大白菜BrCML49基因的分离鉴定及表达特征分析

Identification and Expression Analysis of BrCML49 Gene in Chinese Cabbage

类钙调蛋白(Calmodulin-like protein,CML)是植物细胞中一类非常重要的钙感受蛋白,且CML蛋白在花粉萌发和花粉管生长中起到重要作用。该研究以大白菜自交不亲和系‘91-125’自花授粉后柱头为材料,克隆获得大白菜BrCML49基因cDNA全长序列1046 bp,包括960 bp开放阅读框,编码319个氨基酸,包含2个EF-hand保守结构域。序列分析发现,BrCML49蛋白为稳定的亲水蛋白,没有跨膜区。实时荧光定量(qRT-PCR)表达分析显示,BrCML49基因的表达量在‘91-125’的花药中最高,其次是全花和花瓣,在其他组织中表达量较低;BrCML49基因在大白菜自交亲和系‘14S23’的全花和柱头中表达量较高,在其他组织中表达量较低。亚细胞定位分析发现BrCML49主要定位于烟草叶片的细胞膜和细胞核中。该研究结果可为进一步研究大白菜BrCML49基因的功能奠定基础,并为阐明大白菜BrCML49基因参与自交不亲和花粉萌发及花粉管生长的信号机制提供理论依据。

Calmodulin-like protein(CML)is a very important type of calcium-sensing protein in plant cells.Studies have shown that CML proteins play critical roles in pollen germination and pollen tube growth.In this study,using the self-pollinated stigma of the Chinese cabbage self-incompatibility line‘91-125’,we cloned the full-length cDNA sequence of BrCML49 gene and obtained with 1046 bp,including 960 bp open reading frame,encoding 319 amino acids,having 2 conserved EF-hand domains.Sequence analysis found that the BrCML49 protein is a stable hydrophilic protein and does not have the transmembrane region.Real-time quantitative expression analysis(qRT-PCR)showed that BrCML49 gene exhibited the highest expression in the anther of‘91-125’,followed by the whole flower and petal,and the expression level was lower in other tissues.In the Chinese cabbage self-compatibility line‘14S23’,the BrCML49 gene was highly expressed in the whole flower and stigma,and the expression level is lower in other tissues.Subcellular localization analysis revealed that BrCML49 was mainly localized in the cell membrane and nucleus of tobacco leaf.The results of this study can lay a foundation for further study on the function of Chinese cabbage BrCML49 gene,and provide a theoretical basis for elucidating the signaling mechanism of Chinese cabbage BrCML49 gene involved in self-incompatible pollen germination and pollen tube growth.