Browsing by Author "Huang, Li"

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Cellular and molecular characterization of a stem rust resistance locus on wheat chromosome 7AL
(2016-12) Pujol, Vincent; Robles, Jose; Wang, Penghao; Taylor, Jen; Zheng, Peng; Huang, Li; Tabe, Linda; Lagudah, Evans
"BACKGROUND: Wheat stem rust, caused by Puccinia graminis f. sp. tritici, is a major wheat disease which is mainly controlled through the release of resistant cultivars containing one or several resistance genes. Considerable effort has been put into the discovery of new resistance genes, but knowledge of their mechanisms of action is often lacking. In this study, the mechanism of resistance conferred by a recently discovered stem rust resistance locus on wheat chromosome 7AL was investigated through microscopic observations and RNA-sequencing, using the susceptible line Columbus and the independent, backcrossed, resistant lines Columbus-NS765 and Columbus-NS766. RESULTS: Microscopic observations of infected leaves revealed that the resistance conferred by the 7AL resistance locus was initiated 2 days post-inoculation, upon the fungus entry into the plant through the stoma. Resistance was manifested by death of guard and epidermal cells adjacent to an infection site. Occasionally, similar observations were made in the susceptible line, suggesting that the resistance response was the same in all genotypes, but enhanced in the resistant lines. Transcriptomic analysis, combined with assignment of genes to wheat chromosomes, revealed a disproportionately high number of differentially expressed genes were located on chromosomes 7AL and 6A. A number of genes annotated as cysteine-rich receptor-like kinases were located on chromosome 7AL. Closer investigation indicated that the encoded proteins were in fact putative receptor-like cytoplasmic kinases. One of the putative RLCK genes contained a SNP marker previously shown to co-segregate with the 7AL resistance locus. The results also indicated the presence of a large introgression on chromosome 6A in both resistant lines, but whether it has any role in the resistance response is unclear. CONCLUSIONS: This study represents the first investigation on the resistance mechanism conferred by the wheat 7AL stem rust resistance locus. The resistance response was associated with pre-haustorial cell death, and the transcriptome analysis suggested putative receptor-like cytoplasmic kinases as candidate resistance genes for further investigation."
Mapping quantitative trait loci for seed traits in Camelina sativa
(2019-09-09) King, Kevin; Huang, Li; Kang, Jinling; Lu, Chaofu
Camelina (Camelina sativa L. Crantz) is an oilseed crop that has great potential to provide sustainable feedstock for biofuel production and to improve dryland agriculture. A major breeding objective for camelina is to increase seed size and oil content. Understanding the genetics behind variations of seed size and associated traits such as oil content would help breeders develop varieties of increased oil yield that are more robust, easier to plant and harvest, and better for oil processing. In this study, we developed a recombinant inbred population derived from the two camelina accessions, Suneson and Pryzeth, with contrasting traits, especially seed size and oil content. Using 189 lines, a genetic map was constructed containing 2376 single nucleotide polymorphism markers spanning 2034.6 cM of 20 linkage groups with an average density of 1.5 cM per locus. Field trials were conducted for 2 years (2017 and 2018) in two environments (dryland and irrigated) in Bozeman, Montana. The results revealed important correlations of seed size with other associated traits such as oil content, pod size and seed number per pod. Significant QTLs were also discovered for these traits. The results of this study are the first step to isolate genes controlling seed development and oil accumulation and to develop advanced varieties of camelina better adapted to modern agriculture by marker-assisted breeding.
