the recent emergence of field hybrids increased the necessity for understanding their operands and biology. To improve the knowledge of the Peter's Barlipath system, we set three objectives for our study. The first objective is to identify the genetic characterization to genetically characterize the PTT isolates from different continents and identify the main difference of Peter's and its hybrids and investigate genomic regions associated with virulence and leaf symptoms of Peter's using a hybrid population. So in order to genetically characterize PTT from different continents, we collected 250 isolates from Australia, Canada, Hungary and Republic of South Africa and also we included five historical isolates from Canada, Denmark, Japan and Sweden. So once we collected these isolates, we extracted DNA and then we sent this DNA for dark sequencing. So once we obtained our data, we performed a model-based and multivariate cluster analysis to find out the genetic structure of our population. And then we calculated analysis of molecular variants and then we calculated fixation statistics and genetic distances among these populations to identify the genetic difference among these continents. And also then we performed or we constructed a dendrogram and performed form-specific PCR to identify hybrids in our dataset. So this is the dendrogram that we obtained from our... So here you can see the dendrogram that we have obtained from our study. So from this dendrogram, we found out that there are two isolates from Japan and Hungary. They were closely related with the previously identified field hybrid WAC10721 and we suspected that these two isolates were hybrids. So we wanted to confirm whether these isolates were hybrids or not. So because of that, we performed a form-specific PCR amplification and once we performed our form-specific PCR amplification, we confirmed that these two isolates are actually hybrids. So then all of our structure analysis revealed that our population... The structure analysis revealed that the three clusters best described our population. As you can see in this... our principal component analysis figure and out of these three clusters, one cluster contained isolates with at-mix origin. Also we found out that there is a low genetic differentiation among other populations and there are reserve ongoing gene migration. So peteris is a heterotonic fungus. So because of that, it needs to ally from opposite mating types are required for sexual recombination. So these two types are called mating type one and mating type two. So in order to identify the mating preference of pyronephora pterus, we established 15 process and 12 control process. So in this presentation, I'm just going to talk about the mating preference of only PTT and PTM. However, the full research article is available online under this title. So if you are interested in this one, you're more than welcome to have a look. So this table shows examples of process that we perform where each isolate was given the opportunity to mate with the same and the opposite form at the same time. For example, isolate HRS091227, which is a mating type two PTT isolate, was given the opportunity to mate with either a mating type one PTT isolate or a mating type one PTM isolates. And so here you can see the bottom figure shows how we established our crosses in petri dishes with sex, medium and barley straw. So from this study, we found out that some PTT crosses produce escus pores at two time points with the period of no escus pore production, as you can see in the first two crosses. And these PTT isolates mated with the same form during the early escus pore production period and later mated with the opposite form to produce hybrids. So as you can see in the first cross, first two crosses and the PTT isolates did not mate with the isolates of the same form but hybridized with PTT isolates. In conclusion, we found out that PTT isolates seems to have a greater reproduction bigger than PTM. So in order to identify the genomic regions of peterous, peterous genomic regions of peterous associated with virulence, we genotyped 350 unique isolates using that sequencing and we phenotyped these 351 isolates with eight barley genotypes and then we constructed our genetic map and then we found out QTL and then we identified candidate genes underlying this QTL. So from this study, we could found out four QTL for live symptoms. So they are in red color in our map and then four, five QTL for virulence, they are in blue color in the genetic map and then we could found out 68 candidate genes for this nine QTL. So once we phenotyped our population, we found out that some of the isolates were highly virulent across the eight barley genotypes that we phenotyped at the beginning and then we selected these eight isolates which are having high virulence on the tested barley genotypes and then we phenotyped these isolates with another 12 barley genotypes. So these 12 barley genotypes like most of them were commercially available and recommended for net blotch disease, net blotch disease and out of from this phenotypic assessment, we found out that three isolates, as you can see in the bottom figure, they had they were highly virulent across the 20 barley genotypes that we tested with disease reaction score about six as you can see in this figure. So in conclusion, from this study we found evidences for recent ongoing gene migration among Australia, Hungary and Republic of South Africa. We also could found two field hybrids from Hungary and Japan and this Japanese isolate was collected in 1931 and so this isolate was remained undetected as a hybrid for nearly 100 years and the process that we made to identify the mating preference of PTT and PTM revealed that PTT isolates seems to have a great pre-production vigor than PTM isolates and this tells us that the sexual recombinant sexual barriers between PTT and PTM can be overcome under suitable conditions. This is the first QTL mapping study to identify QTL associated with the leaf symptom of peterous. So therefore, we believe that these results form a solid foundation for future studies. So I would like to acknowledge my wonderful supervisors, Anka, Niluva and Lissell, my friends at CCH and Hermitage Research Facility and thank you very much.