All BGRI Abstracts

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Survey of wheat stem rust Puccinia graminis f. sp. tritici in Jordan

BGRI 2018 Poster Abstract
Kholoud Alananbeh The University of Jordan
Ayed Al Abdallat, Monther Tahat

Studies on whet stem rust (WSR) in Jordan are considered to be old. There was only one study conducted in the late 1980's by Abu-Blan and Duwayri (1989) to evaluate the infection of wheat cultivars with black stem rust disease (Puccinia graminis f. sp. tritici). Recently, reports of stem rust were published in Israel and Lebanon in 2010 and first report of Ug99 was reported in Egypt in 2014. The objectives of our research are to: (i) survey wheat growing areas for WSR in Jordan during the years 2017-2020, (ii) identification of WSR races isolated from Jordan morphologically and molecularly, (iii) analyze rust populations in terms of their response to known differential sets, pathotype distribution and diversity, (iv) screening the response of Jordanian wheat germplasm to the identified WSR strains, and (vi) study the population diversity of WSR races using RT-PCR and SNP genotyping. In 2017 a total of 270 fields of wheat and barley in the wheat and barley growing areas in Jordan were surveyed from March-May. The survey covered northern, middle, and southern parts of Jordan (arid and semi-arid regions). Altitude, longitude, and latitude data was recorded. Only few WSR pustules (n=4) were collected because the environmental conditions were not suitable for the disease to develop. On the other hand, wheat stripe rust was very common in the wheat growing areas mainly at the southern parts of the country. Other fungal plant pathogens were also reported including smuts, spots, blotches, powdery mildew, crown rot, fusarium head blight, and flag smut.

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Molecular screening of stem rust resistance genes Sr11, Sr26 and Sr31 in wheat genotypes of Azerbaijan

BGRI 2018 Poster Abstract
Samira Rustamova Institute of Molecular Biology and Biotechnology, Azerbaijan National Academy of Sciences
Shahriyar Sadigov, Alamdar Mammadov, Irada Huseynova

Rust of cereals are considered to be an important disease in many countries, including Azerbaijan. One of these is stem rust caused by Puccinia graminis f. sp. tritici (Pgt). Extensive research on the identification of wheat stem rust resistance genes and effectiveness of these genes in various geographical regions have been conducted. Genetic resistance is one of the most effective ways for controlling stem rust. Sixty-nine stem rust resistance genes (including 45 identified Sr genes and 24 genes with temporary designations) are registered in the Komugi Wheat Genetics Resource Database. It is important to use proper combinations of resistance genes in developing lasting resistance wheat. The main purpose of the study was to identify lines caring Sr11, Sr26 and Sr31 genes, which are effective to the predominant Pgt races in Azerbaijan. Durum and bread wheat genotypes differing in their disease resistance, productivity and other physiological traits were chosen from the wheat gene bank of the Research Institute of Crop Husbandry (Baku, Azerbaijan) for analysis. DNA extraction was carried out according to standard CTAB protocol. RT-PCR performed using KASP markers (KASP_6BL_BS0074288_51 and KASp_6BL_Tdurum contig55744_822) identified nine durum genotypes (out of 34 genotypes) and seven wheat genotypes (out of ten genotypes), caring Sr11. Using the dominant STS marker (Sr26#43) a diagnostic 207 bp amplicon for Sr26 gene, was observed in 11 of the 42 wheat genotypes tested. In eight of the 42 wheat genotypes tested, the diagnostic 1,110 bp amplicon was observed using the Lr26-Sr31-Yr9 locus specific marker iag95, characteristic of Sr31 gene located at 1BL.1RS translocation. For the first time, wheat germplasm in Azerbaijan was analyzed using KASP genotyping technology and genetic resources, and resulted in the identification of wheat lines with effective resistant to Puccinia graminis f. sp. tritici race TKTTF.

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Histopathological characterization of R-gene mediated resistance to stripe rust in wheat

BGRI 2018 Poster Abstract
Kamran Saleem Department of Agroecology, Aarhus University, Denmark
Chris Khadgi,S?rensen, Annemarie Fejer, Justesen, Mogens St?vring, Hovm?ller, , , , , , , , , , , , , , , , , , , , , , , ,

