[Objective] This study was conducted to investigate the pathogenicity of Plasmodiophora brassicae on cabbage grown under different temperature and soil pH conditions. [Method] The pathogenicity of P. brassicae were te...[Objective] This study was conducted to investigate the pathogenicity of Plasmodiophora brassicae on cabbage grown under different temperature and soil pH conditions. [Method] The pathogenicity of P. brassicae were tested at seven different temperatures and at six different soil pH values with the resting spore concentration of lx108 （spores/g） in the soil. The plant survival rate and incidence rate of clubroot were investigated after 90 d. [Result] The incidence rate of clubroot on cabbage among the different temperature sets varied in a descending order as follows： 30 ℃〉25 ℃〉20 ℃〉35 ℃〉15 ℃〉10 ℃〉5 ℃ at soil pH value of 6, indicating that the pathogenicity of P. brassicae was weak at 5 and 10 ~（3. The incidence rate increased with soil temperature increasing from 15 to 30 ℃, but decreased at 35 ℃. The incidence rates of clubroot were 80.36%, 100%, 65%, 10.77%, 3.23% and 0% at soil pH 4, 5, 6, 7, 8 and 9 at 25 ℃, respectively. The growth of cabbage was inhibited and the survival rate was reduced at pH 4.The incidence rates of clubroot were low at pH value of 7 and 8, and was 0% at pH 9. The Chinese cabbage grew better at pH value of 5 and 6, but had high incidence rates of clubroot. [Conclusion] The results revealed that the incidence rate of clubroot on cabbage was closely related to the temperature and soil pH.展开更多
Research progress was reviewed on the differential systems for physiologic races of Plasmodiophora brassicae Woron,including Williams,differential system and European clubroot differential（ECD） set.The existing prob...Research progress was reviewed on the differential systems for physiologic races of Plasmodiophora brassicae Woron,including Williams,differential system and European clubroot differential（ECD） set.The existing problems and countermeasures of the different differential systems were discussed,and a research status quo on the molecular identification and detection of clubroot pathogen in crucifers were introduced.展开更多
Five commonly-used reference genes： ACT （actin）, UBE （ubiquitin-conjugating enzyme）, RPL2 （ribosomal protein L2）, BRP II （RNA polymerase II subunit）, and NADH （nicotinamide adenine dinucleotide） were examin...Five commonly-used reference genes： ACT （actin）, UBE （ubiquitin-conjugating enzyme）, RPL2 （ribosomal protein L2）, BRP II （RNA polymerase II subunit）, and NADH （nicotinamide adenine dinucleotide） were examined using geNorm software as reference genes for RT-qPCR. Among the tested reference genes, ACT and UBE were the most stable in all samples. In parallel, expression analysis of nitrilases in Brassica juncea var. tumida, was performed to preliminarily investigate the molecular interactions between nitrilase and clubroot development at 10, 15, 20, 25, 30, and 40 d postinoculation （dpi） with a suspension of resting spores of Plasmodiophora brassicae. The results showed that different gene expressions of nitrilases were regulated during the initial periods of clubroot development. The expression level of BjNIT1 increased sharply from 20 to 40 dpi in infected roots while there were no remarkable changes in healthy roots. From 15 to 30 dpi, the expression levels of BjNIT2 and BjNIT4 in infected roots were lower than those in non-infected roots. Finally, BjNIT2 in treatment was down approximately to control at 40 dpi. Our results suggest that BjNIT1, which promoted overproductions of auxin, might be involved in P. brassicae infection of B. juncea.展开更多
A SYBR Green I real-time PCR assay was developed to detect and quantify Plasmodiophora brassicae ribosomal DNA（rDNA） and internal transcribed spacer（ITS）.A pair of primers PBF1/PBR1 was designed based on the conse...A SYBR Green I real-time PCR assay was developed to detect and quantify Plasmodiophora brassicae ribosomal DNA（rDNA） and internal transcribed spacer（ITS）.A pair of primers PBF1/PBR1 was designed based on the conservative region of rDNA-ITS of P.brassicae.The positive plasmid pB12 was obtained and used as the template to create standard curve.The specificity,sensitivity,and reproducibility of real-time PCR were evaluated respectively.Naturally and artificially infested soil samples containing different concentrations of P.brassicae were detected.The results demonstrated that standard curve established by recombinant plasmid was shown a fine linear relationship between threshold cycle and template concentration.The melting curve was specific with the correlation coefficient of 0.995 and that the amplification efficiency was 93.8%.The detection limit of P.brassicae genomic DNA was approximately 40 copies per 25 μL.The sensitivity of the assay was at least 100-fold higher than conventional PCR.Only DNA from P.brassicae could be amplified and detected using this assay,suggesting the highly specific of this assay.The coefficient of variation was less than 3%,indicating the PCR method revealed high reproducibility.The detection limit in soil samples corresponded to 1 000 resting spores g-1soil.Bait plants were used to validate the real-time PCR assay.This developed real-time PCR assay allows for fast and sensitive detection of P.brassicae in soil and should be useful in disease management and pest interception so as to prevent further spread of P.brassicae.展开更多
