Puccinia triticina(Pt), as the causal agent of wheat leaf rust, employs a plethora of effector proteins to modulate wheat immunity for successful colonization. Understanding the molecular mechanisms underlying Pt effe...Puccinia triticina(Pt), as the causal agent of wheat leaf rust, employs a plethora of effector proteins to modulate wheat immunity for successful colonization. Understanding the molecular mechanisms underlying Pt effector-mediated wheat susceptibility remains largely unexplored. In this study, an effector Pt_21 was identified to interact with the apoplast-localized wheat thaumatin-like protein TaTLP1 using a yeast two-hybrid assay and the Pt_21-TaTLP1 interaction was characterized. The interaction between Pt_21 and TaTLP1 was validated by in vivo co-immunoprecipitation assay. A TaTLP1 variant,TaTLP1C71A, that was identified by the site-directed mutagenesis failed to interact with Pt_21. Pt_21was able to suppress Bax-mediated cell death in leaves of Nicotiana benthamiana and inhibit TaTLP1-mediated antifungal activity. Furthermore, infiltration of recombinant protein Pt_21 into leaves of transgenic wheat line overexpressing TaTLP1 enhanced the disease development of leaf rust compared to that in wild-type leaves. These findings demonstrate that Pt_21 suppresses host defense response by directly targeting wheat TaTLP1 and inhibiting its antifungal activity, which broadens our understanding of the molecular mechanisms underlying Pt effector-mediated susceptibility in wheat.展开更多
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
The gene postulation, cluster analysis and pedigree analysis of 20 wheat cultivars to 28 physiological races of yellow rust (Puccinia striiformis) were conducted under greenhouse conditions, The spectrum of their re...The gene postulation, cluster analysis and pedigree analysis of 20 wheat cultivars to 28 physiological races of yellow rust (Puccinia striiformis) were conducted under greenhouse conditions, The spectrum of their resistance were compared each other. None of the 20 cultivars were resistant to all the test pathogens. The cultivars containing resistance genes Yr5 and Yr24 were not find and genes Yr8, Yrl9 and Yr27 could not be postulated from cultivars tested due to the susceptibility to all isolates used. A total of 6 probale seedling yellow rust resistance genes or gene combinations (Yrl, Yr2, Yr2 + YrHVII, Yr3 + unknown, Yr3 + Yr4, YrAlba) were postulated in the wheat cultivars (Atou, Flanders, Maris Huntsman, Bouquet, Holdfast, Elite Lepeuple, and Vilmorin 27). The gene combination Yr2+YrHVII with the highest frequency (35%) was present in 7 cultivars. The Yr genes present in some wheat cultivars could not be postulated because of non-matching virulence combinations with any of known genes. Cluster result showed that Yr2 and Yr3 are the most important genes in the cultivars. The 13 cultivars are believed to have the pedigree of Noe, which was selected from South Russian wheat. These results will be useful for wheat breeding and provide information about genetic control of wheat yellow rust.展开更多
White rust caused by Puccinia horiana is a destructive disease of chrysanthemum plants.To better understand the resistance mechanisms of composite species to this disease,the leaf cuticular traits,antioxidant and defe...White rust caused by Puccinia horiana is a destructive disease of chrysanthemum plants.To better understand the resistance mechanisms of composite species to this disease,the leaf cuticular traits,antioxidant and defensive enzymes activities of immune(Chrysanthemum makinoi var.wakasaense)and highly susceptible(Ajania shiwogiku var.kinokuniense)species were compared.Trichome density of two species was markedly different,negatively associated with plant resistance to P.horiana.Total wax load in C.makinoi var.wakasaense was two times more than that in A.shiwogiku var.kinokuniense.The wax composition in immune one was abundant in esters and primary alcohols.Superoxide dismutase(SOD,EC 1.15.1.1),peroxidase(POD,EC 1.11.1.7),polyphenoloxidase(PPO,EC 1.14.18.1 or EC 1.10.3.2)and phenylalanine ammonia lyase(PAL,EC 4.3.1.5)activitieswere investigated.In C.makinoi var.wakasaense,the activity of SOD and POD increased rapidly after inoculation,whichmight be non-host induced reactive oxygen species(ROS)activated antioxidant enzymes,however SOD and POD remained a low and steady level in the highly susceptible one after inoculation.Quick increase in PPO activities after inoculation was observed in both species,however it remained higher in C.makinoi var.wakasaense at the late period of inoculation.PAL in C.makinoi var.wakasaense was induced after pathogen inoculation,but not in A.shiwogiku var.kinokuniense,suggesting that these two enzymes might contribute to the resistance to P.horiana.展开更多
