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目的:克隆丹参中的苯丙氨酸裂解酶基因(Smpal),研究其表达特征及其对丹参中酚酸类成分的影响。方法:通过cDNA末端快速扩增(RACE)方法克隆了Smpal全长基因,分别使用甲基茉莉酸(MeJA)、水杨酸(SA)、赤霉素(GA3)诱导Smpal的表达。代谢物含量通过液质联用仪检测。结果:Smpal全长cDNA序列为2354bp,包含一个2133bp的开放阅读框,编码711个氨基酸残基的蛋白,含有长度为21bp的5’非编码区,171bp的3’非编码区。Smpal在根茎叶中都有表达,在根中表达量最多,其次是茎和叶。结论:Smpal与其他物种中pal基因具有62.4%-93%的同源性,Southern杂交表明Smpal属多基因家族。丹参中迷迭香酸(RA)和丹酚酸B(LAB)的含量与Smpal的表达相关。
OBJECTIVE: To clone the phenylalanine lyase gene (Smpal) in Salvia miltiorrhiza and study its expression characteristics and its influence on the phenolic acids in salvia miltiorrhiza. Methods: The full length Smpal gene was cloned by rapid amplification of cDNA ends (RACE). The expression of Smpal was induced by methyl jasmonate (MeJA), salicylic acid (SA) and gibberellin (GA3) respectively. Metabolite content was detected by LC-MS. RESULTS: The full length cDNA of Smpal was 2354bp and contained a 2133bp open reading frame (ORF) encoding a protein of 711 amino acids. It contained a 5 ’noncoding region of 21bp in length and a 171bp 3’ noncoding region. Smpal was expressed in rhizome leaves and most expressed in roots, followed by stems and leaves. Conclusion: Smpal shares 62.4% -93% homology with pal genes in other species. Southern hybridization indicates that Smpal is a multi-gene family. The contents of rosmarinic acid (RA) and salvianolic acid B (LAB) in Salvia miltiorrhiza Bge are related to the expression of Smpal.