A complete cDNA of potato Phytophthora infestans-induced hypersensitive response-related protein gene (POTHR-1) was cloned using rapid amplification of cDNA ends (RACE) strategy according to a fragment sequence which ...A complete cDNA of potato Phytophthora infestans-induced hypersensitive response-related protein gene (POTHR-1) was cloned using rapid amplification of cDNA ends (RACE) strategy according to a fragment sequence which we had cloned using suppression subtractive hybridization (SSH) technique. The potato POTHR-1 gene encodes a protein of 225 amino acids, which shares 81% identity with tobacco hin1 gene-enoded protein (harpin-induced protein). Southern blot revealed that there are two to three copies of POTHR-1 in potato genome. The POTHR-1 gene expression in potato leaves showed that its transcripts accumulated remarkably in leaves after 36 h inoculation with P. infestans. Mechanical wounding and jasmonic acid (JA) could induce the POTHR-1 gene expression and osmotic stress just induce a slight accumulation of POTHR-1 gene mRNA, while salicylic acid (SA) had no detectable function on the induction accumulation of POTHR-1 gene transcripts. The potato POTHR-1 gene may preferentially associate with hypersensitive response (HR) or biotic cell death during interaction between host and pathogen.展开更多
Late blight caused by Phytophthora infestans is the most serious disease of tomato production in China. Studies on the genetics of resistance and identification of molecular markers are very useful for breeding late b...Late blight caused by Phytophthora infestans is the most serious disease of tomato production in China. Studies on the genetics of resistance and identification of molecular markers are very useful for breeding late blight resistant varieties. The objective of this paper was to study the inheritance of late blight resistance and identify simple sequence repeat （SSR） markers associated with resistance allele in tomato （Lycopersicon esculentum Mill）. The results came from an F2 progeny of 241 plants derived from a cross between 5~ inbred line that is susceptible to late blight and a resistant accession CLN2037E. The late blight responses of F2 plants were tested by artificially inoculation of detached-leaflets in plate and natural infection assayed under greenhouse conditions. Both methods showed that the resistance is dominant and inherited as monogenic trait. Genetic mapping and linkage analysis showed that the late blight resistance gene Ph-ROL was located on chromosome 9 with a genetic distance of 5.7 cM to the SSR marker TOM236.展开更多
Potato late blight caused by Phytophthora infestans is one of the most destructive plant diseases that threaten global food security. Early and effective diagnosis of P. infestans is required before disease management...Potato late blight caused by Phytophthora infestans is one of the most destructive plant diseases that threaten global food security. Early and effective diagnosis of P. infestans is required before disease management decisions are made. Here, we developed a quick protocol to detect P. infestans based on a loop-mediated isothermal amplification(LAMP) assay. The P. infestans specific multiple copy DNA sequences(PiSMC), a transposon-like element, provides an ideal target for molecular detection of this pathogen. We designed highly specific and sensitive primers allowing effective LAMP detection of the pathogen at 64℃ in 70 min. In the validation assay, all 15 P. infestans isolates collected from China, Europe and South America could be positively detected, but results of the other 9 Phytophthora species infecting different plants, fungal and bacterial plant pathogens tested were negative. The detection limit of this assay is 1 pg P. infestans DNA. Moreover, the LAMP-PiSMC assay is able to detect P. infestans from infected leaves, tubers and soil. Taken together, this study reports the development of a specific and sensitive LAMP-PiSMC assay for early diagnosis of potato late blight.展开更多
Using a subtractive hybridization （SH）/cDNA-AFLP combinational approach, differentially expressed genes involved in the potato-Phytophthora infestans interaction were identified. These included genes potentially con...Using a subtractive hybridization （SH）/cDNA-AFLP combinational approach, differentially expressed genes involved in the potato-Phytophthora infestans interaction were identified. These included genes potentially controlling pathogenesis or avr genes in P. infestans as well as those potentially involved in potato resistance or susceptibility to this pathogen. Forty-one differentially expressed transcript, derived fragments （TDFs）, resulting from the interaction, were cloned and sequenced. Two TDFs, suggested as potential pathogenicity factors, have sequence similarity to N-succinyl diaminopimelate aminotransferase and a transcriptional regulator, TetR family gene, respectively. Two other TDFs, suggested as potential avr genes, have sequence similarity to an EST sequence from Avr41Cf.41Avr91Cf- 9 and a P. infestans avirulence-associated gene, respectively. Genes＇ expression and origin were confirmed using Southern blots, Northern blots and qRT-PCR, he., potential resistance gene DL81 was induced at 12 hpi in the moderately resistant cultivar, whereas it was down-regulated as early as 6 hpi in the susceptible cultivar. On the other hand, DL21 was induced at 6 hpi （3.38-fold） in response to the highly aggressive isolate （US8） and strongly up-regulated thereafter （25.13-fold at 120 hpi.）, whereas it was only slightly up-regulated in response to the weakly aggressive isolate US11 （3.82-fold at 96 hpi）, suggesting its potential involvement as a susceptibility gene.展开更多
