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.
National Agronomic Research Institute of Algeria
Among the many biotic constraints to wheat production in Algeria, rusts and in particular yellow rust (Puccinia striiformis), are among the most prevalent diseases that occur mostly all over the northern part of the country. Yellow rust has become now sporadic due to the exploitation of effective resistant genes in different forms and combinations (from CIMMYT and ICARDA). Earlier, durable resistance was probably due to many genes, such Yr18, Yr9, Yr27 and Yr1.
Yellow rust appeared as epidemic in 2004, over 600 000 ha of bread wheat ; severity exceeded 70%. Yields from affected fields of Hidhab a susceptible variety did not exceed 5.2 q/ha, while resistant cultivars yielded up to 48 q/ha. Monitoring of the pathogen virulence factors and their changes provides basic information for the development of an early warning system. This experiment was carried out in 5 Eastern Algeria locations. 30 lines of a standard set of yellow rust and 14 near-isogenic lines from ICARDA were sown in 2-m rows in 2014 and 2015. According to the results, virulence on Heines Kolben (Yr2), Kalyansona (Yr2), Lee (Yr7), Avocet R (YrA), Federation*4/Kavkaz (Yr9), Yr6/6*Avocet ?S?, Yr7/6*Avocet ?S?, Yr9/6*Avocet "S", Yr17/6*Avocet "S", TP1295 (Yr25) and YrSU was common during those two seasons. The frequency of virulence on plants with Yr2, Yr6, Yr7, Yr9 or YrA and Yr27 was up to 80%. No virulence was observed on plants with Yr1, Yr3, Yr4, Yr5, Yr8, Yr10, Yr15 and Yr18 genes. This material was extensively used in our breeding programs and several new cultivars are in the on farm trials where a participatory selection approach is used. All resistant and performing new varieties are being spread for replacement of most old susceptible ones.
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.
Department of Botany, Faculty of Biology, Government College Murree, Pakistan
Barkat Ali, Muhammad Fayyaz, Atiq ur Reman Rattu, Abdul Samad Mumtaz, Muhammad Imtiaz
The new arrival of wheat rust pathotypes through migration during wheat cropping season requires regular monitoring to secure wheat production. In the present study, we collected leaf rust (Puccinia triticina Eriks.) infected wheat leaves from three major wheat growing provinces of Pakistan in the year 2014 to assess the haplotype diversity of P. triticina (Pt) isolates. The rDNA ITS sequence data of collected isolates was used in NCBI BLAST analysis. The blast hits showed best matches with Pt accessions EU014050 (Iran), JN120331 (Iran), JX533577 (Iran), AY956549 (Iran), DQ417412 (Czech Republic), DQ417418 (Israel), DQ417413 (Slovakia) and AF511083 (Louisiana). However, in cluster analysis, the Pakistani isolates showed strong bootstrap support with only Iranian and Indian (races 77-5 & 104-4) accessions that indicated eastward migratory mode of Pt pathotypes in Pakistan through westerly wind patterns. The predominant genotype DQ417412 (similar in alignment with AY956549 from Iran) overcome the resistance of top Pakistan mega varieties Seher06, Inqilab91, Kiran95, SKD1, TJ83 and NIFA-Batoor. Hence, the ITS based information remains a rapid molecular tool for pathogen surveillance across countries and continents.
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.
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.
