Changes in morphological phenotypes and withanolide composition of Ri-transformed roots of Withania somnifera
Changes in morphological phenotypes and withanolide composition of Ri-transformed roots of Withania somnifera
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DOI：
10.1007/s00299-006-0260-0
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发表时间：
2007-05-01
影响因子：
6.2
通讯作者：
Tepfer, David
中科院分区：
文献类型：
--
作者：
Bandyopadhyay, Maumita;Jha, Sumita;Tepfer, David
利用发根农杆菌的遗传转化将发育变异性引入睡茄中，目的是随着小行星的产生而改变。接种W. somnifera与A.发根菌株LBA 9402和A4产生典型的转化根系、转化愈伤组织系和同时具有根脱分化和再分化的生根愈伤组织系。这些形态上不同的转化系在T-DNA含量、生长速率和星状积累方面存在差异。所有具有典型转化根形态的株系都含有T-L T-DNA，并且90%的株系携带T-R T-DNA，与用于感染的菌株无关。醉茄素A的积累在转化的根系WSKHRL-1中最大（0.44%干重）。这是首次在W.嗜睡。菌株A4诱导的生根愈伤组织均含有T-L和T-R-DNA。相反，用菌株LBA 9402获得的根愈伤组织系中有50%仅含有T-R T-DNA。所有生根愈伤组织系都积累了withaferin A和withaferin D。LBA 9402诱导的愈伤组织不含T-LT-DNA基因，而A4菌株诱导的愈伤组织均含有T-LT-DNA基因。这些愈伤组织系中有4个产生醉茄素A（0.15-0.21%干重）和醉茄素D（0.08-0.11%干重），并且它们比转化的根系生长更快。这是首次报道在W.嗜睡。
Developmental variability was introduced into Withania somnifera using genetic transformation by Agrobacterium rhizogenes, with the aim of changing withasteroid production. Inoculation of W. somnifera with A. rhizogenes strains LBA 9402 and A4 produced typical transformed root lines, transformed callus lines, and rooty callus lines with simultaneous root dedifferentiation and redifferentiation. These morphologically distinct transformed lines varied in T-DNA content, growth rates, and withasteroid accumulation. All of the lines with the typical transformed root morphology contained the T-L T-DNA, and 90% of them carried the T-R T-DNA, irrespective of the strain used for infection. Accumulation of withaferin A was maximum (0.44% dry weight) in the transformed root line WSKHRL-1. This is the first detection of withaferin A in the roots of W. somnifera. All of the rooty callus lines induced by strain A4 contained both the T-L and the T-R-DNAs. In contrast, 50% of the rooty-callus lines obtained with strain LBA 9402 contained only the T-R T-DNA. All the rooty callus lines accumulated both withaferin A and withanolide D. The callusing lines induced by LBA 9402 lacked the T-L T-DNA genes, while all the callusing lines induced by strain A4 contained the T-L DNA. Four of these callus lines produced both withaferin A (0.15-0.21% dry weight) and withanolide D (0.08-0.11% dry weight), and they grew faster than the transformed root lines. This is the first report of the presence of withasteroids in undifferentiated callus cultures of W. somnifera.