All BGRI Abstracts

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Existence of divergent lineages, virulence phenotypes and DNA methylation in the Canadian Puccinia striiformis population

BGRI 2018 Poster Abstract
Gurcharn Singh Brar Crop Development Centre/Department of Plant Science, University of Saskatchewan, Saskatoon, Canada
Sajid,Ali, Dinah, Qutob, Steve, Ambrose, Ron, Maclachlan, Kun, Lou, Curtis, Pozniak, Yong-Bi, Fu, Andrew, Sharpe, Randy, Kutcher, , , , , , , , , , , ,

Puccinia striiformis f. sp. tritici (Pst), the cause of wheat stripe rust, is one of the most important pathogens of wheat. Attempts have been made in the past to characterize the worldwide genetic structure of Pst populations, excluding Canada. Characterization of 59 isolates identified 33 races with three most common races representing half of the population and subtle differences in races of eastern and western prairies. For molecular characterization, 48 isolates were sequenced to obtain SNPs and genotyped with Pst-specific SSR markers. Isolates that were suspected of recombination based on SNP data were examined for their telia production ability as a proxy for sexual recombination. The study revealed that the majority of the population was clonal, however, not exclusively clonal, with the existence of four genetic lineages. Two lineages previously reported were identified: PstS0, representing an old northwestern-European and PstS1, an invasive warmer-temperature adapted lineage. Additionally, two new lineages, PstPr and PstS1-related, were detected that have not been reported previously. The PstPr and PstS1-related lineages produced more telia than the other lineages and had double the number of unique recombination events compared to PstS0 and PstS1. PstPr was concluded to be a sexual recombinant and an exotic incursion, which was closely associated with PstS5, PstS7 (Warrior), and PstS8 (Kranich) lineages, all of which arose by sexual recombination in the center of diversity - the Himalayan region. The total phenotypic variation in the population could not be explained solely by molecular genotypes, and a hypothesis on existence of epigenetic machinery in the Pst genome was tested. Homologs of the DNMTases class (DNMT1) were identified, providing compelling evidence of a role for DNA methylation. As a first report of DNA methylation, an average of ~5%, 5-methyl cytosine (5-hmC) in the Puccinia epigenome indicated the possibility of epigenetic regulation, which merits further investigation.

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Virulence evolution of Puccinia striiformis f.sp. tritici on wheat in Kenya between 1970 to 1992 and 2009 to 2014

BGRI 2018 Poster Abstract
Mercy Wamalwa Egerton University Njoro, Kenya
Ruth,Wanyera, James, Owuoche, Julian, Rodriguez, Annemarie, Justesen, Lesley, Lesley, Sridhar, Bhavani, Cristobal, Uauy, Mogens, Hovm?ller, , , , , , , , , , , , , ,

Emergence of new virulent races of Puccinia striiformis f. sp. tritici (Pst) to stripe (yellow) rust resistance genes in wheat (Triticum aestivum L.) has historically resulted in severe yield losses worldwide. We conducted a study to characterize the virulence profiles of Pst races prevalent in Kenya from historic (1970-1992) and recent collections (2009-2014). Pst isolates collected during surveys in Kenya were characterized at the Global Rust Research Centre (GRRC), Denmark. Yellow rust differential sets (wheat lines with known Yr resistance genes), and strain-specific sequence-characterized-amplified-region (SCAR) markers were used to group the Pst isolates as Pst1 or Pst2. Virulence to Yr1, Yr2, Yr3,Yr6, Yr7, Yr8, Yr9, Yr17, Yr25, Yr27, and the seedling resistance in AvocetS were detected. A total of 12 virulence profiles /races were detected in isolates obtained during 1970 to 1992, while six races were detected from samples collected between 2009 to 2014. In both periods, races with virulence profiles Yr2, Yr6, Yr7, Yr8, Yr25, Yr27, Avs and Yr2, Yr6, Yr7, Yr8, Yr17, Yr25, AvS were common. The SCAR results revealed that both Pst1 and Pst2 strains were present in the Pst isolates tested, Pst1 even in isolates from the 1970s. Additional isolates were also identified with neither Pst1 nor Pst2 profiles. From our findings, race analysis is key to understand the race diversity and pre-breeding efforts for effective resistance gene deployment.

