菠萝叶刺形成相关基因AcSPL14克隆与载体构建

Cloning and Vector Construction of Pineapple Leaf Thorn Related Gene AcSPL14

本研究以菠萝叶片为材料,从中克隆到一个可能与菠萝叶刺形成相关的基因AcSPL14 (Ananas comosus squamosa promoter-binding-like protein 14)。该基因cDNA全长为1 330 bp,开放阅读框为1 086 bp,编码362个氨基酸。结构域分析结果显示AcSPL14蛋白序列具有SPL蛋白家族共有的功能结构域——SBP(SQUAMOSA promoter binding protein-like)结构域,且与石刁柏(Asparagus officinalis)、铁皮石斛(Dendrobium catenatum)、月季(Rosa chinensis)和麻风树(Jatropha curcas)的SBP序列相似性分别为53%、51%、47%和46%。在此基础上,本研究进一步构建了p BI121-AcSPL14过表达载体,获得了工程菌。本研究为AcSPL14在菠萝中的生物学功能鉴定提供了理论依据,对深入研究菠萝叶刺发生的分子机理具有重要意义。

In this study, gene AcSPL14(Ananas comosus squamosa promoter-binding-like protein 14) was cloned from the leaves of Ananas comosus, which was considerably related to the phenotype of leaf spine in pineapple.The cDNA full length of AcSPL14 was 1 330 bp, with the open reading frame(ORF) of 1 086 bp, and encoded a protein consisted of 362 amino acid. The domain analysis showed that the AcSPL14 protein sequence possessed a common functional domain of the SPL protein family, containing a typical SBP(SQUAMOSA promoter binding protein-like) domain. Phylogenetic analysis indicated that protein sequences of AcSPL14 was 53%, 51%, 47% and 46% homologous with those of Asparagus officinalis, Dendrobium catenatum, Rosa chinensis and Jatropha curcas,respectively. Furthermore, a pBI121-AcSPL14 overexpression vector was constructed in this study and the engineering bacteria was obtained. This study could provide a theoretical basis for the identification of the biological function of AcSPL14 in pineapple, and was of great significance for the in-depth study of the molecular mechanism of pineapple leaf thorn.