Jianping Zhang, Peng Zhang, Robert Park, Narayana Upadhyaya, Robert McIntosh, Sambasivam Periyannan, Brande Wulff, Burkhard Steuernagel, Evans Lagudah
Evolution of rust pathogens continues to pose challenges to global wheat production. Major resistance (R) genes, which encode proteins of the NBS-LRR (Nucleotide-binding site, leucine-rich repeat) family, have been a valuable resource for breeders to minimise yield losses from infection. Many wheat varieties harbor numerous R genes that could be identified and cloned in order to engineer more sustainable disease control. The advent of targeted gene enrichment and next-generation sequencing (NGS) has allowed rapid cloning of specific R genes, thus enhancing efforts to pyramid these genes and investigate their underlying resistance mechanisms. Several R genes present different phenotypes in certain genetic backgrounds, and cloning them would be an important step towards uncovering their interactions. Hybrid necrosis is one such phenotype observed in crosses of wheat genotypes involving the R gene Lr13 and complementary genes, Ne1 and Ne2, occurring in different allelic forms. It was recently concluded that Lr13 and an allele of Ne2 are actually the same gene based on genetic and mutational studies. The capability of Lr13 to confer both leaf rust resistance and hybrid necrosis cannot be answered without first cloning it. The lack of tightly linked markers coupled with the proximal 2BS chromosomal location of Lr13 does not make it easily amenable to map-based cloning. The NGS-based pipeline MutRenSeq (mutagenesis and R-gene enrichment sequencing) was used on EMS (Ethyl methanesulfonate) induced, susceptible Lr13 mutants along with support from comparative genomics to ascertain candidate gene sequences for Lr13, which are at advanced stages of screening and confirmation. Definite proof that a single gene is involved will only come with transformation studies when the cloned Lr13 candidate transformed into a susceptible line confers both a resistance phenotype in the transgenic line and a necrotic phenotype in the offspring of crosses between the transgenic line and a line possessing Ne1.
University of Mohammed V/ICARDA
HAFSSA,KABBAJ, AYED, AL ABDALLAT, GREGOR, GORJANC, JESSE, POLAND, MIKAEL, MILOUDI NACHIT, AHMED, AMRI, BOUCHRA, BELKADI, KARIM, FILALI MALTOUF, FILIPPO, BASSI MARIA, , , , , , , , , , , ,
Durum wheat (Triticum durum Desf.) is a major stable crop and it represents a base of the Mediterranean diet. This region is subject to a Mediterranean climate, which is extremely unpredictable with severe changes in moisture and temperature occurring each crop season. This unpredictability is summarized by breeders as GxE and the identification of traits controlling this interaction is quintessential to ensure stability in production season after season. To study the genetics of yield stability, four RILs populations derived from elite x elite crosses were assessed for yield and 1,000-kernel weights across five diverging environments in Morocco and Lebanon. These 550 RILs were characterized with 4,909 polymorphic SNPs via genotyping by sequencing. A consensus map was derived by merging the individual genetic maps of each population. Finally, imputation was used to fill all the missing haplotypes and reach a reduction of missing data to below 8%. Several significant QTLs were identified to be linked to TKW, grain yield and a stability index, namely AMMI wide adaptation index (AWAI). A second approach to identify loci controlling stability was the use of a global panel of 288 elites, accessions and landraces tested in 15 diverging environment. Multi-locations data were compiled via GxE models to derive the AWAI stability index. In addition, this panel was characterized with 8,173 polymorphic SNPs via Axiom 35K array. Significant associations were identified for all traits, including QTLs unique to AWAI. The sum of the identified QTLs can now be pyramid via marker assisted selection and molecular designed crosses in order to obtain very stable cultivars.
