摘要
【目的】克隆茉莉花磷酸甲羟戊酸激酶(PMK)基因(JsPMK),对其进行生物信息学分析并检测其表达时间节律性,为深入研究茉莉花萜类化合物生物合成途径及其调控机制提供理论参考。【方法】以茉莉花花瓣为材料,采用RT-PCR扩增JsPMK基因全长,利用生物信息学软件对其编码蛋白进行预测分析,并采用qRT-PCR检测JsPMK基因在茉莉花开放过程中的表达情况。【结果】克隆获得的JsPMK基因(GenBank登录号为MH311042)全长1951 bp,包含1518 bp完整的开放阅读框(ORF),共编码505个氨基酸,其编码蛋白定位于细胞质,为稳定的非分泌蛋白。JsPMK蛋白含有GHMP激酶N端保守区和C端保守区,且在N端保守区含一个ATP结合位点Gly-X-Gly-XX-Ala。JsPMK蛋白与野生油橄榄、芝麻、丹参和番茄的PMK蛋白氨基酸序列相似性均较高(在80%以上),其中与野生油橄榄亲缘性最近,表明不同植物PMK蛋白高度保守。JsPMK基因在未开放(18∶00)时表达量最低,随着茉莉花逐渐开放,JsPMK基因表达量极显著上升(P<0.01,下同),在20∶00达最大值,随后表达量极显著下降。【结论】JsPMK基因表达可能受生物钟调控,在茉莉花萜类化合物合成和香气释放中发挥重要的调控作用。
【Objective】Phosphomevalonate kinase(PMK)gene(JsPMK)was cloned from Jasminum sambac(L.)Ait,and its bioinformatics was analyzed,its expression rhythmicity was detected to provide theoretical reference for studying the synthesis path of terpenoid in J.sambac and its regulation mechanism.【Method】J.sambac petal was used as material.The full length of gene JsPMK was amplified by RT-PCR,and its encoded protein was predicted by bioinformatics software.Expressions of JsPMK in opening process of J.sambac was detected by qRT-qPCR.【Result】A total of 1951 bp fulllength of JsPMK(GenBank accession number MH311042)was obtained,including 1518 bp open reading frame(ORF)and encoding a protein of 505 amino acids which was located in the cytoplasm and belonged to non-secretory protein.The JsPMK protein contained two typical conserved domains-GHMP_kinases_N domain and GHMP_kinases_C domain,and associated with ATP binding sites Gly-X-Gly-XX-Ala in the N-terminal.The phylogenetic tree indicated that amino acids of JsPMK protein had high affinity with PMK protein of Olea europaea,Sesamum indicum and Salvia miltiorrhiza(over 80%).In addition,the similarity between JsPMK protein and O.europaea was the highest.The results indicated that PMK protein of different plants were highly conserved.Gene JsPMK performed the lowest expression when J.sambac was not open(at 18:00),and ascented extremely(P<0.01,the same below)as it started to bloom and reached to the maximum at 20:00,then presented extremely downward trend.【Conclusion】Gene JsPMK may be controlled by biological block,and play an important regulation effects in synthesis of terpenoid and aroma releasing in J.sambac.
作者
陈笛
王鹏杰
郑玉成
郑知临
陈桂信
叶乃兴
CHEN Di;WANG Peng-jie;ZHENG Yu-cheng;ZHENG Zhi-lin;CHEN Gui-xin;YE Nai-xing(College of Horticulture,Fujian Agriculture and Forestry University/Key Laboratory of Tea Science at Universities in Fujian,Fuzhou 350002,China)
出处
《南方农业学报》
CAS
CSCD
北大核心
2018年第10期1909-1916,共8页
Journal of Southern Agriculture
基金
福建省自然科学基金项目(2016J01110)
福州市科技计划项目(2017N0018)
