金针菇tps1基因序列分析及不同温度下tps1、tps2基因定量表达研究

Sequence analysis of tps1 and relative expression of tps1 and tps2 under different temperature in Flammulina velutipes

研究探讨金针菇海藻糖合成相关酶基因及其功能,为进一步揭示其生长发育过程中的变化规律奠定基础。本研究对金针菇6-磷酸海藻糖合成酶基因tps1的序列进行分析,并以金针菇孢子单核体菌株Dan3为实验材料,采用实时荧光定量PCR技术比较了tps1、tps2基因在不同温度变化下金针菇中的相对表达量差异。结果表明,金针菇tps1基因c DNA序列包含1455 bp的ORF,编码1个由484个氨基酸残基组成的蛋白质;对其起始密码子上游2000 bp调控区域进行分析,发现具有典型的热激反应元件(HSE,C4T)和压力反应元件(STRE,AG4)等调控元件;实时荧光定量PCR结果显示,相对于对照组21℃处理,37℃高温及4℃低温处理2 h后,tps1、tps2基因相对表达量均有显著(p<0.05)升高。说明tps1、tps2基因在转录水平上受到温度调控。

The study aimed at discussing trehalose synthesis-related enzyme genes of Flammulina velutipes and their functions ,which could lay the foundation for revealing the changing rule during growth and development process. The sequence of tps1 gene was analyzed and relative quantitative expression changes of tps1 and tps2 in mycelia of Flammulina velutipes strain Dan 3 under temperature variations were measured by using real-time PCR. Results showed that open reading frame of tps1 gene contained 1455 bp encoding a polypeptide of 484 amino acid residues. After analysis of the section of 2000 bp upstream of initiator codon,it was found that the promoter contained typical regulatory elements,STRE（C4T） and HSE（AG4）. The real-time PCR analysis showed that the relative expression of tps1 gene and tps2 gene both significantly（p〈0.05） increased after 2 hours management of relatively high temperature 37 ℃ and relatively low temperature 4 ℃. The results confirmed that the expression of the tps1 and tps2 gene was regulated by temperature at transcriptional level.