Ravi P Singh, Julio Huerta-Espino
Aphids are major pests of wheat, able to cause up to 40% yield reduction solely due to direct feeding and up to 60% when feeding is combined with the transmission of viral diseases. Wheat resistance to aphids has proven to be effective in protecting yields and also in reducing the transmission rate of viral diseases. Moreover, aphid resistance is fundamental to reduce the negative impacts that the indiscriminate use of insecticides have on the environment and human health. In this study we report the results derived from the evaluation of 326 synthetic hexaploid wheat (SHW) derived lines against the greenbug (Schizaphis graminum [Rondai]). Primary SHWs were crossed with CIMMYT elite lines and further selected in the breeding pipeline. Therefore, such lines have acceptable agronomic characteristics for its further use in breeding programs. The 326 SHW derived lines were evaluated at seedling stage, in five augmented incomplete blocks, arranged in split-plots, with two treatments (infested vs. non-infested) and with resistant and susceptible checks replicated 16 times. The measured variables were chlorophyll content with a SPAD meter and a visual damage score in a scale 0-100 was also taken. Measurements were recorded when the susceptible check was dead due to aphid feeding. The evaluations were repeated two times for confirmation. Our results indicate the presence of genetic variation for S. graminum resistance. We identified about 4 % of the lines to carry high levels of resistance against this aphid. These lines are currently used in CIMMYT's bread wheat breeding program to incorporate the resistance in elite germplasm.
Umesh Rosyara, Suchismita Mondal, Ravi Singh, Susanne Dreisigacker
Grain yield is the most important economic trait in wheat breeding. The detailed understanding of the genetic architecture of grain yield is crucial and the determining factor to optimize genomics-assisted selection strategies in wheat. First, we performed a marker and haplotype-based genome-wide association study (GWAS) for grain yield (GY) and yield stability coefficient (Pi) on 4,302 advanced breeding lines from five CIMMYT international bread wheat trails grown in multiple (optimally irrigated and stress) environments. All lines were genotyped using genotyping-by-sequencing. A haplotype map was built based on linkage disequilibrium between markers. Twenty-nine markers and 16 haplotypes were associated with GY and Pi across two and three germplasm trials with allelic effects ranging from 2 to 11% across environments. Secondly, we performed genomic prediction, testing eight different prediction models incorporating single markers (base model), haplotypes, epistatic interactions, and significant markers/haplotypes identified in GWAS. Initial results show that by including haplotypes and epistatic interactions among haplotypes (main effect and genome-wide), prediction accuracies range between 0.33-0.49 for GY, a 3 to 22.5% improvement over the base model. Despite the identification of significant marker/haplotype trait associations across traits and environments in GWAS, accounting for these markers in genomic prediction does not improve the prediction models. Our results suggest that the haplotype-based approach can increase prediction ability, but that the knowledge of the genetic architecture of grain yield might not have significant consequence on genomic-assisted selection.
Mohammad Kassem, Ghinwa Lababedi, Naim Al-Husien
Leaf rust is the most common rust in wheat production areas of Syria and causes significant annual yield losses. Using genotypes with durable resistance is the most economical way of controlling the disease. One of the best-known leaf rust resistance genes is Lr46 that confers a slow rusting type of adult plant resistance. The main objective of this study was to identify Lr46 in durum wheat genotypes using morphological and molecular markers. Thirty-two durum wheat genotypes were evaluated for response to leaf rust at the seedling and adult plant stages. Twelve genotypes (37.5%) were resistant (R), 10 (31.25%) were moderately resistant (MR), seven (21.87%) were moderately susceptible (MS), and three (9.37%) were susceptible (S). Molecular marker analyses using SSR marker wmc44 showed that 16 genotypes (50%) carried Lr46/Yr29. The genotypes possessing the marker linked to Lr46/Yr29 could be used for selection of Lr46/Yr29 in breeding for slow rusting resistance in durum.
