State Key Laboratory of Crop Stress Biology for Arid Areas and College of Life Sciences, Northwest A&F University, Yangling
Liyang Chen, Baoyu Huai, Shoujun Hu, Lijing Pang, Pu Yuan, Zhensheng Kang
Infection of pathogens in plants induces production and accumulation of reactive oxygen species (ROS). ROS are not only involved in plant defense responses, but directly restrict or kill pathogens. To counteract this attack, it is necessary for pathogens to remove host-produced ROS. However, the mechanisms protecting pathogens against host-derived oxidative stress are little known. In this study, a superoxide dismutase (SOD) gene, PsSOD2, was cloned from Puccinia striiformis f. sp. tritici (Pst). Quantitative reverse transcription PCR (qRT-PCR) analysis indicated that PsSOD2 is an in-planta induced gene active in the early stage of Pst infection. Prokaryotic expression and biochemical characterization revealed that PsSOD2 encoded a Cu-only SOD. The predicted signal peptide for protein secretion was functional in an invertase-mutated yeast strain. Transient expression in Nicotiana benthamiana suggested that PsSOD2 is localized in plasma membrane and dependent on glycophosphatidyl inositol (GPI) anchor at the C terminus. Furthermore, Size exclusion chromatography and bimolecular fluorescence complementation validated dimerization of PsSOD2. Overexpression of PsSOD2 in N. benthamiana significantly decreased ROS production triggered by flg22. Knockdown of PsSOD2 using a host-induced gene silencing (HIGS) system reduced the virulence of Pst, which was correlated to ROS accumulation in HIGS plants. These results suggest that PsSOD2 is a pivotal virulence factor that is localized in hyphal plasma membrane to promote Pst infection by scavenging host-derived ROS.
Northwest A&F University
Xiaoguo Zhu, Zhensheng Kang
RNA interference (RNAi) is a powerful genetic tool to accelerate research in plant biotechnology and to control biotic stresses by manipulating target gene expression. However, the potential of RNAi in wheat to efficiently and durably control the devastating stripe rust fungus Puccinia striiformis f. sp. tritici (Pst), remained largely under explored, so far. To address this issue, we generated transgenic wheat lines expressing double-stranded RNA targeting PsFUZ7 transcripts of Pst. We analyzed expression of PsFUZ7 and related genes, and resistance traits of these transgenic wheat lines. We show that PsFUZ7 is an important pathogenicity factor that regulates infection and development of Pst. A PsFUZ7 RNAi construct stably expressed in two independent transgenic lines of wheat confers strong resistance to Pst. Pst hyphal development is strongly restricted, and necrosis of cells in plant resistance responses was induced significantly. We conclude that trafficking of RNA molecules from wheat plants to Pst may lead to a complex molecular dialogue between wheat and the rust pathogen. Moreover, we confirm the RNAi-based crop protection approaches can be used as a novel control strategy against rust pathogens in wheat.
Wheat Research Institute, Faisalabad, Pakistan
Ghulam Mahboob Subhani, Javed Ahmad, Abid Mahmood
Global warming affects the environmental parameters of agro-based countries like temperature increase, melting of glaciers, floods, erratic rains, low temperature, frost and high temperature. As a result agriculture is becoming more vulnerable to global environmental shifts. In case of wheat, erratic or low rains badly affect the wheat crop of rainfed areas of the country along with high temperature at seedling or juvenile stage. Similarly, frost affects the early sown wheat crop in irrigated areas of Punjab. Lesser availability of irrigation water from water reservoirs also reduces the wheat crop productivity. Sudden increase in temperature (>30?C) during the month of March adversely affect the grain filling. High temperature during grain filling stage interferes with the photosynthetic activities of the plant due to enhanced maturity, grain become shriveled and results in low grain yield. The threat of these environmental changes can only be overcome through breeding with specific objectives which is cost effective once obtained.
Hence development of wheat varieties for frost, drought and heat tolerance is the only feasible solution to combat these stresses which is being used at Wheat Program of Ayub Agricultural Research Institute, Faisalabad, Pakistan. New emphasis is also being given to develop frost resistant wheat varieties due to changing scenario of last few years. The institute is actively involved for the development of heat, drought and frost tolerant wheat varieties. During working for tolerance against any of these stresses plant types to be breed are physiologically and morphologically modeled in such a way that they should be capable of tolerating respective stress. In addition to breeding work an extensive research is also being done at Wheat Research Institute, AARI., Faisalabad to investigate best agronomic strategies to make wheat crop best adapted to environmental stress conditions.
