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目的研究镰刀菌诱导结香对白木香倍半萜合成酶(sesqui-TPS)表达的影响与倍半萜类化合物含量的动态变化。方法小孔滴注法进行镰刀菌诱导造香实验并割取造香前后12月内7个时间点的结香部位(沉香树脂),采用实时荧光定量PCR法(q PCR)测定sesqui-TPS基因表达水平,气相色谱-质谱联用(GC-MS)技术分析倍半萜类化合物含量的动态变化。结果镰刀菌诱导白木香形成沉香2~12个月内sesqui-TPS基因相对表达量分别为10.85,0.793 1,6.484,611.4,5 800,4 211;人工诱导2个月前未检出倍半萜类次生代谢产物,4~12个月内共检出14个倍半萜类化合物,其相对百分含量分别为21.40%,25.52%,36.44%,32.40%和55.70%。结论镰刀菌诱导白木香结香过程中sesqui-TPS基因主要在后期响应伤害胁迫,其表达水平对促进倍半萜类成分生物合成具有滞后性,为进一步研究白木香结香过程中功能基因对萜类成分的生物合成途径调控作用奠定基础。
Objective To study the effect of Fusarium oxysporum L. on the expression of sesqui-TPS and the dynamic changes of sesquiterpene compounds. Methods Flos-rose shoots were induced by foul-drip method and the anthesis resin (aloe vera resin) at seven time points within 12 months before and after harvesting was harvested. The real-time quantitative PCR (q PCR) was used to determine the sesqui-TPS gene The expression levels of sesquiterpene compounds were analyzed by gas chromatography-mass spectrometry (GC-MS). Results The relative expression levels of sesqui-TPS gene in 10 ~ 25 months after Fusarium spp. Inducing the formation of alopecu copula were 10.85,0.793, 1,6.484,611.4 and 5,800,4211 respectively. Sesquiterpene was not detected two months ago after artificial induction A total of 14 sesquiterpenoids were detected within 4 ~ 12 months. The relative percentages of them were 21.40%, 25.52%, 36.44%, 32.40% and 55.70%, respectively. Conclusion The sesqui-TPS gene induced by Fusarium solani mainly responds to injury stress in the late stage. The expression of sesqui-TPS gene lags behind the biosynthesis of sesquiterpenoids. In order to further study the effect of functional genes on terpenoid Class composition of the biosynthetic pathway to lay the foundation for the regulation.