Michel E. Ghanem, Sarrah Ben M'Barek, Gustavo Azzimonti, Silvia Pereyra, Silvia Germán, Felix Marza, Amor Yahyaoui, Pawan Singh, Michael Baum, Hans-Joachim Braun
Based on a global network of wheat partners, precision field-based wheat phenotyping platforms are being developed with the support of the CGIAR Research Program on Wheat and co-investing national agricultural research institutes. This collaboration strategy aims to i) strengthen the quality of phenotypic data to fully exploit the potential of genomic data, ii) strategic prioritization of activities based on trait screening capacities and regional needs, iii) sharing knowledge and germplasm to accelerate superior germplasm development and dissemination, iv) development of capacities. Phenotyping activities are being conducted for wheat blast (Magnaporthe oryzae) in Bolivia, Septoria tritici blotch (STB) in durum wheat in Tunisia, and for multiple diseases (leaf rust, Fusarium head blight, and STB) in bread wheats in Uruguay. Subject to further funding, additional platforms will be implemented, to contribute to a faster development of broad genetic based resistant, high yielding wheat varieties, and complementing evaluations currently performed for diseases and heat, drought and yield potential (Kenya, Ethiopia, Turkey, Mexico).
Institute of Agricultural Environment and Resources, Yunnan Academy of Agricultural Sciences, China
Xianming Chen, Anmin Wan, Jiasheng Chen, Mingliang Ding
Wheat stripe rust (Puccinia striiformis f. sp. tritici, Pst) is the most destructive disease of wheat worldwide. Breeding and planting resistant cultivars is the most economic, effective, as well as environmental methods to control the disease. Yunnan is a severe epidemic zone in China, which provides new incursions for other parts of China. Study on virulence of the Pst population and effectiveness of resistance genes, will provide information for breeding and rational use of resistance genes. One hundred and thirty-six136 isolates collected from 9 regions of Yunnan were tested using a set of 18 Yr NILs with genes Yr1, Yr5, Yr6, Yr7, Yr8, Yr9, Yr10, Yr15, Yr17, Yr24, Yr27, Yr32, Yr43, Yr44, YrSP, YrTr1, YrExp2, YrTyTye. Stripe rust races were named by octal code. The results showed that the Pst population in Yunnan is highly variable in races and virulence. A total of 64 races were identified and the top two most frequent races were 550273 (Virulence/Avirulence formula: 1, 6, 7, 9, 27, 43, 44, SP, Exp2, Tye / 5, 8, 10, 15, 17, 24, 32, Tr1 and 550073(Virulence/Avirulence Formula: 1, 6, 7, 9, 43, 44, SP, Exp2, Tye / 5, 8, 10, 15, 17, 24, 27, 32,Tr1), with frequency of 28.68% and 11.76%, respectively. The remaining races had frequencies less than 5.0%. No virulence were found for Yr5, Yr10, Yr15, and Yr32. The frequencies of virulence to Yr24, YrTr1, Yr8, and Yr17 ranged from 0.74% to 11.76%. The frequency of virulence to Yr27 was 52.94%; and virulence to Yr1, Yr6, Yr7, Yr9, Yr43, Yr44, YrSP, YrExp2, and YrTye ranged from 79.94% to 91.91%. The results will guide the breeding and wheat production. (This study was supported by National Natural Science Foundation of China, Grant No. 31260417 and 31560490)
Leonardo,Crespo-Hererra, Julio, Huerta, Ravi, Singh, , , , , , , , , , , , , , , , , , , , , , , ,
Malnutrition affects more than 2 billion people across the globe, particularly zinc and iron deficiency causes major health problem in developing world. The biofortified staple food crops such as wheat, is an important channel to contribute to the hidden hunger problem in low income countries. Breeding for enhanced zinc concentration in wheat was initiated by crossing high zinc sources identified among synthetic wheats, T. dicoccum, T. spelta and landraces. These crosses have resulted in wheat varieties with competitive yields and enhanced grain zinc were adapted by farmers in South Asia. CIMMYT-derived early-maturity wheat cultivar 'Zinc-Shakti' with about 40% increased zinc (+14 ppm), is now grown in eastern India through public-private partners. The two CIMMYT-derived biofortified varieties: 'WB2' and 'HPBW01' released in 2016 for northwestern plains zone of India. In Pakistan, 'Zincol' was released in 2016. The first high zinc wheat variety (Bari-Gom 33) with better resistance to wheat blast have been released in Bangladesh for commercial cultivation in 2017. Targeted crosses with increased population sizes were used to obtain superior progeny lines that have high zinc levels in combination with other essential traits. This has resulted in the incorporation of several novel alleles for grain zinc and iron in elite, high-yielding germplasm. High zinc and iron are under quantitative genetic control and further progress is possible as multiple QTL are pyramided in high yielding wheats. High-throughput, non-destructive phenotyping for grain zinc and iron using the X-ray fluorescence (XRF) analysis has facilitated the selection dramatically. Gene discovery and mapping studies leading to the utilization of markers to further improve the breeding efficiency. Rapid adoption of high zinc wheat varieties in South Asia and beyond is expected with the second wave of high zinc wheat lines with superior yield, heat and drought tolerance and resistance to rusts and other foliar diseases.