CREA Research Centre for Genomics and Bioinformatics
,International Durum Wheat Genome Sequencing Consortium
The domestication of wild emmer wheat ~10,000 years ago by early agrarian societies have led to the selection of domesticated emmer and subsequently of durum wheat through a process of selection for non-brittle rachis and free-threshing forms. Durum wheat and became established as a prominent crop only ~1,500-2,000 years ago. We have completed the 10.45 Gb assembly of the 14 chromosomes of the modern DW cultivar 'Svevo' and provides, via comparison with the wild emmer assembly, an account of the genome-wide modifications imposed by 10,000 years of selection and breeding on the genome architecture of tetraploid wheat. A number of regions that were under selection during the domestication of wild emmer or the subsequent selection of durum wheat have been identified. Furthermore, we have projected on the durum wheat genome about 1,500 QTLs for morphological phenological and quality traits, grain yield components and disease resistance reported from published biparental mapping or GWAS. NBS-LRR genes are prominently involved in signaling and plant disease resistance. The durum wheat genome contains more than 66,000 genes and among them we annotated about 1,500 complete NBS-LRR genes. A similar number was found in the wild emmer genomes, nevertheless the comparison of the two genomes has identified some NBS-LRR genes specific for durum wheat. The availability of the complete genome of durum wheat will speed up the identification and the isolation of new resistance genes as well as the breeding for high-yielding and more resilient cultivars.
Agriculture Botany Division, Nepal Agricultural Research Council
Baidya Nath,Mahto, Sarala, Sharma, Madan Raj, Bhatta, Mahesh, Subedi, Deepak, Pandey, Nutan Raj, Gautam, Suraj, Baidya, Roshan, Basnet, Rudra, Bhattarai, Ajaya, Karkee, Suk Bahadur, Gurung, Prem Bahadur, Magar, Sunita, Adhikari, Bhagarathi, Shahi, Basistha, Acharya
A total of 41 bread wheat (Triticum aestivum L.) varieties have been released so far in Nepal since 1960. Farmers have been gradually adopting newly released varieties due to disease and lodging resistance, better yield performance and good taste. In Nepal, wheat area coverage, production and productivity have been increased by almost seven, sixteen and two folds, respectively in the last 56 years. Performance of varieties varies from one region to another. Yellow rust is the major problems in hills while leaf rust is the primary issue on the plains. Stem rust is sporadic in localized areas of Nepal. Wheat research program in Nepal has released 9 wheat varieties resistant to Ug99 namely Vijaya, Tilottama, Banganga, Gaura, Dhaulagiri, Danphe, Sworgadwari, Munal and Chyakhura. Vijay, Tilottama and Banganga are also resistant to leaf rust while, Dhaulagiri, Danphe, Sworgadwari, Munal and Chyakhura are resistant to yellow rust. Since the release of Vijay, the first Ug99 resistant variety in Nepal during 2010, source seed production of rust resistant varieties has been increasing significantly each year with present coverage under these varieties being around 40%. WK 1204 has been occupied 35% area in hills of Nepal. Seed production and distribution of such high yielding disease resistant varieties through public-private partnership is leading to quality seed supply for varietal diversity and better food security in the country.
Agricultural Biotechnology Research Institute, AARI, Faisalabad PAKISTAN
Shahid Nazir, Muhammad Waqas, Jamil Imran, Habib Muhammad, Zaffar Iqbal
Wheat is a major staple food in Pakistan and its production is subject to many yield limiting factors. Among biotic stresses, rusts have been the most devastating. Hence, the development of rust resistant genotype is the ultimate solution. The traditional approach of transferring resistant genes from wheat related species is time-consuming and laborious. It is complicated by the need to perform inoculation tests on plants in segregating populations, also requiring the application of appropriate races. Molecular markers could tag the presence of important resistance genes and allow breeders to identify the resistance genes rapidly and accurately. Therefore, use of molecular markers can help breeder in developing resistant wheat cultivars to minimize yield losses. To harvest the beauty of this system, 60 candidate wheat candidate varieties (included in provincial wheat yield trial) were screened against rusts using linked DNA markers for genes i.e. Lr-34/Yr-18, Lr-46/Yr-29, Lr-28, Lr-19, Sr-2 and Sr-32. Total genomic DNA was isolated and used as template in PCR for the verification of rust resistant genes. The gene Lr-34/Yr-18 was found present in one genotype and absent in 54 genotypes whereas one genotype was observed as heterozygote with respect to this gene. 49 candidate varieties for Lr-46/Yr-29, 03 for Lr-28, 56 for Lr-19, 38 for Sr-2 and 54 for Sr-32 were found positive showing presence of these genes in the new varieties. Missing entries were tested twice but no resistant gene(s) was detected. This information was shared with respective breeding institute to design the future research program. Furthermore, this molecular information was used for rust resistant gene pyramiding work to develop the durable resistance in wheat against rusts and crosses were attempted utilizing high yielding genotypes and genotypes carrying maximum rust resistance genes.