基金Supported by Science and Technology Project of Yunnan Province(2014RA061)Special Fund for Modern Agriculture Research System for Rape of Yunnan Province~~
文摘[Objective] This study was conducted to investigate the pathogenicity of Plasmodiophora brassicae on cabbage grown under different temperature and soil pH conditions. [Method] The pathogenicity of P. brassicae were tested at seven different temperatures and at six different soil pH values with the resting spore concentration of lx108 （spores/g） in the soil. The plant survival rate and incidence rate of clubroot were investigated after 90 d. [Result] The incidence rate of clubroot on cabbage among the different temperature sets varied in a descending order as follows： 30 ℃〉25 ℃〉20 ℃〉35 ℃〉15 ℃〉10 ℃〉5 ℃ at soil pH value of 6, indicating that the pathogenicity of P. brassicae was weak at 5 and 10 ~（3. The incidence rate increased with soil temperature increasing from 15 to 30 ℃, but decreased at 35 ℃. The incidence rates of clubroot were 80.36%, 100%, 65%, 10.77%, 3.23% and 0% at soil pH 4, 5, 6, 7, 8 and 9 at 25 ℃, respectively. The growth of cabbage was inhibited and the survival rate was reduced at pH 4.The incidence rates of clubroot were low at pH value of 7 and 8, and was 0% at pH 9. The Chinese cabbage grew better at pH value of 5 and 6, but had high incidence rates of clubroot. [Conclusion] The results revealed that the incidence rate of clubroot on cabbage was closely related to the temperature and soil pH.
基金Supported by the National Science and Technology Program of the Ministry of Science and Technology of China(2010BAD01B04)Research Fund of Department of Science and Technology of Sichuan Province(2008NG0003)the Genetic Engineering Fund of Department of Finance of Sichuan Province(2011JYGC06)~~
文摘Research progress was reviewed on the differential systems for physiologic races of Plasmodiophora brassicae Woron,including Williams,differential system and European clubroot differential（ECD） set.The existing problems and countermeasures of the different differential systems were discussed,and a research status quo on the molecular identification and detection of clubroot pathogen in crucifers were introduced.
基金financially supported by the Natural Science Foundation of Chongqing Science and Technology Commission,China (2008BB1370)Fuling Agricultural Science Institute of Chongqing,China
文摘Five commonly-used reference genes： ACT （actin）, UBE （ubiquitin-conjugating enzyme）, RPL2 （ribosomal protein L2）, BRP II （RNA polymerase II subunit）, and NADH （nicotinamide adenine dinucleotide） were examined using geNorm software as reference genes for RT-qPCR. Among the tested reference genes, ACT and UBE were the most stable in all samples. In parallel, expression analysis of nitrilases in Brassica juncea var. tumida, was performed to preliminarily investigate the molecular interactions between nitrilase and clubroot development at 10, 15, 20, 25, 30, and 40 d postinoculation （dpi） with a suspension of resting spores of Plasmodiophora brassicae. The results showed that different gene expressions of nitrilases were regulated during the initial periods of clubroot development. The expression level of BjNIT1 increased sharply from 20 to 40 dpi in infected roots while there were no remarkable changes in healthy roots. From 15 to 30 dpi, the expression levels of BjNIT2 and BjNIT4 in infected roots were lower than those in non-infected roots. Finally, BjNIT2 in treatment was down approximately to control at 40 dpi. Our results suggest that BjNIT1, which promoted overproductions of auxin, might be involved in P. brassicae infection of B. juncea.
基金supported by the emarked fund for Moden Agro-Industry Technology Research System, China (CARS25)the National Natural Science Foundation of China (31201473)the Key Laboratory of Biology and Genetic Improvement of Horticulture Crops, Ministry of Agriculture, China
文摘A SYBR Green I real-time PCR assay was developed to detect and quantify Plasmodiophora brassicae ribosomal DNA（rDNA） and internal transcribed spacer（ITS）.A pair of primers PBF1/PBR1 was designed based on the conservative region of rDNA-ITS of P.brassicae.The positive plasmid pB12 was obtained and used as the template to create standard curve.The specificity,sensitivity,and reproducibility of real-time PCR were evaluated respectively.Naturally and artificially infested soil samples containing different concentrations of P.brassicae were detected.The results demonstrated that standard curve established by recombinant plasmid was shown a fine linear relationship between threshold cycle and template concentration.The melting curve was specific with the correlation coefficient of 0.995 and that the amplification efficiency was 93.8%.The detection limit of P.brassicae genomic DNA was approximately 40 copies per 25 μL.The sensitivity of the assay was at least 100-fold higher than conventional PCR.Only DNA from P.brassicae could be amplified and detected using this assay,suggesting the highly specific of this assay.The coefficient of variation was less than 3%,indicating the PCR method revealed high reproducibility.The detection limit in soil samples corresponded to 1 000 resting spores g-1soil.Bait plants were used to validate the real-time PCR assay.This developed real-time PCR assay allows for fast and sensitive detection of P.brassicae in soil and should be useful in disease management and pest interception so as to prevent further spread of P.brassicae.