Molecular Mechanisms of the Co-Evolution of Wheat and Rust Pathogens
(MDPI, 2023-04) Annan, Emmanuel N.; Huang, Li
Wheat (Triticum spp.) is a cereal crop domesticated >8000 years ago and the second-most-consumed food crop nowadays. Ever since mankind has written records, cereal rust diseases have been a painful awareness in antiquity documented in the Old Testament (about 750 B.C.). The pathogen causing the wheat stem rust disease is among the first identified plant pathogens in the 1700s, suggesting that wheat and rust pathogens have co-existed for thousands of years. With advanced molecular technologies, wheat and rust genomes have been sequenced, and interactions between the host and the rust pathogens have been extensively studied at molecular levels. In this review, we summarized the research at the molecular level and organized the findings based on the pathogenesis steps of germination, penetration, haustorial formation, and colonization of the rusts to present the molecular mechanisms of the co-evolution of wheat and rust pathogens.
Optimized High Throughput Ascochyta Blight Screening Protocols and Immunity to A. pisi in Pea
(MDPI AG, 2023-03) Annan, Emmanuel N.; Nyamesorto, Bernard; Yan, Qing; McPhee, Kevin; Huang, Li
Ascochyta blight (AB) is a destructive disease of the field pea (Pisum sativum L.) caused by necrotrophic fungal pathogens known as the AB-disease complex. To identify resistant individuals to assist AB resistance breeding, low-cost, high throughput, and reliable protocols for AB screening are needed. We tested and optimized three protocols to determine the optimum type of pathogen inoculum, the optimal development stage for host inoculation, and the timing of inoculation for detached-leaf assays. We found that different plant development stages do not affect AB infection type on peas, but the timing of inoculation affects the infection type of detached leaves due to wound-induced host defense response. After screening nine pea cultivars, we discovered that cultivar Fallon was immune to A. pisi but not to A. pinodes or the mixture of the two species. Our findings suggest that AB screening can be done with any of the three protocols. A whole-plant inoculation assay is necessary for identifying resistance to stem/node infection. Pathogen inoculation must be completed within 1.5 h post-detachment to avoid false positives of resistance for detach-leaf assays. It is essential to use a purified single-species inoculum for resistant resource screenings to identify the host resistance to each single species.
Stem rust resistance in wheat is suppressedby a subunit of the mediator complex
(2020-02) Hiebert, Colin W.; Moscou, Matthew J.; Hewitt, Tim; Steuernagel, Burkhard; Hernandez-Pinzon, Inma; Green, Phon; Pujol, Vincent; Zhang, Peng; Rouse, Matthew N.; Jin, Yue; McIntosh, Robert A.; Upadhyaya, Narayana; Zhang, Jianping; Bhavani, Sridhar; Vrana, Jan; Karafiatova, Miroslava; Huang, Li; Fetch, Tom; Dolezel, Jaroslav; Wulff, Brande B. H.; Lagudah, Evans; Spielmeyer, Wolfgang
Stem rust is an important disease of wheat that can be controlled using resistance genes. The gene SuSr-D1 identified in cultivar ‘Canthatch’ suppresses stem rust resistance. SuSr-D1 mutants are resistant to several races of stem rust that are virulent on wild-type plants. Here we identify SuSr-D1 by sequencing flow-sorted chromosomes, mutagenesis, and map-based cloning. The gene encodes Med15, a subunit of the Mediator Complex, a conserved protein complex in eukaryotes that regulates expression of protein-coding genes. Nonsense mutations in Med15b.D result in expression of stem rust resistance. Time-course RNAseq analysis show a significant reduction or complete loss of differential gene expression at 24 h post inoculation in med15b.D mutants, suggesting that transcriptional reprogramming at this time point is not required for immunity to stem rust. Suppression is a common phenomenon and this study provides novel insight into suppression of rust resistance in wheat.
