Plant-associated microbes represent a key determinant of plant fitness through acquiring nutrients,promoting growth,and resisting to abiotic and biotic stresses.However,an extensive characterization of the bacterial a...Plant-associated microbes represent a key determinant of plant fitness through acquiring nutrients,promoting growth,and resisting to abiotic and biotic stresses.However,an extensive characterization of the bacterial and fungal microbiomes present in different plant compartments of soybean in field conditions has remained elusive.In this study,we investigated the effects of four niches(roots,stems,leaves,and pods),four genotypes(Andou 203,Hedou 12,Sanning 16,and Zhonghuang 13),and three field locations(Jining,Suzhou,and Xuzhou)on the diversity and composition of bacterial and fungal communities in soybean using 16S and internal transcribed spacer rRNA amplicon sequencing,respectively.The soybean microbiome significantly differed across organs.Host genotypes explained more variation in stem bacterial community composition and leaf fungal community composition.Field location significantly affected the composition of bacterial communities in all compartments and the effects were stronger in the root and stem than in the leaf and pod,whereas field location explained more variation in stem and leaf fungal community composition than in the root and pod.The relative abundances of potential soybean fungal pathogens also differed among host organs and genotypes,reflecting the niches of these microbes in the host and probably their compatibility to the host genotypes.Systematic profiling of the microbiome composition and diversity will aid the development of plant protection technologies to benefit soybean health.展开更多
We developed one species-specific PCR assays for rapid and accurate detection of the pathogenic fungi Verticilliumalbo-atrum in diseased plant tissues and soil. Based on differences in internal transcribed spacer (ITS...We developed one species-specific PCR assays for rapid and accurate detection of the pathogenic fungi Verticilliumalbo-atrum in diseased plant tissues and soil. Based on differences in internal transcribed spacer (ITS) sequences ofVerticillium spp., a pair of species-specific primers, Vaa1/Vaa2, was synthesized. After screening 17 isolates of V. albo-atrum, 121 isolates from the Ascomycota, Basidiomycota, Deuteromycota, and Oomycota, the Vaa1/Vaa2 primers amplifiedonly a single PCR band of approximately 330 bp from V. albo-atrum. The detection sensitivity with primers Vaa1/Vaa2 was10 fg of genomic DNA. Using ITS1/ITS4 as the first-round primers, combined with Vaa1/Vaa2, the nested PCR procedureswere developed, and the detection sensitivity increased 1 000-fold to 10 ag. The detection sensitivity for the soil pathogenswas 100-conidia g-1 soil. The PCR-based methods developed here could simplify both plant disease diagnosis and pathogenmonitoring as well as guide plant disease management.展开更多
Soybean root diseases are associated with numerous fungal and oomycete pathogens;however,the community dynamics and interactions of these pathogens are largely unknown.We performed 13 loop-mediated isothermal amplific...Soybean root diseases are associated with numerous fungal and oomycete pathogens;however,the community dynamics and interactions of these pathogens are largely unknown.We performed 13 loop-mediated isothermal amplification(LAMP)assays that targeted specific soybean root pathogens,and traditional isolation assays.A total of 159 samples were collected from three locations in the Huang-Huai-Hai region of China at three soybean growth stages(30,60,and 90 days after planting)in 2016.In LAMP results,we found that pathogen communities differed slightly among locations,but changed dramatically between soybean growth stages.Phytophthora sojae,Rhizoctonia solani,and Fusarium oxysporum were most frequently detected at the early stage,whereas Phomopsis longicolla,Fusarium equiseti,and Fusarium virguliforme were most common in the later stages.Most samples(86%)contained two to six pathogen species.Interestingly,the less detectable species tended to exist in the samples containing more detected species,and some pathogens preferentially co-occurred in diseased tissue,including P.sojae–R.solani–F.oxysporum and F.virguliforme–Calonectria ilicicola,implying potential interactions during infection.The LAMP detection results were confirmed by traditional isolation methods.The isolated strains exhibited different virulence to soybean,further implying a beneficial interaction among some pathogens.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
The aim of the study was to establish a set of differential strains and to identify soybean resistant genes to Phytophthora root rot and then to apply those strains for analysis of the resistant genes Rps1a,Rps1c,and ...The aim of the study was to establish a set of differential strains and to identify soybean resistant genes to Phytophthora root rot and then to apply those strains for analysis of the resistant genes Rps1a,Rps1c,and Rps1k that soybean cultivars or lines may carry.Virulence formula of 125 Phytophthora sojae isolates were determined using the hypocotyls inoculation technique,the strains,which includ 6 isolates with different virulence formulas,were applied to identify the resistance of 55 soybean cultivars or lines and resistant genes were analyzed using the gene postulating procedure.Eighteen reaction types occurred in 55 cultivars or lines and results of gene postulation indicated that 2 cultivars or lines probably carried gene Rps1c and no cultivar may carry genes Rps1a or Rps1k.A few of soybean cultivars or lines from Huanghuai Region carry Rps genes Rps1a,Rps1c and Rps1k and tend to infect by P.sojae,so resistant cultivars or lines need to be bred and popularized actively.展开更多
基金supported by grants from the earmarked fund for China Agriculture Research System(CARS004-PS14)the National Key R&D Program of China(2018YFD0201000)the Special Fund for Agroscientific Research in the Public Interest,China(201303018)。
文摘Plant-associated microbes represent a key determinant of plant fitness through acquiring nutrients,promoting growth,and resisting to abiotic and biotic stresses.However,an extensive characterization of the bacterial and fungal microbiomes present in different plant compartments of soybean in field conditions has remained elusive.In this study,we investigated the effects of four niches(roots,stems,leaves,and pods),four genotypes(Andou 203,Hedou 12,Sanning 16,and Zhonghuang 13),and three field locations(Jining,Suzhou,and Xuzhou)on the diversity and composition of bacterial and fungal communities in soybean using 16S and internal transcribed spacer rRNA amplicon sequencing,respectively.The soybean microbiome significantly differed across organs.Host genotypes explained more variation in stem bacterial community composition and leaf fungal community composition.Field location significantly affected the composition of bacterial communities in all compartments and the effects were stronger in the root and stem than in the leaf and pod,whereas field location explained more variation in stem and leaf fungal community composition than in the root and pod.The relative abundances of potential soybean fungal pathogens also differed among host organs and genotypes,reflecting the niches of these microbes in the host and probably their compatibility to the host genotypes.Systematic profiling of the microbiome composition and diversity will aid the development of plant protection technologies to benefit soybean health.
