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质合成等。 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1800 bp,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 development and cuticular wax sunthesis of male gametophytes. In this study, LbMS2-2 gene encoding fatty acyl reductase was isolated using RACE(rapid-amplification of cDNA ends) method from Ningxia wolfberry ( Lycium barbarum L.). LbMS2-2 consisted of an open reading frame (ORF) with length 1800 bp, which encoded a protein of 599 amino acid residues, and its isoelectric point was 9.00. Bioinformation analysis indicated that LbMS2-2 protein was located in the chloroplast. The amino acid sequence of LbMS2-2 shared high homology with fatty acyl reductase from Capsicum annuum、Nicotiana tabacum and Solanum tuberosum. Real-time fluorescence quantitative PCR results revealed that LbMS2 expressed highly in tetrad stage、single nucleus pollen stage and double nucleus pollen stage of anther development. The results of subcellular localization and in situ hybridization further confirmed that LbMS2-2 gene mainly detected in the tapetum and microspores of anther, and its protein was predominantly localized in chloroplast. As the result, the fatty acyl reductase is an important gene in the organ development of Lycium barbarum L.
石晶,管翠萍,郑蕊,等.枸杞脂肪酰基还原酶基因的克隆及表达分析[J].植物遗传资源学报,2018,19(2):351-360.
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