Wheat stem rust, caused by Puccinia graminis f. sp. tritici(Pgt), is a potentially devastating fungal disease of wheat worldwide. The present study was to evaluate the resistance of 42 wheat monogenic lines with known...Wheat stem rust, caused by Puccinia graminis f. sp. tritici(Pgt), is a potentially devastating fungal disease of wheat worldwide. The present study was to evaluate the resistance of 42 wheat monogenic lines with known stem rust resistance(Sr) genes and 69 wheat cultivars to three new Pgt races(34C0MRGQM, 34C3MKGQM, and 34C6MTGSM)identified from aeciospores at the seedling and adult-plant stages. The phenotyping results revealed that monogenic lines harboring resistance genes Sr9e, Sr17, Sr21, Sr22, Sr26, Sr30, Sr31, Sr33, Sr35, Sr36, Sr37, Sr38, Sr47, SrTmp,and SrTt3 were effectively resistant to all three Pgt races at the seedling and adult-plant stages. In contrast, monogenic lines containing Sr5, Sr6, Sr7b, Sr9a, Sr9d, Sr9f, Sr9g, Sr9b, Sr16, Sr24, Sr28, and Sr39 were highly susceptible to these races at both seedling and adult-plant stages. The other lines with Sr8a, Sr10, Sr11, Sr13, Sr14, Sr15, Sr18, Sr20,Sr19, Sr23, Sr25, Sr27, Sr29, Sr32, and Sr34, displayed variable levels of resistance to one or two of the tested races.Seedling infection types(ITs) and adult-plant infection responses(IRs) indicated that 41(59.4%) of the wheat cultivars showed high resistance to all the three races. Molecular marker analysis showed that four wheat culitvars likely carried Sr2, 20 wheat culitvars likely carried Sr31, 9 wheat culitvars likely carried Sr38, and none of the cultivars carried Sr24,Sr25, and Sr26. Our results provide a scientific basis for rational utilization of the tested Sr genes and wheat cultivars against these novel Pgt races.展开更多
Wheat powdery mildew, caused by Blumeria graminis f. sp. tritici, is an important disease in China. To characterize the virulence and diversity of the pathogen, 1 082 isolates were obtained from 8 major wheat-growing ...Wheat powdery mildew, caused by Blumeria graminis f. sp. tritici, is an important disease in China. To characterize the virulence and diversity of the pathogen, 1 082 isolates were obtained from 8 major wheat-growing regions during the spring growing season in 2011. The virulence test was performed by inoculation on detached leaves of 22 differential lines with known Pm genes. Frequencies of virulence on these genotypes ranged from 0 to 97.4%. None of the 1 082 isolates was compatible to Pm21 and less than 20.0%were virulent to the genotype carrying Pm13. In contrast, the virulence frequencies of each population was more than 50.0%to differentials carrying Pm1a, Pm3b, Pm3c, Pm3f, Pm5a, Pm6 and Pm8. In total, 1 028 pathotypes were detected, of which 984 were unique. Phenotypic diversity indices revealed a high level of diversity within populations. Genetic distance between different populations correlated signiifcantly with geographical distance (R2=0.494, P 0.001). In addition, isolates from Xinjiang appear to form a separate group. Signiifcant positive or negative associations between alleles at pairs of virulence loci were detected in 57 allele pairs to Pm genes. Virulence and diversity of the 8 populations suggested that varieties with effective resistance gene combinations should be developed at a regional level.展开更多
Puccinia striiformis Westend.f.sp.tritici Erikss.(Pst)infects wheat and causes stripe rust.The rust is heteroecious with wheat as the primary uredinial and telial host and barberry(Berberis spp.)as the alternate pycni...Puccinia striiformis Westend.f.sp.tritici Erikss.(Pst)infects wheat and causes stripe rust.The rust is heteroecious with wheat as the primary uredinial and telial host and barberry(Berberis spp.)as the alternate pycnial and aecial host.More than 40 Berberis species have been identified as alternate hosts for Pst,and most of these are Chinese Berberis species.However,little is known about Berberis species or their geographic distributions in the Yunnan-Guizhou plateau in southwestern China.The Yunnan-Guizhou plateau is considered to be an important and relatively independent region for the evolution of the wheat stripe rust pathogen in China because the entire disease cycle can be completed within the region.In this study,we conducted a survey of barberry plants in the Yunnan-Guizhou plateau and identified the eight Pst-susceptible Berberis species under controlled conditions,including B.julianae,B.tsienii,B.veitchii,B.wilsonae,B.wilsonae var.guhtzunica,B.franchetiana,B.lepidifolia and B.pruinosa.These species are reported here for the first time to serve as alternate hosts for the wheat stripe rust pathogen under controlled conditions.展开更多
Puccinia striiformis f. sp. tritici (Pst) is one of the pathogenic fungi on wheat, caused stripe rust that is a great threat for wheat production all over the world. Intensive efforts have been made to study genetics ...Puccinia striiformis f. sp. tritici (Pst) is one of the pathogenic fungi on wheat, caused stripe rust that is a great threat for wheat production all over the world. Intensive efforts have been made to study genetics of wheat resistance to this disease, but few on avirulence of the pathogen due mainly to the nature of obligate biotrophism and the lack of systems for studying its genetics and molecular manipulations. To overcome these limitations, a natural Pst population comprising 352 isolates representative of a diverse virulence spectrum was genotyped using 97 secreted protein-single nucleotide polymorphism (SP-SNP) markers to identify candidate avirulence genes using association analysis. Among avirulence genes corresponding to 19 resistance genes, significantly associated SP-SNP markers were detected for avirulence genes AvYr1, AvYr2, AvYr6, AvYr7, AvYr8, AvYr44, AvYrExp2, AvYrSP, and AvYrTye. These results indicate that association analysis can be used to identify markers for avirulence genes. This study has laid the foundation for developing more SP-SNPs for mapping avirulence genes using segregating populations that can be generated through sexual reproduction on alternate hosts of the pathogen.展开更多
