Xiue Wang

Serine/threonine kinase gene Stpk-V, a key member of powdery mildew resistance gene Pm21, confers powdery mildew resistance in wheat

Powdery mildew resistance gene Pm21, located on the chromosome 6V short arm of Haynaldia villosa and transferred to wheat as a 6VS·6AL translocation (T6VS·6AL), confers durable and broad-spectrum resistance to wheat powdery mildew. Pm21 has become a key gene resource for powdery mildew resistance breeding all over the world. In China, 12 wheat varieties containing Pm21 have been planted on more than 3.4 million hectares since 2002. Pm21 has been intractable to molecular genetic mapping because the 6VS does not pair and recombine with the 6AS. Moreover, all known accessions of H. villosa are immune to powdery mildew fungus. Pm21 is still defined by cytogenetics as a locus. In the present study, a putative serine and threonine protein kinase gene Stpk-V was cloned and characterized with an integrative strategy of molecular and cytogenetic techniques. Stpk-V is located on the Pm21 locus. The results of a single cell transient expression assay showed that Stpk-V could decrease the haustorium index dramatically. After the Stpk-V was transformed into a susceptible wheat variety Yangmai158, the characterized transgenic plants showed high and broad-spectrum powdery mildew resistance similar to T6VS·6AL. Silencing of the Stpk-V by virus-induced gene silencing in both T6VS·6AL and H. villosa resulted in their increased susceptibility. Stpk-V could be induced by Bgt and exogenous H2O2, but it also mediated the increase of endogenous H2O2, leading to cell death and plant resistance when the plant was attacked by Bgt.

QTLs for fusarium head blight response in a wheat DH population of Wangshuibai/Alondra's'

SummaryA doubled haploid (DH) wheat population derived from the cross Wangshuibai/Alondra‘s’ was developed through chromosome doubling of haploids generated by anther culture of hybrids. Fusarium head blight (FHB) was evaluated for three years from 2001 to 2003 in Jianyang, Fujian Province, China, where epidemics of FHB have been consistently severe. After 307 pairs of simple sequence repeat (SSR) primers were screened, 110 pairs were polymorphic between Wangshuibai and Alondra`s’, and used to construct a genetic linkage map for detection of quantitative trait loci (QTLs). A stable QTL for low FHB severity was detected on chromosomes 3B over all three years, and QTLs on chromosomes 5B, 2D, and 7A were detected over two years. Additional QTLs on chromosomes 3A, 3D, 4B, 5A, 5D, 6B and 7B showed marginal significance in only one year. Six QTLs were detected when phenotypic data from three years were combined. In addition, significant additive-by-additive epistasis was detected for a QTL on 6A although its additive effect was not significant. Additive effects (A) and additive-by-additive epistasis (AA) explained a major portion of the phenotypic variation (76.5%) for FHB response. Xgwm533-3B and Xgwm335-5B were the closest markers to QTLs, and have potential to be used as selectable markers for marker-assisted selection (MAS) in wheat breeding programs.

Development and characterization of wheat- Leymus racemosus translocation lines with resistance to Fusarium Head Blight

Wheat scab (Fusarium Head Blight, FHB) is a destructive disease in the warm and humid wheat-growing areas of the world. Finding diverse sources of FHB resistance is critical for genetic diversity of resistance for wheat breeding programs. Leymus racemosus is a wild perennial relative of wheat and is highly resistant to FHB. Three wheat- L. racemosus disomic addition (DA) lines DA5Lr#1, DA7Lr#1 and DALr.7 resistant to FHB were used to develop wheat- L.racemosus translocation lines through irradiation and gametocidal gene-induced chromosome breakage. A total of nine wheat-alien translocation lines with wheat scab resistance were identified by chromosome C-banding, GISH, telosomic pairing and RFLP analyses. In line NAU614, the long arm of 5Lr#1 was translocated to wheat chromosome 6B. Four lines, NAU601, NAU615, NAU617, and NAU635, had a part of the short arm of 7Lr#1 transferred to different wheat chromosomes. Four other lines, NAU611, NAU634, NAU633, and NAU618, contained translocations involving Leymus chromosome Lr.7 and different wheat chromosomes. The resistance level of the translocation lines with a single alien chromosome segment was higher than the susceptible wheat parent Chinese Spring but lower than the alien resistant parent L. racemosus. At least three resistance genes in L. racemosus were identified. One was located on chromosome Lr.7, and two could be assigned to the long arm of 5Lr#1 and the short arm of 7Lr#1.

Disease Resistance Gene Analogs (RGAs) in Plants.

