以蛹虫草为材料,研究了不同浓度乙烯利和乙烯受体抑制剂1-甲基环丙烯(1-methylcyclopropene, 1-MCP)对蛹虫草子实体后期成熟产生虫草素的影响。根据不同处理方法将栽培蛹虫草分为乙烯组,1-MCP组和对照组。通过实时定量PCR(real-time quantitative PCR,qRT-PCR)测定各组蛹虫草子实体中腺苷酸琥珀酸合成酶(Adss)、腺苷酸琥珀酸裂解酶(Adsl)、腺苷酸激酶 (AK)、嘌呤核苷磷酸化酶(PNP)等虫草素合成基因的表达量;高效液相色谱法测量子实体中虫草素含量。结果表明,当乙烯利质量浓度为100 mg/L可以显著提高PNP、Adss、AK、Adsl基因的表达量,乙烯组虫草素的质量浓度为(25.37±0.22)μg/mL而对照组为(10.04±0.17)μg/mL;1.0 mg/L的1-MCP则对子实体的虫草素含量以及PNP、Adss、AK、Adsl基因表达有明显的抑制作用,1-MCP组虫草素的质量浓度为(0.52±0.12)μg/mL。由此可知,添加一定浓度的外源乙烯能够促进虫草素代谢相关基因表达量上调,后期成熟子实体中虫草素的含量也显著提高。而乙烯受体抑制剂可以显著降低子实体中虫草素合成相关基因的表达量及其含量。
In this study, the effects of ethephon and ethylene receptor inhibitor 1-methylcyclopropene (1-MCP) on cordycepin production in the fruit body of Cordyceps militaris were studied. The fruit bodies were treated with ethylene, 1-MCP respectively. Real-time quantitative PCR was performed to detect the gene expression of adenyl-succinic acid synthase (Adss), adenyl-succinic acid lyase (Adsl), adenyl-kinase (AK) and purine nucleoside phosphorylase (PNP) among different groups. The content of cordycepin in the fruiting body was measured by HPLC. The results showed that the expression levels of PNP、Adss、AK and Adsl genes were significantly increased when the concentration of ethephon was 100 mg/L. The content of cordycepin in the ethylene group and control group were (25.37±0.22) μg/mL and (10.04±0.17) μg/mL respectively. And 1-MCP (1.0 mg/L) could significantly inhibit the cordycepin content in the fruit body and the expression of PNP、Adss、AK and Adsl genes. The cordycepin content in 1-MCP group was (0.52±0.12) μg/mL. In conclusion, the addition of exogenous ethylene at a certain concentration could promote the up-regulation of cordycepin metabolism-related gene expression, and the content of cordycepin in mature fruit bodies at a later stage was also significantly increased. Ethylene receptor inhibitors could significantly reduce the expression and content of cordycepin biosynthesis-related genes in the fruiting body.
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