The incompatible combinations between races of Puccinia striiformis and wheat cultivars with low reaction type resistance were examined by means of fluorescent microscopy, differential interference contrast microscopy...The incompatible combinations between races of Puccinia striiformis and wheat cultivars with low reaction type resistance were examined by means of fluorescent microscopy, differential interference contrast microscopy and electron microscopy. The incompatible combinations consisted of wheat cultivar Niuzhute plus CY28, Hybrid46 plus CY29 and Tianxuan882 plus CY29, while the compatible combination was wheat cultivar Huixianhong plus CY28. The observation revealed a striking difference in the fungal development and the host responses between susceptible and resistant wheat cultivars following infection by the pathogen. The main histological manifestation of the pathogen development in the resistant wheat cultivars include inhibition of hyphal growth, delay of hyphal branching and colony formation, decrease of formation of haustorial mother cells and haustoria, and occurrence of host cell necrosis. The observation by electron microscopy demonstrated that a series of abnormal changes occurred in intercellular hyphae, haustorial mother cells and haustoria during pathogen development in the resistant wheat cultivars. The cytoplasm became more electron-dense and vacuoles in the cytoplasm increased in number and size. The cell walls of hyphae, haustorial mother cells were thickened irregularly. The organelles were disorderly distributed in the cytoplasm and the haustorial mother cells and haustoria lost their physiological function. In the final stage of the pathogen development, the intercellular hyphae, haustorial mother cells and haustoria became necrotic and collapsed. The structural defense reactions such as formation of cell wall apposition, collar and encasement of haustorium were essentially more pronounced in the infected wheat leaves of the resistant cultivars than in the susceptible one. In addition, the relationship between the host resistance expression and the histological and cytological features occurred in the incompatible combination was discussed in this paper.展开更多
A high degree of virulence diversity has been maintained in the population of Puccinia graminis f. sp. tritici (Pgt) in northwestern United States. Although Berberis vulgaris is present in the region and Pgt has bee...A high degree of virulence diversity has been maintained in the population of Puccinia graminis f. sp. tritici (Pgt) in northwestern United States. Although Berberis vulgaris is present in the region and Pgt has been isolated from aecial infections on B. vulgaris, the population is too diverse to be explained by the limited presence of B. vulgaris alone. Since 2008, we have isolated P. graminis from aecial infections on fruits of Mahonia repens and Mahonia aquifolium from northwestern United States. These two native woody shrub species, widely distributed in western North America, were once classified as resistant to P. graminis based on artificial inoculations. By isolating P. graminis from aecia, we established that M. repens and M. aquifolium along with B. vulgaris (albeit infrequent) serve as the alternate hosts ofP. graminis in the region. The isolates of P. graminis from Mahonia of North America had diverse virulence patterns and most of the isolates could be differentiated on Morocco, Line E, Chinese Spring, Little Club, LMPG-6, Rusty, and other genotypes that are considered to be universally susceptible to most Pgt isolates. This discovery explained the persistence of virulence diversity of Pgt observed in isolates derived from uredinia on cereal crops in the region. In addition to cereal crops, uredinial stage of the P. graminis population is sustained by wild grasses, especially Elymus glaucus, a native grass sharing the same habitat with the rusted Mahonia spp. Although virulence to some important stem rust resistance genes was observed in some isolates derived from Mahonia of North America when tested against single stem rust resistance gene stocks, the overall virulence is very limited in these isolates. This is likely a result of limited selection pressure on the rust population. In contrast to northwestern United Sates, the Pgt population in east of the Rocky Mountains of North America has declined steadily with a single race, QFCSC, being predominant in the last decade. This decline is likely due to a combination of factors, of which a lack of sexual recombination in the region is perhaps the most important one.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
