《植物生理学报》 2018, 54(4): 627-635

忽地笑LaABCG3转运蛋白基因的克隆与表达分析

韩小康, 王蓉, 刘彦彤, 夏冰, 汪仁^*

江苏省中国科学院植物研究所, 南京210014

通信作者：汪；E-mail: jswangren@aliyun.com

摘　要：

石蒜科植物体内的生物碱具有重要的药用价值, 尽管部分生物碱的生物合成途径已被发现, 但与其相关的转运机制尚未报道。本研究在忽地笑(Lycoris aurea)经茉莉酸甲酯(MeJA)处理后的差异转录组测序数据的基础上, 发现了一个ABC转运蛋白基因, 并命名为ABCG3。采用cDNA末端快速扩增(RACE)技术克隆了该基因的编码区全长2 157 bp, 编码719个氨基酸, 包含6个跨膜结构域。LaABCG3的组织分布与生物碱积累组织相重合。此外, 在MeJA处理下, LaABCG3的表达量和石蒜科生物碱, 包括加兰他敏、石蒜碱、水仙环素的含量均受到显著诱导。本研究为探索忽地笑中石蒜科生物碱的转运和积累机制打下了基础。

关键词：忽地笑; 基因克隆; ABC转运蛋白; 茉莉酸甲酯; 石蒜科生物碱

收稿：2017-11-01   修定：2018-03-20

资助：国家自然科学基金(31700271和31600074)、江苏省自然科学基金(BK20170617)、江苏省省属公益院所科研条件与能力建设项目(BM2015019)、海洋公益性行业科研专项(201505023)。

Cloning and expression analysis of LaABCG3 transporter gene from Lycoris aurea

HAN Xiao-Kang, WANG Rong, LIU Yan-Tong, XIA Bing, WANG Ren^*

Institute of Botany, Jiangsu Province and Chinese Academy of Sciences, Nanjing 210014, China

Corresponding author: WANG Ren; E-mail: jswangren@aliyun.com

Abstract:

Amaryllidaceae alkaloids have been reported to be of important medical value. Although the biosynthetic pathway of some Amaryllidaceae alkaloids had been described in plants, their transport mechanism has not been illuminated. Based on the comparative transcriptome sequencing data of Lycoris aurea in response to methyl jasmonate (MeJA) treatment, an ATP-binding cassette (ABC) transporter gene, named as LaABCG3, was isolated. By using rapid amplification of cDNA ends (RACE) method, we cloned its full-length cDNA containing a 2 157-bp open reading frame (ORF) encoding a protein of 719 amino acids with 6 transmembrane domains. The expression pattern of LaABCG3 was consistent with the accumulation pattern of Amaryllidaceae alkaloids. Besides, under MeJA treatment, both the expression level of LaABCG3 and the contents of some Amaryllidaceae alkaloids including galantamine, lycoramine and narciclasine were significantly induced. This study will provide a theoretical basis for investigating Amaryllidaceae alkaloid transport and accumulation mechanism in L. aurea.

Key words: Lycoris aurea; gene cloning; ABC transporters; methyl jasmonate; Amaryllidaceae alkaloids