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Application of Yr52 Gene in Wheat Improvement for Stripe Rust Resistance
Scientia Agricultura Sinica 2022, 55(11): 2077-2091
Published: 01 June 2022
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【Objective】

The objective of this study is to evaluate the application value of the high temperature adult plant (HTAP) resistance gene Yr52 in wheat production for improving stripe rust resistance. And wheat lines with good agronomic characters and high disease resistance were developed and selected. It laid a foundation for making full use of the existing HTAP resistance resources and improving the yield related traits.

【Method】

The stripe rust resistance gene Yr52 was introgressed to Lunxuan 987 (LX987), Bainong Aikang 58 (AK58) and Han 6172 (H6172) by backcrossing and self-crossing combined with marker-assisted selection breeding. Adult-plant resistance of donor parent, receptor cultivars and their progeny lines were evaluated in the disease nursery fields at Mianyang, Sichuan and Yangling, Shaanxi by mixed endemic physiological races CYR32, CYR33 and CYR34. In comparison to the Chinese Spring reference genome, the flanking SSR markers Xcfa2040 (6.8 cm-Yr52) and Xbarc182 (1.2 cm-Yr52) of Yr52 were combined to search for markers of 35K SNP chip in the physical interval of target genes, and developed into derived cleaved amplified polymorphic sequences (dCAPS) and kompetitive allele specific PCR (KASP) markers. The resistance gene Yr52 was detected in BC2F5:6 progeny lines.

【Result】

The evaluation of adult plant resistance and agronomic traits indicated that nineteen BC2F5:6 lines with LX987 background was obtained: Among of them, 11 were high resistance (IT=0-3, DS=1%-20%), 8 were moderate resistance (IT=4-6, DS=15%-30%), the average thousand kernel weight (TKW), kernels per spike (KPS), productive tiller number per line (PTN), plant height (PH) and spike length (SL) was 45.33 g, 46, 7, 113.26 cm and 10.05 cm, respectively. Four BC2F5:6 families with AK58 background: all showed high resistance (IT=0-3, DS=5%-25%); the average TKW, KPS, PTN, PH and SL were 44.67 g, 48, 7, 96.54 cm and 10.17 cm, respectively. Five BC2F5:6 lineages with H6172 background showed high resistance to stripe rust (IT=0-3, DS=5%-20%). The average TKW, KPS, PTN, PH and SL were 43.74 g, 49, 8, 109.72 cm and 10.06 cm, respectively. The detection rate of three simple sequence repeat (SSR) molecular markers Xbarc182, Xcfa2040 and Xwmc557 linked to Yr52, in offspring population were 78.57%, 66.67% and 66.67%, respectively. One dCAPS marker Xdcaps-Yr52-1 and one KASP marker Xkasp-Yr52-1 were successfully developed, and the detection rates were 73.68% and 41.67%, respectively. The agronomic traits of lines with high (IT=0-3) and medium (IT=4-6) resistance levels were compared. The results showed that the average TKW (P>0.05), PTN (P>0.05) and KPS (P<0.05) of lines with IT=0-3 were higher than those of families with moderate resistance level lines (IT=4-6). Five lines with disease resistance and stable agronomic traits were selected by evaluated of PH=80-105 cm, PTN≥6, KPS≥45, TKW≥42 g, SL≥8 cm.

【Conclusion】

Yr52 was found to be resistant to all of the present predominant races in the adult plant stage. After introgression Yr52 into the main susceptible Chinese wheat varieties, the progeny lines with good disease resistance and agronomic characters could be used for breeding resistance varieties with multi-gene polymerization, it is enriching for the diversity of disease resistance genes and achieving durable utilization. The development of molecular markers will facilitate detect the utilization of Yr52 gene in resistance identification of germplasm in the future.

Issue
Population Genetic Analysis of Puccinia striiformis tritici in Main Winter-Increasing Areas Based on Virulent Phenotypes and Genotypes
Scientia Agricultura Sinica 2023, 56(14): 2629-2642
Published: 16 July 2023
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【Objective】

To clarify the virulence structure and genetic diversity of Pst populations in the major winter-increasing areas of China, and to provide reference for the prevention and control of Pst and the rational layout of wheat resistance genes in the winter-increasing areas and the wheat production in Huang-huai-hai.