Institute of Agricultural Sciences, Banaras Hindu University, Varanasi 221005, India
Punam Singh Yadav, Ramesh Chand, Vinod Kumar Mishra, Uttam Kumar, Arun Kumar Joshi
The Sr2 gene has been used extensively in bread wheat improvement for durable stem rust resistance. Interestingly, the resistance of Sr2, associated with the pleotrophic gene Pbc expressed as pseudo-black chaff (PBC), is tightly linked with Yr30/Lr27/Pm genes conferring multiple disease resistance. The linkage map of chromosome 3BS revealed that Sr2 is 0.43cM away from lesion mimic (lm) locus. The RIL population (Yangmai#6 ? Sonalika) of 88 lines including parents where Sonalika carries Sr2 and lm while Yangmai#6 is deficient to both was evaluated for three years (2013-2016). The objective was to determine if this fragment is inherited as one unit and provides resistance to multiple diseases. Twenty four SSR markers distributed between 0.00 to 7.09cM on 3BS covering both Sr2 (5.57cM) and lm locus (6.0cM) were studied in the RIL population. Phenotyping was done for Sr2 associated PBC and lesion mimic along with disease severity for leaf rust, and spot blotch. Positive and significant correlations were observed between leaf stem rust resistance with Sr2 carrying PBC and lm. However, lines with lm either alone or with Sr2 (showing PBC) exhibited spot blotch susceptibility. The reverse situation does not hold not true where genotypes carrying Sr2 alone showed no correlation with spot blotch resistance. This indicates that the Sr2 complex is inherited as a single unit. Use of 24 SSR also suggest that Sr2 and lm loci are tightly linked and inherited together. The co-inheritance of Sr2 and lm ensures the stability and durability of rust resistance. However, the discouraging observation of spot blotch susceptibility due to lm gene suggests a limitation in achieving multiple disease resistance in environments where spot blotch is important. We identified two transgressive segregates in the population showing least expression of lm despite the presence of Sr2 and lm together.
University of the Free State, South Africa
Nelzo Ereful, Botma Visser, Lesley Boyd, Zakkie Pretorius
Adult plant resistance (APR) to stem rust, caused by Puccinia graminis f. sp. tritici (Pgt), is often conferred by multiple minor genes and has the potential to be durable. A preceding project identified two Kenyan wheat lines (W1406 and W6979) from the Genome Resource Unit (Norwich, UK) that exhibit APR to Pgt. The aim of this study was to investigate the APR response to Pgt race PTKST in W1406 and W6979 compared to 37-07, a susceptible control line. Histological investigation of inoculated flag leaf sheaths indicated a significant and quantifiable decrease in Pgt colony size in the APR lines at 120 hours post inoculation (hpi). Molecular analysis supported the observed fungal biomass decrease in the APR lines at 120 hpi. RNAseq analysis identified 169 transcripts differentially expressed in W1406 and 166 transcripts in W6979 when comparing 24 and 72 hpi to 0 hpi. In W1406 transcripts encoding putative pectinesterases, lipid-transfer proteins and leucine-rich repeat-like proteins were induced at 72 hpi. In W6979 only a corresponding putative pectinesterase encoding transcript was identified. Although the induced defence response in the two APR lines exhibited some dissimilarity, it potentially involves cell wall modification in both lines. Two independent sets of peroxidases were induced at 24 and 72 hpi in both lines, suggesting independent signalling events. Expression analysis suggests the occurrence of two phases of gene expression, one at 24 hpi and another at 72 hpi; the latter seeming to correspond to the inhibition of Pgt growth, manifesting as the observed APR phenotype.