Ultrastructural changes in both pathogen and host cells in the interaction between Puccinia striiformis and wheat cultivar (Libellula) with slow-rusting resistance were observed by transmission electron microscopy. ...Ultrastructural changes in both pathogen and host cells in the interaction between Puccinia striiformis and wheat cultivar (Libellula) with slow-rusting resistance were observed by transmission electron microscopy. Observations revealed marked changes in ultrastructure of both pathogen and host cells. In the pathogen respect, there were many vesicles appeared in the intercellular hyphae and gradually fused into bigger vacuoles, a number of fat drops and electron-dense granules accumulated, mitochondria became swollen and some of them degraded into vesicles, and the plasmalemma of intercellular hyphae became dark. In the haustoria, the cytoplasm degraded gradually and developed a vacuole in the center, fat drops increased, the extrahaustorial matrix widened with a great amount of electron-dense fibrillar and granular materials, and most of the haustoria died with in conjunction with the disappearance of fat drops and other organelles. Structural defense of the host, including formation of cell wall apposition, collar and papilla, occurred in the host respect. Host resistance expression and cytological features occurring in the slow-rusting resistance were discussed.展开更多
Wheat leaf rust caused by Puccinia triticina, is the most common and widely distributed wheat rust in the world. In order to study the genetic structure of leaf rust population 14 pairs of AFLP and 6 pairs of FAFLP pr...Wheat leaf rust caused by Puccinia triticina, is the most common and widely distributed wheat rust in the world. In order to study the genetic structure of leaf rust population 14 pairs of AFLP and 6 pairs of FAFLP primers evaluated on 86 isolates samples collected in Iran during spring of 2009. Results showed that almost all investigated isolates were genetically different and special pattern of AFLP allele’s that confirm high genetic diversity within leaf rust population was observed. Analyses showed, all provinces were classified into three major groups particularly similar clusters were found between then neighboring provinces. Rust spore can follow the migration pattern in short and long distances to neighbor in provinces. Results indicated that the greatest variability was revealed by 97% of genetic differentiation within leaf rust populations and the lesser variation of 3% was observed between the rust populations. These results suggested that each population was not completely identical and high gene flow has occurred among the leaf rust population of different provinces. The highest differentiation and genetic distance among the Iranian leaf rust populations was detected between leaf rust population in Sistan and Baluchistan and highest similarity was observed between in Ardabil provinces. The high pathogenic variability of leaf rust races in Ardabil and Northern Khorasan may be an indication that these two regions are the center of origin of pathogenic arability. Present study shows that leaf rust population in Iran is highly dynamic and variable.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