Wheat yellow (stripe) rust is a recurrent problem throughout the world, and resistant varieties are an efficient means of managing the disease. Therefore, characterization of diverse sources of resistance is of prime importance for wheat breeding. The objective of the study was to investigate variation in host response in incompatible interactions conferred by different R-genes. Epifluorescence and confocal microscopic methods were utilized for histopathological investigation of six yellow rust R-genes (Yr1, Yr5, Yr6, Yr15, Yr17 and Yr27) in Avocet S background, with Avocet S as the control. Fungal colony size and area of hypersensitive response (HR) were assessed for each interaction at 4, 8 and 16 days post inoculation (dpi). The pattern for Avocet Yr15 was distinct, because HR arrested the pathogen very early and rapidly restricted pathogen growth. Avocet Yr1 and Avocet Yr5 showed a less rapid HR and restriction of pathogen growth, but most colonies were completely surrounded by HR at 8 and 16 dpi. In Avocet Yr6 the size of colonies and the extent of HR were highly variable with continuous change up to 16 dpi. More extensive pathogen growth was observed in Avocet Yr17 and Avocet Yr27, where HR induction was delayed, resulting in large intermingled colonies at 16 dpi. All interactions were clearly different from the susceptible control. Thus each R-gene produced a different temporal and spatial distribution of fungal colonies and HR response. Colony size distributions and HR response patterns are potential parameters for characterization of host resistances with different modes of action in wheat against Pst. The results also expand our comprehension of host resistance in wheat against P. striiformis.

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Wheat stem rust pathogen (Pgt) Identification and Characterization in Egypt using Single Nucleotide Polymorphism (SNP) markers.

BGRI 2018 Poster Abstract
Samar Mohamed Esmail Wheat Dis. Res. Dept., Plant Pathol. Res. Inst., A.R.C., Sakha, Egypt
Les John Szabo

Stem rust, caused by Puccinia graminis f. sp. tritici (Pgt), is one of the most serious disease of wheat worldwide. The discovery of new Pgt races in Africa, Ug99 and its variants, brings a new threat to global wheat production. In this study, 50 single pustule stem rust samples, were collected during 2015-2016 from the International Stem rust Trap Nursery (ISRTN) and commercial wheat fields in Sakha, the most important wheat growing region in Egypt. SNP-genotyping was carried out at USDA-ARS Cereal Disease Laboratory. Infection and genotype data confirmed that none of these samples belonged to the Pgt Ug99 race group. Forty-five samples were successfully genotyped consisting of 12 multi-locus genotypes (MLGs). The majority (86.7%) of the samples belonged to three clades: 10 samples, clade III-B (MLG.04, race TTRTF) collected from Misr 3, Sakha 95 and Sids 14 wheat lines; 12 samples, clade IV-A.2 (MLG.06, race TKTTF) collected from Sr 5, Sr6, Sr7a, Sr7b, Sr8b, Sr9a, Sr9e, Sr10, Sr11, Sr15, Sr16 and Sr17 wheat lines; 17 samples, clade IV-E.2 (MLG.11, race TKKTF) from Sr13, Sr14, Sr19, SrMcN, Sr24, Misr 1, Misr 2, Sakha95 and Sids 12 wheat lines. Pgt samples belonging to clades IV-A.2 and IV-E.2 have been observed from Europe to the Middle East, and samples from clade III-B from the southern Caucasus Mountains, Middle East to northeast Africa. The remaining six samples collected from Sr12, Sr18, Sr20, Sr21, Sr22 and Sr25 wheat lines represent two new genotypes (MLG.14 and MLG.17) that have not been assigned to clades. MLG.14 was also observed in samples from Azerbaijan, Iraq and Eritrea. In contrast, this represents the first detection of MLG.17. These results suggest continued variability of the Pgt population in Egypt therefore, emphasizing the importance regularly monitoring to timely identify new races, and utilize this information in screening and identification of effective sources of resistance.

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Molecular and field based characterization of yellow rust resistance in wheat germplasm across locations in Pakistan

BGRI 2018 Poster Abstract
Aamir Iqbal The University of Agriculture, Peshawar, Pakistan
Muhammad Khan, Muhammad Ismail, Sher Nawab, Abdullah Jalal, Muhammad Imtiaz, Sajid Ali

Rust disease response is used to assess the resistance status of breeding lines, which is required to be tested across location and complemented with molecular markers. The current study was designed to characterize yellow rust resistance in 29 introduced advanced CIMMYT wheat lines along with three check varieties across three contrasting wheat growing regions (Peshawar, Mansehra and Lakki-Marwat) during wheat season 2015-16. A high disease pressure was observed across all three locations as favorable cold and wet climatic conditions prevailed during 2015-16. The maximum disease was recorded at Mansehra (up to 90%) followed by Peshawar (up to 50%) and Lakki-Marwat (up to 45%). There was a significant variability amongst the tested wheat lines for yellow rust severity and in yield potential. Among the advanced lines, W-SA-104, W-SA-115 and W-SA-118 had better grain and biological yield. Based on disease and yield parameters, cluster analysis of 29 wheat lines along with three checks grouped wheat lines into four clusters. None of these wheat lines showed resistance at every location (Average coefficient of infection "ACI" = 0). The maximum co-efficient of infection (55) was recorded at Mansehra whereas the minimum (0) was recorded at Peshawar and Lakki-Marwat. Twenty-six of these wheat lines were identified to possess partial resistance to yellow rust (with ACI < 20). Genotyping for the presence of resistance gene markers STS-7 (linked with Yr5), SC-Y15 (linked with Yr17) and Xwmc-44 (linked with Yr29) revealed the highest frequency of Yr17 (90.60%), followed by Yr29 (87.5%) and Yr5 (50%). The three resistant genes together were present only in 15 wheat lines (46.87%). Our results thus revealed the presence of variation in resistance response based on both field testing and molecular markers which could be utilized in wheat breeding to develop better resistance varieties to be exploited at field level.