Alternative splicing(AS)of pre-mRNAs increases transcriptome and proteome diversity,regulates gene expression through multiple mechanisms,and plays important roles in plant development and stress responses.However,the...Alternative splicing(AS)of pre-mRNAs increases transcriptome and proteome diversity,regulates gene expression through multiple mechanisms,and plays important roles in plant development and stress responses.However,the prevalence of genome-wide plant AS changes during infection and the mechanisms by which pathogens modulate AS remain poorly understood.Here,we examined the global AS changes in tomato leaves infected with Phytophthora infestans,the infamous Irish famine pathogen.We show that more than 2000 genes exhibiting significant changes in AS are not differentially expressed,indicating that AS is a distinct layer of transcriptome reprogramming during plant-pathogen interactions.Furthermore,our results show that P.infestans subverts host immunity by repressing the AS of positive regulators of plant immunity and promoting the AS of susceptibility factors.To study the underlying mechanism,we established a luminescence-based AS reporter system in Nicotiana benthamiana to screen pathogen effectors modulating plant AS.We identified nine splicing regulatory effectors(SREs)from 87 P.infestans effectors.Further studies revealed that SRE3 physically binds U1-70K to manipulate the plant AS machinery and subsequently modulates AS-mediated plant immunity.Our study not only unveils genome-wide plant AS reprogramming during infection but also establishes a novel AS screening tool to identify SREs from a wide range of plant pathogens,providing opportunities to understand the splicing regulatory mechanisms through which pathogens subvert plant immunity.展开更多
Over 170 years after the infamous Irish Potato Famine,potato late blight(PLB)caused by Phytophthora infestans remains the single most devastating disease of global potato production,causing up to 10 billion USD in yie...Over 170 years after the infamous Irish Potato Famine,potato late blight(PLB)caused by Phytophthora infestans remains the single most devastating disease of global potato production,causing up to 10 billion USD in yield loss and management costs.Through decades of research,growers and agronomists in the field as well as laboratory scientists have made significant progress in understanding the molecular pathogenesis process of this critical pathosystem and effective management strategies to control PLB.Yet,the need to feed an ever-increasing global population under changing climate demands continued improvement in efficient and sustainable PLB management schemes that can be implemented across a broad economic spectrum.In this review,we briefly summarize the current understanding of the molecular interaction between P.infestans and its host plants,highlight the current integrated pest management strategy to control PLB on local and continental scales,and discuss the potential of further improvement of sustainable PLB control through genetic enhancement of crop resistance and emerging crop protection technologies.展开更多
Phosphite (Phi)-based fungicides are used to control the oomycete Phytophthora infestans which causes late blight disease, the most devastating disease in potatoes. In order to examine the effects of Phi-based fungici...Phosphite (Phi)-based fungicides are used to control the oomycete Phytophthora infestans which causes late blight disease, the most devastating disease in potatoes. In order to examine the effects of Phi-based fungicides on potato tubers through foliar or post-harvest application, a metabolite profiling approach based on gas chromatography coupled to mass spectrometry (GC-MS) has been established. A total of 132 metabolites were detected using the GC-MS approach. Among these, 34 metabolites were identified after normalization and annotated with a compound name with standard mass spectral library. Metabolomic analysis of Phi-treated plants showed significant differences in the levels of many metabolites especially amino acids. Multivariate statistical approaches, such as principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA), were employed to explore the relationships between metabolites to detect group differences. A good discrimination between the control and the Phi-treated plants was observed, which demonstrated that significant changes in the metabolite profile have been caused by the two different Phi applications (foliar or post-harvest). This finding suggests that the alteration of specific metabolite levels by accumulation of Phi can lead to resistance against the pathogen.展开更多
The isolates of Phytophthora infestans on tomato in the Guangxi Zhuang Autonomous Region, China, were determined for the sensitivities to metalaxyl, cymoxanil and dimethomorph to give the basic information for integra...The isolates of Phytophthora infestans on tomato in the Guangxi Zhuang Autonomous Region, China, were determined for the sensitivities to metalaxyl, cymoxanil and dimethomorph to give the basic information for integrating disease management. Sensitivities were tested by measuring the radial growth on agar medium amended with fungicide, compared with the floating-leaf-disk method. 239 isolates were collected from eight tomato growing areas during 2000-2006. The testing results indicated that the frequencies of sensitive, intermediate, and resistant isolates to metalaxyl were 42.26, 35.98, and 23.53%, respectively. Variations in sensitivities amongst isolates from different areas or different years were very high for metalaxyl. All isolates from Tianlin and Wuxuan were sensitive to metalaxyl, but the metalaxyl-resistant isolates predominated in Tianyang, with the frequency of 51.35%. The EC50 values of certain isolates from Tianyang were higher than 500 μg·mL^-1 and their resistance levels were over 100 000 folds. Cymoxanil has been used for nearly 10 years in Guangxi, and dimethomorph has been used for 5-6 years. However, there was no decrease in sensitivity of P. infestans populations and the sensitivities of the pathogen were nearly normally distributed. Hence, their mean ECs0 value [cymoxanil （0.1647 ±0.0255） μg·mL^-1, dimethomorph （0.0970 ± 0.0052） μg·mL^-1 could be used as the baseline sensitivities for monitoring the field resistance development. The comparison with the floating-leaf-disk method indicates that both the techniques provided equivalent results. These studies suggested that metalaxyl can be continuously applied in Tianlin, Wuxuan, and Nanning due to the resistant isolates that have not been found, while for those areas with resistant isolate, the use of metalaxyl should be reduced or alternated, and cymoxanil or dimethomorph was recommended for controlling late blight disease of tomato.展开更多
