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目的研究低温胁迫对人参皂苷生物合成途径的3个基因家族3-羟基-3-甲基戊二酰CoA还原酶(PgHMGR)、鲨烯合酶(PgSS)、鲨烯环氧酶(PgSE)的影响,探讨各个家族成员响应低温的机制,并推测出响应低温的关键家族基因。方法将3周大的新鲜人参愈伤组织放置在5℃的冰箱中,分别在0、0.5、1、2、3 d后进行取样,分别记作CK、D1、D2、D3、D4,进行RNA提取和反转录实验,实时荧光定量PCR检测低温胁迫处理下不同基因的表达量。结果 PgHMGR1的表达量在D3时期达到对照组的1.3倍,PgHMGR2的表达量在D1时期达到峰值,为对照组的3.8倍;PgSS1的表达量在D3时期达到对照组的1.7倍;PgSE1和PgSE2的表达量均在D1时期分别达到对照组的6.9倍和6倍。其他家族基因(PgHMGR3、PgSS2、PgSE3)的表达量均无显著性变化。结论人参皂苷生物合成途径各个基因家族积极响应低温胁迫处理,推测PgHMGR1、PgHMGR2、PgSS1、PgSE1和PgSE2是人参愈伤组织响应低温胁迫时的关键家族基因。
Objective To study the effects of chilling stress on the expression of 3 genes, PgHMGR, PgSS, Squalene cyclooxygenase (PgSE) in the ginsenoside biosynthesis pathway Influence, investigate the mechanisms by which individual family members respond to hypothermia, and speculate on the key family genes that respond to hypothermia. Methods Three-week-old fresh ginseng callus were placed in a refrigerator at 5 ℃, and were sampled after 0, 0.5, 1, 2 and 3 d respectively and labeled as CK, D1, D2, D3 and D4 Extraction and reverse transcription experiments, real-time fluorescence quantitative PCR detection of different genes under low temperature stress. Results The expression of PgHMGR1 was 1.3-fold higher than that of control at D3. The expression level of PgHMGR2 peaked at D1, which was 3.8-fold that of the control group. The expression level of PgSS1 was 1.7-fold that of the control group at D3 stage. The expression levels in the D1 were 6.9 times and 6 times the control group respectively. There was no significant change in the expression of other family genes (PgHMGR3, PgSS2, PgSE3). Conclusion All gene families of ginsenoside biosynthesis pathway respond positively to low temperature stress. It is speculated that PgHMGR1, PgHMGR2, PgSS1, PgSE1 and PgSE2 are the key family genes in response to low temperature stress.