Puccinia striiformis, the causal agent of stripe rust in wheat, barley, and various wild grasses, produces urediniospores and teliospores on these primary or auxiliary hosts. Telial formation, which stops producing in...Puccinia striiformis, the causal agent of stripe rust in wheat, barley, and various wild grasses, produces urediniospores and teliospores on these primary or auxiliary hosts. Telial formation, which stops producing infectious urediniospores, is affected by various host and environmental conditions. However, it is not clear if variation exists among different isolates in the United States. To determine the differences in the pathogen population, 1423 isolates collected in the United States from 2013 to 2016 were tested for telial formation in the seedling stage at a diurnal temperature profile of 4°C - 20°C. The percentages of telial formation varied greatly among the isolates. Of the 1423 isolates, 62.97% produced telia with the percentages of telial formation ranging from 1% to 95% under the test conditions. The formation of telia was significantly affected by the year and the month when the isolates were collected. The epidemiological regions or states, host plants (wheat, barley, and grasses), and races of the isolates did not significantly affect telial formation. However, significant effects on telial formation were observed by interactions between year and region, year and race, month and region and among year, month, and region, as well as between year and month. The results showed that telial formation is a complex trait under the genetic control of the pathogen isolates for adaptation to different environments. Further studies are needed to identify genes involved in the formation of telia and the relationship of telial formation to the survival, aggressiveness, fitness, and evolution of the pathogen.展开更多
Puccinia triticina, the causal agent of wheat leaf rust, is one of the most devastating rust fungi attacking wheat worldwide. Seventy-six isolates of the wheat leaf rust pathogen from Yunnan, Sichuan, Gansu and Henan ...Puccinia triticina, the causal agent of wheat leaf rust, is one of the most devastating rust fungi attacking wheat worldwide. Seventy-six isolates of the wheat leaf rust pathogen from Yunnan, Sichuan, Gansu and Henan provinces, China, were tested on wheat leaf rust differentials and the population structure was analyzed using four presumably neutral partial sequence markers such as elongation factor-1α(EF-1α), glyceraldehyde-3-phosphate dehydrogenase(GAPDH), β-tubulin(TUB) and the second largest RNA polymerase subunit(RPB2). The phenotypic diversity of Yunnan and Sichuan populations was higher than that of Gansu and Henan populations. The four populations were separated into two clusters based on the pathogenic data. A total of 12 single nucleotide polymorphisms(SNPs) and 32 haplotypes were identified among the four sequences. The 32 haplotypes were divided into two clusters in a neighbor-joining tree. Bayesian analyses also identified two clusters. Pairwise FST between populations in different regions were significantly different(P<0.05). Analysis of molecular variance(AMOVA) indicated that 68% of the total genetic variation was within populations.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
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
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
基金supported by the National Natural Science Foundation of China (32172384 and 31501623)the Natural Science Foundation of Hebei (C2020204028)+1 种基金the Key Research and Development Project of Hebei Province (20326505D)the “Hundred Talents Program” for the Introduction of High-level Overseas Talents in Hebei Province (E2020100004)。
文摘Puccinia triticina(Pt), as the causal agent of wheat leaf rust, employs a plethora of effector proteins to modulate wheat immunity for successful colonization. Understanding the molecular mechanisms underlying Pt effector-mediated wheat susceptibility remains largely unexplored. In this study, an effector Pt_21 was identified to interact with the apoplast-localized wheat thaumatin-like protein TaTLP1 using a yeast two-hybrid assay and the Pt_21-TaTLP1 interaction was characterized. The interaction between Pt_21 and TaTLP1 was validated by in vivo co-immunoprecipitation assay. A TaTLP1 variant,TaTLP1C71A, that was identified by the site-directed mutagenesis failed to interact with Pt_21. Pt_21was able to suppress Bax-mediated cell death in leaves of Nicotiana benthamiana and inhibit TaTLP1-mediated antifungal activity. Furthermore, infiltration of recombinant protein Pt_21 into leaves of transgenic wheat line overexpressing TaTLP1 enhanced the disease development of leaf rust compared to that in wild-type leaves. These findings demonstrate that Pt_21 suppresses host defense response by directly targeting wheat TaTLP1 and inhibiting its antifungal activity, which broadens our understanding of the molecular mechanisms underlying Pt effector-mediated susceptibility in wheat.