Institut National de la Recherche Agronomique INRA Morocco
Kumarse Nazari, David Hodson, Tine Thach, Julian Rodriguez Algaba, Mogens Støvring Hovmøller
Wheat rusts, notably yellow rust, are amongst the most damaging diseases on wheat in Morocco. The objective of this survey was to assess the incidence and severity of wheat rust diseases across Morocco. The survey was carried out during April-May 2017 where growth stage of wheat ranged from anthesis to physiological maturity. The severity and response rating for the adult plant field reaction to rusts were based on the modified Cobb scale. A total of 117 bread wheat fields were inspected. The survey revealed that the most prevalent disease was yellow rust (96 out of 117 fields). Leaf rust, SLD (Septoria Like Diseases) and to some extent root rot complex were less prevalent. Leaf rust was only observed in 8 out of 117 inspected fields and exhibited low severity. Stem rust was observed in only one field. Following the drought of 2016, the 2017 growing season was an epidemic year for yellow rust in Morocco. It was detected across all regions and 50% of inspected fields were highly infected. Those that were lightly or not infected were sprayed with fungicides up to two times. Almost all commercial bread wheat cultivars in Morocco are highly susceptible to yellow rust. Appearance of new virulent races is leading to the breakdown of resistance in major cultivars e.g., Arrihan, which had very few pustules of yellow rust in 2013 was highly susceptible in the last three years. Samples of yellow rust from 2016 revealed a new virulent race in all samples, temporarily designated Pst (new) [virulence pattern: [Yr-,2,3,-,-,6,7,8,9,-,-,17,-,25,-,32,Sp,AvS,-]. Thirty-four samples submitted to GRRC in 2017 were all of the same genotype, identical to the new race already detected in 2016. The results demonstrate that surveillance and genotyping/race phenotyping of samples may be important for early-warning and anticipatory breeding strategies.
University Mentouri of Constantine, Algeria
Wheat is the world's most widely grown food crop. New races of pathogens frequently overcome current resistant varieties. To address this issue Algeria has strategies for immediate action, medium term protection and long-term research efforts to develop new resistant wheat varieties. Yellow rust is a very important disease of wheat in Algeria where 60% of the wheat crop is grown under cooler high elevation climate conditions (2?C ? 15?C). Crop losses reached 80% during the 2004/2005 epidemics. Strategies adopted to reduce the risk of wheat rust are ongoing yearly surveillance, awareness, and early warning systems to farmers; and breeding and developing new varieties with high yield potential and durable resistance. Several highly resistant varieties (Tiddis, Boumerzoug, Massine, Akhamokh and Yacine) were selected and promoted following seed multiplication and commercial release. They are also widely used in crosses to improve local varieties. The newly released varieties are being distributed to farmers that grow susceptible varieties. This gene deployment will provide a natural barrier between eastern to western Algeria to intercept the major direction of air flow. Fungicide control is now routinely applied soon after rust detection or even preemptively. The level of awareness for wheat rusts across Algeria is now very high. Training among farmers for visual detection is widely promoted by plant protection and extension services. These strategies have been very effective in mitigating the threat of wheat stripe rust such that losses have not exceeded 10% over the last five years.
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 the Free State and Seed-Co
Vicky Coetzee, Cornelia M. Bender, Renée Prins, Zacharias A. Pretorius
Notwithstanding the re-emergence and importance of wheat stem rust caused by Puccinia graminis f. sp. tritici (Pgt), the degree of protection provided by different types of resistance has not been carefully investigated in contemporary studies. Seven wheat entries were exposed to stem rust infection and fungicide response in a split-plot field experiment over two seasons. Severe epidemics of Pgt race PTKST, generated by frequent inoculation of spreader rows within and around the trial, developed in both years. By comparing grain yield in rusted and fungicide sprayed plots, varieties SC Nduna (Sr31) and SC Stallion (Sr2+Sr31) sustained mean yield losses of 28.8% and 20.7%, respectively. From entries with adult plant resistance (APR), Kingbird recorded a loss of 10.1% as compared to W1406 (19.5%) and W6979 (15.4%). Grain yield of SC Sky which exhibits all stage resistance (ASR) was reduced by 6.4% over the two seasons. The highest yield loss (47.9%) was measured for Line 37, the susceptible control. A significant linear relationship occurred between percentage yield loss and AUDPC in both seasons (R2=0.99 and 0.83). This study showed that not all sources of APR to stem rust provided the same level of protection under severe disease pressure. In the absence of virulence for SC Sky, ASR conferred the most protection.