Strawberry anthracnose, caused by Colletotrichum spp., is a major disease of cultivated strawberry. This study identifies 31 isolates of Colletotrichum spp. which cause strawberry anthracnose in Zhejiang Province and ...Strawberry anthracnose, caused by Colletotrichum spp., is a major disease of cultivated strawberry. This study identifies 31 isolates of Colletotrichum spp. which cause strawberry anthracnose in Zhejiang Province and Shanghai City, China. Eleven isolates were identified as C. acutatum, 10 as C. gloeosporioides and 10 as C. fragariae based on morphological characteristics, phylogenetic and sequence analyses. Species-specific polymerase chain reaction （PCR） and enzyme digestion further confirmed the identification of the Colletotrichum spp., demonstrating that these three species are currently the causal agents of strawberry anthracnose in the studied regions. Based on analysis of rDNA internal transcribed spacers （ITS） sequences, sequences of all C. acutatum were identical, and little genetic variability was observed between C. fragariae and C. gloeosporioides. However, the conservative nature of the Mvnl specific site from isolates of C. gloeosporioides was confirmed, and this site could be used to differentiate C. gloeosporioides from C. fragariae.展开更多
Although the phosphate 1（PHO1）gene family has been implicated in inorganic phosphate transport and homeostasis,the underlying mechanism of this gene in the strawberry has not yet been revealed.In the present study,w...Although the phosphate 1（PHO1）gene family has been implicated in inorganic phosphate transport and homeostasis,the underlying mechanism of this gene in the strawberry has not yet been revealed.In the present study,we analyzed the expression of the PHO1;H9 gene in the strawberry（Fragaria×ananassa）,revealing the involvement of this gene in the regulation of phosphorus（P）content.The coding sequence（CDS）of the PHO1;H9 gene,was isolated from the cultivated strawberry‘Sachinoka’and named as Fa PHO1;H9.The full-length CDS of this gene was 2 292 bp,encoding 763 amino acids,and the protein contained both SYG1/Pho81/XPR1（SPX）and ERD1/XPR1/SYG1（EXS）domains,which were involved in phosphate（Pi）signaling.Real-time reverse transcription-polymerase chain reaction（RT-PCR）data suggested that the level of Fa PHO1;H9 expression was consistent with the P content in different organs,except for the petiole.Particularly,its expression level was also correlated with P content in fruits of different developmental stages.The expression of Fa PHO1;H9 was also consistent with P content in leaves under different concentrations of P fertilizer application.Furthermore,transgenic Arabidopsis lines were generated,and the P content in Arabidopsis plants over-expressing Fa PHO1;H9was significantly higher than that in wild-type plants.Therefore,we proposed that Fa PHO1;H9 functions in P transport.展开更多
草莓Fra a是一类过敏原蛋白。为研究Fra a蛋白致敏机理,本研究根据已知的栽培草莓(Fragaria×ananassa)Fra a序列设计引物,从"红颜"草莓中克隆得到三个Fra a基因序列,其ORF长度分别为483 bp、483 bp和480 bp,各自编码160...草莓Fra a是一类过敏原蛋白。为研究Fra a蛋白致敏机理,本研究根据已知的栽培草莓(Fragaria×ananassa)Fra a序列设计引物,从"红颜"草莓中克隆得到三个Fra a基因序列,其ORF长度分别为483 bp、483 bp和480 bp,各自编码160个、160个和159个氨基酸,分别命名为Fa Fra a 1、Fa Fra a 2和Fa Fra a 3。生物信息学预测表明其均具有bet v 1家族保守结构域,预测蛋白均无跨膜结构域和信号肽。Fa Fra a 1与桃的氨基酸相似度为81%,李、杏为80%;Fa Fra a 2与月季的氨基酸相似度为82%,苹果、樱桃为81%;Fa Fra a 3与月季的氨基酸相似度为92%,苹果为83%。Fra a与蔷薇科植物的进化关系更近。构建原核表达载体pET32afraa1、pET32a-fraa2和pET32a-fraa3,转化到大肠杆菌BL-21(DE3)p Lys S菌株并诱导融合蛋白表达,结果表明目的蛋白Fra a 1、Fra a 3在原核表达系统中能顺利表达,Fra a 1在上清、沉淀中均有大量分布,Fra a 3主要存在于沉淀中。未得到Fra a 2目的蛋白。本研究为进一步了解Fra a的功能和制备抗体提供了理论依据。展开更多