To gain more precise information about molecular genetic variation for wild populations of Blumeria graminis f. sp. tritici from Qinghai Province, China, 38 single-colony isolates were purified from samples collected ...To gain more precise information about molecular genetic variation for wild populations of Blumeria graminis f. sp. tritici from Qinghai Province, China, 38 single-colony isolates were purified from samples collected from Haidong District, Xining City and Hainan Tibetan Autonomous Prefecture in 2010. The virulence of 21 isolates among them was tested at seedling stage on 34 wheat cultivars(lines) carrying known powdery mildew(Pm) resistant genes. The results showed that V1 a, V3 a, V3 c, V3 e, V5 a, V6, V7, V8 and V19 had high virulence frequencies(〉75%), indicating a wide distribution; and V1 c, V5 b, V12, V13, V16, V21, VXBD, V2+6, V2+Mld and V4+8, with less distribution, appeared to be lower in frequencies(0-20%). The Nei's gene diversity(H), Shannon's information index(I) and the percentage of polymorphic loci(P) were 0.23, 0.35 and 67.65%, respectively, which revealed a virulent diversity. The results from single nucleotide polymorphisms(SNPs) of 38 isolates showed that three housekeeping genes were found to contain a total of 9 SNP sites. 10 haplotypes(H1-H10) were inferred from the concatenated sequences, with 1 haplotype(H1) comprising of over 55% of Qinghai population. Phylogenic analysis did not show obvious geographical subdivision between the isolates. A multilocus haplotype network presented a radial structure, with H1 in the central as an inferred ancestor. Using analysis of molecular variance(AMOVA), we found 1.63% of the total variation was among populations and 98.37% within populations, with a low fixations index(FST=0.01634, P〈0.05). This revealed a relatively high genetic diversity but a low genetic divergence in Qinghai population. Moreover, the molecular data on gene flow(Nm=6.32) confirmed the migration of pathogen populations among areas in Qinghai Province.展开更多
Stripe rust, caused by Puccinia striiformis f. sp. tritici, is an important wheat disease in China, seriously threatening wheat production. Understanding the winter survival of the fungus is a key for predicting the s...Stripe rust, caused by Puccinia striiformis f. sp. tritici, is an important wheat disease in China, seriously threatening wheat production. Understanding the winter survival of the fungus is a key for predicting the spring epidemics of the disease, which determines the crop loss. Estimation of P. striiformis f. sp. tritici winter survival requires processing a large number of samples for sensitive detection of the pathogen in wheat leaf tissue using real-time quantitative reverse transcription PCR (qRT-PCR). A bottleneck for the analysis is the acquisition of a good yield of high quality RNA suitable for qRT-PCR to distinguish dead and alive fungal hyphae inside leaves. Although several methods have been described in the literatures and commercial kits are available for RNA extraction, these methods are mostly too complicated, expensive and inefficient. Thus, we modified three previously reported RNA extraction methods with common and low-cost reagents (LiCI, SDS and NaCI) to solve the problems and selected the best to obtain high quality and quantity RNA for use in qRT-PCR. In the three improved methods, the NaCI method was proven to be the best for extracting RNAfrom urediniospores of and wheat leaves infected by P. striiformis f. sp. tritici, although the modified LiCI and SDS methods also increased yield of RNA compared to the previous methods. The improved NaCI method has the following advantages: 1) Complete transfer of urediniospores of P. striiformis f. sp. tritici from the mortar and pestle can ensure the initial amount of RNA for the qRT-PCR analysis; 2) the use of low-cost NaCI to replace more expensive Trizol can reduce the cost; 3) the yield and quality of RNA can be increased; 4) the improved method is more suitable for a large number and high quantity of samples from fields. Using the improved NaCI method, the amount of RNA was increased three times from urediniospores of P. striiformis f. sp. tritici compared from the extraction kit. Approximately, 10.11 IJg total RNA of high quality was obtained from 100 mg of infected leaves, which was 8.8, 6.5, 3.4 and 2.1 folds of the amounts obtained from the previous LiCI, SDS, NaCI and traditional Trizol methods, respectively. The method could be used to study the overwintering rates of R striiformis f. sp. tritici over a large region of wheat production for predicting epidemic levels by determining pathogen survival levels after winter. The method can alsobe used in any studies which need a large number of high quality RNA samples.展开更多
Simple sequence repeats (SSR) have been widely used as molecular markers due to their abundance and high polymorphism, However, up to now, the SSR markers had not been developed in the obligate biotrophic phytopatho...Simple sequence repeats (SSR) have been widely used as molecular markers due to their abundance and high polymorphism, However, up to now, the SSR markers had not been developed in the obligate biotrophic phytopathogenic fungus, Blumeria graminis f.sp. tritici. From (AC)10 and (AG)10 enriched genomic libraries for Bgt, 25 primer pairs were designed using the FIASCO (fast isolation by AFLP of sequences containing repeats) protocol. Five primer pairs exhibited polymorphism with allelic diversity from two to seven alleles and produced 29 alleles in a survey of 90 isolates collected from six provinces (cities) in China, while the others displayed monomorphic. Levels of observed heterozygosity ranged from 0.000-0.044 (mean 0.025) and expected heterozygosity ranged from 0.297-0.816 (mean 0.538). These molecular markers provide a novel source to genetic diversity assays and to genetic and physical mapping ofBgt. SSR markers of Bgt need to be further explored.展开更多