Plant pathogenic fungi produce mycotoxins which may lead host plants to produce disease symptoms and may have a significant positive role in the course of disease development.The putative Phytophthora infestans toxins...Plant pathogenic fungi produce mycotoxins which may lead host plants to produce disease symptoms and may have a significant positive role in the course of disease development.The putative Phytophthora infestans toxins were prepared by culturing in liquid medium（60 g rye and 10% tomato juice L-1） for 1 mon at 17℃,filtering through four layers of cheesecloth and being precipitated by ammonium sulfate.The resulted putative toxin solutions were used to study the effects on 3 potato varieties（both leaf and tuber tissues）.The results show that potato leaves and tubers exhibit symptoms similar to the late blight resulted from P.infestans infection.Potato varieties reacting differentially to both the toxin dilutions and toxins produced by different P.infestans isolates suggested the presence of toxin-mediated specificities between potato and P.infestans.Potato leaf and tuber tissues have different and contrary reactions to the toxin.Toxin-mediated specificities are likely present between potato and P.infestans.展开更多
Ninety isolates collected from five different provinces and cities (Hebei, Yunan, Sichuan, Chongqing, Inner Mongolia ) during 1997 - 1998 were tested with potato clones possessed 8 dominant mono-genes R1, R3, R4, R6, ...Ninety isolates collected from five different provinces and cities (Hebei, Yunan, Sichuan, Chongqing, Inner Mongolia ) during 1997 - 1998 were tested with potato clones possessed 8 dominant mono-genes R1, R3, R4, R6, R7, R9, R10, R11 and the combination of R1-R4 of the international late blight differential host. It concluded that: (1) in the 90 isolates tested 21 different races were detected, of which the 1.3.4. 7. 9. 10. 11 was very common with frequency of 32. 2%, followed by 1. 3. 4. 6. 7. 9. 10. 11 and 3.4.7. 9. 11 with frequency of 10.0%; (2) all tested resistant genes were compatible to the virulent gene of isolates of Phytophthora infestans assayed, which indicated that the virulent genes were very common in the tested population of Phytophthora infestans.展开更多
Abstract [ Objective ] This study aimed to investigate the phenotypic characteristics of Phytophthora infestans from Guizhou Province. The metalaxyl resistance, mating type and isozyme genotype of 292 P. infestans st...Abstract [ Objective ] This study aimed to investigate the phenotypic characteristics of Phytophthora infestans from Guizhou Province. The metalaxyl resistance, mating type and isozyme genotype of 292 P. infestans strains isolated from Guizhou Province were analyzed. [ Result] Metalaxyl-resistant P. infestans strains were found in different areas of Guizhou Province. Resistant, moderately resistant and susceptible strains accounted for 35.62% , 33.90% and 30.48% , re- spectively. Both mating types A1 and A2 were found, which accounted for 65.38% and 34.62%, respectively. P. infestans strains isolated from Guizhou Province mainly belonged to Gpi genotypes 100/100 and 100/100/111, accounting for 50.68% and 43.84%, respectively. [ Conclusion] This study provides the theoretical basis for the prevention and control of potato late blight in Guizhou Province.展开更多
The occurrence of sexual reproduction accelerates the population genetic variation of Phytophthora infestans and makes it more difficult to control.The systematic analysis of the differentiation of phenotype(mating ty...The occurrence of sexual reproduction accelerates the population genetic variation of Phytophthora infestans and makes it more difficult to control.The systematic analysis of the differentiation of phenotype(mating type and metalaxyl sensitivity)and genotype(mtDNA haplotype and SSR genotype)of 65 single oospore strains of P.infestans was carried out in this article.Five test strains were isolated from Heilongjiang Province and Mongolia Autonomous Region.The experiment results showed that the isolation ratio of metalaxyl resistance(MR:HR)of single oospore strains produced through the cross of medium resistance and high resistant parents was 18:13;the isolation ratio of the metalaxyl resistance(S:MR:HR)of single oospore strains produced through the cross of sensitive and high resistant parents was 4:12:7.The progenies of single oospore strains produced through self-fertility parents with medium resistance were all of the medium resistance.The mating types A1:A2 was greater than 1:1 in single oospore strains of the progenies,which did not conform to the Mendel's law of inheritance.All single oospore strains of the progenies inherited mitochondrial DNA fragments from only one parent.Sexual recombination of single oospore strains was verified by using two pairs of SSR primers(Pi4B and Pi4G).At the locus of Pi4B and Pi4G in the cross of KS-37 and KS-25,the separation frequencies of allele were 19:12 and 14:17,respectively.They produced two new genotype strains.This study could provide a basis for formulating disease control strategies.展开更多
Pathogen effectors target diverse subcellular organelles to manipulate the plant immune system.Although the nucleolus has emerged as a stress marker and several effectors are localized in the nucleolus,the roles of nu...Pathogen effectors target diverse subcellular organelles to manipulate the plant immune system.Although the nucleolus has emerged as a stress marker and several effectors are localized in the nucleolus,the roles of nucleolar-targeted effectors remain elusive.In this study,we showed that Phytophthora infestans infection of Nicotiana benthamiana results in nucleolar inflation during the transition from the biotrophic to the necrotrophic phase.Multiple P.infestans effectors were localized in the nucleolus:Pi23226 induced cell death in N.benthamiana and nucleolar inflation similar to that observed in the necrotrophic stage of infection,whereas its homolog Pi23015 and a deletion mutant(Pi23226DC)did not induce cell death or affect nucleolar size.RNA immunoprecipitation and individual-nucleotide-resolution UV crosslinking and immunoprecipitation sequencing analysis indicated that Pi23226 bound to the 30 end of 25S rRNA precursors,resulting in accumulation of unprocessed 27S pre-rRNAs.The nucleolar stress marker NAC082 was strongly upregulated under Pi23226-expressing conditions.Pi23226 subsequently inhibited global protein translation in host cells by interacting with ribosomes.Pi23226 enhanced P.infestans pathogenicity,indicating that Pi23226-induced ribosome malfunction and cell death were beneficial for pathogenesis in the host.Our results provide evidence for the molecular mechanism underlying RNA-binding effector activity in host ribosome biogenesis and lead to new insights into the nucleolar action of effectors in pathogenesis.展开更多
