不同发根农杆菌菌株对黄花蒿毛状根诱导效果的改进

S. Hassan, Zahra Belb Belbasi
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引用次数: 12

摘要

自古以来,植物就被认为是药物的重要来源。来自植物的次生代谢物已显示出治疗特性。青蒿素及其衍生物是菊科中最著名的萜类化合物。目前,为了提高这类萜类化合物的产量,人们已经进行了大量的生物技术试验,其中大多数是利用发根农杆菌(A4、A7、Ar15834、Ar9534、Ar9402、Ar318和Ar50)进行毛状根培养。在本研究中,我们试图研究不同菌株对黄花苜蓿毛状根诱导频率的影响。以黄花蒿叶片为材料制备外植体。然后将其浸泡在根状芽孢杆菌溶液中进行感染。在MS培养基中培养14 d。为了减少植物坏死,在培养基中加入0.1%和0.5%两种不同浓度的抗氧化剂。毛状根诱导鉴定采用PCR法。在1 / 2 MS培养基上,植株有明显的毛状根。用PCR方法研究了不同毛状根转化系中T-DNA的存在。结果表明,添加0.5%抗坏血酸的MS培养基可显著降低植株褐变率(约80%)。另外,A7和Ar318侵染黄花苜蓿叶片外植体的转化率最高,分别为79%和24%。综上所述,上述菌株均能在抗坏血酸作为植物褐变抑制剂的帮助下诱导黄花蒿毛状根。由于黄花蒿毛状根具有激素自养特性,且毛状根侧枝较大,可用于提高制药工业中青蒿素的产量。关键词:黄花蒿;农杆菌属rhizogenes;青蒿素;毛状根。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
IMPROVEMNET OF HAIRY ROOT INDUCTION IN Artemisia annua BY VARIOUS STRAINS OF Agrobacterium rhizogenes
Plants have been considered as a significant source of drugs since ancient times. Secondary metabolites derived from plants have revealed therapeutic properties. Artemisinin and its derivatives are the most famous terpenoids in the Asteraceae family. Nowadays, numerous attempts have been done to improve the production rate of these kinds of terpenoids in biotechnological procedures which most of them are allocated to hairy root cultures using Agrobacterium rhizogenes (A4, A7, Ar15834, Ar9534, Ar9402, Ar318 and Ar50). In this study, we tried to investigate the effect different strains of A. rhizogenes on frequency of hairy root induction in A. annua. Explants were prepared from leaves of A. annua. Then they were soaked in A. rhizogenes solution to be infected. They were cultured in MS medium for 14 days. For decreasing plant necrosis, two different concentrations of 0.1 and 0.5 % were added to the medium as anti– oxidant. For hairy root induction approve, PCR method was applied. Hairy roots were obvious in plantlets on 1⁄2 MS medium culture. The presence of T–DNA in different transformed lines of hairy roots was investigated by PCR. The results indicated that making 0.5 % ascorbic acid MS Medium will lead to a dramatic decrease (about 80 %) in plantlet’s browning. In addition, the highest and lowest transformation rate of 79 % and 24 % were yielded, infecting the leaf explants of A. annua by A7 and Ar318 strains respectively. To sum it up, it was shown that all mentioned strains are capable to induce hairy root in A. annua by the help of ascorbic acid as an inhibitor for plantlet’s browning. Because of hormone–autotrophic feature and great lateral branches of hairy roots, the induced hairy roots of A. annua can be applied to increase artemisinin production in pharmaceutical industry. Keyword: Artemisia annua; Agrobacterium rhizogenes; artemisinin; hairy root.
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