Take-all is a serious disease found in wheat across the world. Haynaldia villosa is considered to be resistant to take-all at a high level. TH3 was an amphiploid (2n =42, AABBVV) between Triticum durum and Haynaldia...Take-all is a serious disease found in wheat across the world. Haynaldia villosa is considered to be resistant to take-all at a high level. TH3 was an amphiploid (2n =42, AABBVV) between Triticum durum and Haynaldia viUosa with significant resistance to take-all fungus isolated from China. In greenhouse experiment, the derivatives of the hybrid between wheat and TH3 showed better resistance to take-all than that of the wheat control. One of the derivatives named HW918-5 was selected for further analysis. Cytological and genomic in situ hybridization (GISH) analysis indicated that a monotelosome originated from H. villosa existed in the genome of the offspring of the line HW918-5. The monotelosome with promising resistant gene for take-all was located on the 3V chromosome of H. villosa in the further PCR-based molecular analysis.展开更多
Fimbrin, a regulator of actin cytoskeletal dynamics that participates in numerous physiological and biochemical processes, controls multiple developmental processes in a variety of tissues and cell types. However, the...Fimbrin, a regulator of actin cytoskeletal dynamics that participates in numerous physiological and biochemical processes, controls multiple developmental processes in a variety of tissues and cell types. However, the role of fimbrin in pathogen defense of wheat and the mechanisms have not been well studied. Here, we investigated that the expression of TaFIM1 gene of wheat was significantly induced in response to avirulent race of Puccinia striiformis f. sp. tritici(Pst) and silencing of TaFIM1 by virus-induced gene silencing method. The results show that silencing of TaFIM1 resulted in a reduction of resistance against the stripe rust indicated by both phenotypes and a histological examination of Pst growth. Additionally, the expression level of Ta FIM1 gene was up-regulated under abiotic stresses. These findings suggest that Ta FIM1 functions as a positive regulator of pathogen resistance of wheat plants and response to abiotic stress. Our work may show new light on understanding the roles of fimbrin in wheat.展开更多
The paper was to study the effects of different cultivation patterns( mix cultivation and monocultivation) of wheat on population structure of Puccinia striiformis West. f. sp. tritici in the fields. Five race-specifi...The paper was to study the effects of different cultivation patterns( mix cultivation and monocultivation) of wheat on population structure of Puccinia striiformis West. f. sp. tritici in the fields. Five race-specific-markers( CY32,CY31,CY29,CY23 and Shuiyuan pathotype) were used to survey 113 infected samples collected from two cultivation patterns. The results indicated that frequency of race-specific-markers under monocultivation was higher than that under mix cultivation; the dominant race-specific-markers were CY32 and CY29 under monocultivation,and the frequency of detection were 81. 5% and 78. 5%,respectively. The dominant race-specific-markers were CY29 and Shuiyuan pathotype under mix cultivation,and the frequency of detection are 41. 7% and 18. 8%,respectively.Several race-specific-markers were detected in single infected leaf,and 41. 7% of infected single leaf were detected with more than two race-specific-markers,58. 3% of infected single leaf were detected with one race-specific-marker under mix cultivation pattern,while there were 75. 0% infected leaves with more than two race-specific-markers and 25. 0% infected single leaf detected with one race-specific-marker under monocultivation pattern. The results indicated that mix cultivation pattern of wheat can reduce races on single leaf,affect the distribution of races in infected leaves,and suppress the occurrence frequency of dominant races of P. striiformis in the fields significantly,subsequently reduced severity and prevalence of the disease.展开更多
Wheat stripe rust,caused by Puccinia striiformis f.sp.tritici(Pst),is one of the most destructive fungal diseases of wheat,and seriously threatens safe production of the crop worldwide.In China,new races historically ...Wheat stripe rust,caused by Puccinia striiformis f.sp.tritici(Pst),is one of the most destructive fungal diseases of wheat,and seriously threatens safe production of the crop worldwide.In China,new races historically appeared and rapidly developed to be predominant races and have resulted in ineffectiveness and replacement of wheat resistance cultivars as well as massive reduction in yield.In the present study,the relative parasitic fitness of the two newlyemerged Yr5-virulent races(TSA-6 and TSA-9)were compared with those of four currently predominant Chinese races(CYR31,CYR32,CYR33,and CYR34)based on evaluation on 10 Chinese wheat cultivars.As a result,there were significant differences in the relative parasitic fitness parameters among overall tested races based on multiple comparison(LSD)analysis(P<0.05).The principal component analysis(PCA)of overall parasitic fitness parameters indicated that the sporulation ability,infection and spore survivability,expansion capacity,and potential pathogenicity were the most important parasitic fitness attributes of the tested races.Based on the establishment of extracted three principal components and a comprehensive factor score mathematical models,evaluations of the parasitic fitness attributes of tested races showed that the level of relative parasitic fitness of the tested six races was:CYR32(1.15)>TSA-9(0.95)>TSA-6(0.92)>CYR34(0.29)>CYR31(–1.54)>CYR33(–1.77).The results indicated that two Yr5-virulent races TSA-9 and TSA-6 possessed relative parasitic fitness higher than races CYR34,CYR31,and CYR33,but lower than race CYR32,and have potential risks in developing to be predominant races.Therefore,continual monitoring of both Yr5-virulent races,and their variants is needed.The use of wheat cultivars(lines)with Yr5 resistance gene singly in wheat breeding is essential for being avoided,and is suggested to combine with other effective stripe rust resistance genes.展开更多
