摘要
脂肪酰基还原酶基因广泛参与植物的脂类代谢过程,影响植物雄配子体花药发育以及表皮蜡酯合成等。本研究利用RACE方法从宁夏枸杞(宁杞1号)花药中克隆脂肪酰基还原酶LbMS2-2基因,开放阅读框全长1800bp,编码599个氨基酸,等电点为9.00。生物信息学分析表明,LbMS2-2蛋白定位于叶绿体中,该蛋白序列与茄科植物甜辣椒、烟草和马铃薯中的脂肪酰基还原酶表现出较高的序列相似性;实时荧光定量PCR显示,LbMS2-2基因在枸杞花器官中表达,且在枸杞花药发育的四分体时期、单核花粉时期和双核花粉时期表达量较高。原位杂交结果证实该基因只在花药绒毡层和小孢子中表达。亚细胞定位结果进一步验证LbMS2-2基因的叶绿体定位。以上结果表明,枸杞脂肪酰基还原酶基因是枸杞花器官发育过程中的重要基因。
Fatty acyl reductase are widely involved in the process of lipid metabolism, affecting anther develop-ment and cuticular wax sunthesis of male gametophytes. In this study, LbMS2-2 gene encoding fatty acyl reduetasewas isolated using RACE （ rapid-amplification of cDNA ends） method from Ningxia wolfberry （Lycium barbarum L. ）.LbMS2-2 contains an open reading frame（ORF） with length of 1800 bp, encoding a deduced protein of 599 aminoacid residues, with its isoelectric point of 9.00. Bioinformatic analysis indicated that LbMS2.2 protein was located tothe chloroplast. The amino acid sequence of LbMS2-2 shared high homology with fatty acyl xeductase from Capsicumannuum L. ,Nicotiana tabacum L. and Solanum tuberosum L.. Real-time fluorescence quantitative PCR results re-vealed that LbMS2-2 expressed highly in tetrad stage ,single nucleus pollen stage and double nucleus pollen stage ofanther development. The results of subcellular localization and in situ hybridization further confirmed that the LbMS2-2 gene mainly detected in the tapetum and microspores of anther, and its protein was predominantly localized in chlo-roplast. As the result,the fatty acyl reductase is an important gene in the organ development of Lycium barbarum L.
作者
石晶
石鑫强
管翠萍
郑蕊
杨亚珺
王丽娟
梁文裕
SHI Jing, SHI Xin-qiang, GUAN Cui-ping, ZHENG Rui, YANG Ya-jun, WANG Li-juan, LIANG Wen-yu(College of Life Science ,Ningxia University, Yinchuan 750021)
出处
《植物遗传资源学报》
CAS
CSCD
北大核心
2018年第2期351-360,共10页
Journal of Plant Genetic Resources
基金
国家自然科学基金(31401444
31260065
31360361)
关键词
枸杞
花药发育
脂肪酰基还原酶
LbMS2-2
表达分析
wolfberry（Lycium barbarum L. ）
anther development
fatty acyl reductase
LbMS2-2
expression analysis
