Burkholderia glumae causing seedling rot and grain rot of rice was listed as a plant quarantine disease of China in 2007. It＇s quite necessary to set up effective detection methods for the pathogen to manage further ...Burkholderia glumae causing seedling rot and grain rot of rice was listed as a plant quarantine disease of China in 2007. It＇s quite necessary to set up effective detection methods for the pathogen to manage further dispersal of this disease. The present study combined the real-time PCR method with classical PCR to increase the detecting efficiency, and to develop an accurate, rapid and sensitive method to detect the pathogen in the seed quarantine for effective management of the disease. The results showed that all the tested strains of B. glumae produced about 139 bp specific fragments by the real-time PCR and the general PCR methods, while others showed negative PCR result. The bacteria could be detected at the concentrations of 1×10^4 CFU/mL by general PCR method and at the concentrations below 100 CFU/mL by real-time fluorescence PCR method. B. glumae could be detected when the inoculated and healthy seeds were mixed with a proportion of 1：100.展开更多
A survey on isolation and detection of the casual organism of bacterial grain rot of rice was conducted during 1997-2006. In 2006, six pathogenic bacterial strains were isolated from two symptomless seed samples of ri...A survey on isolation and detection of the casual organism of bacterial grain rot of rice was conducted during 1997-2006. In 2006, six pathogenic bacterial strains were isolated from two symptomless seed samples of rice （Oryza sativa L.） originally produced in Hainan Province and then planted in Zhejiang Province, China. They were identified as Burkholderia glumae which is the causal organism of bacterial grain rot of rice by physiological characteristics, colony morphology, pathogenicity test, Biolog, fatty acid methyl ester （FAME） analysis and RAPD-PCR compared with the four standard reference strains. It is confirmed that there is the infection of B. glumae in so-called ＇health looking seeds＇.展开更多
Bacterial panicle blight caused by Burkholderia glumae is one of the most severe seed-borne bacterial diseases of rice in the world. Currently, this disease has affected many countries of Asia, Africa, South and North...Bacterial panicle blight caused by Burkholderia glumae is one of the most severe seed-borne bacterial diseases of rice in the world. Currently, this disease has affected many countries of Asia, Africa, South and North America. It is a typical example of the shifting from minor plant disease to major disease due to the changes of environmental conditions. Some virulent factors of B. glumae have been identified, including toxoflavins and lipases, whose productions are dependent on the Tof I/Tof R quorum-sensing system, and type III effectors. In spite of its economic significance, neither effective control measure for this disease nor resistant rice variety is currently available. In recent years, genomics, transcriptomics and other molecular methods have provided useful information for better understanding the molecular mechanisms underlying B. glumae virulence and the rice defence mechanisms against pathogens. For the prevention of this pathogen, our laboratory has developed a rapid and sensitive multiplex PCR assay for detecting and distinguishing B. glumae from other Burkholderia species. This improved understanding of B. glumae will shed new light on bacterial panicle blight disease management.展开更多
The present study combined the fluoride ion-selective electrode （ISE） method with classical PCR to increase the detection efficiency of Burkholderia glumae. The four antisera of rabbit anti whole cell of B. glumae h...The present study combined the fluoride ion-selective electrode （ISE） method with classical PCR to increase the detection efficiency of Burkholderia glumae. The four antisera of rabbit anti whole cell of B. glumae had been successfully obtained by polyclonal antibody technique, which were ASBgl4, ASBg32, ASBg26 and ASBg04, respectively. The titers of four antisera reached 1：32 in ODD test and 1：5 120 in agglutination reaction. All antisera were identified to be qualified antibodies through combined method. The specificity of ASBgl4 and ASBg32 was very high, but that of ASBg26 and ASBg04 was not very satisfying. The results showed that all tested strains of B. glumae produced 500 bp specific fragments by direct PCR and immunocapture PCR detection, while others showed nega- tive PCR result. Comparing the sensitivity of these two detection methods, direct PCR could detect about 1 x 105 cfu/mL of bacterial suspension while immunocap- ture PCR could detect as few as 1× 103 cfu/mL, 102 times higher than direct PCR in sensitivity. In the detecting experiments of artificially inoculated seeds, at least two seeds were required in direct PCR, while only one seed was enough for immunocapture PCR detection.展开更多
Horizontal gene transfer(HGT)has been proved a major driving force in prokaryotic evolution.However,the molecular functions of these transferred genes in pathogenic bacteria especially plant pathogenic bacteria are st...Horizontal gene transfer(HGT)has been proved a major driving force in prokaryotic evolution.However,the molecular functions of these transferred genes in pathogenic bacteria especially plant pathogenic bacteria are still not fully investigated.In this study,the whole-genome in silico analysis was performed and found a syringopeptin synthetase(syp)homolog in Burkholderia glumae,which can cause bacterial panicle blight in rice,was predicted to be horizontally transferred from Pseudomonas ancestor with solid confidence by phylogenetic analysis.The comprehensive molecular experiments were performed to study the potential role of this gene in B.glumae.Inoculation of rice panicles with the syp mutant resulted in 60%lower disease index compared with the wild type(WT)parent strain,suggesting the requirement of syp for the full virulence of B.glumae.Chromatography analysis of exudates from B.glumae showed suppression of synthesis of metabolites analogous to syringopeptin in the mutants.All these data raise the possibility of HGT phenomenon in shaping the virulence and adaptation of B.glumae over evolutionary time.展开更多
