漾濞大泡核桃病程相关蛋白10基因JsPR10-1的克隆与表达分析

病程相关蛋白（pathogenesis related protein, PR） 10的激活与积累在植物抗逆境胁迫中有非常重要的作用。根据漾濞大泡核桃（Juglans sigillata）编码PR10的EST（expressed sequence tag）序列设计引物,利用快速扩增cDNA末端技术,克隆得到PR10基因的全长cDNA序列,并命名为JsPR10-1。JsPR10-1全长cDNA为776 bp,含有483 bp的开放阅读框、74 bp 5′-非编码区以及219 bp 3′-非编码区,编码含有160个氨基酸的蛋白质。全长基因序列中含有1个124 bp的内含子。JsPR10-1编码的蛋白质与栎树（Quercus suber）、欧洲山毛榉（Fagus sylvatica）以及欧洲板栗（Castanea sativa）的PR10相似性较高,并且在PR10的系统进化树中与双子叶植物聚为一支。qRT-PCR分析结果表明,植物信号分子水杨酸、茉莉酸、乙烯以及过氧化氢处理均可诱导JsPR10-1表达。在接种胶孢炭疽菌（Colletotrichum gloeosporioides）后,JsPR10-1的表达量迅速上升并在接种8 h时达到最大值,表明JsPR10-1参与漾濞大泡核桃对胶孢炭疽菌的防卫反应。本研究为揭示漾濞大泡核桃抗性机制奠定了理论依据。

Cloning and Expression Analysis of a Pathogenesis-related Protein 10 Gene from Juglans sigillata

The activation and accumulation of pathogenesis-related protein （PR）10 play predominant roles in the defense response against pathogens. Based on a Juglans sigillata expressed sequence tag lEST） encoding PR10, gene-specific primers were designed and used to amplify the full-length cDNA of a novel PR10 gene by rapid amplification of cDNA ends （RACE）. This gene was named as JsPR10-1. JsPR10-1 was 776 bp in length with an open reading frame （ORF） of 483 bp, a 5＇-untranslated region （UTR） of 74 bp, and a 3＇-UTR of 219 bp, and the ORF encoded a protein with 160 amino acid residues. There was an intron of 124 bp in the genomic sequence of JsPRIO-1. Homology analysis indicated that JsPR10-1 was homologous with PR10s from Quercus suber, Fagus sylvatica and Castanea sativa; moreover, JsPR10-1 clustered together with the PR10s from dicots in the phylogenetic tree. qRT-PCR analysis showed that the expression levels of JsPR10-1 were induced by four different plant signaling molecules, including salicylic acid, jasmonic acid, ethylene, and H202. In addition, after inoculation with Colletotrichum gloeosporioides, JsPR10-1 was sharply up-regulated with the highest expression level at 8 h. The JsPRIO-1 gene was involved in the defense response against C. gloeosporioides. This experiment lays a theoretical foundation for revealing the mechanism of resistance in J. sigillata.