关键词: 茉莉酸甲基转移酶, 紫花苜蓿, 基因克隆, 序列分析, 不育系

Abstract: To explore the effects of Jasmonates (JAs) on the fertility of alfalfa (Medicago sativa L.) under the background of per-laboratory BSA-Seq analysis on sterile lines, this study successfully cloned a 1 047 bp gene from alfalfa genome by PCR method. The function is to control jasmonate O-methyl transferase synthesis gene, named MsJMT. Analysis of the expression pattern showed that the gene encoded 348 amino acids, and the encoding protein was an unstable non-secretory hydrophilic protein. Subcellular localization results showed nuclear and cytoplasmic expression. The amino acid sequence of the gene was 99% similar to that of five leguminous plants. qPCR was used to analyze the spatio-temporal expression differences in the bud fertility of alfalfa. The results showed that the expression levels of sterile lines were up-regulated and significantly higher than those of maintainers in stage Ⅲ at the 0.01 level. However, the Jasmonate (JA) content of flower buds in stages Ⅰ to Ⅳ was opposite. The results showed contrasted that the JA content in the sterile line was lower than that in the maintainer line during the whole period, and the highest was in the maintainer line Ⅲ. The buds of stage Ⅲ were treated by spraying with exogenous Methyl jasmonate (MeJA). The results showed that the anther degenerated in advance at 0.5 mmol·L^-1 and 1 mmol·L^-1levels. The conclusion was that MsJMT may affect alfalfa fertility through the JA pathway.

Key words: Jasmonate methyl transferase (JMT), Medicago sativa L., Gene cloning, Sequence analysis, Sterile line