美洲南瓜 WRKYⅡ 家族鉴定及应答白粉菌侵染的表达分析

摘要

WRKY 家族基因在多种植物抵抗白粉病菌侵染过程中发挥重要作用。WRKY Ⅱ 亚族是 WRKY 转录因子家族的重要组成部分,但其美洲南瓜抗性白粉病研究较少。利用生物信息学方法,鉴定美洲南瓜 WRKY Ⅱ 家族基因共 67 个成员,编号为 CpWRKY01 ~ CpWRKY67 ,分布在美洲南瓜 20 条染色体上。其中,60 个基因为片段复制基因。 CpWRKYs 编码的氨基酸数目为 70~621 个,分子量为 8 126.05~67 051.06 ku ,等电点为 4.94~9.96 ,脂肪族氨基酸指数为 45.60~74.36 ,平均亲水性均为负值,且亚细胞定位分析表明,大部分家族成员定位在细胞核。 CpWRKYs 均包含 WRKY 或 WRKY superfamily 结构域,14 个转录因子还包含 plant zn clast 结构域；2 个转录因子包含 bZIP superfamily 结构域。CpWRKYs 蛋白包含 1~9 个基序,大部分成员含有基序 1 、基序 2 和基序 5。 CpWRKYs 成员启动子区域含有多个防御反应相关作用元件和植物激素相关元件。系统发育分析表明,美洲南瓜 WRKY Ⅱ 蛋白分为 5 个亚族。根据转录组和 qRT-PCR 分析白粉菌侵染不同时间的 CpWRKYs 表达量,共检测到 53 个成员表达量。其中, CpWRKY3 、 CpWRKY11 、 CpWRKY30 、 CpWRKY54 表达量在接种白粉菌后上调,且在美洲南瓜抗病近等基因系“ NIL-PM10.1 ”的表达量显著高于感病材料“ JIN234 ”。研究为 WRKY 家族基因在美洲南瓜抵抗白粉菌侵染过程的调控作用提供坚实基础,为白粉病抗性育种提供丰富的基因资源。

关键词

美洲南瓜 / WRKY 转录因子 / 基因家族 / 白粉病 / 基因表达

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基金资助

龙江科技英才春雁支持计划项目(CYQN24016)

黑龙江省重点研发计划项目(GA23B007)