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 project from the National Basic Research Program of China (2006CB100200)China Postdoctoral Science Foundation (20080430052)+1 种基金the National Natural Science Foundation of China(30471131)the National Key Technology R&D Program of China(2006BAD08A05,2006BAD02A16)
文摘The gene postulation, cluster analysis and pedigree analysis of 20 wheat cultivars to 28 physiological races of yellow rust (Puccinia striiformis) were conducted under greenhouse conditions, The spectrum of their resistance were compared each other. None of the 20 cultivars were resistant to all the test pathogens. The cultivars containing resistance genes Yr5 and Yr24 were not find and genes Yr8, Yrl9 and Yr27 could not be postulated from cultivars tested due to the susceptibility to all isolates used. A total of 6 probale seedling yellow rust resistance genes or gene combinations (Yrl, Yr2, Yr2 + YrHVII, Yr3 + unknown, Yr3 + Yr4, YrAlba) were postulated in the wheat cultivars (Atou, Flanders, Maris Huntsman, Bouquet, Holdfast, Elite Lepeuple, and Vilmorin 27). The gene combination Yr2+YrHVII with the highest frequency (35%) was present in 7 cultivars. The Yr genes present in some wheat cultivars could not be postulated because of non-matching virulence combinations with any of known genes. Cluster result showed that Yr2 and Yr3 are the most important genes in the cultivars. The 13 cultivars are believed to have the pedigree of Noe, which was selected from South Russian wheat. These results will be useful for wheat breeding and provide information about genetic control of wheat yellow rust.
基金supported by Jiangsu Agriculture Science and Technology Innovation Fund[Grant No.CX(18)2020]the National Natural Science Foundation of China(Grant Nos.31672192 and 31700620)+1 种基金the National Key Research and Development Program of China(Grant No.2018YFD1000402)the Fundamental Research Funds for the Central Universities(Grant No.KJQN201812).
文摘White rust caused by Puccinia horiana is a destructive disease of chrysanthemum plants.To better understand the resistance mechanisms of composite species to this disease,the leaf cuticular traits,antioxidant and defensive enzymes activities of immune(Chrysanthemum makinoi var.wakasaense)and highly susceptible(Ajania shiwogiku var.kinokuniense)species were compared.Trichome density of two species was markedly different,negatively associated with plant resistance to P.horiana.Total wax load in C.makinoi var.wakasaense was two times more than that in A.shiwogiku var.kinokuniense.The wax composition in immune one was abundant in esters and primary alcohols.Superoxide dismutase(SOD,EC 1.15.1.1),peroxidase(POD,EC 1.11.1.7),polyphenoloxidase(PPO,EC 1.14.18.1 or EC 1.10.3.2)and phenylalanine ammonia lyase(PAL,EC 4.3.1.5)activitieswere investigated.In C.makinoi var.wakasaense,the activity of SOD and POD increased rapidly after inoculation,whichmight be non-host induced reactive oxygen species(ROS)activated antioxidant enzymes,however SOD and POD remained a low and steady level in the highly susceptible one after inoculation.Quick increase in PPO activities after inoculation was observed in both species,however it remained higher in C.makinoi var.wakasaense at the late period of inoculation.PAL in C.makinoi var.wakasaense was induced after pathogen inoculation,but not in A.shiwogiku var.kinokuniense,suggesting that these two enzymes might contribute to the resistance to P.horiana.
基金supported by National Natural Science Foundation of China(30070496,30125031)Research Program of Shaanxi Province(2001K02-G4).
