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作物学报 ›› 2021, Vol. 47 ›› Issue (7): 1275-1296.doi: 10.3724/SP.J.1006.2021.04192
苏亚春1,2, 李聪娜1, 苏炜华1, 尤垂淮3, 岑光莉1, 张畅1, 任永娟1, 阙友雄1,2,*(
)
SU Ya-Chun1,2, LI Cong-Na1, SU Wei-Hua1, YOU Chui-Huai3, CEN Guang-Li1, ZHANG Chang1, REN Yong-Juan1, QUE You-Xiong1,2,*()
摘要：
类甜蛋白(thaumatin-like protein, TLP)在植物生长发育和抵抗逆境胁迫中发挥重要作用。为全面了解甘蔗TLP家族基因的结构和功能特性, 本研究利用甘蔗野生种割手密基因组数据库和生物信息学方法, 首先筛选获得122个SsTLP家族基因并进行蛋白理化性质、基因染色体分布、基因结构和系统进化等分析; 其次, 从我国大陆主栽甘蔗品种ROC22中克隆到1条SsTLP87的同源基因ScTLP1, 并对其基因表达模式、亚细胞定位、瞬时表达和转录自激活活性进行鉴定。结果表明, 122个SsTLP家族基因分布在28条割手密染色体上, 且存在多基因聚集现象, 每条染色体上有1~13个SsTLP基因, 多数SsTLP蛋白被预测为定位于细胞质的酸性疏水性不稳定蛋白。SsTLP家族基因结构类型多样, 其内含子数目(1~30)、内含子相位(0~2)和Motif (1~5)存在多种类型和分布。系统进化树分析显示, SsTLP基因编码蛋白分别聚类在TLP家族的I~X分支上, 其中具有抑菌潜力的第V类SsTLP数量最多(36个)。从甘蔗栽培种ROC22克隆获得的ScTLP1基因编码227个氨基酸残基, 属于第V类进化聚类组, 定位于细胞膜, 不具有转录自激活活性, 其与割手密SsTLP87蛋白的氨基酸序列相似性高达99.56%。qRT-PCR分析表明, ScTLP1基因在甘蔗不同组织部位组成型表达, 且在蔗芽部位的表达量最高。ScTLP1基因在脱落酸(abscisic acid, ABA)和水杨酸(salicylic acid, SA)处理下的表达量无显著变化, 但受甘蔗黑穗病菌(Sporisorium scitaminea)、茉莉酸甲酯(methyl jasmonate, MeJA)、氯化钠(sodium chloride, NaCl)、聚乙二醇(polyethylene glycol, PEG)和4℃低温诱导上调表达。瞬时过表达ScTLP1基因后, 本氏烟(Nicotiana benthamiana)叶片中的DAB染色加深, 乙烯合成相关基因NtEFE26和茉莉酸代谢途径相关基因NtPR3上调表达, 且接种烟草青枯菌(Pseudomonas solanacearum)和烟草茄病镰刀菌蓝色变种(Fusarium solani var. coeruleum)的ScTLP1基因瞬时过表达叶片发病情况较对照组轻, 表明ScTLP1基因可以诱发本氏烟的过敏反应, 参与乙烯和茉莉酸信号传导途径, 增强本氏烟对病原菌的防御效应。研究结果为深入探究甘蔗TLP家族基因的结构和功能奠定了良好的基础。
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