Emel Ozer, Mehmet Karaman, Mozaffar Roustaii, Jalal Kamali
Erratic weather patterns associated with climate change pose unique challenges for wheat breeders playing a key part in the fight to ensure global food security. Within the rainfed winter wheat areas of Turkey and Iran this erratic weather patterns may prevent attaining maximum potential increases in winter wheat genetic gains. This is primarily related with the fact that the ranking of tested varieties may greatly change from one year to the other. Erratic weather patterns may interfere with breeders decision on the ideotype(s) they should aim for during selection. To support breeding decisions, this study aimed at optimizing major traits through modelling different combinations of environments and defining probabilities of the range of variation of traits (phenology and pant height) that maximized grain yields. Optimal phenology was found to be highly related with the temperatures at which the winter wheat varieties were exposed at around heading time (20 days before and after heading). Specifically later winter wheat varieties were exposed to higher temperature both before and after heading and this exposure had a negative effect on grain filling duration and final grain yield. Finally, the use of at least five different wheat varieties in one production field (with different phenology and plant height) was compared to a field with monoculture to test for improved resilience. It was concluded that by selecting one best wheat variety in a wide range of environments it was possible to maximize grain yield and that using a set of diverse varieties was not beneficial.
University of Bologna, Italy/ International Center for Agricultural Research in the Dry Areas, Morocco
Hafssa Kabbaj, khaoula El hassouni, Elisabetta Frascaroli, Angelo Petrozza, Stephan Summerer, Marco Maccaferri, Miguel Sanchez-Garcia, Roberto Tuberosa, Filippo M. Bassi
Global food security is faced with many threats including population growth and changing climate. To cope with these threats a new paradigm shift is required to ensure sufficient and sustainable crop production. Hybrid technology could represent a partly strategic solution for durum wheat, but the understanding of its heterotic behavior is very limited. In this study, 53 F1 plants were produced via half diallel scheme and North Carolina design II, using as parental elite lines selected on the basis of their genetic distance. These hybrids along with their parents were evaluated for different physiological and root traits on a precision phenotyping platform (Lemnatec) at different levels of water stress. Additionally, a second root test was conducted in near field condition via a basket method to determine shallow or deep rooting behavior. Hybrids with the most heterotic combinations in terms of above and below ground biomass were identified. However, in order to ensure adequate pollination between heterotic parents, their flowering time must overlap. To identify good matching partners, a GWAS study was conducted to identify genomic regions associated with the control of flowering time in durum wheat. A total of 384 landraces and modern germplasm were assessed at 13 environments with different temperatures and day length throughout the season. Genotyping was conducted by 35K Axiom array to generate 8,173 polymorphic SNPs. In total, 12 significant QTLS for landraces and 17 QTLs for modern germplasm were identified consistently across environments. These two results when combined will allow to predict the best parental partners for hybrid production via markers screening on the basis of their genetic similarity to the most heterotic groups, and with matching flowering times.
Ethiopian Institute of Agricultural Research (EIAR)
Stripe rust caused by Puccinia striiformis f.sp.tritici, is one of the major diseases of wheat in the world. Experiments were carried out at two sites in Ethiopia (Kulumsa and Meraro) during the 2015 cropping season to evaluate the response of 198 elite bread wheat genotypes and two checks to the prevailing races of stripe rust at adult plant and seedling stage. The genetic profile of these genotypes was assessed using 13006 SNP markers and an association mapping was explored to determine marker?trait association. About 72.5% and 42.5% of the lines exhibited resistance at Kulumsa and Meraro, respectively. Out of 198 genotypes tested in the greenhouse, 31% exhibited common resistance for Kubsa and mixed stripe rust isolate. Only 8966 of the SNPs were polymorphic, only these were used for association mapping analysis. These markers spanned an average density of 3.47 cM per marker, with the poorest density on the D genome. Almost half of these markers were on known chromosomes, but had no position on the consensus map of bread wheat. Analysis of population structure revealed the existence of three clusters and the estimated genomic wide Linkage Disequilibrium (LD) decay in this study ranged from 0 to 50 cM. 53 SNPs in ten genomic regions located on wheat chromosome 1AL, 2AL, 2BL, 2DL, 3BL, 4BL, 4DL, 5AS, 7AL and 7BL were identified. Thirty nine SNP markers in five genomic regions at Kulumsa and 14 SNP markers in six genomic regions at Meraro explained more than 25.5% and 35.1% of phenotypic variability respectively. For seedling stage, 21 markers in ten genomic regions located on wheat chromosomes 1B, 2A, 2B, 3A, 3B, 4B, 4D, 5A, 6B and 7B were associated with resistant. These loci may be useful for choosing parents and incorporating new resistance genes into locally adapted cultivars.
ICAR-Indian Institute of Wheat and Barley Research, Karnal
Satish Kumar, Ratan Tiwari, Gyanendra Pratap Singh
Stripe rust, is a major constraint to wheat production in the more than 12.8 m ha region of the Northern Hills and North Western Plains zones in India. The previously deployed resistance genes Yr9 and Yr27 are no longer effective. New sources of resistance (Yr5, Yr10, Yr15, Yr24) became available under the umbrella of an Indo-Australian collaborative project. A set of advanced backcross derivative lines out yielded the checks in preliminary evaluations and were promoted to station-level (16 lines) and national (5 lines) trials. A new cohort of resistance genes (Yr47, Yr51, Yr57) are now available and are being used in the breeding program. Resistance genes Yr17, Yr18, Yr31, Yr36, Yr40, Yr53, YrC591, and Yr70 are also being used. The recent progress in development of high yielding, stripe rust resistant lines will help to address future threats from stripe rust.