[Objective] The paper was to improve polyacrylamide gel electrophorus in Phytophthora infeatans SSR Marker.[Method] With the disease sample of P.infeatans collected from Guyuan in Ningxia Province in 2009 as test mate...[Objective] The paper was to improve polyacrylamide gel electrophorus in Phytophthora infeatans SSR Marker.[Method] With the disease sample of P.infeatans collected from Guyuan in Ningxia Province in 2009 as test material,its DNA was extracted and amplified with PCR,and its products were carried out polyacrylamide gel electrophoresis.[Result] 12% polyacrylamide gel electrophoresis was used to detect primers D13,G11 and PI02,and 8% polyacrylamide gel electrophoresis was used to detect primers PI4B,PI63,SSR4,SSR8 and SSR11,then 0.1% silver nitrate was used to stain,and an ideal electrophoresis and staining effect was obtained.[Conclusion] The electrophoresis and staining method suitable for P.infeatans SSR Marker established in the study had the characteristics of high sensitivity,simple operation and clear bands,which was an effective,simple and quick detection method.展开更多
Resistance(R) genes against plant pathogens often have age-related resistance(ARR) effects. However, the mechanism involved in this phenomenon remains unknown. In this paper, Solanum lycopersicum ‘CLN2037B' and S...Resistance(R) genes against plant pathogens often have age-related resistance(ARR) effects. However, the mechanism involved in this phenomenon remains unknown. In this paper, Solanum lycopersicum ‘CLN2037B' and S. pimpinellifolium ‘L3708' harboring the Ph-3 gene, as well as S. habrochaites ‘LA2099', ‘LA1777' and ‘LA1033' harboring quantitative trait loci(QTLs), were tested to investigate age-related resistance against late blight(LB; caused by Phytophthora infestans) in the three-leaf stage of the plants. The results demonstrated that the QTL-related LB resistance showed the same age-related resistance as the Ph-3-mediated resistance at the six- and nine-leaf stages compared with the three-leaf stage. This indicated that there is a common defense mechanism in tomatoes against P. infestans via ARR. In addition, we combined ethylene(ET), salicylic acid(SA) and jasmonic acid(JA) mutants with virus-induced gene silencing(VIGS) to study the Ph-3-dependent resistance signaling pathway. The results showed that ethylene and salicylic acid, but not jasmonic acid, are involved in the LB resistance mediated by the Ph-3 gene.展开更多
Potato late blight, caused by the oomycete Phytophthora infestans, is one of the most devastating diseases in the agricultural sector around the world. Many genes (R genes) conferring resistance to late blight have be...Potato late blight, caused by the oomycete Phytophthora infestans, is one of the most devastating diseases in the agricultural sector around the world. Many genes (R genes) conferring resistance to late blight have been identified in various potato species and most of these R genes have been used in potato breeding. The aim of this study was to develop and validate PCR-based assays for the R genes Rpi-blb1, Rpi-blb2, Rpi-blb3 and Rpi-bt1, to distinguish between late blight resistant and late blight susceptible potato progeny in the given breeding background. A total of 100 breeding progeny were screened for the presence of these R genes and tested for resistance against P. infestans mating type A2, genotype US-8 strain, using detached leaf and tuber rot assays. PCR products for the Rpi-blb1 and Rpi-bt1 resistance genes were identified in the resistant progeny but were absent in the susceptible ones;therefore these PCR assays could differentiate between late blight resistant and susceptible plants. Genotypic data from the DNA markers derived from the Rpi-blb1 and Rpi-bt1 genes was found to correlate with the phenotypic data for foliar late blight but not with data for tuber rot. Our results demonstrate that markers derived from these two R genes could be useful for marker-assisted selection (MAS) for foliar late blight resistance in potato breeding programs.展开更多
Late blight (LB), caused by the oomycete Phytophthora infestans, is one of the most devastating diseases of tomato. Three major genes Ph-1, Ph-2 and Ph-3 conferring resistance to LB have been identified and mapped to ...Late blight (LB), caused by the oomycete Phytophthora infestans, is one of the most devastating diseases of tomato. Three major genes Ph-1, Ph-2 and Ph-3 conferring resistance to LB have been identified and mapped to the chromosomes 7, 10 and 9, respectively. However, PCR-based molecular markers associated with these genes are limited. Molecular markers are extremely useful in the screening and selection of tomato lines for the development of LB resistant genotypes. The objective of this study was to identify molecular markers associated with Ph-3 gene conferring LB resistance in tomato. Four co-dominant markers were found to be associated with Ph-3, all of which were sequence characterized amplified region (SCAR) type. Breeding lines and cultivars were inoculated with a field isolate of Phytophthora infestans to collect phenotypic data on disease resistance. Genotypic data from molecular markers associated with Ph-3 were in close agreement with the phenotypic data for the lines tested. With the verification of genotypic data from novel molecular markers in known genotypes supported by phenotypic data, the novel molecular markers may be useful in screening tomato populations aiming to develop LB resistant genotypes or cloning the LB resistant genes.展开更多
文摘A complete cDNA of potato Phytophthora infestans-induced hypersensitive response-related protein gene (POTHR-1) was cloned using rapid amplification of cDNA ends (RACE) strategy according to a fragment sequence which we had cloned using suppression subtractive hybridization (SSH) technique. The potato POTHR-1 gene encodes a protein of 225 amino acids, which shares 81% identity with tobacco hin1 gene-enoded protein (harpin-induced protein). Southern blot revealed that there are two to three copies of POTHR-1 in potato genome. The POTHR-1 gene expression in potato leaves showed that its transcripts accumulated remarkably in leaves after 36 h inoculation with P. infestans. Mechanical wounding and jasmonic acid (JA) could induce the POTHR-1 gene expression and osmotic stress just induce a slight accumulation of POTHR-1 gene mRNA, while salicylic acid (SA) had no detectable function on the induction accumulation of POTHR-1 gene transcripts. The potato POTHR-1 gene may preferentially associate with hypersensitive response (HR) or biotic cell death during interaction between host and pathogen.
