University of Queensland, St. Lucia
Adnan Riaz, Jonathan Powell, Timothy Fitzgerald, Kemal Kazan, Neena Mitter, Evans Lagudah, Lee T. Hickey
The Lr34/Yr18/Sr57/Pm38/Ltn1 multi-resistance locus has been deployed and remained effective in wheat cultivars for more than 100 years. The durability and pleiotropic nature makes Lr34 a unique and highly valuable resource for rust resistance breeding. Despite its functional annotation as an ABC transporter, the mode of action is unknown. Considering this, we aimed to decipher molecular factors and signaling components essential for Lr34 function using RNA-seq of Chara resistant (Lr34) and Chara mutant (heavy ion irradiation, HII) susceptible wheat lines. Screening of Chara and Chara HII lines with Lr34-specific markers confirmed the integrity of Lr34 in both lines; however, phenotyping confirmed rust and powdery mildew susceptibility in the Chara HII lines. Plants were grown under controlled conditions and infected with Puccinia triticina pathotype 76-1,3,5,7,9,10,12,13+Lr37 at the flag leaf stage. Flag leaves were sampled at 0, 24, 48, 72, 96 and 168 hours post inoculation (hpi) from mock and infected plants. Based on real-time PCR analysis of basal defense genes and the Lr34 gene, we selected 72 hpi for RNA-seq with four biological replicates per condition. The samples were sequenced on an Illumina Hiseq 4000 at the Beijing Genomics Institute, China. A total of 9.0 Gb of sequence (2.25 Gb/library) from 16 libraries for four conditions was obtained. Differential expression analysis was performed using the Tuxedo analysis pipeline with standard parameters. Analysis revealed deletion of DNA fragments with collinear gene order on chromosomes 1A, 2D, 5A, 5B, 5D and 7D of Chara HII mutants. To determine the significance of the deletions we performed bulk segregant analyses on segregating F2 populations of Chara ? Chara HII crosses. Analyses revealed key genomic regions associated with Lr34-functional resistance and we are in the process of validating candidate genes using qPCR.
The University of Georgia (UGA)
Suraj Sapkota, James Buck, Jerry Johnson, John Youmans
Leaf rust disease, caused by the fungal pathogen Puccinia triticina, is a major biotic constraint of wheat production worldwide. Genetic resistance is the most effective, economic, and environmentally safe method to control and reduce losses caused by this disease. More than 70 leaf rust resistance genes have been identified and mapped to specific chromosomes; however, continuous evolution of new leaf rust races requires constant search for new sources of resistance with novel QTL/genes. The objectives of this study were to identify sources of resistance, and to map genomic loci associated with leaf rust resistance using genome wide association study (GWAS) approach. Phenotypic evaluation of 297 spring wheat genotypes against a prevalent race of leaf rust in Georgia revealed that most of the genotypes were susceptible, and only 24 genotypes were found resistant. Furthermore, GWAS detected 10 markers on chromosomes 2A, 2B, 6A, 7A, and 7B significantly associated with leaf rust resistance. A marker on chromosome 7AS was identified revealing a novel genomic region associated with leaf rust resistance. The new identified sources of resistance and QTL could be used in wheat breeding programs to improve leaf rust resistance.
National Research Council of Canada (NRC)-Saskatoon
Kerry Boyle, Tammy Francis, Peng Gao, Brittany Polley, Christine Sidebottom, Brent McCallum, Harpinder Randhawa, Tom Fetch, Randy Kutcher, Sylvie Cloutier, Pierre R. Fobert
Most rust resistant genes in wheat are race-specific (R), with relatively few genes conferring resistance only at the adult stage that have been described as slow rusting genes (APR). Pyramiding multiple R, APR or APR+R genes has been used successfully over many years to achieve durable rust resistance. To further enhance this strategy, a genetic genomics approach was exploited to identify genes with different resistant mechanisms and the most effective gene pyramids.
