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本研究以香水柠檬幼嫩叶片为材料,在前期通过De Novo技术获得的F-box基因片段基础上设计引物,利用RT-PCR技术成功克隆出长度为585 bp的F-box基因,命名为LFB1,该基因编码194个氨基酸,该氨基酸序列在N端具有F-box结构域,生物信息学分析表明:该蛋白分子量为48.94 k D,等电点:5.2。进化分析发现其与克里曼丁桔(Citrus clementina)聚在一起。q RT-PCR分析表明:该基因在植株的根、茎、嫩叶、老叶、花蕾、花苞、幼果、成熟果中均有表达,LFB1基因在组织中的表达量是:茎>老叶>根>花蕾>成熟果>嫩叶>幼果;在花的不同器官的表达分析,发现该基因在雄蕊中的表达量最高,其它组织则相对较低,在雄蕊中基因的表达量是其它组织的10倍左右;在逆境胁迫下,该基因的表达量发生明显变化,表明该基因与植株的生长发育、花粉的发育以及环境胁迫可能有着一定的关系。
Based on the F-box gene fragment obtained by De Novo technology in the early stage, primers were designed based on the young leaves of Perfume and Lemon, and the F-box gene with a length of 585 bp was successfully cloned by RT-PCR and named as LFB1 This gene encodes 194 amino acids. The amino acid sequence has an F-box domain at the N-terminus. Bioinformatics analysis shows that the protein has a molecular weight of 48.94 kD and an isoelectric point of 5.2. The evolutionary analysis found it to be clustered with Citrus clementina. q RT-PCR analysis showed that the gene was expressed in roots, stems, young leaves, old leaves, buds, buds, young fruits and mature fruits. The expression level of LFB1 gene in the tissues was: stem> old leaves > Roots> buds> mature fruit> young leaves> young fruit. The expression analysis of different organs in flower showed that the expression level of the gene in stamens was the highest while that in other tissues was relatively low. The expression level of genes in stamens was other Under the stress of stress, the expression of this gene changed significantly, indicating that the gene may have a certain relationship with plant growth and development, pollen development and environmental stress.