Levent Ozturk, Ismail Cakmak
Zinc (Zn) deficiency is an important health problem worldwide, affecting about two billion people, especially children and women. Zinc deficiency related diseases are more prevailing in developing countries because populationa rely on cereals (i.e., wheat, rice and maize) as a staple food which are inherently low in micronutrients. Zinc concentration in cereal grains can be improved by genetic or agronomic biofortification. Optimized applications of soil and foliar Zn fertilizers has been found effective for cereals like wheat and rice but not significantly in maize. Current study focuses to elucidate the physiological reasons behind the poor response of maize to foliar applications compared to wheat. Experiments with stable isotope of Zn (70Zn) revealed the differences in leaf uptake, root and shoot translocation of foliar-applied Zn in wheat and maize. The results suggested that wheat has greater capacity for leaf absorption and translocation of foliarly applied Zn compared to maize. The increased leaf Zn uptake and localization in wheat was confirmed by a visual demonstration using Zn-responsive fluorescent dye Zinpyr and fluoresce microscopy. This study provides valuable information to maximize the uptake and deposition of foliarly applied Zn to cereal grains.
CREA-Research Centre for Cereal and Industrial Crops
Elisabetta Mazzucotelli, Oadi Matny, Antonietta Saccomanno, Raffaella Battaglia, Francesca Desiderio, Agata Gadaleta, Nicola Pecchioni, Pasquale De Vita, Giovanni Laido, Luigi Cattivelli, Brian Steffenson
The recent emergence of new widely virulent and aggressive strains of rusts (particularly stripe and stem rust) is threatening Italian durum wheat (Triticum turgidum L. var. durum) production, especially under the trend of higher temperature and humidity. A big effort has been undertaken to explore the genetic variability for resistance to these fungal pathogens and discovering novel resistance genes. In particular, a wide set of tetraploid wheat lines was genotyped with several thousands of SNP markers and used for association mapping. This large collection consisted of a group of durum wheat cultivars, produced from the beginning of the last century up to now, a collection of wild emmer wheats (T. dicoccoides), and lines belonging to other wild and domesticated tetraploid subspecies, as a large untapped source of genetic diversity. In a tight cooperation with the University of Minnesota, this collection was evaluated for reaction to several races of stem and stripe rust pathogens in both controlled greenhouse and field conditions. Among the genotypes belonging to the collection are parents of segregating populations which were used for the validation of mapping results. Novel resistance loci were identified, that can be incorporated into new durum varieties through breeding programs. The QTLs found in this study, together with those available in literature, were projected to the recently sequenced durum wheat genome in order to define more precisely the chromosome regions and candidate genes involved in resistance to rusts. Lines which were resistant to multiple races of rust pathogens were also found among both T. dicoccoides and durum wheat cultivars as a source of resistance genes, whose cloning will be undertaken based on the results here obtained.
This study was supported by the Italian Ministry of Foreign Affairs and International Cooperation, with the special grant RES-WHEAT.
Chris K. Sørensen, Rodrigo Labouriau, Annemarie Justesen, Mogens Hovmøller
Host vernalisation and temperature strongly affect the susceptibility of winter crops to pathogenic fungi. However, how the interaction of these environmental factors influence host susceptibility to Puccinia striiformis, the yellow (stripe) rust fungus, is poorly understood. An experimental system was developed to examine the effect of vernalisation, temperature regime (standard; 18 day/12 night °C and low; 12 day/6 night °C) and plant growth (seedling and adult plant stages) on changes in susceptibility of agronomically important winter wheat and triticale genotypes to P. striiformis races ('Warrior' and 'Kranich') highly predominant in several European countries. Host genotypes exposed to prolonged periods of low temperature, termed vernalisation, reduced disease susceptibility on specific winter host genotypes, although its effect differed considerably by the temperature regime and the P. striiformis race deployed. The influence of vernalisation on host susceptibility was more apparent at low temperature for the 'Warrior' race and at standard temperature for the 'Kranich' race. Triticale genotypes inoculated with the 'Kranich' race were particularly affected by the influence of vernalisation and temperature regime by displaying a shift towards reduced susceptibility at standard temperature. The effect of plant growth stage, i.e., vernalised seedlings versus adult plants, was most evident for the 'Warrior' race at standard temperature and at low temperature for the 'Kranich' race by revealing a lower infection type at the adult plant stage. The research findings presented here contributed to a better understanding of the role of environmental factors in host susceptibility. This, in fact, will aid in the development of more efficient early-warning systems and disease management strategies to the yellow rust fungus and in the decision making for the deployment of winter wheat and triticale genotypes.
