Farkya Sunita, Bisaria Virendra Swarup
Department of Biochemical Engineering and Biotechnology, Indian Institute of Technology Delhi, Hauz Khas, New Delhi 110016, India.
J Biosci Bioeng. 2008 Feb;105(2):140-6. doi: 10.1263/jbb.105.140.
A hairy root line of Linum album LYR2i was obtained via the genetic transformation of a cotyledon segment of the plant by cocultivating it with Agrobacterium rhizogenes and culturing it in hormone-free B5 Gamborg liquid medium. Characteristic changes in root morphology associated with variations in biosynthetic capabilities were achieved by adding different combinations of phytohormones to the basal medium. A reversible system between hairy roots and its cultures with a diversified morphology was established by including and excluding phytohormones in the basal medium. The medium containing indole 3 acetic acid (IAA) at a concentration of 3 mg/l (MI3) induced thicker root tips and that containing 2,4-dichlorophenoxy acetic acid (2,4-D) at a concentration of 1 mg/l (MD1) induced compact green callus. Podophyllotoxin and 6-methoxypodophyllotoxin content increased by 1.86-fold and 1.45-fold as a result of adding IAA (MI3) and 2,4-D (MD1) to the basal medium, respectively. After regeneration, the hairy roots regained their original morphology and biosynthetic capability on hormone-free basal medium. The transformation status of the regenerated hairy roots was confirmed by PCR analysis.
通过将亚麻（Linum album LYR2i）子叶节段与发根农杆菌共培养，并在无激素的B5 Gamborg液体培养基中培养，获得了亚麻的毛状根系。通过向基础培养基中添加不同组合的植物激素，实现了与生物合成能力变化相关的根形态特征变化。通过在基础培养基中添加和去除植物激素，建立了毛状根与其具有多样化形态的培养物之间的可逆系统。含有浓度为3 mg/l的吲哚-3-乙酸（IAA）的培养基（MI3）诱导根尖变粗，含有浓度为1 mg/l的2,4-二氯苯氧乙酸（2,4-D）的培养基（MD1）诱导形成紧密的绿色愈伤组织。由于分别向基础培养基中添加了IAA（MI3）和2,4-D（MD1），鬼臼毒素和6-甲氧基鬼臼毒素的含量分别增加了1.86倍和1.45倍。再生后，毛状根在无激素的基础培养基上恢复了其原始形态和生物合成能力。通过PCR分析确认了再生毛状根的转化状态。