文摘Late blight caused by Phytophthora infestans is the most serious disease of tomato production in China. Studies on the genetics of resistance and identification of molecular markers are very useful for breeding late blight resistant varieties. The objective of this paper was to study the inheritance of late blight resistance and identify simple sequence repeat （SSR） markers associated with resistance allele in tomato （Lycopersicon esculentum Mill）. The results came from an F2 progeny of 241 plants derived from a cross between 5~ inbred line that is susceptible to late blight and a resistant accession CLN2037E. The late blight responses of F2 plants were tested by artificially inoculation of detached-leaflets in plate and natural infection assayed under greenhouse conditions. Both methods showed that the resistance is dominant and inherited as monogenic trait. Genetic mapping and linkage analysis showed that the late blight resistance gene Ph-ROL was located on chromosome 9 with a genetic distance of 5.7 cM to the SSR marker TOM236.
基金funded by the Fok Ying Tung Education Foundation of China(151025)the National Natural Science Foundation of China(31772144)the Development and Reform Commission of Shandong Province,China for funding the project。
文摘Potato late blight caused by Phytophthora infestans is one of the most destructive plant diseases that threaten global food security. Early and effective diagnosis of P. infestans is required before disease management decisions are made. Here, we developed a quick protocol to detect P. infestans based on a loop-mediated isothermal amplification(LAMP) assay. The P. infestans specific multiple copy DNA sequences(PiSMC), a transposon-like element, provides an ideal target for molecular detection of this pathogen. We designed highly specific and sensitive primers allowing effective LAMP detection of the pathogen at 64℃ in 70 min. In the validation assay, all 15 P. infestans isolates collected from China, Europe and South America could be positively detected, but results of the other 9 Phytophthora species infecting different plants, fungal and bacterial plant pathogens tested were negative. The detection limit of this assay is 1 pg P. infestans DNA. Moreover, the LAMP-PiSMC assay is able to detect P. infestans from infected leaves, tubers and soil. Taken together, this study reports the development of a specific and sensitive LAMP-PiSMC assay for early diagnosis of potato late blight.
基金supported by a grant of the Natural Sciences and Engineering Research Council of Canada (NSERC) to F. Daayf
文摘Using a subtractive hybridization （SH）/cDNA-AFLP combinational approach, differentially expressed genes involved in the potato-Phytophthora infestans interaction were identified. These included genes potentially controlling pathogenesis or avr genes in P. infestans as well as those potentially involved in potato resistance or susceptibility to this pathogen. Forty-one differentially expressed transcript, derived fragments （TDFs）, resulting from the interaction, were cloned and sequenced. Two TDFs, suggested as potential pathogenicity factors, have sequence similarity to N-succinyl diaminopimelate aminotransferase and a transcriptional regulator, TetR family gene, respectively. Two other TDFs, suggested as potential avr genes, have sequence similarity to an EST sequence from Avr41Cf.41Avr91Cf- 9 and a P. infestans avirulence-associated gene, respectively. Genes＇ expression and origin were confirmed using Southern blots, Northern blots and qRT-PCR, he., potential resistance gene DL81 was induced at 12 hpi in the moderately resistant cultivar, whereas it was down-regulated as early as 6 hpi in the susceptible cultivar. On the other hand, DL21 was induced at 6 hpi （3.38-fold） in response to the highly aggressive isolate （US8） and strongly up-regulated thereafter （25.13-fold at 120 hpi.）, whereas it was only slightly up-regulated in response to the weakly aggressive isolate US11 （3.82-fold at 96 hpi）, suggesting its potential involvement as a susceptibility gene.
基金the Chinese National Science Fund(31901862,31772144,31721004)Natural Science Foundation of Jiangsu Province(SBK2019040604)+1 种基金China Postdoctoral Science Foundation(2018M640494)the Fundamental Research Funds for the Central Uni-versities(JCQY201904,KYXK202010).