Burkholderia glumae presumably induces a grain rot symptom of rice that is threatening to rice production in most rice producing states of the USA. The present study was to identify the causal agent of bacteria panicl...Burkholderia glumae presumably induces a grain rot symptom of rice that is threatening to rice production in most rice producing states of the USA. The present study was to identify the causal agent of bacteria panicle blight (BPB), virulence based on hypersensitive reactions and distribution of the pathogen within a plant. 178 rice panicles samples were analyzed with semi-selective media (CCNT), polymerase chain reaction (PCR) with bacterial DNA gyrase (gyrB) specific markers, and hypersensitive reactions on tobacco leaves. A total of 73 samples out of 178 produced a yellow bacterial colony with similar morphology on CCNT medium suggesting they were bacterial panicle diseases. However, with PCR reactions we only determined that 45 of 73 were due to B. glumae, and the causal agent for the remaining samples was undetermined. Within the 45 samples, 31 highly, 6 moderately, and 5 weakly virulent isolates were grouped based on lesion sizes of the hypersensitive reactions. Pathogenicity variability among the 45 B. glumae detected suggests that different degrees of pathogenicity exist. To determine the existence of bacteria in different plant tissues, naturally infected plant parts were examined with CCNT media and PCR analysis. B. glumae was again isolated from seeds followed by stems and sheaths from light yellow pigmented CCNT media. In contrast, roots and leaves show no visible yellow pigment on CCNT. Consistent PCR products were produced from the stem, sheath, and seed, but not from the root and leaves. These findings suggest that B. glumae is distributed in the stem, sheath, and seed, and not in the leaf and root. Together this study demonstrated the usefulness of artificial culture media, tobacco reactions, and DNA test with PCR for characterization of BPB, and distribution of bacteria in plants. These findings will help to understand the mechanism of bacteria translocation in plants.展开更多
基金supported by National Natural Science Foundation of China(Grant No.30671397 and No.30871655)the Public Beneficial Research Project of Agricultural Ministry,China(Grant No.nyhyzx07-056)
文摘Burkholderia glumae causing seedling rot and grain rot of rice was listed as a plant quarantine disease of China in 2007. It＇s quite necessary to set up effective detection methods for the pathogen to manage further dispersal of this disease. The present study combined the real-time PCR method with classical PCR to increase the detecting efficiency, and to develop an accurate, rapid and sensitive method to detect the pathogen in the seed quarantine for effective management of the disease. The results showed that all the tested strains of B. glumae produced about 139 bp specific fragments by the real-time PCR and the general PCR methods, while others showed negative PCR result. The bacteria could be detected at the concentrations of 1×10^4 CFU/mL by general PCR method and at the concentrations below 100 CFU/mL by real-time fluorescence PCR method. B. glumae could be detected when the inoculated and healthy seeds were mixed with a proportion of 1：100.
基金the National Natural Science Foundation of China (Grant No. 30671397)
文摘A survey on isolation and detection of the casual organism of bacterial grain rot of rice was conducted during 1997-2006. In 2006, six pathogenic bacterial strains were isolated from two symptomless seed samples of rice （Oryza sativa L.） originally produced in Hainan Province and then planted in Zhejiang Province, China. They were identified as Burkholderia glumae which is the causal organism of bacterial grain rot of rice by physiological characteristics, colony morphology, pathogenicity test, Biolog, fatty acid methyl ester （FAME） analysis and RAPD-PCR compared with the four standard reference strains. It is confirmed that there is the infection of B. glumae in so-called ＇health looking seeds＇.
基金supported by the Special Fund for Agro-Scientific Research in the Public Interest of China(Grant No.201303015)the National Natural Science Foundation of China(Grant No.30871655)
文摘Bacterial panicle blight caused by Burkholderia glumae is one of the most severe seed-borne bacterial diseases of rice in the world. Currently, this disease has affected many countries of Asia, Africa, South and North America. It is a typical example of the shifting from minor plant disease to major disease due to the changes of environmental conditions. Some virulent factors of B. glumae have been identified, including toxoflavins and lipases, whose productions are dependent on the Tof I/Tof R quorum-sensing system, and type III effectors. In spite of its economic significance, neither effective control measure for this disease nor resistant rice variety is currently available. In recent years, genomics, transcriptomics and other molecular methods have provided useful information for better understanding the molecular mechanisms underlying B. glumae virulence and the rice defence mechanisms against pathogens. For the prevention of this pathogen, our laboratory has developed a rapid and sensitive multiplex PCR assay for detecting and distinguishing B. glumae from other Burkholderia species. This improved understanding of B. glumae will shed new light on bacterial panicle blight disease management.