The devastating wheat stripe(yellow)rust pathogen,Puccinia striiformis f.sp.tritici(Pst),is a macrocyclic and heteroe-cious fungus.Pst produces urediniospores and teliospores on its primary host,wheat,and pycniospores...The devastating wheat stripe(yellow)rust pathogen,Puccinia striiformis f.sp.tritici(Pst),is a macrocyclic and heteroe-cious fungus.Pst produces urediniospores and teliospores on its primary host,wheat,and pycniospores and aeciospores are produced on its alternate hosts,barberry(Berberis spp.)or mahonia(Mahonia spp.).Basidiospores are developed from teliospores and infect alternate hosts.These five spore forms play distinct roles in Pst infection,disease development,and fungal survival,etc.However,the specific genes and mechanisms underlying these functional differences are largely unknown.In this study,we performed,for the first time in rust fungi,the deep RNA sequencing to examine the transcriptomic shift among all five Pst spore forms.Among a total of 29,591 identified transcripts,951 were specifically expressed in basidiospores,whereas 920,761,266,and 110 were specific for teliospores,pycniospores,aeciospores,and urediniospores,respectively.Additionally,transcriptomes of sexual spores,namely pycniospores and basidiospores,showed significant differences from those of asexual spores(urediniospores,teliospores,and aeciospores),and transcriptomes of urediniospores and aeciospores were more similar to each other than to the three other spore forms.Especially,the basidiospores and pycniospores which infected the berberis shows wide differences in the cell wall degrading-enzymes and mating and pheromone response genes.Besides,we also found that there are 6234 differential expressed genes between the urediniospores and pycniospores,while only have 3 genes have alternative splicing enents,suggesting that differential genes expression may make more contribution than AS.This comprehensive transcriptome profiling can substantially improve our understanding of the developmental biology of the wheat stripe rust fungus.展开更多
Background:Septoria tritici blotch(STB)(Zymoseptoria tritici)is a major disease of durum wheat,an economic crop grown in the highlands of Wollo in Ethiopia.Methods:To determine the status of this disease,we conducted ...Background:Septoria tritici blotch(STB)(Zymoseptoria tritici)is a major disease of durum wheat,an economic crop grown in the highlands of Wollo in Ethiopia.Methods:To determine the status of this disease,we conducted surveys in five districts of Wollo(Meket,Woreilu,Wadila,Jama,and Dessie Zuria)during the 2015 cropping season.We visited 75 farm plots to determine the prevalence,incidence,and severity of STB.Results:STB prevalence varied among locations,genotypes,planting dates,growth stages,previous crops,plant population,weed population,and soil types.Similarly,disease intensity also varied along all independent variables.The level of incidence was high in all the visited districts,and the level severity ranged from 9.9 to 59.3%while the incidence varied from 50 to 100%.The mean differences in incidence and severity within the districts’variable classes,altitude,varieties,growth stage,plant population,planting date,previous crop,weed population,and soil type were high.The independent variables,districts,altitude,varieties,growth stage,plant population,planting date,previous crops,weed population,and soil type,were significantly associated with high incidence and severity of STB as single predictor in the logistic regression model.A reduced multiple variable model was fitted using districts,altitude,varieties,growth stage,plant population,planting date,previous crop,weed population,and soil type as independent variables.High incidence(>50%)and severity(>25%)had a high probability of association to all independent variables,except previous crop.Low disease incidence(≤50%)and low disease severity(≤25%)had high probability of association to the previous crop.Conclusion:Environmental variables,cultivation practice,and responses were important for the development of STB.Therefore,these factors must be considered in designing strategies for the effective management of STB.展开更多
Evaluation with the pot assay at seedling stage in greenhouse showed that oat (Arena saliva) was highly resistant to take-all disease to which, however, wheat (Triticum aestivum) was extremely susceptible. The oat roo...Evaluation with the pot assay at seedling stage in greenhouse showed that oat (Arena saliva) was highly resistant to take-all disease to which, however, wheat (Triticum aestivum) was extremely susceptible. The oat roots were shown to be inhibitory to the invasion and spread of take-all causing fungus G. graminis var. tritici by the following criteria: (i) less infection sites were observed (about 1/7 of those in wheat); (ii) the ectotrophic growth of G. graminis var. tritici on oat roots was much slower than that on those of wheat, and the runner hyphae appeared as kidney- or fork-shaped hyphopodia on the surface of oat roots which could not be discerned on that of wheat roots; (iii) the period from inoculation to penetration into the epidermis of oat roots was about 2.9 times as long as that of wheat; (iv) the infection hyphae were hindered substantially when it was about to penetrate into the epidermis of oat roots with the mycelium deformed; and (v) the cortical layer of oat roots was revealed to be展开更多