Plants have developed effective mechanisms to recognize and respond to infections caused by pathogens. Plant resistance gene analogs (RGAs), as resistance (R) gene candidates, have conserved domains and motifs that play specific roles in pathogens’ resistance. Well-known RGAs are nucleotide binding site leucine rich repeats, receptor like kinases, and receptor like proteins. Others include pentatricopeptide repeats and apoplastic peroxidases. RGAs can be detected using bioinformatics tools based on their conserved structural features. Thousands of RGAs have been identified from sequenced plant genomes. High-density genome-wide RGA genetic maps are useful for designing diagnostic markers and identifying quantitative trait loci (QTL) or markers associated with plant disease resistance. This review focuses on recent advances in structures and mechanisms of RGAs, and their identification from sequenced genomes using bioinformatics tools. Applications in enhancing fine mapping and cloning of plant disease resistance genes are also discussed.

Development and characterization of a Triticum aestivum-Haynaldia villosa translocation line T4VS⋅4DL conferring resistance to wheat spindle streak mosaic virus

Previous studies showed that a T. aestivum-H. villosa disomic substitution line DS4V(4D) showed a high level of resistance to wheat spindle streak mosaic virus (WSSMV). By crossing DS4V(4D) with the common wheat variety Yangmai #5, plants were obtained that were double monosomic for chromosomes 4V and 4D. Univalents are prone to misdivision at the centromere, and fusion of the derived telocentric chromosomes leads to the production of Robertsonian whole-arm translocations. We screened the progenies of such double monosomic plants by C-banding and genomic in situ hybridization and identified one compensating translocation where the short arm of 4V was translocated to the long arm of 4D of wheat, T4VS⋅4DL. RFLP analysis using the group-4 specific probe BCD110 was used to confirm the translocation. The T4VS⋅4DL translocation stock, accessioned as NAU413, is highly resistant to WSSMV and is also of good agronomic type. The WSSMV resistance gene located on 4VS was designated Wss1.

Effects of the 6VS.6AL translocation on agronomic traits and dough properties of wheat

Nineteen common wheat cultivars and advanced lines carrying a 6VS.6AL translocation and five parents were sown at two locations in Jiangsu in 2004–05 season to assess the effects of the translocation on grain yield and dough properties. In general, there were no significant differences between 6VS.6AL lines and their recurrent parents in agronomic, mixograph and starch pasting traits, including grain yield, grains/spike, grain weight/spike, mixing time and peak viscosity. 6VS.6AL lines showed slightly but significantly higher thousand-kernel weight and plant height, and small negative effects on test weight, flour yield and flour colour. However, significant variation occurred for all traits among sister lines from the same cross, indicating that additional selection could lead to further improvement. It was concluded that the 6VS.6AL translocation can be used in wheat breeding programs as a donor of resistance to powdery mildew with no obvious undesirable effects on agronomic and quality traits.

Assessment of genetic diversity of Yunnan, Tibetan, and Xinjiang wheat using SSR markers.

A total of 206 SSR (Simple Sequence Repeats) primer pairs were used to detect genetic diversity in 52 accessions of three unique wheat varieties of western China. A total of 488, 472, and 308 allelic variants were detected in 31 Yunnan, 15 Tibetan and 6 Xinjiang wheat accessions with an average of PIC values 0.2764, 0.3082, and 0.1944, respectively. Substantial differences in allelic polymorphisms were detected by SSR markers in all the 21 chromosomes, the 7 homoeologous groups, and the three genomes (A, B, and D) in Yunnan, Tibetan, and Xinjiang wheat. The highest and lowest allelic polymorphisms in all the 21 chromosomes were observed in 3B and 1D chromosomes, respectively. The lowest and highest allelic polymorphisms among the seven homoeologous groups was observed in 6 and 3 homoeologous groups, respectively. Among the three genomes, B genome showed the highest, A the intermediate, and D the lowest allelic polymorphism. The genetic distance (GD) indexes within Yunnan, Tibetan, and Xinjiang wheat, and between different wheat types were calculated. The GD value was found to be much higher within Yunnan and Tibetan wheat than within Xinjiang wheat, but the GD value between Yunnan and Tibetan wheat was lower than those between Yunnan and Xinjiang wheat, and between Tibetan and Xinjiang wheat. The cluster analysis indicated a closer relationship between Yunnan and Tibetan wheat than that between Yunnan and Xinjiang wheat or between Tibetan and Xinjiang wheat.