基金We thank Dr Chen Qinghe,Dr Ko Wenhong,Dr Ho Hanhin,Dr Hu Baishi,Dr Peng Jinghuo and China General Microbiological Culture Collection Center(CGMCC)for providing some isolates.This work was supported by the China National 863 Program(2003AA249020).
文摘We developed one species-specific PCR assays for rapid and accurate detection of the pathogenic fungi Verticilliumalbo-atrum in diseased plant tissues and soil. Based on differences in internal transcribed spacer (ITS) sequences ofVerticillium spp., a pair of species-specific primers, Vaa1/Vaa2, was synthesized. After screening 17 isolates of V. albo-atrum, 121 isolates from the Ascomycota, Basidiomycota, Deuteromycota, and Oomycota, the Vaa1/Vaa2 primers amplifiedonly a single PCR band of approximately 330 bp from V. albo-atrum. The detection sensitivity with primers Vaa1/Vaa2 was10 fg of genomic DNA. Using ITS1/ITS4 as the first-round primers, combined with Vaa1/Vaa2, the nested PCR procedureswere developed, and the detection sensitivity increased 1 000-fold to 10 ag. The detection sensitivity for the soil pathogenswas 100-conidia g-1 soil. The PCR-based methods developed here could simplify both plant disease diagnosis and pathogenmonitoring as well as guide plant disease management.
基金supported by the grants to Prof.Zheng Xiaobo and Prof.Wang Yuanchao from the National Key R&D Program of China(2018YFD0201000)the earmarked fund for China Agriculture Research System(CARS-004-PS14)+1 种基金the National Natural Science Foundation of China(31721004)by the grant to Associate Prof.Ye Wenwu from the National Natural Science Foundation of China(31772140)。
文摘Soybean root diseases are associated with numerous fungal and oomycete pathogens;however,the community dynamics and interactions of these pathogens are largely unknown.We performed 13 loop-mediated isothermal amplification(LAMP)assays that targeted specific soybean root pathogens,and traditional isolation assays.A total of 159 samples were collected from three locations in the Huang-Huai-Hai region of China at three soybean growth stages(30,60,and 90 days after planting)in 2016.In LAMP results,we found that pathogen communities differed slightly among locations,but changed dramatically between soybean growth stages.Phytophthora sojae,Rhizoctonia solani,and Fusarium oxysporum were most frequently detected at the early stage,whereas Phomopsis longicolla,Fusarium equiseti,and Fusarium virguliforme were most common in the later stages.Most samples(86%)contained two to six pathogen species.Interestingly,the less detectable species tended to exist in the samples containing more detected species,and some pathogens preferentially co-occurred in diseased tissue,including P.sojae–R.solani–F.oxysporum and F.virguliforme–Calonectria ilicicola,implying potential interactions during infection.The LAMP detection results were confirmed by traditional isolation methods.The isolated strains exhibited different virulence to soybean,further implying a beneficial interaction among some pathogens.
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funded by the Soybean Industrial Science and Technology System of China (Nycytx-004)
文摘The aim of the study was to establish a set of differential strains and to identify soybean resistant genes to Phytophthora root rot and then to apply those strains for analysis of the resistant genes Rps1a,Rps1c,and Rps1k that soybean cultivars or lines may carry.Virulence formula of 125 Phytophthora sojae isolates were determined using the hypocotyls inoculation technique,the strains,which includ 6 isolates with different virulence formulas,were applied to identify the resistance of 55 soybean cultivars or lines and resistant genes were analyzed using the gene postulating procedure.Eighteen reaction types occurred in 55 cultivars or lines and results of gene postulation indicated that 2 cultivars or lines probably carried gene Rps1c and no cultivar may carry genes Rps1a or Rps1k.A few of soybean cultivars or lines from Huanghuai Region carry Rps genes Rps1a,Rps1c and Rps1k and tend to infect by P.sojae,so resistant cultivars or lines need to be bred and popularized actively.