Agharakar Research Institute Pune
yashavanthakumar,Kakanur, Vijendra, Baviskar, Ajit, Chavan, Vilas, Surve, Vijay, Khade, Juned, Bagwan, Vitthal, Gite, Shrikant, Khairanar, Sameer, Raskar, Deepak, Bankar, Satish, Misra, , , , , , , ,
MACS 3949 is a durum wheat variety developed at Agharkar Research Institute, Pune derived through selection method from 39th IDYN (CIMMYT). The variety was identified by 55th All India Wheat and Barley workshop CCS HAU, Hissar and subsequently notified by Central Sub Committee on Crop Standards, India. On the basis of mean of three years (2013-14, 2014-15 and 2015-16) data from All India coordinated experiment, grain yield of MACS 3949 (43.98 q/ha) was higher to all the checks Viz., NIDW 295 (39.70 q/ha) and UAS 428 (41.78 q/ha). Overall, MACS 3949 showed a yield advantage of about 10.78 % over NIDW 295 and 5.24 % over UAS 428. The important morphological traits of the variety described as, semi dwarf with average plant height around 81 (78-83) cm, medium sized strong waxy semi erect green leaves, parallel dense spikes with long spreading awns. Grains were amber colored, bold lustrous, semi hard, elliptical in shape with short brush, soft threshing at maturity and1000-grain weight was about 47 (42-53) gm. The variety has shown resistance to leaf rusts, in particularly the seedling resistance to race 77-complex of leaf rust, stem rust, leaf blight, powdery mildew, flag smut and karnal bunt under both natural and artificial screening conditions. It has high protein content (12.9 %), better nutritional quality (Zinc 40.6 ppm, Iron 38.6 ppm) with good milling quality (Test weight 81.4 kg/hl) and best cooking quality for pasta product having highest overall acceptability 7.25. The newly developed durum wheat variety MACS 3949 released for cultivation at Peninsular Zone in India, which is having rich source of nutritional pasta quality with high zinc and iron content will be a promising one for future potential of export at international market.
Northwest A&F University
Dejun Han, Jia Guo, Manuel Spannagl, Jianhui Wu, Aizhong Cao, Peidu Chen, IWGSC, Lili Huang, Jun Guo, Klaus Mayer, Zhensheng, Kang
Wheat cultivation in many regions faces threats by devastating fungal infections. However, wheat cultivar 92R137 shows resistance to Puccinia striiformis infection. To isolate the stripe rust resistance gene Yr26, an integrated transcriptomic and comparative genomics approach was undertaken. Near-isogenic lines of wheat (carrying Yr26 or not) infected with two Puccinia striiformis f. sp. tritici (Pst) (Virulence or avirulence to Yr26) were analysed in a dual detailed time series RNA-seq study. The emerging IWGSC refseq v1.0 genome assembly sequence serves as a valuable template and was also used for comparative genomics studies of the gene candidate region with the genome sequences of close relatives and wheat progenitors. Using bulked segregant analysis (BSA) to identify polymorphic SNPs between parent and resistant DNA (R-bulk) and susceptible DNA (S-bulk), flanking markers for Yr26 were identified. These two markers were mapped to the Chinese spring reference genome sequence, spanning a region of about 250 kb. The synteny analysis of this candidate region in CS chr1B with chr1A, chr1D, Wild Emmer Wheat (Td_chr1A and Td_chr1B) and Barley (chr1H) identified three candidate Yr26 genes. To detect gene candidates a dual time series RNA-seq analysis was performed. Genes differently expressed between rust susceptible (NIL-S) host lines and rust resistant (NIL-R) lines, carrying the Yr26 candidate gene were analysed. Both lines were inoculated with Pst carrying different avirulence factors (Pst-CYR32 and Pst-V26), compatible or incompatible with the corresponding wheat lines. Differential gene expression analysis (DEG) between compatible and incompatible interaction revealed DEGs in the wheat genome and in the Pst genome. From a network analysis of both wheat and Pst genes, we inferred connected co-expressed modules. Resulting modules showed particular enrichments for disease resistance, defense response to fungus and cell wall components.