【Method】

A total of 148 Pst isolates were collected and isolated from the major winter-increasing areas such as Sichuan Basin, Hubei and southern Henan, and the virulence phenotype was identified by using Chinese differentials and single-gene lines, and 17 pairs of KASP-SNP primers were used to mark the isolates and complete the genotype analysis.

【Result】

Based on the Chinese differentials, 14 known races and 63 unknown pathotypes were identified, among which CYR34 (16.2%), G22-14 (12.2%), CYR32 (6.8%), CYR33 (5.4%) were the dominant races (pathotypes); based on the single-gene lines, 113 races (pathotypes) were identified, among which race1 (7.4%), race2 (3.4%), race3 (3.4%) were the dominant races (pathotypes). The Guinong 22 group was the largest epidemic group of Pst population in China’s winter-increasing area, and all tested Pst isolates did not infect single-gene lines varieties carrying Yr5 and Yr15. The virulence phenotype and genotype of CYR34 and G22-14 showed diversification by single-gene lines virulence identification and molecular marker, indicating that there was high differentiation within these two dominant races. The clustering based on the virulence data of two sets of differentials showed that the Pst populations in Sichuan Basin and southern Hubei were similar, while the Pst populations in northwestern Hubei and southern Henan were similar; the genetic clustering based on KASP-SNP molecular data showed that there was genotype differentiation between the Pst populations in Sichuan Basin, southern Hubei and northwestern Hubei, southern Henan; Structure analysis showed that Sichuan Basin, southern Hubei population mainly had two genetic backgrounds, northwestern Hubei, southern Henan population mainly had one genetic background; population genetic differentiation analysis showed that Sichuan Basin Pst population and southern Henan Pst population had the largest Fst value, which was 0.118, with the largest genetic difference and obvious genetic differentiation; northwestern Hubei population and southern Henan population had the smallest degree of genetic differentiation, Fst value was 0.010; gene flow analysis obtained Nm value between northwestern Hubei population and southern Henan population was 25.236, Nm>4, there was a high-level gene flow between them, northwestern Hubei and southern Henan population and Sichuan Basin population had Nm values of 2.923 and 1.864 respectively, both had a low-level gene flow; genetic diversity analysis results showed that Sichuan Basin, southern Hubei region Pst population had a high-level of genetic diversity, northwestern Hubei, southern Henan Pst population had a low-level of genetic diversity. The above conclusions all support that Sichuan Basin, southern Hubei population has genetic differentiation with northwestern Hubei, southern Henan population.

【Conclusion】

Single-gene lines can accurately identify Chinese Pst races; Pst populations in China’s major winter-increasing areas have different sources.

Issue
Molecular evidence of the west-to-east dispersal of Puccinia striiformis f. sp. tritici in central Shaanxi and the migration of the inoculum from Gansu
Journal of Integrative Agriculture (JIA) 2025, 24(6): 2251-2265
Published: 21 October 2023
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Downloads:10

Wheat stripe rust, caused by Puccinia striiformis f. sp. tritici (Pst), is an airborne disease. In China, it frequently develops initially in central Shaanxi and southwestern Gansu, and from there, inoculum spreads to the eastern wheat production regions. Field investigations have suggested that Pst could spread from the west to the east within central Shaanxi and that Gansu could serve as the inoculum source for central Shaanxi, but there is no direct evidence for this hypothetical dispersal route. In the current study, 321 Pst isolates collected from central Shaanxi and Gansu in the 2019–2020 and 2020–2021 winter wheat cropping seasons were genotyped using 23 pairs of KASP-SNP markers. The dispersion among subpopulations was analyzed using several approaches, and overall, the populations were found to exhibit high levels of genetic diversity. There was little genetic divergence (0.05>FST>0) within central Shaanxi. However, significant gene flow (Nm>4) driven by wind-oriented dispersal from west (Baoji) to east (Weinan) occurred. There was also gene flow among the 4 Gansu subpopulations of Tianshui, Longnan, Pingliang, and Qingyang. Migration of the pathogen occurred between central Shaanxi and Gansu. Migration from Gansu to central Shaanxi was major compared with that from central Shaanxi to Gansu that was minor. Genetic variation occurred among isolates, instead of among subpopulations and within isolates. Linkage disequilibrium revealed that there was strong genetic recombination in the subpopulations from Gansu and central Shaanxi. Therefore, the present study provides molecular evidence that Pst spread from west to east in central Shaanxi and showed that Gansu (especially Longnan and Tianshui) was one of the major origins of the pathogen inoculum of wheat stripe rust in central Shaanxi. The results revealed the west-to-east transmission route of wheat stripe rust in central Shaanxi, being used to guide integrated management of the disease.