Several new combinations of rust genes were created and tested in the Thatcher background, revealing synergistic ("booster") effects involving Lr21 with Lr16. With QTL mapping approach, we found that genes combined from 7D, 1B and 7B conferred an almost immune response to leaf rust, while genes from 7D, 1B and 3B provided an almost immune response to stripe rust. With a genomics approach, a large scale transcriptome analysis was conducted on key rust resistant genes including six R genes, three APR genes and one gene pyramid with Lr34+Lr16 over a time series during the infection process of both seedlings and adult plants. Detailed transcriptome analysis of gene expression associated with different major and minor leaf rust genes, alone or in combination, identified common and unique aspects of defense responses. For example, Lr9 is different from the other three leaf rust genes tested, with resistance triggered at a very early stage, consistent with pre-haustorial resistance. R genes Lr21 and Lr16 were also significantly different compared to other R and APR genes. With gene co-expression network analysis, a shared unique gene module mediated by Lr34 and Lr67 was also identified. This large transcriptome dataset also allowed the development of a rust-wheat interactome atlas for rust functional genomics research in wheat.
Research centre SELTON
Alena Hanzalova, Jaroslav Matyk, Pavel Bartos
In the Czech Republic all three rust species on wheat occur. Leaf rust (Puccinia triticina) can be found almost everywhere, and it can cause yield losses up to 40% mainly in warmer parts of the country in South Moravia.
Yellow rust, typical for cooler climate, occurred in relatively long time intervals. However in 2013 new pathotypes tolerating higher temperatures occurred and caused yield losses. In 2016 yellow rust incidence was lower, being still important in Moravia, where yellow rust occurred already in previous years.
Stem rust incidence was very rare in the last years. However in Germany, outbreaks and new pathotypes (e.g. Digalu) of stem rust in 2013 were recorded and comeback of stem rust to Central Europe can be expected.
Rust control consists of chemical control and especially of breeding for resistance, that aims at combined resistance to all three rusts. On the scale 9 high resistance, 1 high susceptibility average 4 year rating (2013-2016) of the tested cultivars was 6.4 for yellow rust, 5.7 for leaf rust and 6.2 for stem rust.
"Triple rust resistance" was recorded in spring wheat LOTTE and winter wheat line SG-S 1684 13, high resistance to yellow rust and stem rust in the cultivar Steffi. Resistance to all three rusts of 14 winter wheat cultivars and 12 breeding lines from the Plant Breeding Station-Stupice is summarized on separate tables and described in the text.
Department of Plant Molecular Biology, University of Delhi South Campus
L-myo-inositol phosphate synthase (MIPS; EC 126.96.36.199) have been involved in abiotic stress tolerance and its disruption leads to spontaneous cell death and enhanced tolerance to pathogen. However, its molecular mechanism underlying role of MIPS in growth, immunity and abiotic stress tolerance remains unknown. To delve deeper into the conserved molecular mechanism of MIPS action during growth and stress condition, we characterized the overexpression transgenic of TaMIPS and mutant lines of AtMIPS1. Subsequent, transcriptome analysis revealed the activation of ET/JA dependent immune response in transgenic and SA defense response in mutant. Pull-down analysis revealed the interaction of TaMIPS2 with ethylene synthesis (ACO) and signaling protein (CTR1) component. Due to the established role of ethylene during the skotomorphogenesis, we investigated the effect of myo-inositol phosphate synthase role in ethylene response during hook formation. Our results thus suggest the requirement of MIPS for ethylene response and regulating the growth and immunity.