文摘The incompatible combinations between races of Puccinia striiformis and wheat cultivars with low reaction type resistance were examined by means of fluorescent microscopy, differential interference contrast microscopy and electron microscopy. The incompatible combinations consisted of wheat cultivar Niuzhute plus CY28, Hybrid46 plus CY29 and Tianxuan882 plus CY29, while the compatible combination was wheat cultivar Huixianhong plus CY28. The observation revealed a striking difference in the fungal development and the host responses between susceptible and resistant wheat cultivars following infection by the pathogen. The main histological manifestation of the pathogen development in the resistant wheat cultivars include inhibition of hyphal growth, delay of hyphal branching and colony formation, decrease of formation of haustorial mother cells and haustoria, and occurrence of host cell necrosis. The observation by electron microscopy demonstrated that a series of abnormal changes occurred in intercellular hyphae, haustorial mother cells and haustoria during pathogen development in the resistant wheat cultivars. The cytoplasm became more electron-dense and vacuoles in the cytoplasm increased in number and size. The cell walls of hyphae, haustorial mother cells were thickened irregularly. The organelles were disorderly distributed in the cytoplasm and the haustorial mother cells and haustoria lost their physiological function. In the final stage of the pathogen development, the intercellular hyphae, haustorial mother cells and haustoria became necrotic and collapsed. The structural defense reactions such as formation of cell wall apposition, collar and encasement of haustorium were essentially more pronounced in the infected wheat leaves of the resistant cultivars than in the susceptible one. In addition, the relationship between the host resistance expression and the histological and cytological features occurred in the incompatible combination was discussed in this paper.
文摘A high degree of virulence diversity has been maintained in the population of Puccinia graminis f. sp. tritici (Pgt) in northwestern United States. Although Berberis vulgaris is present in the region and Pgt has been isolated from aecial infections on B. vulgaris, the population is too diverse to be explained by the limited presence of B. vulgaris alone. Since 2008, we have isolated P. graminis from aecial infections on fruits of Mahonia repens and Mahonia aquifolium from northwestern United States. These two native woody shrub species, widely distributed in western North America, were once classified as resistant to P. graminis based on artificial inoculations. By isolating P. graminis from aecia, we established that M. repens and M. aquifolium along with B. vulgaris (albeit infrequent) serve as the alternate hosts ofP. graminis in the region. The isolates of P. graminis from Mahonia of North America had diverse virulence patterns and most of the isolates could be differentiated on Morocco, Line E, Chinese Spring, Little Club, LMPG-6, Rusty, and other genotypes that are considered to be universally susceptible to most Pgt isolates. This discovery explained the persistence of virulence diversity of Pgt observed in isolates derived from uredinia on cereal crops in the region. In addition to cereal crops, uredinial stage of the P. graminis population is sustained by wild grasses, especially Elymus glaucus, a native grass sharing the same habitat with the rusted Mahonia spp. Although virulence to some important stem rust resistance genes was observed in some isolates derived from Mahonia of North America when tested against single stem rust resistance gene stocks, the overall virulence is very limited in these isolates. This is likely a result of limited selection pressure on the rust population. In contrast to northwestern United Sates, the Pgt population in east of the Rocky Mountains of North America has declined steadily with a single race, QFCSC, being predominant in the last decade. This decline is likely due to a combination of factors, of which a lack of sexual recombination in the region is perhaps the most important one.
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.
基金supported by Provincial Key Scientific and Technological Project of Guizhou,China ([2007] 5003)Guizhou Province Scientific and Technological Research,China ([2007] 2051)
文摘Ultrastructural changes in both pathogen and host cells in the interaction between Puccinia striiformis and wheat cultivar (Libellula) with slow-rusting resistance were observed by transmission electron microscopy. Observations revealed marked changes in ultrastructure of both pathogen and host cells. In the pathogen respect, there were many vesicles appeared in the intercellular hyphae and gradually fused into bigger vacuoles, a number of fat drops and electron-dense granules accumulated, mitochondria became swollen and some of them degraded into vesicles, and the plasmalemma of intercellular hyphae became dark. In the haustoria, the cytoplasm degraded gradually and developed a vacuole in the center, fat drops increased, the extrahaustorial matrix widened with a great amount of electron-dense fibrillar and granular materials, and most of the haustoria died with in conjunction with the disappearance of fat drops and other organelles. Structural defense of the host, including formation of cell wall apposition, collar and papilla, occurred in the host respect. Host resistance expression and cytological features occurring in the slow-rusting resistance were discussed.