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Combining ability estimation for yield and yield related traits in Triticum aestivum

BGRI 2018 Poster Abstract
Nusrat Parveen Vegetable Research Institute AARI, Faisalabad, Pakistan.
Etlas,Amin, , , , , , , , , , , , , , , , , , , , , , , , , , , ,

In the present study five bread wheat genotypes (9797, 9801, 9802, Chakwal-50 and Chakwal-86) were tested in a 5?5 full diallel analysis for the estimation of combining ability for yield and its related traits. In randomized complete block design (RCBD) twenty F1s along with their parents were planted in field with three replications in the research area of Department of Plant Breeding and Genetics, University of Agriculture, during 2014-15. Plant height, No. of grains/spike, spike length, No. of productive tillers/plant, flag leaf area, No. of spikelets/spike, 1000 grain weight and grain yield per plant were studied. Except spike length mean squares due to GCA were highly significant for all the traits. All the characters showed highly significant mean squares for SCA and RCA. SCA variance was lower than GCA variance for number of grains/spike and spike length presenting the major role of additive gene action in the inheritance of these traits. While for plant height, flag leaf area, number of spikelets/spike, number of fertile tillers/plant, 1000 grain weight and grain yield/plant the value of GCA variance was lower than the value of SCA variance exhibiting non-additive gene action. Chakwal-50 was the best general combiner for plant height, spike length, number of spikelets/spike, number of grains/spike and grain yield/plant. The best specific combination for most of the traits was 9802?Chakwal-86. In future wheat breeding research programmes, good specific and general combiners can be exploited.

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MicroRNAs and their mega effects on gene expression in response to leaf rust in wheat (Triticum aestivum L.)

BGRI 2018 Poster Abstract
Summi Dutta Department of BioEngineering, Birla Institute of Technology, Mesra, Ranchi, India
Manish Kumar, Kunal Mukhopadhyay

Bread wheat (Triticum aestivum L.) being the world's most popular edible cereal, plays a major role in global economy. Rust in wheat leaves, caused by Puccinia triticina, affects grain quality and severely retards its production worldwide. Micro(mi)RNAs are considered major components of gene silencing and so have a great role to play during stress. The present study focuses on identification of miRNAs, produced by host to suppress pathogen as well as delivered by pathogens to encounter host defence mechanism. Therefore, these miRNAs may be called as leaf rust responsive microRNAs. Small RNA and degradome libraries were prepared from a pair of near isogenic lines of wheat (HD2329, HD2329+Lr24), one set was mock inoculated while the other set was inoculated with urediniospores of leaf rust pathogen. Using these libraries as input a vast number of miRNAs rather a population of miRNAs were identified derived from wheat that were targeting genes mostly involved in functions like defense response, signal transduction, development, metabolism, and transcriptional regulation.
When reads specifically produced under pathogen inoculation were taken as input with Puccinia triticina genome sequences as reference, only three putative miRNA precursor loci were detected and the molecules produced were called miRNA-like molecules as their precursors lacked one or two criteria essential for a true miRNA precursor. The identified miRNAs were targeting genes like F-box protein, MAP kinase, calmodulin and susceptible antioxidant protein. We further identified the presence of argonaute and dicer like domains in Puccinia proteome available at FungiEnsembl which strengthens presence of RNAi-like activities in Puccinia.
In addition, differential expression of wheat as well as Puccinia small RNAs using stem loop RT-PCR under varying time points of disease progression (0-168 hpi) revealed their direct connection with stress responses.

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Introgression of Sr50 and SrWeb genes in hexaploid wheat using molecular markers for enhancing stem rust resistance

BGRI 2018 Poster Abstract
Punam Singh Yadav Institute of Agricultural Sciences, Banaras Hindu University, Varanasi 221005, India
Vinod Kumar Mishra, Uttam Kumar, Ramesh Chand, Akhilesh Mishra, Arun Joshi