Open Access Research Article Issue
A small knottin-like peptide negatively regulates in wheat to stripe rust resistance during early infection of wheat
The Crop Journal 2023, 11(2): 457-467
Published: 13 October 2022
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Downloads:15

Although Blufensins (Bln) have important functions in the response of plants to biotic stress the precise functioning of Bln in wheat remains largely unknown. Here we isolated a Bln gene (TaBln4) from Suwon 11 infected by Puccinia striiformis f. sp. tritici (Pst). Expression of TaBln4 increased in host plants at the early stage of infection with a virulent Pst race (CYR31) but was unchanged in response to infection by an avirulent race (CYR23). Transcription levels of TaBln4 were also regulated by hormone and abiotic stresses. Expression of TaBln4 in tobacco leaves suppressed Bax-induced programmed cell death. Knockdown of TaBln4 by virus-induced gene silencing inhibited colonization of race CYR31 by increasing the accumulation of H2O2 and formation of hypersensitive responses (HR). Transient overexpression of TaBln4 by a transient overexpression system (BSMV-VOX) increased the susceptibility of wheat to CYR31. Results from bimolecular fluorescence complementation and pull-down assays demonstrated that TaBLN4 interacted with calmodulin. Taken together, our results suggest that TaBln4 negatively regulates resistance in wheat to Pst in a reactive oxygen species (ROS)- and HR-dependent manner.

Open Access Research paper Issue
SNP-based linkage mapping for validation of adult plant stripe rust resistance QTL in common wheat cultivar Chakwal 86
The Crop Journal 2019, 7(2): 176-186
Published: 11 January 2019
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Wheat crops in China are constantly challenged by stripe rust. Deployment of cultivars with diverse resistances is the best strategy to control the disease. A recombinant inbred line (RIL) population derived from a cross between the resistant cultivar Chakwal 86 and susceptible landrace Mingxian 169 was studied in multiple environments to examine the underlying genetics and to identify quantitative trait loci (QTL) for stripe rust resistance. One hundred and twenty-eight RILs were genotyped with wheat 35K SNP array and a genome-wide linkage map with 1480 polymorphic SNP markers, or bins, was constructed. Two major QTL on chromosomes 1BL and 3BS, and one minor QTL on 6BS had significant effects in reducing stripe rust severity. The QTL were validated using composite interval mapping (CIM) and inclusive composite interval mapping (ICIM). These methods explained 59.0%–74.1% of the phenotype variation in disease response. The QTL on chromosome 1BL was confirmed to be Yr29/Lr46 and the one on 3BS was the resistance allele identified in CIMMYT germplasm but was not Yr30/Sr2. The QTL on 6BS probably corresponded to previously known QTL. RILs with combined QTL were more resistant than those with single or no QTL. Kompetitive allele-specific PCR (KASP) assays for the QTL with largest effect QTL on chromosome 3BS were performed on a subset of RILs and 150 unrelated wheat lines. The QTL on 3BS with its linked KASP markers can be used in marker-assisted selection to improve stripe rust resistance in breeding programs.

Open Access Review Issue
Understanding the lifestyles and pathogenicity mechanisms of obligate biotrophic fungi in wheat: The emerging genomics era
The Crop Journal 2018, 6(1): 60-67
Published: 12 December 2017
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Obligate biotrophic fungi cause serious and widespread diseases of crop plants, but are challenging to investigate because they cannot be cultured in vitro. The two economically important groups of biotrophic fungi parasitizing wheat are the rust and powdery mildew pathogens, but their obligate biotrophic lifestyles and pathogenicity mechanisms are not well understood at the molecular level. With the advent of next generation sequencing technology, increasing numbers of pathogen genomes are becoming available. Research in plant pathology has entered a new genomics era. This review summarizes recent progress in understanding the biology and pathogenesis of biotrophic fungal pathogens attacking wheat based on pathogen genomics. We particularly focus on the three wheat rust and the powdery mildew fungi in regard to genome sequencing, avirulence gene cloning, effector discovery, and pathogenomics. We predict that coordinated study of both wheat and its pathogens should reveal new insights in biotrophic adaptation, pathogenicity mechanisms, and population dynamics of these fungi that will assist in development of new strategies for breeding wheat varieties with durable resistance.

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