Rice blast, caused by Magnaporthe oryzae, is one of the most devastating diseases. Using map-based strategy and in silico approach we isolated a new rice （Oryza sativa L.） blast resistance allele of Pid3, designated...Rice blast, caused by Magnaporthe oryzae, is one of the most devastating diseases. Using map-based strategy and in silico approach we isolated a new rice （Oryza sativa L.） blast resistance allele of Pid3, designated Pi25, from a stable blast resistance cultivar Gumei2. Overexpression analysis and complementation test showed that Pi25 conferred blast resistance to M. oryzae isolate js001-20. Sequence analysis showed that Pi25 was an intronless gene of 2772 nucleotides with single nucleotide substitution in comparison to Pid3 at the nucleotide position 459 and predicatively encoded a typical coiled coil-nucleotide binding site-leucine rich repeat （CC-NBS-LRR） protein of 924 amino acid residuals with 100% identity to Pid3 putative protein. The susceptible allele pi25 in Nipponbare contained a nonsense mutation at the nucleotide position 2209 resulting in a truncated protein with 736 amino acid residuals. In addition, 14 nucleotide substitutions resulting in 10 amino acid substitutions were identified between Pi25 and pi25 upstream the premature stop codon in the susceptible allele. Although the mechanism of Pi25/Pid3-mediated resistance needs to be further investigated, the isolation of the allele would facilitate the utilization of Pi25/Pid3 in rice blast resistance breeding program via transgenic approach and marker assisted selection.展开更多
NBS-LRR(nucleotide-binding site and leucine-rich-repeat)是植物中最大类抗病基因家族之一。番茄基因组测序完成为全基因组水平上分析NBS-LRR抗病基因家族提供了机遇。利用生物信息学方法对番茄NBS-LRR抗病基因家族成员数目进行鉴定...NBS-LRR(nucleotide-binding site and leucine-rich-repeat)是植物中最大类抗病基因家族之一。番茄基因组测序完成为全基因组水平上分析NBS-LRR抗病基因家族提供了机遇。利用生物信息学方法对番茄NBS-LRR抗病基因家族成员数目进行鉴定,并对其染色体定位和系统发育关系进行了分析。随后,将番茄和马铃薯NBS-LRR抗病基因进行了比较基因组学分析。结果表明:番茄基因组共包括252个NBS-LRR抗病基因,分布于番茄12条染色体上;63.5%的基因成簇存在,大部分为串联重复;系统发育关系分析表明番茄CC-NBS-LRR(CNL)亚家族较其他亚家族扩展程度大;同线性分析发现番茄中共79个NBS-LRR抗病基因与马铃薯基因具有同源关系。结果将为番茄NBS-LRR抗病基因家族的深入研究提供依据,同时也为利用番茄NBS-LRR基因进行基因定位以及相关抗病基因克隆等奠定基础。展开更多
MicroRNA （miRNA） has emerged as an important regulator of gene expression in plants. 146 miRNAs were identified from apple （Malus domestica cv. Golden Delicious） by bioinformatic analysis and RNA library sequencin...MicroRNA （miRNA） has emerged as an important regulator of gene expression in plants. 146 miRNAs were identified from apple （Malus domestica cv. Golden Delicious） by bioinformatic analysis and RNA library sequencing. From these, 135 were conserved and 11 were novel miRNAs. Target analysis predicted one of the novel miRNAs, Md-miRLn11 （Malus domestica microRNA Ln11）, targeted an apple nucleotide-binding site （NBS）-Ieucine-rich repeat （LRR） class protein coding gene （Md-NBS）. 5/ RACE assay confirmed the ability of Md-miRLn11 to cleave Md-NBS at the 11-12-nt position. Analysis of the expression of Md-miRLn11 and Md-NBS during the optimum invasion period in 40 apple varieties showed that the expression of Md-NBS gene in resistant varieties is higher than in susceptible varieties, with an inverse pattern for Md-miRLn11. Seedlings from the resistant apple variety ＇JiGuan＇ were used to carry out an Agrobacterium infiltration assay, and then inoculated with the apple leaf spot disease. The result showed a clear decline of disease resistance in JiGuan apples. In contrast, the susceptible variety ＇FuJi＇ infiltrated with the Md-NBS gene showed a significant increase in disease resistance. Based on the above results, we propose that Md-miRLn11 regulates Md-NBS gene expression in particular under the condition of pathogen infection, and that the Md-miRLn11 targeting P-loop site may regulate many NBS-LRR protein class genes in woody plants.展开更多