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Genetic variability of drought sdaptive traits in nepalese wheat (Triticum aestivum L.) germplasm

BGRI 2018 Poster Abstract
Dipendra Pokharel Department of Agriculture, Sunsari, Nepal
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Wheat (Triticum aestivum L.) is one of the major cereal crops vital for global food supply. Most of the wheat crop in developing world including that of Nepal is either grown with limited irrigation or under rainfed conditions and thus face moisture stress at one or more growth stages limiting grain yield. An experiment was carried out at the Institute of Agriculture and Animal Science, Rampur to evaluate the genetic variability of selected drought adaptive traits in Nepalese wheat germplasm. The wheat genotypes evaluated comprised of Nepalese landraces and commercial cultivars, CIMMYT (International Center for Maize and Wheat Improvement) derived advanced introduction lines and three checks with differential drought adaptability. The wheat genotypes were grown in pots (single plant) arranged in a replicated split plot design in greenhouse under two contrasting moisture regimes, optimum and moisture stressed. The genotypes were evaluated for water use, water use efficiency, relative leaf water content and biomass production. The ANOVA (Analysis of Variance) revealed significant variation between environments and among the wheat genotypes for most of the traits studied. A wide range of variability was observed for water use, water use efficiency, biomass yield and relative leaf water content in moisture stressed and non-stressed environments. Nepalese cultivar Gautam showed a number of favorable drought adaptive traits, whereas, Bhrikuti was average in this respect. Based on the scores of drought adaptive traits recently released Cultivar (cv). Vijay was characterized as drought sensitive. A number of landraces and advanced breeding lines showed high level of water use efficiency and other positive traits for drought adaptation.

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First report of virulence to resistance genes Yrsp, Yr1 and Yr3 by wheat yellow rust pathogen (Puccinia striiformis f. sp. triti

BGRI 2018 Poster Abstract
Safarali Safavi Crop and Horticultural Science Research Department, Ardabil Agricultural and Natural Resources Research and Education Center, Ag
Farzad Afshari

Yellow (stripe) rust caused by Puccinia striiformis f. sp. tritici is the most devastating disease of bread wheat (Triticum aestivum) in the world. A wide range of virulent yellow rust pathotypes is evolving in different regions of the world causing the breakdown of widely utilized sources of resistance in wheat. Hence, the knowledge of virulence factors of pathogen and determining of effective resistance genes in the region will enable breeders to target those useful genes in their breeding programs. During cropping seasons of 2015-2016 and 2016-2017, virulence of the wheat yellow rust was investigated by planting differential cultivars and isogenic lines in a yellow rust trap nursery in Ardabil, northwest of Iran . Results showed stripe rust infections on some cultivars carrying Yr genes such as Yr1, Yr3, and Yrsp previously known to be resistant. The virulence spectrum of race population in Ardabil was identical to the Warrior race or its variants which is different from characterized races in Ardabil by carrying virulence combination for Yr1, Yr3, Yr17, Yr32, and YrSP and is avirulent on Yr8 and Yr27. Except for Yr8, Yr17 and Yr27, the common races in Ardabil are generally avirulent on Yr1, Yr3, and YrSP. This is the first report of race population in Ardabil (Iran) which is similar to the Warrior race or its variants.

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Genetic variability in bread wheat (Triticum Aestivum L. ) accessions using functional and random DNA Markers

BGRI 2018 Poster Abstract
Kachalla Kyari Mala Lake Chad Research Institute, Maiduguri, Borno State-Nigeria
Dattijo Aminu, Zakari Goji Silas Turaki, Fatima Henkrar, Udupa Sripada

The research was conducted at ICARDA, Rabat. Twenty-four accessions were obtained from LCRI for marker analysis. Wizard Genomic DNA Purification Kit was used for DNA extraction. DNA was extracted by CTAB method and quantified using 1.0 % (w/v) agarose gels. Total of 12 loci, 5 functional and 7 linked random DNA markers to the traits of interest were used. PowerMarker and DARwin software were used to calculate the No. of alleles and values of genetic diversity, PIC, genetic distance, and NJ dendrogram. The total No. of detected alleles was 39; and mean No. of alleles was 3.25. No. of alleles range from 1 (Dreb-B1) to 9 (Xgwm577). Genetic diversity index ranged from 0.0000 in Dreb-B1 to 0.8471 in Xgwm577. The PIC value was also varied from 0.0000 (Dreb-B1) to 0.8296 (Xgwm577). The frequency of biotic resistance linked random DNA marker allele at Xgwm144 and Xwmc44, associated with yellow and leaf rust gene was 25% each. Marker alleles Xgwm577 and Xgwm533 linked to Stb2 and Stb8 at 150 and 120bp have frequencies of 21 and 4%. The frequency of abiotic resistance showed 50% of accessions had 1R segment (1BL.1RS translocation) and 58% of accessions showed presence of 120bp allele of Xwmc89, associated with QTL for drought tolerant. Functional marker alleles of Dreb-B1 associated with drought tolerant genes showed alleles frequency in all accessions. Linked marker allele Xgwm111 linked to heat tolerant gene showed 17% allele frequency at 220bp. Rht1 and Rht2, the allele frequencies were 92 and 4%. 92% of the cultivars had photoperiod insensitive allele at Ppd-D1 locus. VrnA1a and VrnA1c primer pair amplified at 965, 876, and 484bp, allele frequency of 13 and 87%. Cluster analysis had grouped the accessions into 5 at a genetic distance level 0.15.