文摘Alternative splicing(AS)of pre-mRNAs increases transcriptome and proteome diversity,regulates gene expression through multiple mechanisms,and plays important roles in plant development and stress responses.However,the prevalence of genome-wide plant AS changes during infection and the mechanisms by which pathogens modulate AS remain poorly understood.Here,we examined the global AS changes in tomato leaves infected with Phytophthora infestans,the infamous Irish famine pathogen.We show that more than 2000 genes exhibiting significant changes in AS are not differentially expressed,indicating that AS is a distinct layer of transcriptome reprogramming during plant-pathogen interactions.Furthermore,our results show that P.infestans subverts host immunity by repressing the AS of positive regulators of plant immunity and promoting the AS of susceptibility factors.To study the underlying mechanism,we established a luminescence-based AS reporter system in Nicotiana benthamiana to screen pathogen effectors modulating plant AS.We identified nine splicing regulatory effectors(SREs)from 87 P.infestans effectors.Further studies revealed that SRE3 physically binds U1-70K to manipulate the plant AS machinery and subsequently modulates AS-mediated plant immunity.Our study not only unveils genome-wide plant AS reprogramming during infection but also establishes a novel AS screening tool to identify SREs from a wide range of plant pathogens,providing opportunities to understand the splicing regulatory mechanisms through which pathogens subvert plant immunity.
基金supported by the National Natural Science Foundation of China (32130088)the Guangdong Major Project of Basic and Applied Basic Research,China(2021B0301030004) to Dong Suomengthe Agricultural Science and Technology Innovation Program of Chinese Academy of Agricultural Sciences (CAAS-ASTIP) to Zhou Shaoqun
文摘Over 170 years after the infamous Irish Potato Famine,potato late blight(PLB)caused by Phytophthora infestans remains the single most devastating disease of global potato production,causing up to 10 billion USD in yield loss and management costs.Through decades of research,growers and agronomists in the field as well as laboratory scientists have made significant progress in understanding the molecular pathogenesis process of this critical pathosystem and effective management strategies to control PLB.Yet,the need to feed an ever-increasing global population under changing climate demands continued improvement in efficient and sustainable PLB management schemes that can be implemented across a broad economic spectrum.In this review,we briefly summarize the current understanding of the molecular interaction between P.infestans and its host plants,highlight the current integrated pest management strategy to control PLB on local and continental scales,and discuss the potential of further improvement of sustainable PLB control through genetic enhancement of crop resistance and emerging crop protection technologies.
文摘Phosphite (Phi)-based fungicides are used to control the oomycete Phytophthora infestans which causes late blight disease, the most devastating disease in potatoes. In order to examine the effects of Phi-based fungicides on potato tubers through foliar or post-harvest application, a metabolite profiling approach based on gas chromatography coupled to mass spectrometry (GC-MS) has been established. A total of 132 metabolites were detected using the GC-MS approach. Among these, 34 metabolites were identified after normalization and annotated with a compound name with standard mass spectral library. Metabolomic analysis of Phi-treated plants showed significant differences in the levels of many metabolites especially amino acids. Multivariate statistical approaches, such as principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA), were employed to explore the relationships between metabolites to detect group differences. A good discrimination between the control and the Phi-treated plants was observed, which demonstrated that significant changes in the metabolite profile have been caused by the two different Phi applications (foliar or post-harvest). This finding suggests that the alteration of specific metabolite levels by accumulation of Phi can lead to resistance against the pathogen.
文摘The isolates of Phytophthora infestans on tomato in the Guangxi Zhuang Autonomous Region, China, were determined for the sensitivities to metalaxyl, cymoxanil and dimethomorph to give the basic information for integrating disease management. Sensitivities were tested by measuring the radial growth on agar medium amended with fungicide, compared with the floating-leaf-disk method. 239 isolates were collected from eight tomato growing areas during 2000-2006. The testing results indicated that the frequencies of sensitive, intermediate, and resistant isolates to metalaxyl were 42.26, 35.98, and 23.53%, respectively. Variations in sensitivities amongst isolates from different areas or different years were very high for metalaxyl. All isolates from Tianlin and Wuxuan were sensitive to metalaxyl, but the metalaxyl-resistant isolates predominated in Tianyang, with the frequency of 51.35%. The EC50 values of certain isolates from Tianyang were higher than 500 μg·mL^-1 and their resistance levels were over 100 000 folds. Cymoxanil has been used for nearly 10 years in Guangxi, and dimethomorph has been used for 5-6 years. However, there was no decrease in sensitivity of P. infestans populations and the sensitivities of the pathogen were nearly normally distributed. Hence, their mean ECs0 value [cymoxanil （0.1647 ±0.0255） μg·mL^-1, dimethomorph （0.0970 ± 0.0052） μg·mL^-1 could be used as the baseline sensitivities for monitoring the field resistance development. The comparison with the floating-leaf-disk method indicates that both the techniques provided equivalent results. These studies suggested that metalaxyl can be continuously applied in Tianlin, Wuxuan, and Nanning due to the resistant isolates that have not been found, while for those areas with resistant isolate, the use of metalaxyl should be reduced or alternated, and cymoxanil or dimethomorph was recommended for controlling late blight disease of tomato.