基金supported by the Zhejiang Natural Science Foundation(No.R307131 and No.Y3080528)the National High Technology Research and Development Program of China(No.2009AA101101)
文摘Rice blast, caused by Magnaporthe oryzae, is one of the most devastating diseases. Using map-based strategy and in silico approach we isolated a new rice （Oryza sativa L.） blast resistance allele of Pid3, designated Pi25, from a stable blast resistance cultivar Gumei2. Overexpression analysis and complementation test showed that Pi25 conferred blast resistance to M. oryzae isolate js001-20. Sequence analysis showed that Pi25 was an intronless gene of 2772 nucleotides with single nucleotide substitution in comparison to Pid3 at the nucleotide position 459 and predicatively encoded a typical coiled coil-nucleotide binding site-leucine rich repeat （CC-NBS-LRR） protein of 924 amino acid residuals with 100% identity to Pid3 putative protein. The susceptible allele pi25 in Nipponbare contained a nonsense mutation at the nucleotide position 2209 resulting in a truncated protein with 736 amino acid residuals. In addition, 14 nucleotide substitutions resulting in 10 amino acid substitutions were identified between Pi25 and pi25 upstream the premature stop codon in the susceptible allele. Although the mechanism of Pi25/Pid3-mediated resistance needs to be further investigated, the isolation of the allele would facilitate the utilization of Pi25/Pid3 in rice blast resistance breeding program via transgenic approach and marker assisted selection.
文摘NBS-LRR(nucleotide-binding site and leucine-rich-repeat)是植物中最大类抗病基因家族之一。番茄基因组测序完成为全基因组水平上分析NBS-LRR抗病基因家族提供了机遇。利用生物信息学方法对番茄NBS-LRR抗病基因家族成员数目进行鉴定,并对其染色体定位和系统发育关系进行了分析。随后,将番茄和马铃薯NBS-LRR抗病基因进行了比较基因组学分析。结果表明:番茄基因组共包括252个NBS-LRR抗病基因,分布于番茄12条染色体上;63.5%的基因成簇存在,大部分为串联重复;系统发育关系分析表明番茄CC-NBS-LRR(CNL)亚家族较其他亚家族扩展程度大;同线性分析发现番茄中共79个NBS-LRR抗病基因与马铃薯基因具有同源关系。结果将为番茄NBS-LRR抗病基因家族的深入研究提供依据,同时也为利用番茄NBS-LRR基因进行基因定位以及相关抗病基因克隆等奠定基础。
基金ACKNOWLEDGMENTS We thank the Fruit Tree Research Institute of the Chinese Academy of Agricultural Sciences for providing plant materials and apple Alternaria leaf spot （Alternaria alternata f.sp.mali）. This work was supported by the Doctoral Program Special Fund of the Ministry of Education of China （20100008110036）, the National Natural Science Foundation of China （31071784）, and the Beijing Natural Science Foundation （6102017）. No conflict of interest declared.
文摘MicroRNA （miRNA） has emerged as an important regulator of gene expression in plants. 146 miRNAs were identified from apple （Malus domestica cv. Golden Delicious） by bioinformatic analysis and RNA library sequencing. From these, 135 were conserved and 11 were novel miRNAs. Target analysis predicted one of the novel miRNAs, Md-miRLn11 （Malus domestica microRNA Ln11）, targeted an apple nucleotide-binding site （NBS）-Ieucine-rich repeat （LRR） class protein coding gene （Md-NBS）. 5/ RACE assay confirmed the ability of Md-miRLn11 to cleave Md-NBS at the 11-12-nt position. Analysis of the expression of Md-miRLn11 and Md-NBS during the optimum invasion period in 40 apple varieties showed that the expression of Md-NBS gene in resistant varieties is higher than in susceptible varieties, with an inverse pattern for Md-miRLn11. Seedlings from the resistant apple variety ＇JiGuan＇ were used to carry out an Agrobacterium infiltration assay, and then inoculated with the apple leaf spot disease. The result showed a clear decline of disease resistance in JiGuan apples. In contrast, the susceptible variety ＇FuJi＇ infiltrated with the Md-NBS gene showed a significant increase in disease resistance. Based on the above results, we propose that Md-miRLn11 regulates Md-NBS gene expression in particular under the condition of pathogen infection, and that the Md-miRLn11 targeting P-loop site may regulate many NBS-LRR protein class genes in woody plants.
