论文部分内容阅读
自交不亲和性是甘蓝在长期进化过程中形成的防止自交衰败、促进杂交优势的一种复杂而完善的遗传机制。克隆自交不亲和性相关基因对甘蓝自交不亲和性的深入研究和利用有重要意义。本研究通过挖掘0~60 min自花和异花授粉的甘蓝柱头转录组数据,筛选到一个受自花授粉诱导上调表达的基因,命名为BoSPI。BoSPI开放阅读框534 bp,编码177个氨基酸,理论等电点为4.21,不包含信号肽和跨膜区,含有4个保守的EF-hand结构域。BoSPI基因起始密码子上游2000 bp的核苷酸序列中含有真菌诱导响应、代谢调节以及器官形成等应答元件。BoSPI基因在大肠杆菌中可诱导表达为17 kD的蛋白。BoSPI在柱头中表达量最高,在花瓣、萼片、叶片、雄蕊表达量较低。BoSPI蛋白被定位在细胞膜和细胞质。自花授粉30 min后对BoSPI基因的诱导表达显著增强。表明BoSPI参与了柱头响应自花花粉刺激的分子过程,可能是实现甘蓝自交不亲和性相关的某种新功能基因。
Self-incompatibility is a complex and complete genetic mechanism formed in the long-term evolution of cabbage to prevent self-fertility decline and promote the advantages of hybridization. It is of great importance to further study and utilize self-incompatibility of cloned self-incompatibility related genes in cabbage. In this study, a self-pollination-induced up-regulated gene was screened by digging the transcriptome of self-pollination and cross-pollination of Brassica oleracea from 0 to 60 min and named as BoSPI. The BoSPI open reading frame is 534 bp in length and encodes 177 amino acids with a theoretical isoelectric point of 4.21. It contains no signal peptide and transmembrane domain and contains four conserved EF-hand domains. The nucleotide sequence of 2000 bp upstream of the start codon of BoSPI gene contains response elements such as fungus-induced response, metabolism regulation and organ formation. The BoSPI gene induces a protein of 17 kD expressed in E. coli. The expression level of BoSPI was the highest in stigma, but lower in petals, sepals, leaves and stamens. BoSPI protein is localized in the cell membrane and cytoplasm. After 30 min self-pollination, the expression of BoSPI gene was significantly enhanced. This indicated that BoSPI is involved in the stigma response to self-pollen-stimulating molecular processes and may be a new functional gene related to the self-incompatibility of cabbage.