文摘Wheat leaf rust caused by Puccinia triticina, is the most common and widely distributed wheat rust in the world. In order to study the genetic structure of leaf rust population 14 pairs of AFLP and 6 pairs of FAFLP primers evaluated on 86 isolates samples collected in Iran during spring of 2009. Results showed that almost all investigated isolates were genetically different and special pattern of AFLP allele’s that confirm high genetic diversity within leaf rust population was observed. Analyses showed, all provinces were classified into three major groups particularly similar clusters were found between then neighboring provinces. Rust spore can follow the migration pattern in short and long distances to neighbor in provinces. Results indicated that the greatest variability was revealed by 97% of genetic differentiation within leaf rust populations and the lesser variation of 3% was observed between the rust populations. These results suggested that each population was not completely identical and high gene flow has occurred among the leaf rust population of different provinces. The highest differentiation and genetic distance among the Iranian leaf rust populations was detected between leaf rust population in Sistan and Baluchistan and highest similarity was observed between in Ardabil provinces. The high pathogenic variability of leaf rust races in Ardabil and Northern Khorasan may be an indication that these two regions are the center of origin of pathogenic arability. Present study shows that leaf rust population in Iran is highly dynamic and variable.
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.
文摘Puccinia striiformis, the causal agent of stripe rust in wheat, barley, and various wild grasses, produces urediniospores and teliospores on these primary or auxiliary hosts. Telial formation, which stops producing infectious urediniospores, is affected by various host and environmental conditions. However, it is not clear if variation exists among different isolates in the United States. To determine the differences in the pathogen population, 1423 isolates collected in the United States from 2013 to 2016 were tested for telial formation in the seedling stage at a diurnal temperature profile of 4°C - 20°C. The percentages of telial formation varied greatly among the isolates. Of the 1423 isolates, 62.97% produced telia with the percentages of telial formation ranging from 1% to 95% under the test conditions. The formation of telia was significantly affected by the year and the month when the isolates were collected. The epidemiological regions or states, host plants (wheat, barley, and grasses), and races of the isolates did not significantly affect telial formation. However, significant effects on telial formation were observed by interactions between year and region, year and race, month and region and among year, month, and region, as well as between year and month. The results showed that telial formation is a complex trait under the genetic control of the pathogen isolates for adaptation to different environments. Further studies are needed to identify genes involved in the formation of telia and the relationship of telial formation to the survival, aggressiveness, fitness, and evolution of the pathogen.
基金financial supports by the National Natural Science Foundation of China (31671967)the National Key Research and Development Program from the Ministry of Science and Technology, China (2016YFD0300705)+1 种基金the National GMO New Variety Breeding Project, China (2014ZX0801101B)the earmarked fund for China Agriculture Research System (CARS-3)
文摘Puccinia triticina, the causal agent of wheat leaf rust, is one of the most devastating rust fungi attacking wheat worldwide. Seventy-six isolates of the wheat leaf rust pathogen from Yunnan, Sichuan, Gansu and Henan provinces, China, were tested on wheat leaf rust differentials and the population structure was analyzed using four presumably neutral partial sequence markers such as elongation factor-1α(EF-1α), glyceraldehyde-3-phosphate dehydrogenase(GAPDH), β-tubulin(TUB) and the second largest RNA polymerase subunit(RPB2). The phenotypic diversity of Yunnan and Sichuan populations was higher than that of Gansu and Henan populations. The four populations were separated into two clusters based on the pathogenic data. A total of 12 single nucleotide polymorphisms(SNPs) and 32 haplotypes were identified among the four sequences. The 32 haplotypes were divided into two clusters in a neighbor-joining tree. Bayesian analyses also identified two clusters. Pairwise FST between populations in different regions were significantly different(P<0.05). Analysis of molecular variance(AMOVA) indicated that 68% of the total genetic variation was within populations.
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 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 (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.