TaRar1 Is Involved in Wheat Defense against Stripe Rust Pathogen Mediated by YrSu
(2017-02) Wang, Xiaojing; Wang, Yaru; Liu, Peng; Ding, Tan; Mu, Xiaoqian; Liu, Xiping; Wang, Xiaojie; Zhao, Mengxin; Huai, Baoyu; Huang, Li; Kang, Zhensheng
RAR1 is a eukaryotic zinc-binding protein first identified as required for race-specific resistance to powdery mildew in barley. To study the function of TaRAR1 involvement in wheat (Triticum aestivum L.) defense against the infection of stripe rust pathogen Puccinia striiformis f. sp. tritici (Pst), we identified and cloned three wheat homeologous genes highly similar to the barley HvRar1, designated as TaRar1-2A, TaRar1-2B, and TaRar1-2D. The three TaRAR1 proteins all contain two conserved cysteine-and histidine-rich domains (CHORD-I and -II) shared by known RAR1-like proteins. Characterization of TaRar1 expression revealed that the expression was tissue-specific and up-regulated in wheat during stripe rust infection. Moreover, the transcription of TaRar1 was induced by methyl jasmonate, ethylene, and abscisic acid hormones. The same results were observed with drought and wound treatments. After TaRar1 was silenced in wheat cultivar Suwon11 containing the stripe rust resistance gene YrSu, the endogenous salicylic acid (SA) level, the hydrogen peroxide (H2O2) accumulation and the degree of hypersensitive response (HR) were significantly decreased, and the resistance to the avirulent pathotype of stripe rust was compromised. Meanwhile, the expression of catalase, an enzyme required for H2O2-scavenging, was up-regulated. Taken together, we concluded that TaRar1 is involved in wheat defense against stripe rust mediated by YrSu, and the defense was through SA to influence reactive oxygen species accumulation and HR.
A transcriptomic-guided strategy used in identification of a wheat rust pathogen target and modification of the target enhanced host resistance to rust pathogens
(Frontiers Media SA, 2022-09) Nyamesorto, Bernard; Zhang, Hongtao; Rouse, Matthew; Wang, Meinan; Chen, Xianming; Huang, Li
Transcriptional reprogramming is an essential feature of plant immunity and is governed by transcription factors (TFs) and co-regulatory proteins associated with discrete transcriptional complexes. On the other hand, effector proteins from pathogens have been shown to hijack these vast repertoires of plant TFs. Our current knowledge of host genes' role (including TFs) involved in pathogen colonization is based on research employing model plants such as Arabidopsis and rice with minimal efforts in wheat rust interactions. In this study, we begun the research by identifying wheat genes that benefit rust pathogens during infection and editing those genes to provide wheat with passive resistance to rust. We identified the wheat MYC4 transcription factor (TF) located on chromosome 1B (TaMYC4-1B) as a rust pathogen target. The gene was upregulated only in susceptible lines in the presence of the pathogens. Down-regulation of TaMYC4-1B using barley stripe mosaic virus-induced gene silencing (BSMV-VIGS) in the susceptible cultivar Chinese Spring enhanced its resistance to the stem rust pathogen. Knockout of the TaMYC4-1BL in Cadenza rendered new resistance to races of stem, leaf, and stripe rust pathogens. We developed new germplasm in wheat via modifications of the wheat TaMYC4−1BL transcription factor.
Wheat mutant MNR220 delays haustoria formation during leaf rust pathogenesis at the seedling stage
(2016-09) Talajoor, Mina R.; Wang, Xiaojing; Zhang, Hongtao; Zhang, Peng; Bhavani, Sridhar; Tabe, Linda; Lagudah, Evans; Huang, Li
This study reports further characterization of the defence mechanism of an EMS-induced mutant (MNR220) that displays enhanced resistance to leaf rust, stem rust, stripe rust and powdery mildew diseases. The broad-spectrum resistance of the mutant was characterized by a microscopy study and additional race screenings of two rust pathogens. The mutant displayed less disease severity in response to all the races tested, when compared with the wild-type. The microscopy study of the pathogenesis of a leaf rust race PBJJG on the mutant revealed that the resistance was first evident at the pre-haustorial stage; formation of haustoria was significantly delayed or inhibited in the mutant compared with the wild-type. The mutant displayed microscopic level of cell death even in the absence of any pathogens. These findings suggest that MNR220 mediates a spontaneous cell death that delays the infection process by delaying haustoria formation of the pathogen.