Ug99 is a devastating race of Puccinia graminis f.sp. tritici possessing virulence against resistant genes Sr31 and Sr24. This race is highly adoptive and has spread quite rapidly with 13 known variants covering 13 different countries. For reducing the vulnerability of wheat in south Asia to the Ug99, breeding durable resistant varieties is important. India, second largest wheat producer, falls in the predicted pathway of Ug99. Most of the Indian germplasm possesses Sr31 and Sr24 in their background. HUW468, a well adopted variety of north eastern plains zone (NEPZ) of India, carries durable resistance gene Sr2. To strengthen it, a MABB program was initiated to introgressed two major genes (Sr50 and SrWeb) using a donor line PMBWIR4 from CIMMYT. The foreground selection was performed with Xgwm47 for SrWeb and IB267 for Sr50 followed by the background selection by using 128 polymorphic SSR markers covering all chromosomes. Backcross progenies of HUW468 were screened in the field condition by using of Pgt race 21A-2 at IARI, Regional Station, Indore located in the central India. Superior selected lines from BC2F4:5 generation was planted at three locations in India namely; Varanasi, Indore and Dharwad. HUW468-09-25-47-09 and HUW468-09-25-47-56 were selected from BC2F5 generation having Sr50 and SrWeb along with Sr2 gene, superior agronomic performance and with 93.5% and 92.7% genome recovery, respectively. These two lines also possess 6-10 % yield superiority over the recipient parent HUW468. These lines have been submitted for registration in NBPGR (National Bureau of Plant Genetic Resources), India.

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Study of the effect of planting date on the severity of yellow rust disease on bread wheat in northeastern Syria

BGRI 2018 Poster Abstract
Omran Youssef University of Hohenheim, Germany
Afrem,Issa, Helim, Youssef, Nawzad, Suleiman, Abdul Rahman, Issa, , , , , , , , , , , , , , , , , , , , , ,

Wheat is grown in Syria during the November-December. Wheat is exposed to many strains that negatively affect its productivity especially rust diseases, which was reported on wheat in Syria for many years and the most severe in 2010, Therefore, we studied the effect of planting dates on the severity and development of yellow rust disease. Where the field trials of the 2010-2011 season were carried out at the two locations in northeastern of Syria: Al-Qamishli Research Center and Yanbouh Research Station in Al-Malekia. By cultivating the susceptible bread wheat Cham 8, where six dates were planted starting from 02.10. 2010, a difference of 15 days. The results showed there was a difference in the severity of the yellow rust disease according to the dates of cultivation and thus the stages of growth in the plant and this was evident in the Yanbouh location where the onset of the onset of injury on 08.04.2011 in the all dates and developed the infection to 40S degrees and 30%. Also, on the 24. 04. 2011, the infection was recorded at the Qamishli location only on the third and fourth dates. The disease did not develop more than 10S and 10% due to climatic conditions due to rain and high temperature during the season. The results showed a positive correlation between the evolution of the disease and vegetative growth of plants, where the growth of plants was more active at the site of Yanbouh, especially in the second, third and fourth dates in the development of infection on plants in the rest of the dates because of weak and slow growth of plants.

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Wheat rusts status and population structure across Pakistan during wheat growing seasons 2015-16 and 2016-17

BGRI 2018 Poster Abstract
Sajid Ali The University of Agriculture, Peshawar, Pakistan
Muhammad,Khan, Safi, Kathi, Zahoor, Swati, Manzoor, Hussain, Annemarie, Justesen, Muhamamd, Imtiaz, , , , , , , , , , , , , , , , , ,

Considering the importance of wheat rust diseases in Pakistan and the recent identification of yellow rust pathogen (Puccinia striiformis f. sp. tritici) centre of diversity in Pakistan, the present study was designed to assess the status of three wheat rusts across the country during 2015-16 and 2016-17 and analyze the population structure of P. striiformis f. sp. tritici . A total of 451 fields (from 68 districts) were surveyed during 2016 and 480 fields (from 69 districts) during 2017. A high yellow rust pressure was present during 2016 throughout Pakistan, while it was predominant only in the Northern half during 2017. Leaf rust was present in the central part of the country, while stem rust was only found in the south. In Sindh province (located in the south), yellow rust was reported unexpectedly with high severity (>60%) on varieties like Kiran and Galaxy during both the years. A set of 513 samples of P. striiformis were genotyped with microsatellite markers to assess the population diversity and spatial structure. and infer on the cause of epidemics in the Sindh province. Population genetics analyses confirmed a recombinant population structure across all locations except the Sindh province, where relatively lower diversity and lack of recombination signature was revealed. At least five genetic groups were identified in the overall population, which were found across all locations, except Sindh province where one of the genetic groups was predominant. The P. striiformis population from Sindh province with low diversity that caused unexpected epidemics in a relatively warmer region needs to be further investigated for specific adaptation traits. Our results confirmed the high diversity across Pakistan, which lies in the Himalayan centre of diversity of the pathogen. This high diversity was present in locations without the presence of alternate host (Berberis spp.) and could potentially be associated with regular migrants from the Berberis zone into the whole country.

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