作物学报 ›› 2023, Vol. 49 ›› Issue (9): 2472-2484.doi: 10.3724/SP.J.1006.2023.24244
玉泉馨1(), 杨宗桃1, 张海1, 程光远1, 周营栓1, 焦文迪1, 曾康1, 罗廷绪1, 黄国强1, 张木清2,*(), 徐景升1,*()
YU Quan-Xin1(), YANG Zong-Tao1, ZHANG Hai1, CHENG Guang-Yuan1, ZHOU Ying-Shuan1, JIAO Wen-Di1, ZENG Kang1, LUO Ting-Xu1, HUANG Guo-Qiang1, ZHANG Mu-Qing2,*(), XU Jing-Sheng1,*()
摘要:
植物囊泡膜蛋白相关蛋白PVA12 (plant vesicle-associated membrane protein (VAMP)-associated proteins homolog 12)属于VAP27家族蛋白, 在细胞中介导内质网囊泡运输以及膜融合。甘蔗(Saccharum spp. hybrid) PVA12应答甘蔗花叶病毒(Sugarcane mosaic virus, SCMV)侵染尚未见报道。本研究从栽培种新台糖22号(ROC22)中克隆了PVA12基因, 命名为ScPVA12。该基因开放读码框(open reading frame, ORF)的长度为735 bp, 其编码长度为244 aa的蛋白。生物信息学分析表明, ScPVA12是一种不稳定的亲水性脂溶蛋白, C端具有跨膜结构域; 二级结构中无规则卷曲占比最高; 进化树分析表明, 该蛋白在单子叶和双子叶植物中存在明显分化。酵母双杂交(yeast two-hybrid, Y2H)和双分子荧光互补(bimolecular fluorescence complementation, BiFC)试验表明, ScPVA12与SCMV-P3N-PIPO蛋白互作。亚细胞定位试验表明ScPVA12定位于内质网。共定位试验表明ScPVA12与SCMV-P3N-PIPO共定位于内质网。实时荧光定量PCR分析发现, ScPVA12基因在甘蔗各组织中均有表达, 在第8节间中的表达量最低, 在正七叶中的表达量最高; SCMV侵染对ScPVA12基因表达影响显著, 在SCMV胁迫下ScPVA12基因先下调表达, 后期恢复正常水平。
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