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Accelerated Cloning and Characterization of Adult Plant Resistance Genes in Wheat

BGRI 2018 Poster Abstract
Sreya Ghosh John Innes Centre
Burkhard,Steuernagel, Caixia, Lan, Miroslava, Karafi?tov?, Ksenia, Krasileva, Jaroslav, Dole?el, Evans, Lagudah, Ravi, Singh, Brande, Wulff, , , , , , , , , , , , , ,

Adult Plant Resistance (APR) genes are broad-spectrum, partial-resistance genes that have the potential to contribute to sustainable control of wheat rust diseases. However, their isolation and characterization are complicated by the lack of precise molecular markers required for their identification, and therefore their use in plant breeding programs has been limited. Recent developments including the falling cost of sequencing and the increasing use of sequence capture methods to reduce genome complexity have enabled previously intractable methods such as mutational genomics to clone genes in wheat. Despite their increasing ease of use, many of these approaches require prior knowledge of the gene space and, in some cases, the gene family of the target gene to be cloned. As the APRs cloned so far do not belong to any common gene family, it is not possible to use general features of these identified APRs to conduct biased searches for novel APRs. This project aims to use an unbiased gene isolation technique called MutChromSeq, which combines chromosome flow-sorting and mutational genomics, and is independent of fine mapping, to rapidly clone the recently discovered APR gene Lr68 (Leaf Rust 68). Cloning APRs allows breeders to trace genes cheaply and quickly using gene-specific markers, enabling them to build effective and durable resistance gene pyramids. It also allows us to elucidate any common mechanism of action they have, helping researchers and breeders understand better the basis of their durable resistance. At the same time, the generation time of wheat has become one of the major limiting factors for the response time of breeders to rust epidemics. Thus, this project also aims to combine marker-assisted selection with accelerated generation advancement ('speed breeding') for rapid germplasm structuring and field performance evaluation.

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Mining of hulled wheat species for yield gain in bread wheat (Triticum aestivum L.)

BGRI 2018 Poster Abstract
Vikas Venu Kumaran ICAR-Indian Agricultural Research Institute, Regional Station, Wellington, The Nilgiris, Tamilnadu, India
SIVASAMY,MURUGASAMY, JAYAPRAKASH, PARAMASIVAM, RAJESH KUMAR, MEENA, , , , , , , , , , , , , , , , , , , , , , , ,

Emmer wheat (Triticum dicoccum L.), tetraploid species (AABB) and spelt wheat (Triticum spelta L.), hexaploid species (AABBDD) are old world hulled wheat species cultivated centuries ago in different parts of the world. These species were later replaced by higher yielding bread and durum wheat in the last centuries. Grain yield is influenced by grain number per unit area and grain size which correlates positively with grain weight. Increasing the grain number was extensively and intensively explored in the past 100 years of wheat breeding which has nearly reached to saturation and leaves little room for further yield increase due to grain number?grain size trade off. Grain size/grain weight is believed to be major driving force for further improvement of wheat yield. Both the species have been characterised with larger grain size and higher grain weight; therefore an ideal source to improve the grain size/grain weight while maintaining the grain number per spike in the cultivated bread wheat. A total of 25 accessions each of emmer and spelt wheat with good grain size and weight were crossed with 5 elite bread wheat lines. In the F2 generation, recombinant lines with good grain size, higher grain weight and grains number were further backcrossed with bread wheat. Stable lines with free threshing were obtained at BC4F4 generations and were analysed for quality. Thousand grain weight (TGW) and harvest index (HI) ranged from 46-55g and 0.47-0.58 in stable lines respectively. Stable lines yielded 16-21% than the high yielding check while number of grains per spike was maintained as that of check. Stable lines involving spelt crosses have higher grain size, TGW and HI than emmer wheat crosses. Stable lines could be released directly as cultivar or else used as one of the parents in the wheat improvement programme.