Ethiopian Institute of Agricultural Research
Information about changes associated with advances in crop breeding is essential for understanding yield-limiting factors and developing new strategies for future breeding programmes. Thirty-six durum wheat varieties released since 1966 were evaluated in three replications of the Randomized Complete Design at Debre Zeit and Akaki, Ethiopia during the 2016 cropping season to estimate the amount of genetic gain made over time in grain yield potential, yield-associated traits and in protein content. Analysis of variance revealed significant differences among varieties for all 16 quantitative traits, protein content and protein harvest in Kg ha-1 at each of the locations. Grain yield varied between 1.66t ha-1 for Arendato released in 1966 to 3.90t ha-1 for Megenagna released in 2012 with mean of 2.952t ha-1 at Debre Zeit. At Akaki yield range was between 2.45 and 5.04t ha-1 with mean of 3.992t ha-1. 25 varieties surpassed Arendato (3.754t ha-1) at this location. In the combined ANOVA significant difference between the varieties was observed only for spike length, spikelets spike-1, grains spikelet, grains spike-1, plant height, days to flowering, thousand grain weight and hectoliter weight. Varieties specifically adapted to only one of the locations, widely adapted varieties and varieties not adapted to any of the locations were identified. Regression analysis revealed that grain yield has increased by 22kg ha-1 year-1 since 1966; an increase of 40.6% over yield in 1966. This was accompanied with a significant decline of 11.4% in spike length, 6.7% in spikelets spike-1, 17.9% in protein content and 31.2% in protein yield ha-1 and a significant increase of 41.1% in grains spikelet-1, 32.9% in number of grains spike-1, 22.3% in thousand grain weight, 17.8% in grain filling period, 23.9% in seed growth rate, 40.1% in grain yield production rate, 7.9% in harvest index.
Seed and Plant Improvement Institute, Agricultural Research, Education and Extension Organization, Karaj, Iran.
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Stripe rust of wheat, caused by Puccinia striiformis f. sp. trirtici (Pst) is an important disease in many parts of Iran. Over last two decades several epidemics have occurred in Iran causing the breakdown of widely utilized sources of resistance in wheat cultivars. Fifty isolates were collected from different parts of Iran during 2017. Eight isolates of Pst. from 2017 have been processed to date for race analysis. Infection types were assessed on a 0-9 scale 16 and 18 days after inoculation using a scale similar to that described by McNeal et al. (1971). Infection types (ITs) 7 to 9 were regarded as virulent (susceptible) and lower than 7 were avirulent. Pathotypes 102E158A+,Yr27; 6E158A+,Yr27; 102E158A+,Yr27; 166E154A+,Yr27; 38E174A+; 38E158A+,Yr27; 238E190A+,Yr27 and 38E190A+,Yr27 were identified. Pathotype 238E190A+,Yr27+ (from West of Iran) was more aggressive during this study. Plants with Yr1, Yr4, Yr5, Yr10, Yr15, Yr24 and YrSP were resistant to all pathotypes. Pathotypes with virulence on plant with gene/s Yr2, Yr6, Yr7, Yr8, Yr9, Yr25, Yr26, Yr27, Yr32, YrSD, YrSU, YrND and YrA were more common. Seedling tests of Iranian wheat cultivars to race 238E190A+,Yr27+ showed that the new released cultivars that included Parsi, Baharan, Bahar, Pishgam, Zareh, Urom, Maihan, Dena, Haydarei and Shabrang were resistant to the new aggressive race with virulence on plants with Yr27.
