Response 5-phosphomevalonate Kinase Enzymes to Low Doses of Gamma Radiation and Its Related to the Production of Terpenoids Indole Alkaloids from Catharanthus rouses

I. Salama, G. Ali, M. K. A. el-Megid
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Abstract

GAMMA rays effect, on different dose levels on the production of ajmalicine as well as 5-phosphomevalonate kinase enzyme activety, was studied in Catharanthus roseus cell suspension cultures. The seedlings were taken from the young leaf puds and grown on MS solid medium for 7 weeks. Calli individual serially sub cultured by transfing to maintenance medium consisting of Gamborg’s B5 medium. Cell suspensions were exposed to gamma rays after 4 weeks at dose levels of 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5 and 8Gy. Ajmalicine quantities were analyzed on VIS–spectrophotometer; also, the 5-phosphomevalonate kinase enzyme was particularly purified by ultrafiltration and activety analayzed, in addition, native PAGE electrophoresis was determined. 5-phosphomevalonate kinase enzyme bands and ajmalicine accumulation quantities were changed after the exposure to low doses from 1 to 6Gy, but the higher doses were lethal. The appropriated doses to activety of 5-phosphomevalonate kinase enzyme and Ajmalicine production were from 1 to 5Gy. Other dose levels (5.5 and 6Gy) did not affect on the 5-phosphomevalonate kinase enzyme activety and Ajmalicine production.
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5-磷酸戊酸激酶对低剂量γ辐射的响应及其与花楸萜类吲哚类生物碱生产的关系
研究了不同剂量γ射线对长春花悬浮培养细胞中ajmalicine生成及5-磷酸戊酸激酶活性的影响。从幼嫩的叶浆中取出幼苗,在MS固体培养基上培养7周。愈伤组织个体通过转移到由甘堡氏B5培养基组成的维持培养基上进行连续继代培养。4周后,细胞悬液暴露于1、1.5、2、2.5、3、3.5、4、4.5、5、5.5、6、6.5、7、7.5和8Gy剂量水平的伽马射线中。用紫外分光光度计分析杨木碱的含量;对5-磷酸戊酸激酶进行了超滤纯化和活性分析,并进行了PAGE电泳测定。1 ~ 6Gy低剂量暴露后,5-磷酰戊酸激酶酶谱带和ajmalicine积累量发生变化,但高剂量暴露致死。5-磷酰戊酸激酶活性和Ajmalicine产量适宜剂量为1 ~ 5Gy。其他剂量水平(5.5 gy和6Gy)对5-磷酰戊酸激酶活性和Ajmalicine的产生没有影响。
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