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Harnessing the predictive power of epidemiological modelling for wheat yellow rust disease

BGRI 2018 Poster Abstract
Vanessa Bueno-Sancho John Innes Centre
Christopher,Judge, Francesca, Minter, Nik, Cunniffe, Richard, Morris, Diane, Saunders, , , , , , , , , , , , , , , , , , , ,

Wheat yellow rust is a disease caused by the fungus Puccinia striiformis f. sp tritici (PST) that is a significant threat to wheat production worldwide. Recently, a novel approach called "Field Pathogenomics" was developed that allows acquisition of genotypic data from field samples of PST-infected wheat. This has enabled us to study the re-emergence of this pathogen in the UK and understand the different races that form the current PST population. However, the dynamics of pathogen transmission and dispersal still remain unknown and understanding this is essential for designing effective surveillance. The objective of this project is to develop a spatially-explicit model for the spread of PST that can contribute to better management of the disease and be used as a warning system for wheat yellow rust infection in the UK. The first aim is to study how PST spreads at the field level and determine whether there are differences between PST races in terms of disease dynamics. To this end, a set of markers have been designed that can be used to genotype field-collected isolates and determine which race they belong to. Field trials were also undertaken across the UK using wheat varieties that are known to be susceptible to the disease, with PST-infected wheat samples collected during the 2015-2016 and 2016-2017 seasons. These samples will be genotyped to study the prevalence of different PST races and determine whether PST genotypes identified early in the season are predictive of dominant genotypes found later in the season. Understanding PST dynamics within a field is key to build an epidemiological model that can predict how this disease behaves. This would improve disease management, targeting of chemical sprays and optimize pathogen surveillance.

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Identification and Analysis of RNA Editing Sites in the Chloroplast Transcripts of Aegilops tauschii L.

BGRI 2018 Poster Abstract
Mengxing Wang State Key Laboratory of Crop Stress Biology in Arid Areas, Northwest A&F University, Yangling 712100, Shaanxi, China
Hui,Liu, Guangwei, Xing, Xiaojun, Nie, Song, Weining, , , , , , , , , , , , , , , , , , , , , ,

RNA editing is an important way to convert cytidine (C) to uridine (U) at specific sites within RNA molecules at a post-transcriptional level in the chloroplasts of higher plants. Although it has been systematically studied in many plants, little is known about RNA editing in the wheat D genome donor Aegilops tauschii L. Here, we investigated the chloroplast RNA editing of Ae. tauschii and compared it with other wheat relatives to trace the evolution of wheat. Through bioinformatics prediction, a total of 34 C-to-U editing sites were identified, 17 of which were validated using RT-PCR product sequencing. Furthermore, 60 sites were found by the RNA-Seq read mapping approach, 24 of which agreed with the prediction and six were validated experimentally. The editing sites were biased toward tCn or nCa trinucleotides and 50-pyrimidines, which were consistent with the flanking bases of editing sites of other seed plants. Furthermore, the editing events could result in the alteration of the secondary structures and topologies of the corresponding proteins, suggesting that RNA editing might impact the function of target genes. Finally, comparative analysis found some evolutionarily conserved editing sites in wheat and two species-specific sites were also obtained. This study is the first to report on RNA editing in Aegilops tauschii L, which not only sheds light on the evolution of wheat from the point of view of RNA editing, but also lays a foundation for further studies to identify the mechanisms of C-to-U alterations.

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Breeding of high yielding, rusts resistance and Zn-enriched wheat varieties for different agro-ecological zones of Pakistan

BGRI 2018 Poster Abstract
Maqsood Qamar Wheat Program, National Agricultural Research Center (NARC) Islamabad
Sikander,Khan Tanveer, Muhammad, Sohail, Muhammad, Shahzad Ahmed, Sayed, H. Abbass, Sundas, Wagar, Atiq, Rattu, Muhammad, Imtiaz, , , , , , , , , , , , , , , ,

Wheat plays a vital role in multifaceted farming system of Pakistan. Like other many other countries, Pakistan's sustainable wheat production is also continuously threatened by a number of biotic and abiotic stresses. Among the biotic stresses, three rust diseases of wheat have been the most devastating. Stem rust was effectively controlled with adoption of the semi-dwarf spring wheats of the Green Revolution. However, the threat of the evolution of Ug99 race of stem rust in East Africa and its migration to Iran cannot be neglected. The Chance of of Ug99 migrating from Iran into Pakistan, coupled with the presence of dangerous new races of stripe and leaf rusts invites enormous efforts for development of rust resistant varieties for sustainable production of the wheat in the country. In this regard the Wheat Program, NARC, Pakistan initiated an intensive breeding program with financial and technical support of USDA and CIMMYT. Diverse sources of resistance to the three rusts particularly to the stem rust race Ug99 were introduced from CIMMYT. Through the rigorous selection procedure, four rusts resistant wheat varieties (NARC 2011, Pakistan 2013, Zincol 2016 and Borlaug 2016) have been released. These varieties are also resistant to Ug99. The varieties i.e. NARC 2011, Borlaug 2016 and Zincol 2016 are performing well in irrigated areas whereas Pakistan 2013 is suitable for rainfed conditions. The variety Zincol 2016 has high Zn content (35 ppm) in grain as compared to national standard check variety (25 ppm). These varieties are not only higher yielding but also possess good grain quality and other desirable traits. A considerable quantity of seed of the varieties is already present in the national seed system and will reduce the risk of Ug99 threat.

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