基金supported by the grant from the Agricultural Scientific Achievements Transform Foundation,China (02EFN215300561)the Ministry of Science and Technology of Yunnan Province,China (2002NG09)the Modern Agriculture Industrial Technology System in Yunnan Province,China
文摘Plant pathogenic fungi produce mycotoxins which may lead host plants to produce disease symptoms and may have a significant positive role in the course of disease development.The putative Phytophthora infestans toxins were prepared by culturing in liquid medium（60 g rye and 10% tomato juice L-1） for 1 mon at 17℃,filtering through four layers of cheesecloth and being precipitated by ammonium sulfate.The resulted putative toxin solutions were used to study the effects on 3 potato varieties（both leaf and tuber tissues）.The results show that potato leaves and tubers exhibit symptoms similar to the late blight resulted from P.infestans infection.Potato varieties reacting differentially to both the toxin dilutions and toxins produced by different P.infestans isolates suggested the presence of toxin-mediated specificities between potato and P.infestans.Potato leaf and tuber tissues have different and contrary reactions to the toxin.Toxin-mediated specificities are likely present between potato and P.infestans.
文摘Ninety isolates collected from five different provinces and cities (Hebei, Yunan, Sichuan, Chongqing, Inner Mongolia ) during 1997 - 1998 were tested with potato clones possessed 8 dominant mono-genes R1, R3, R4, R6, R7, R9, R10, R11 and the combination of R1-R4 of the international late blight differential host. It concluded that: (1) in the 90 isolates tested 21 different races were detected, of which the 1.3.4. 7. 9. 10. 11 was very common with frequency of 32. 2%, followed by 1. 3. 4. 6. 7. 9. 10. 11 and 3.4.7. 9. 11 with frequency of 10.0%; (2) all tested resistant genes were compatible to the virulent gene of isolates of Phytophthora infestans assayed, which indicated that the virulent genes were very common in the tested population of Phytophthora infestans.
基金Supported by Agricultural Science and Technology Project of Guizhou Province in 2011([2011]3022)
文摘Abstract [ Objective ] This study aimed to investigate the phenotypic characteristics of Phytophthora infestans from Guizhou Province. The metalaxyl resistance, mating type and isozyme genotype of 292 P. infestans strains isolated from Guizhou Province were analyzed. [ Result] Metalaxyl-resistant P. infestans strains were found in different areas of Guizhou Province. Resistant, moderately resistant and susceptible strains accounted for 35.62% , 33.90% and 30.48% , re- spectively. Both mating types A1 and A2 were found, which accounted for 65.38% and 34.62%, respectively. P. infestans strains isolated from Guizhou Province mainly belonged to Gpi genotypes 100/100 and 100/100/111, accounting for 50.68% and 43.84%, respectively. [ Conclusion] This study provides the theoretical basis for the prevention and control of potato late blight in Guizhou Province.
基金Supported by Heilongjiang Province Natural Science Foundation(C2016019)Cooperative Innovation System of Potato Modern Agricultural Industry Technology in Heilongjiang Province(HNWJZT201801)。
文摘The occurrence of sexual reproduction accelerates the population genetic variation of Phytophthora infestans and makes it more difficult to control.The systematic analysis of the differentiation of phenotype(mating type and metalaxyl sensitivity)and genotype(mtDNA haplotype and SSR genotype)of 65 single oospore strains of P.infestans was carried out in this article.Five test strains were isolated from Heilongjiang Province and Mongolia Autonomous Region.The experiment results showed that the isolation ratio of metalaxyl resistance(MR:HR)of single oospore strains produced through the cross of medium resistance and high resistant parents was 18:13;the isolation ratio of the metalaxyl resistance(S:MR:HR)of single oospore strains produced through the cross of sensitive and high resistant parents was 4:12:7.The progenies of single oospore strains produced through self-fertility parents with medium resistance were all of the medium resistance.The mating types A1:A2 was greater than 1:1 in single oospore strains of the progenies,which did not conform to the Mendel's law of inheritance.All single oospore strains of the progenies inherited mitochondrial DNA fragments from only one parent.Sexual recombination of single oospore strains was verified by using two pairs of SSR primers(Pi4B and Pi4G).At the locus of Pi4B and Pi4G in the cross of KS-37 and KS-25,the separation frequencies of allele were 19:12 and 14:17,respectively.They produced two new genotype strains.This study could provide a basis for formulating disease control strategies.
基金supported by National Research Foundation of Korea(NRF)grants funded by the Korean government(MSIT)(NRF-2018R1A5A1023599[SRC],NRF-2021R1A2B5B03001613,and NRF-2019R1C1C1008698).
文摘Pathogen effectors target diverse subcellular organelles to manipulate the plant immune system.Although the nucleolus has emerged as a stress marker and several effectors are localized in the nucleolus,the roles of nucleolar-targeted effectors remain elusive.In this study,we showed that Phytophthora infestans infection of Nicotiana benthamiana results in nucleolar inflation during the transition from the biotrophic to the necrotrophic phase.Multiple P.infestans effectors were localized in the nucleolus:Pi23226 induced cell death in N.benthamiana and nucleolar inflation similar to that observed in the necrotrophic stage of infection,whereas its homolog Pi23015 and a deletion mutant(Pi23226DC)did not induce cell death or affect nucleolar size.RNA immunoprecipitation and individual-nucleotide-resolution UV crosslinking and immunoprecipitation sequencing analysis indicated that Pi23226 bound to the 30 end of 25S rRNA precursors,resulting in accumulation of unprocessed 27S pre-rRNAs.The nucleolar stress marker NAC082 was strongly upregulated under Pi23226-expressing conditions.Pi23226 subsequently inhibited global protein translation in host cells by interacting with ribosomes.Pi23226 enhanced P.infestans pathogenicity,indicating that Pi23226-induced ribosome malfunction and cell death were beneficial for pathogenesis in the host.Our results provide evidence for the molecular mechanism underlying RNA-binding effector activity in host ribosome biogenesis and lead to new insights into the nucleolar action of effectors in pathogenesis.