Seed and Plant Improvement Institute, Agricultural Research Education and Extension Organization (AREEO), Karaj, Iran
The basidiomycetous fungus, Puccinia graminis f. sp. tritici (Pgt) causes stem rust disease as one of the most destructive wheat pathogens, worldwide. TTKSK and other Pgt races under Ug99 race group are considered as major threats to wheat production in east Africa and CWANA region by defeating the stem rust resistance gene Sr31, while its ineffectiveness was reported in Iran in 2007. Race TKTTF of Pgt caused a severe stem rust epidemic in southern Ethiopia in 2013, and was spread to Europe through 2016 Sicily outbreak. This research describes race identification of Iranian isolates collected during the widespread distribution of stem rust in 2014-16. Purified urediniospores of 123 Pgt isolates were separately inoculated on seedlings of 20 North American differential wheat cultivars carrying different Sr resistance gene/s. Infection types were recorded at 14 days post inoculation (dpi) using Stakman et al. 0-4 scale. Based on the letter code nomenclature, we identified the Pgt races TKTTF, TTTTF, TTKSK, TTKTK, PKTTF, TKSTF, PKSTF, PKTTC, PTRTF, PTTTF, PKSTC, TTRTF, TKSTC and PKRTF in Iran. TKTTF and TTTTF were determined as prevalent Iranian Pgt races. This is the first report of race TTKTK, a new variant of Ug99 race group with virulence on Sr31 and SrTmp resistance genes, in Iran. Since TTKTK primarily occurred in south west of Iran, the migration route for this new race seems to be similar to race TTKSK. The high race variation observed in this study could indicate a high genetic diversity among P. graminis f. sp. tritici populations in Iran, as a wheat center of origin.
Wheat Research Institute, Faisalabad, Pakistan
Javed Ahmad, Abid Mahmood
Wheat is an important cereal crop and staple food in Pakistan. Most of the wheat is cultivated late after cotton, rice and sugarcane. Introduction of long duration Bt cotton varieties further pushed its sowing to late December or even early January. Late sowing of wheat crop results in yield loss in the Punjab province. A study has been conducted in experimental fields of Wheat Research Institute, AARI, Faisalabad, Pakistan to find out the possible reasons of low grain yields in late sown crop. Twelve experimental wheat lines were planted on seven sowing dates starting from 1st November to 30th December with ten days interval. The experimental design was a factorial combination of seven sowing dates as main plots and twelve varieties/ genotypes as subplots in a split-plot design with three replications. Effect of temperature on several crop growth stages was studied. Mean minimum temperature during the month of December, 2016 and January, 2017 remained below 5?C and mean maximum was more than 30?C during March 2017. Weather conditions experienced by the crop at each developmental stage were compared with the optimum conditions required on that specific stage in each sowing date. It was revealed that in late sown crop, different phonological/growth stages of the crop and yield components and grain yield were affected negatively. It was concluded from the study that the late sown crop suffered from two types of temperature stresses. The late sown crop faced low temperature stress at starting phase which result in delayed germination and low tillering. At caryopsis formation and grain filling the same crop face high temperature stress causing reduced grain formation and shriveled grains due to enforced maturity. Sowing of wheat at proper time i.e., by the end of November was recommended to fetch maximum yields.
Crop Diseases Research Institute, PARC Substation, Murree Pakistan
Sufyan,Muhammad, Abid Majeed, Satti, Munir, Anjum, Fayyaz, Muhammad, Atiq ur Rehman, Rattu, Imtiaz, Muhammad, , , , , , , , , , , , , , , , , ,
225 Puccinia striiformis f.sp. tritici isolates collected from wheat growing areas of Pakistan during 2013-2016 were analyzed using 18 near isogenic yellow rust differentials. Seventy eight races were identified among collection in which 20 were common (n > 2). Rest of the races were very rare and encountered only once (n=1). Races 574212, 574232, 474232, 474233, 574213 and 434232 were most frequent (n> 15). Pathogenic diversity analysis of the collection reveal high diversity (H =3.57) of the P. striiformis population of pakistan. On the basis of phenotypic response to yellow rust genes, the most frequent races could be grouped into 5 diverse groups. Distinct grouping was also observed in rarely encountered isolates. Most of the races were highly complex and 80% isolates had complexity ranging from 8 to 11. Virulence frequency for Yr6, Yr7, Yr8, Yr17, Yr27, Yr43 & YrExp2 remained above 80% while that of Yr1, Yr9 and Yr44 remained over 40%. Partial virulence was detected for Yr5, while virulence to Yr10, Yr15, YrSP was found in < 4% isolates. Paper discuss spatial and temporal distribution of P. striiformis races in Pakistan.