文摘Wheat stem rust, caused by Puccinia graminis f. sp. tritici(Pgt), is a potentially devastating fungal disease of wheat worldwide. The present study was to evaluate the resistance of 42 wheat monogenic lines with known stem rust resistance(Sr) genes and 69 wheat cultivars to three new Pgt races(34C0MRGQM, 34C3MKGQM, and 34C6MTGSM)identified from aeciospores at the seedling and adult-plant stages. The phenotyping results revealed that monogenic lines harboring resistance genes Sr9e, Sr17, Sr21, Sr22, Sr26, Sr30, Sr31, Sr33, Sr35, Sr36, Sr37, Sr38, Sr47, SrTmp,and SrTt3 were effectively resistant to all three Pgt races at the seedling and adult-plant stages. In contrast, monogenic lines containing Sr5, Sr6, Sr7b, Sr9a, Sr9d, Sr9f, Sr9g, Sr9b, Sr16, Sr24, Sr28, and Sr39 were highly susceptible to these races at both seedling and adult-plant stages. The other lines with Sr8a, Sr10, Sr11, Sr13, Sr14, Sr15, Sr18, Sr20,Sr19, Sr23, Sr25, Sr27, Sr29, Sr32, and Sr34, displayed variable levels of resistance to one or two of the tested races.Seedling infection types(ITs) and adult-plant infection responses(IRs) indicated that 41(59.4%) of the wheat cultivars showed high resistance to all the three races. Molecular marker analysis showed that four wheat culitvars likely carried Sr2, 20 wheat culitvars likely carried Sr31, 9 wheat culitvars likely carried Sr38, and none of the cultivars carried Sr24,Sr25, and Sr26. Our results provide a scientific basis for rational utilization of the tested Sr genes and wheat cultivars against these novel Pgt races.
基金supported by the National Basic Research Program of China (2013CB127700)the Special Fund for Agro-Scientific Research in the Public Interest, China (201303016)
文摘Wheat powdery mildew, caused by Blumeria graminis f. sp. tritici, is an important disease in China. To characterize the virulence and diversity of the pathogen, 1 082 isolates were obtained from 8 major wheat-growing regions during the spring growing season in 2011. The virulence test was performed by inoculation on detached leaves of 22 differential lines with known Pm genes. Frequencies of virulence on these genotypes ranged from 0 to 97.4%. None of the 1 082 isolates was compatible to Pm21 and less than 20.0%were virulent to the genotype carrying Pm13. In contrast, the virulence frequencies of each population was more than 50.0%to differentials carrying Pm1a, Pm3b, Pm3c, Pm3f, Pm5a, Pm6 and Pm8. In total, 1 028 pathotypes were detected, of which 984 were unique. Phenotypic diversity indices revealed a high level of diversity within populations. Genetic distance between different populations correlated signiifcantly with geographical distance (R2=0.494, P 0.001). In addition, isolates from Xinjiang appear to form a separate group. Signiifcant positive or negative associations between alleles at pairs of virulence loci were detected in 57 allele pairs to Pm genes. Virulence and diversity of the 8 populations suggested that varieties with effective resistance gene combinations should be developed at a regional level.
基金the National Key R&D Program of China(2018YFD0200500)the National Natural Science Foundation of China(31960524,31071641 and 32072358)+1 种基金the Fundamental Research Funds for the Central Universities(2452019046)the Natural Science Basic Research Plan in Shaanxi Province of China(2020JZ-15,2017JM3006)。
文摘Puccinia striiformis Westend.f.sp.tritici Erikss.(Pst)infects wheat and causes stripe rust.The rust is heteroecious with wheat as the primary uredinial and telial host and barberry(Berberis spp.)as the alternate pycnial and aecial host.More than 40 Berberis species have been identified as alternate hosts for Pst,and most of these are Chinese Berberis species.However,little is known about Berberis species or their geographic distributions in the Yunnan-Guizhou plateau in southwestern China.The Yunnan-Guizhou plateau is considered to be an important and relatively independent region for the evolution of the wheat stripe rust pathogen in China because the entire disease cycle can be completed within the region.In this study,we conducted a survey of barberry plants in the Yunnan-Guizhou plateau and identified the eight Pst-susceptible Berberis species under controlled conditions,including B.julianae,B.tsienii,B.veitchii,B.wilsonae,B.wilsonae var.guhtzunica,B.franchetiana,B.lepidifolia and B.pruinosa.These species are reported here for the first time to serve as alternate hosts for the wheat stripe rust pathogen under controlled conditions.
文摘Puccinia striiformis f. sp. tritici (Pst) is one of the pathogenic fungi on wheat, caused stripe rust that is a great threat for wheat production all over the world. Intensive efforts have been made to study genetics of wheat resistance to this disease, but few on avirulence of the pathogen due mainly to the nature of obligate biotrophism and the lack of systems for studying its genetics and molecular manipulations. To overcome these limitations, a natural Pst population comprising 352 isolates representative of a diverse virulence spectrum was genotyped using 97 secreted protein-single nucleotide polymorphism (SP-SNP) markers to identify candidate avirulence genes using association analysis. Among avirulence genes corresponding to 19 resistance genes, significantly associated SP-SNP markers were detected for avirulence genes AvYr1, AvYr2, AvYr6, AvYr7, AvYr8, AvYr44, AvYrExp2, AvYrSP, and AvYrTye. These results indicate that association analysis can be used to identify markers for avirulence genes. This study has laid the foundation for developing more SP-SNPs for mapping avirulence genes using segregating populations that can be generated through sexual reproduction on alternate hosts of the pathogen.
