关键词: 甘蓝型油菜, BnGRP1, 低磷胁迫, 基因编辑

Abstract:

Glycine rich proteins (GRPs) play an important role in plant stress response. In previous study, the candidate gene BnGRP1 in response to low phosphorus stress had been obtained by genome-wide association analysis, but its molecular mechanism was not clear. Sequence analysis showed that the total length of CDS of BnGRP1 was 399 bp, encoding 132 amino acids, of which 62 were glycine, accounting for 47.0% of the total amino acids of the protein. In order to study the biological function of BnGRP1, a double target vector of the target gene was constructed by CRISPR / Cas9 gene editing technology, and was transformed into Brassica napus. Thirteen transgenic positive plants were obtained in the T₀ generation. The mutation sites of Bngrp1 in 5 transgenic plants were analyzed by TA cloning and sequencing technology, and 3 of them showed the sequence variation of Bngrp1, which resulted in the inability of encoding the normal BnGRP1 protein. This study not only provides experimental materials for studying the biological function of BnGRP1, but also provides theoretical and experimental basis for further studying the molecular mechanism of BnGRP1 in response to low phosphorus stress.

Key words: Brassica napus, BnGRP1, low phosphorus stress, gene editing