Kenya Agricultural and Livestock Research Organization
Hanningtone,Wanga, Phelister, Kinyanjui, Sridhar, Bhavani, Thomas, Fetch, , , , , , , , , , , , , , , , , , , , , ,
In 2016 rust surveys were carried out in all the four key wheat growing regions: South Rift (June, July), Mount Kenya (July), North Rift (September) and Central Rift (part of August and September). A total of 304 farms were sampled. Stem rust was detected in 235 (78.3%), yellow rust in twenty-eight (9.3%) and leaf rust in fourteen (4.7%) of the farms. Stem and yellow rust were detected in all the wheat growing regions while leaf rust was detected in South, North and Central Rift. Stem rust infection ranged from TR to 90S with maximum infection in Central Rift (88.3%), Mt. Kenya region (80.3%); South Rift (76.5%) and North Rift (72.4%). Yellow rust infection ranged TR to 60S with maximum infection in Central Rift (16.7%); North Rift(13.3 %) and minimum infection in South Rift( 4.9%),) and Mt. Kenya region ( 1.7%). Leaf rust infection ranged from trace to 50S with maximum infection in North Rift (10.2%) minimum infection in Central Rift (3.3 %) and South Rift (1.2%). Fifty percent of the eight previously released wheat varieties are now susceptible to the Ug99 race. Race analysis results from AAFC Canada suggested the presence of TTKSK which was dominating in North Rift and TTKSK, TTKST and TTTTF were dominant in the screening nursery at Njoro. Yellow rust in the region has increased in the current year owing to the incursion of a probable new race AF2012 which has resulted in increased disease severity on varieties and materials tested in the International nurseries at KALRO, Njoro.
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.
Department of Agroecology, Aarhus University, Denmark
Chris Khadgi,S?rensen, Annemarie Fejer, Justesen, Mogens St?vring, Hovm?ller, , , , , , , , , , , , , , , , , , , , , , , ,
Wheat yellow (stripe) rust is a recurrent problem throughout the world, and resistant varieties are an efficient means of managing the disease. Therefore, characterization of diverse sources of resistance is of prime importance for wheat breeding. The objective of the study was to investigate variation in host response in incompatible interactions conferred by different R-genes. Epifluorescence and confocal microscopic methods were utilized for histopathological investigation of six yellow rust R-genes (Yr1, Yr5, Yr6, Yr15, Yr17 and Yr27) in Avocet S background, with Avocet S as the control. Fungal colony size and area of hypersensitive response (HR) were assessed for each interaction at 4, 8 and 16 days post inoculation (dpi). The pattern for Avocet Yr15 was distinct, because HR arrested the pathogen very early and rapidly restricted pathogen growth. Avocet Yr1 and Avocet Yr5 showed a less rapid HR and restriction of pathogen growth, but most colonies were completely surrounded by HR at 8 and 16 dpi. In Avocet Yr6 the size of colonies and the extent of HR were highly variable with continuous change up to 16 dpi. More extensive pathogen growth was observed in Avocet Yr17 and Avocet Yr27, where HR induction was delayed, resulting in large intermingled colonies at 16 dpi. All interactions were clearly different from the susceptible control. Thus each R-gene produced a different temporal and spatial distribution of fungal colonies and HR response. Colony size distributions and HR response patterns are potential parameters for characterization of host resistances with different modes of action in wheat against Pst. The results also expand our comprehension of host resistance in wheat against P. striiformis.
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.