基金supported by the National Basic Research Program of China(2011CB100403 and 2013CB127704)the Special Fund for Agro-Scientific Research in the Public Interest,China(201303016)
文摘To gain more precise information about molecular genetic variation for wild populations of Blumeria graminis f. sp. tritici from Qinghai Province, China, 38 single-colony isolates were purified from samples collected from Haidong District, Xining City and Hainan Tibetan Autonomous Prefecture in 2010. The virulence of 21 isolates among them was tested at seedling stage on 34 wheat cultivars(lines) carrying known powdery mildew(Pm) resistant genes. The results showed that V1 a, V3 a, V3 c, V3 e, V5 a, V6, V7, V8 and V19 had high virulence frequencies(〉75%), indicating a wide distribution; and V1 c, V5 b, V12, V13, V16, V21, VXBD, V2+6, V2+Mld and V4+8, with less distribution, appeared to be lower in frequencies(0-20%). The Nei's gene diversity(H), Shannon's information index(I) and the percentage of polymorphic loci(P) were 0.23, 0.35 and 67.65%, respectively, which revealed a virulent diversity. The results from single nucleotide polymorphisms(SNPs) of 38 isolates showed that three housekeeping genes were found to contain a total of 9 SNP sites. 10 haplotypes(H1-H10) were inferred from the concatenated sequences, with 1 haplotype(H1) comprising of over 55% of Qinghai population. Phylogenic analysis did not show obvious geographical subdivision between the isolates. A multilocus haplotype network presented a radial structure, with H1 in the central as an inferred ancestor. Using analysis of molecular variance(AMOVA), we found 1.63% of the total variation was among populations and 98.37% within populations, with a low fixations index(FST=0.01634, P〈0.05). This revealed a relatively high genetic diversity but a low genetic divergence in Qinghai population. Moreover, the molecular data on gene flow(Nm=6.32) confirmed the migration of pathogen populations among areas in Qinghai Province.
基金supported by the National Key Basic Research Program of China (2013CB127700)the Na-tional Natural Science Foundation of China (31071640 and 31271985)partially supported by the 111 Project from Education Ministry of China (B07049)
文摘Stripe rust, caused by Puccinia striiformis f. sp. tritici, is an important wheat disease in China, seriously threatening wheat production. Understanding the winter survival of the fungus is a key for predicting the spring epidemics of the disease, which determines the crop loss. Estimation of P. striiformis f. sp. tritici winter survival requires processing a large number of samples for sensitive detection of the pathogen in wheat leaf tissue using real-time quantitative reverse transcription PCR (qRT-PCR). A bottleneck for the analysis is the acquisition of a good yield of high quality RNA suitable for qRT-PCR to distinguish dead and alive fungal hyphae inside leaves. Although several methods have been described in the literatures and commercial kits are available for RNA extraction, these methods are mostly too complicated, expensive and inefficient. Thus, we modified three previously reported RNA extraction methods with common and low-cost reagents (LiCI, SDS and NaCI) to solve the problems and selected the best to obtain high quality and quantity RNA for use in qRT-PCR. In the three improved methods, the NaCI method was proven to be the best for extracting RNAfrom urediniospores of and wheat leaves infected by P. striiformis f. sp. tritici, although the modified LiCI and SDS methods also increased yield of RNA compared to the previous methods. The improved NaCI method has the following advantages: 1) Complete transfer of urediniospores of P. striiformis f. sp. tritici from the mortar and pestle can ensure the initial amount of RNA for the qRT-PCR analysis; 2) the use of low-cost NaCI to replace more expensive Trizol can reduce the cost; 3) the yield and quality of RNA can be increased; 4) the improved method is more suitable for a large number and high quantity of samples from fields. Using the improved NaCI method, the amount of RNA was increased three times from urediniospores of P. striiformis f. sp. tritici compared from the extraction kit. Approximately, 10.11 IJg total RNA of high quality was obtained from 100 mg of infected leaves, which was 8.8, 6.5, 3.4 and 2.1 folds of the amounts obtained from the previous LiCI, SDS, NaCI and traditional Trizol methods, respectively. The method could be used to study the overwintering rates of R striiformis f. sp. tritici over a large region of wheat production for predicting epidemic levels by determining pathogen survival levels after winter. The method can alsobe used in any studies which need a large number of high quality RNA samples.
