作物学报 ›› 2022, Vol. 48 ›› Issue (11): 2934-2944.doi: 10.3724/SP.J.1006.2022.14243
许彬1,2(
), 曹绍玉1,2(
), 苏甜1, 彭梦玲1,2, 吕霞1, 李振林1, 张国平1, 许俊强1,*(
)
XU Bin1,2(
), CAO Shao-Yu1,2(
), SU Tian1, PENG Meng-Ling1,2, LYU Xia1, LI Zhen-Lin1, ZHANG Guo-Ping1, XU Jun-Qiang1,*(
)
摘要:
类钙调蛋白作为钙调蛋白的家族之一, 调控植物生长发育、激素调控及各种胁迫, 对类钙调蛋白对参与花粉萌发及伸长的深入研究具有重要意义。为研究结球甘蓝花粉CMLs与NPG1及相关蛋白互作关系, 探索CMLs在花粉发育及萌发过程的功能, 本研究以结球甘蓝ZG为材料, 克隆到12个花粉发育及萌发相关CMLs基因, 及花粉萌发基因NPG1、NPGR1和NPGR2, 通过酵母双杂交试验检测编码蛋白相互之间的互作关系, 并检测NPG1、NPGR1和NPGR2在不同授粉时间后的表达水平。结果表明, 12个CMLs基因开放阅读框(open reading frame, ORF)在447~1095 bp之间, 其中CML20、CML48、CML49、CML50均含有4个内含子, CML21含有2个内含子, 其余均不含内含子; 编码蛋白均不含信号肽和跨膜域, 均为胞外蛋白, 且均为亲水性蛋白; 分别含有2~4个EF-Hands结构域, 氨基酸数目在148~364个氨基酸之间, 亚细胞定位预测主要定位于细胞膜、液泡及细胞核中; 结球甘蓝NPG1、NPGR1、NPGR2分别聚类到不同的进化枝上, 与油菜、白菜的亲缘关系最近, 与拟南芥亲缘关系较近; 酵母双杂交试验表明, CML13、CML21、CML24、CML42不能与NPG1、NPGR1及NPGR2在体外发生相互作用, CML15、CML20、CML25、CML39、CML41、CML48、CML49均能与NPG1、NPGR1及NPGR2在体外发生相互作用, CML50可与NPGR1、NPGR2互作, 而不能与NPG1发生互作。表达分析表明, NPG1表达量在授粉后的柱头中呈显著上升趋势, NPGR1、NPGR2表达量变化均不显著; CMLs在不同授粉时间均有不同的表达量变化, 其中CML15、CML20、CML39、CML41、CML49均呈现出先升高后下降的趋势。本研究在结球甘蓝中筛选得到能与NPG1、NPGRs互作的CMLs蛋白, 说明多个CMLs能参与花粉萌发及花粉管伸长过程, 期望为Ca2+信号系统参与花粉萌发及花粉管伸长的机制研究提供参考。
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