Molecular cytogenetic characterization of Roegneria ciliaris chromosome additions in common wheat

Abstract The development of alien addition lines is important both for transferring useful genes from related species into common wheat and for studying the relationship between alien chromosomes and those of wheat. Roegneria ciliaris (2n=4x=28, ScScYcYc) is reported to be a potential source of resistance to wheat scab, which may be useful in wheat improvement. The amphiploid common wheat-R. ciliaris and BC1F7 or BC2F6 derivatives were screened by C-banding, genomic in situ hybridization (GISH), fluorescent in situ hybridization (FISH) and restriction fragment length polymorphism (RFLP) for the presence of R. ciliaris chromatin introgressed into wheat. Six lines were identified as disomic chromosome additions (DA), one as a ditelosomic addition (Dt), two as double disomic additions (dDA) and one as a monosomic chromosome addition (MA). RFLP analysis using wheat homoeologous group-specific clones indicated that the R. ciliaris chromosomes involved in these lines belong to groups 1, 2, 3, 5 and 7. The genomic affinities of the added R. ciliaris chromosomes were determined by FISH analysis using the repetitive sequence pCbTaq4.14 as a probe. These data suggest that the R. ciliaris chromosomes in five lines belong to the Sc genome. Based on the molecular cytogenetic data, the lines are designated as DA2Sc#1, Dt2Sc#1L, DA3Sc#1, dDA1Sc#2+5Yc#1, DA5Yc#1, DA7Sc#1, DA7Yc#1 and MA?Yc#1. Based on the present and previous work, 8 of the 14 chromosomes of R. ciliaris have been transferred into wheat.

RLP1.1, a novel wheat receptor-like protein gene, is involved in the defence response against Puccinia striiformis f. sp. tritici

Stripe rust, caused by Puccinia striiformis f. sp. tritici (Pst), is one of the most serious diseases of wheat; therefore, exploring effective resistance-related genes is critical for breeding and studying resistance mechanisms. However, only a few stripe rust resistance genes and defence-related genes have been cloned. Moreover, transgenic wheat with enhanced stripe rust resistance has rarely been reported. Receptor-like proteins (RLPs) are known to be involved in defence and developmental pathways. In this research, a novel RLP gene TaRLP1.1 was characterized as an important stripe rust defence gene. TaRLP1.1 was screened by GeneChip and was found to be induced by Pst specifically in the resistant variety. Knock down of TaRLP1.1 in the stripe rust-resistant plants resulted in increased susceptibility to Pst, and phenolic autofluorogen accumulation at the pathogen–host interaction sites, usually correlated with the hypersensitive response, was decreased dramatically. However, when the TaRLP1.1 gene was transformed into the susceptible wheat variety Yangmai158, the transgenic plants showed highly increased resistance to Pst, and the hypersensitive response was enhanced at the infection sites. Meanwhile, the expression of pathogenesis-related genes decreased in the TaRLP1.1-silenced plants and increased in the TaRLP1.1-overexpressing plants. Thus, it was proposed that TaRLP1.1 greatly contributed to the hypersensitive response during the pathogen–host interaction. Along with the functional analysis, an evolutionary study of the TaRLP1 family was performed. Characterization of TaRLP1.1 may facilitate breeding for stripe rust resistance and better understanding of the evolution of the RLP genes in wheat.

Molecular and cytogenetic characterization of a small alien-segment translocation line carrying the softness genes of Haynaldia villosa

The wheat-alien small segment translocation (SAST) lines carrying the beneficial genes from wild species are useful genetic stocks for wheat improvement. In this study, to introduce the grain hardness-related genes of Haynaldia villosa (L.) Schur. into common wheat (Triticum aesitivum L.), the mature female gametes of whole-arm wheat--H. villosa translocation line T5VS·5DL was irradiated by 60CO-γ ray to develop SAST lines involving 5VS. Among the BC2F2 population, six homozygous SAST lines with different fragment sizes of 5VS were identified by GISH, and the exact fragment sizes were further defined using four 5VS-specific markers and four Ha gene-based markers. The results showed that five lines (NAU5VS-1 to NAU5VS-5) carried the softness gene Dina/Dinb of H. villosa, and that NAU5VS-5 had the smallest alien translocation segment, identified to be a 5VS-6AS·6AL terminal translocation. The translocation chromosome 5VS-6AS·6AL was proved to be stably inherited to the successive generations. In the BC3F2 generation, the individuals having the homozygous 5VS-6AS·6AL translocation chromosomes all showed soft grain texture, with an approximately 50% reduction in the SKCS hardness index compared with that of their backcrossing parent. Both the 5VS-6AS·6AL translocation line and the molecular markers developed in this study will be valuable in wheat breeding for soft grain quality improvement.