基金supported by the National Basic Research Program of China(2006CB100203 and 2011CB100403)the Key Technologies R&D Program of China during the 11th Five-Year Plan period(2006BAD08A05)the Special Fund for Agro-Scientific Research in the Public Interest(3-15),China
文摘Simple sequence repeats (SSR) have been widely used as molecular markers due to their abundance and high polymorphism, However, up to now, the SSR markers had not been developed in the obligate biotrophic phytopathogenic fungus, Blumeria graminis f.sp. tritici. From (AC)10 and (AG)10 enriched genomic libraries for Bgt, 25 primer pairs were designed using the FIASCO (fast isolation by AFLP of sequences containing repeats) protocol. Five primer pairs exhibited polymorphism with allelic diversity from two to seven alleles and produced 29 alleles in a survey of 90 isolates collected from six provinces (cities) in China, while the others displayed monomorphic. Levels of observed heterozygosity ranged from 0.000-0.044 (mean 0.025) and expected heterozygosity ranged from 0.297-0.816 (mean 0.538). These molecular markers provide a novel source to genetic diversity assays and to genetic and physical mapping ofBgt. SSR markers of Bgt need to be further explored.
文摘Take-all is a serious disease found in wheat across the world. Haynaldia villosa is considered to be resistant to take-all at a high level. TH3 was an amphiploid (2n =42, AABBVV) between Triticum durum and Haynaldia viUosa with significant resistance to take-all fungus isolated from China. In greenhouse experiment, the derivatives of the hybrid between wheat and TH3 showed better resistance to take-all than that of the wheat control. One of the derivatives named HW918-5 was selected for further analysis. Cytological and genomic in situ hybridization (GISH) analysis indicated that a monotelosome originated from H. villosa existed in the genome of the offspring of the line HW918-5. The monotelosome with promising resistant gene for take-all was located on the 3V chromosome of H. villosa in the further PCR-based molecular analysis.
基金supported by the National Natural Science Foundation of China (31571960)the NSFC-Xinjiang Joint Fund, China (U1903110)the 111 Project from the Ministry of Education of China (B07049)。
文摘Fimbrin, a regulator of actin cytoskeletal dynamics that participates in numerous physiological and biochemical processes, controls multiple developmental processes in a variety of tissues and cell types. However, the role of fimbrin in pathogen defense of wheat and the mechanisms have not been well studied. Here, we investigated that the expression of TaFIM1 gene of wheat was significantly induced in response to avirulent race of Puccinia striiformis f. sp. tritici(Pst) and silencing of TaFIM1 by virus-induced gene silencing method. The results show that silencing of TaFIM1 resulted in a reduction of resistance against the stripe rust indicated by both phenotypes and a histological examination of Pst growth. Additionally, the expression level of Ta FIM1 gene was up-regulated under abiotic stresses. These findings suggest that Ta FIM1 functions as a positive regulator of pathogen resistance of wheat plants and response to abiotic stress. Our work may show new light on understanding the roles of fimbrin in wheat.
基金Supported by National 973 Project(2013CB127705)Special Fund of Ministry of Agriculture for Wheat Stripe Rust Industry(200903035-7)Key Fund of Yunnan Province of China(2009CC004)
文摘The paper was to study the effects of different cultivation patterns( mix cultivation and monocultivation) of wheat on population structure of Puccinia striiformis West. f. sp. tritici in the fields. Five race-specific-markers( CY32,CY31,CY29,CY23 and Shuiyuan pathotype) were used to survey 113 infected samples collected from two cultivation patterns. The results indicated that frequency of race-specific-markers under monocultivation was higher than that under mix cultivation; the dominant race-specific-markers were CY32 and CY29 under monocultivation,and the frequency of detection were 81. 5% and 78. 5%,respectively. The dominant race-specific-markers were CY29 and Shuiyuan pathotype under mix cultivation,and the frequency of detection are 41. 7% and 18. 8%,respectively.Several race-specific-markers were detected in single infected leaf,and 41. 7% of infected single leaf were detected with more than two race-specific-markers,58. 3% of infected single leaf were detected with one race-specific-marker under mix cultivation pattern,while there were 75. 0% infected leaves with more than two race-specific-markers and 25. 0% infected single leaf detected with one race-specific-marker under monocultivation pattern. The results indicated that mix cultivation pattern of wheat can reduce races on single leaf,affect the distribution of races in infected leaves,and suppress the occurrence frequency of dominant races of P. striiformis in the fields significantly,subsequently reduced severity and prevalence of the disease.
基金supported by the National Natural Science Foundation of China(32072358 and 32272507)the National Key R&D Program of China(2021YFD1401000)+1 种基金the earmarked fund for CARS-03,the Natural Science Basic Research Project in Shaanxi Province of China(2020JZ-15)National“111 Project”of China(BP0719026)。
文摘Wheat stripe rust,caused by Puccinia striiformis f.sp.tritici(Pst),is one of the most destructive fungal diseases of wheat,and seriously threatens safe production of the crop worldwide.In China,new races historically appeared and rapidly developed to be predominant races and have resulted in ineffectiveness and replacement of wheat resistance cultivars as well as massive reduction in yield.In the present study,the relative parasitic fitness of the two newlyemerged Yr5-virulent races(TSA-6 and TSA-9)were compared with those of four currently predominant Chinese races(CYR31,CYR32,CYR33,and CYR34)based on evaluation on 10 Chinese wheat cultivars.As a result,there were significant differences in the relative parasitic fitness parameters among overall tested races based on multiple comparison(LSD)analysis(P<0.05).The principal component analysis(PCA)of overall parasitic fitness parameters indicated that the sporulation ability,infection and spore survivability,expansion capacity,and potential pathogenicity were the most important parasitic fitness attributes of the tested races.Based on the establishment of extracted three principal components and a comprehensive factor score mathematical models,evaluations of the parasitic fitness attributes of tested races showed that the level of relative parasitic fitness of the tested six races was:CYR32(1.15)>TSA-9(0.95)>TSA-6(0.92)>CYR34(0.29)>CYR31(–1.54)>CYR33(–1.77).The results indicated that two Yr5-virulent races TSA-9 and TSA-6 possessed relative parasitic fitness higher than races CYR34,CYR31,and CYR33,but lower than race CYR32,and have potential risks in developing to be predominant races.Therefore,continual monitoring of both Yr5-virulent races,and their variants is needed.The use of wheat cultivars(lines)with Yr5 resistance gene singly in wheat breeding is essential for being avoided,and is suggested to combine with other effective stripe rust resistance genes.