基金Supported by Special Research Project of National Nonprofit Industry(3-20)Funded Projects of Modern Agricultural Technology System(nycytx-15)~~
文摘[Objective] The paper was to improve polyacrylamide gel electrophorus in Phytophthora infeatans SSR Marker.[Method] With the disease sample of P.infeatans collected from Guyuan in Ningxia Province in 2009 as test material,its DNA was extracted and amplified with PCR,and its products were carried out polyacrylamide gel electrophoresis.[Result] 12% polyacrylamide gel electrophoresis was used to detect primers D13,G11 and PI02,and 8% polyacrylamide gel electrophoresis was used to detect primers PI4B,PI63,SSR4,SSR8 and SSR11,then 0.1% silver nitrate was used to stain,and an ideal electrophoresis and staining effect was obtained.[Conclusion] The electrophoresis and staining method suitable for P.infeatans SSR Marker established in the study had the characteristics of high sensitivity,simple operation and clear bands,which was an effective,simple and quick detection method.
基金funded by the National Natural Science Foundation of China(31171975)The Agricultural Science and Technology Innovation Program(CAAS-ASTIP-IVFCAAS)+3 种基金the Key Laboratory of Biology and Genetic Improvement of Horticultural Crops,Ministry of AgricultureP.R.Chinathe Key Laboratory of Biology and Genetic Improvement of Root and Tuber Crops,Ministry of AgricultureP.R.China
文摘Resistance(R) genes against plant pathogens often have age-related resistance(ARR) effects. However, the mechanism involved in this phenomenon remains unknown. In this paper, Solanum lycopersicum ‘CLN2037B' and S. pimpinellifolium ‘L3708' harboring the Ph-3 gene, as well as S. habrochaites ‘LA2099', ‘LA1777' and ‘LA1033' harboring quantitative trait loci(QTLs), were tested to investigate age-related resistance against late blight(LB; caused by Phytophthora infestans) in the three-leaf stage of the plants. The results demonstrated that the QTL-related LB resistance showed the same age-related resistance as the Ph-3-mediated resistance at the six- and nine-leaf stages compared with the three-leaf stage. This indicated that there is a common defense mechanism in tomatoes against P. infestans via ARR. In addition, we combined ethylene(ET), salicylic acid(SA) and jasmonic acid(JA) mutants with virus-induced gene silencing(VIGS) to study the Ph-3-dependent resistance signaling pathway. The results showed that ethylene and salicylic acid, but not jasmonic acid, are involved in the LB resistance mediated by the Ph-3 gene.
文摘Potato late blight, caused by the oomycete Phytophthora infestans, is one of the most devastating diseases in the agricultural sector around the world. Many genes (R genes) conferring resistance to late blight have been identified in various potato species and most of these R genes have been used in potato breeding. The aim of this study was to develop and validate PCR-based assays for the R genes Rpi-blb1, Rpi-blb2, Rpi-blb3 and Rpi-bt1, to distinguish between late blight resistant and late blight susceptible potato progeny in the given breeding background. A total of 100 breeding progeny were screened for the presence of these R genes and tested for resistance against P. infestans mating type A2, genotype US-8 strain, using detached leaf and tuber rot assays. PCR products for the Rpi-blb1 and Rpi-bt1 resistance genes were identified in the resistant progeny but were absent in the susceptible ones;therefore these PCR assays could differentiate between late blight resistant and susceptible plants. Genotypic data from the DNA markers derived from the Rpi-blb1 and Rpi-bt1 genes was found to correlate with the phenotypic data for foliar late blight but not with data for tuber rot. Our results demonstrate that markers derived from these two R genes could be useful for marker-assisted selection (MAS) for foliar late blight resistance in potato breeding programs.
文摘Late blight (LB), caused by the oomycete Phytophthora infestans, is one of the most devastating diseases of tomato. Three major genes Ph-1, Ph-2 and Ph-3 conferring resistance to LB have been identified and mapped to the chromosomes 7, 10 and 9, respectively. However, PCR-based molecular markers associated with these genes are limited. Molecular markers are extremely useful in the screening and selection of tomato lines for the development of LB resistant genotypes. The objective of this study was to identify molecular markers associated with Ph-3 gene conferring LB resistance in tomato. Four co-dominant markers were found to be associated with Ph-3, all of which were sequence characterized amplified region (SCAR) type. Breeding lines and cultivars were inoculated with a field isolate of Phytophthora infestans to collect phenotypic data on disease resistance. Genotypic data from molecular markers associated with Ph-3 were in close agreement with the phenotypic data for the lines tested. With the verification of genotypic data from novel molecular markers in known genotypes supported by phenotypic data, the novel molecular markers may be useful in screening tomato populations aiming to develop LB resistant genotypes or cloning the LB resistant genes.