基金supported by the National Natural Science Foundation of China (No.30571208)the Key Scientific and Technological Project of Hangzhou City (No.200432239),China
文摘Strawberry anthracnose, caused by Colletotrichum spp., is a major disease of cultivated strawberry. This study identifies 31 isolates of Colletotrichum spp. which cause strawberry anthracnose in Zhejiang Province and Shanghai City, China. Eleven isolates were identified as C. acutatum, 10 as C. gloeosporioides and 10 as C. fragariae based on morphological characteristics, phylogenetic and sequence analyses. Species-specific polymerase chain reaction （PCR） and enzyme digestion further confirmed the identification of the Colletotrichum spp., demonstrating that these three species are currently the causal agents of strawberry anthracnose in the studied regions. Based on analysis of rDNA internal transcribed spacers （ITS） sequences, sequences of all C. acutatum were identical, and little genetic variability was observed between C. fragariae and C. gloeosporioides. However, the conservative nature of the Mvnl specific site from isolates of C. gloeosporioides was confirmed, and this site could be used to differentiate C. gloeosporioides from C. fragariae.
基金financially supported by the National Natural Science Foundation of China (31372037)the Program for Excellent Talents in University of Liaoning Province, China (LJQ2014069)
文摘Although the phosphate 1（PHO1）gene family has been implicated in inorganic phosphate transport and homeostasis,the underlying mechanism of this gene in the strawberry has not yet been revealed.In the present study,we analyzed the expression of the PHO1;H9 gene in the strawberry（Fragaria×ananassa）,revealing the involvement of this gene in the regulation of phosphorus（P）content.The coding sequence（CDS）of the PHO1;H9 gene,was isolated from the cultivated strawberry‘Sachinoka’and named as Fa PHO1;H9.The full-length CDS of this gene was 2 292 bp,encoding 763 amino acids,and the protein contained both SYG1/Pho81/XPR1（SPX）and ERD1/XPR1/SYG1（EXS）domains,which were involved in phosphate（Pi）signaling.Real-time reverse transcription-polymerase chain reaction（RT-PCR）data suggested that the level of Fa PHO1;H9 expression was consistent with the P content in different organs,except for the petiole.Particularly,its expression level was also correlated with P content in fruits of different developmental stages.The expression of Fa PHO1;H9 was also consistent with P content in leaves under different concentrations of P fertilizer application.Furthermore,transgenic Arabidopsis lines were generated,and the P content in Arabidopsis plants over-expressing Fa PHO1;H9was significantly higher than that in wild-type plants.Therefore,we proposed that Fa PHO1;H9 functions in P transport.
文摘草莓Fra a是一类过敏原蛋白。为研究Fra a蛋白致敏机理,本研究根据已知的栽培草莓(Fragaria×ananassa)Fra a序列设计引物,从"红颜"草莓中克隆得到三个Fra a基因序列,其ORF长度分别为483 bp、483 bp和480 bp,各自编码160个、160个和159个氨基酸,分别命名为Fa Fra a 1、Fa Fra a 2和Fa Fra a 3。生物信息学预测表明其均具有bet v 1家族保守结构域,预测蛋白均无跨膜结构域和信号肽。Fa Fra a 1与桃的氨基酸相似度为81%,李、杏为80%;Fa Fra a 2与月季的氨基酸相似度为82%,苹果、樱桃为81%;Fa Fra a 3与月季的氨基酸相似度为92%,苹果为83%。Fra a与蔷薇科植物的进化关系更近。构建原核表达载体pET32afraa1、pET32a-fraa2和pET32a-fraa3,转化到大肠杆菌BL-21(DE3)p Lys S菌株并诱导融合蛋白表达,结果表明目的蛋白Fra a 1、Fra a 3在原核表达系统中能顺利表达,Fra a 1在上清、沉淀中均有大量分布,Fra a 3主要存在于沉淀中。未得到Fra a 2目的蛋白。本研究为进一步了解Fra a的功能和制备抗体提供了理论依据。