基金financially supported by the National Key Research and Development Program of China(2021YFD1401000)the National Natural Science Foundation of China(32172380 and 32102175)+1 种基金the 111 Project of the Ministry of Education of China(B0719026)the Open Project Program of State Key Laboratory of Crop Stress Biology for Arid Areas(CSBAAKF2021013).
文摘The devastating wheat stripe(yellow)rust pathogen,Puccinia striiformis f.sp.tritici(Pst),is a macrocyclic and heteroe-cious fungus.Pst produces urediniospores and teliospores on its primary host,wheat,and pycniospores and aeciospores are produced on its alternate hosts,barberry(Berberis spp.)or mahonia(Mahonia spp.).Basidiospores are developed from teliospores and infect alternate hosts.These five spore forms play distinct roles in Pst infection,disease development,and fungal survival,etc.However,the specific genes and mechanisms underlying these functional differences are largely unknown.In this study,we performed,for the first time in rust fungi,the deep RNA sequencing to examine the transcriptomic shift among all five Pst spore forms.Among a total of 29,591 identified transcripts,951 were specifically expressed in basidiospores,whereas 920,761,266,and 110 were specific for teliospores,pycniospores,aeciospores,and urediniospores,respectively.Additionally,transcriptomes of sexual spores,namely pycniospores and basidiospores,showed significant differences from those of asexual spores(urediniospores,teliospores,and aeciospores),and transcriptomes of urediniospores and aeciospores were more similar to each other than to the three other spore forms.Especially,the basidiospores and pycniospores which infected the berberis shows wide differences in the cell wall degrading-enzymes and mating and pheromone response genes.Besides,we also found that there are 6234 differential expressed genes between the urediniospores and pycniospores,while only have 3 genes have alternative splicing enents,suggesting that differential genes expression may make more contribution than AS.This comprehensive transcriptome profiling can substantially improve our understanding of the developmental biology of the wheat stripe rust fungus.
文摘Background:Septoria tritici blotch(STB)(Zymoseptoria tritici)is a major disease of durum wheat,an economic crop grown in the highlands of Wollo in Ethiopia.Methods:To determine the status of this disease,we conducted surveys in five districts of Wollo(Meket,Woreilu,Wadila,Jama,and Dessie Zuria)during the 2015 cropping season.We visited 75 farm plots to determine the prevalence,incidence,and severity of STB.Results:STB prevalence varied among locations,genotypes,planting dates,growth stages,previous crops,plant population,weed population,and soil types.Similarly,disease intensity also varied along all independent variables.The level of incidence was high in all the visited districts,and the level severity ranged from 9.9 to 59.3%while the incidence varied from 50 to 100%.The mean differences in incidence and severity within the districts’variable classes,altitude,varieties,growth stage,plant population,planting date,previous crop,weed population,and soil type were high.The independent variables,districts,altitude,varieties,growth stage,plant population,planting date,previous crops,weed population,and soil type,were significantly associated with high incidence and severity of STB as single predictor in the logistic regression model.A reduced multiple variable model was fitted using districts,altitude,varieties,growth stage,plant population,planting date,previous crop,weed population,and soil type as independent variables.High incidence(>50%)and severity(>25%)had a high probability of association to all independent variables,except previous crop.Low disease incidence(≤50%)and low disease severity(≤25%)had high probability of association to the previous crop.Conclusion:Environmental variables,cultivation practice,and responses were important for the development of STB.Therefore,these factors must be considered in designing strategies for the effective management of STB.
基金This work was supported by theShaanxi Provincial Scientific Research Project (Grant No. 39725033) and the Postdoctoral Science Foundation of China (Grant No. 0208171304) and the Subject Constructional Foundation of Nanjing University (Grant No. 020
文摘Evaluation with the pot assay at seedling stage in greenhouse showed that oat (Arena saliva) was highly resistant to take-all disease to which, however, wheat (Triticum aestivum) was extremely susceptible. The oat roots were shown to be inhibitory to the invasion and spread of take-all causing fungus G. graminis var. tritici by the following criteria: (i) less infection sites were observed (about 1/7 of those in wheat); (ii) the ectotrophic growth of G. graminis var. tritici on oat roots was much slower than that on those of wheat, and the runner hyphae appeared as kidney- or fork-shaped hyphopodia on the surface of oat roots which could not be discerned on that of wheat roots; (iii) the period from inoculation to penetration into the epidermis of oat roots was about 2.9 times as long as that of wheat; (iv) the infection hyphae were hindered substantially when it was about to penetrate into the epidermis of oat roots with the mycelium deformed; and (v) the cortical layer of oat roots was revealed to be