文摘The present study combined the fluoride ion-selective electrode （ISE） method with classical PCR to increase the detection efficiency of Burkholderia glumae. The four antisera of rabbit anti whole cell of B. glumae had been successfully obtained by polyclonal antibody technique, which were ASBgl4, ASBg32, ASBg26 and ASBg04, respectively. The titers of four antisera reached 1：32 in ODD test and 1：5 120 in agglutination reaction. All antisera were identified to be qualified antibodies through combined method. The specificity of ASBgl4 and ASBg32 was very high, but that of ASBg26 and ASBg04 was not very satisfying. The results showed that all tested strains of B. glumae produced 500 bp specific fragments by direct PCR and immunocapture PCR detection, while others showed nega- tive PCR result. Comparing the sensitivity of these two detection methods, direct PCR could detect about 1 x 105 cfu/mL of bacterial suspension while immunocap- ture PCR could detect as few as 1× 103 cfu/mL, 102 times higher than direct PCR in sensitivity. In the detecting experiments of artificially inoculated seeds, at least two seeds were required in direct PCR, while only one seed was enough for immunocapture PCR detection.
基金the National Key R&D Program of China(2018YFD0201202 and 2017YFD0201108)the Agri-X Interdisciplinary Fund of Shanghai Jiao Tong University,China(Agri-X2017010)+3 种基金the State Key Laboratory for Biology of Plant Diseases and Insect Pests of Shanghai Jiao Tong University(SKLOF201802)the Shanghai Committee of Science and Technology(19390743300)the National Natural Science Foundation of China(31200003 and 31770772)Joint Research Funds for Translational Medicine at Shanghai Jiao Tong University(ZH2018ZDA06).
文摘Horizontal gene transfer(HGT)has been proved a major driving force in prokaryotic evolution.However,the molecular functions of these transferred genes in pathogenic bacteria especially plant pathogenic bacteria are still not fully investigated.In this study,the whole-genome in silico analysis was performed and found a syringopeptin synthetase(syp)homolog in Burkholderia glumae,which can cause bacterial panicle blight in rice,was predicted to be horizontally transferred from Pseudomonas ancestor with solid confidence by phylogenetic analysis.The comprehensive molecular experiments were performed to study the potential role of this gene in B.glumae.Inoculation of rice panicles with the syp mutant resulted in 60%lower disease index compared with the wild type(WT)parent strain,suggesting the requirement of syp for the full virulence of B.glumae.Chromatography analysis of exudates from B.glumae showed suppression of synthesis of metabolites analogous to syringopeptin in the mutants.All these data raise the possibility of HGT phenomenon in shaping the virulence and adaptation of B.glumae over evolutionary time.
文摘Burkholderia glumae presumably induces a grain rot symptom of rice that is threatening to rice production in most rice producing states of the USA. The present study was to identify the causal agent of bacteria panicle blight (BPB), virulence based on hypersensitive reactions and distribution of the pathogen within a plant. 178 rice panicles samples were analyzed with semi-selective media (CCNT), polymerase chain reaction (PCR) with bacterial DNA gyrase (gyrB) specific markers, and hypersensitive reactions on tobacco leaves. A total of 73 samples out of 178 produced a yellow bacterial colony with similar morphology on CCNT medium suggesting they were bacterial panicle diseases. However, with PCR reactions we only determined that 45 of 73 were due to B. glumae, and the causal agent for the remaining samples was undetermined. Within the 45 samples, 31 highly, 6 moderately, and 5 weakly virulent isolates were grouped based on lesion sizes of the hypersensitive reactions. Pathogenicity variability among the 45 B. glumae detected suggests that different degrees of pathogenicity exist. To determine the existence of bacteria in different plant tissues, naturally infected plant parts were examined with CCNT media and PCR analysis. B. glumae was again isolated from seeds followed by stems and sheaths from light yellow pigmented CCNT media. In contrast, roots and leaves show no visible yellow pigment on CCNT. Consistent PCR products were produced from the stem, sheath, and seed, but not from the root and leaves. These findings suggest that B. glumae is distributed in the stem, sheath, and seed, and not in the leaf and root. Together this study demonstrated the usefulness of artificial culture media, tobacco reactions, and DNA test with PCR for characterization of BPB, and distribution of bacteria in plants. These findings will help to understand the mechanism of bacteria translocation in plants.
