Induction of Hairy Roots of Scrophularia buergeriana and Its Plant Regeneration

doi:10.11983/CBB16246

Abstract

Abstract: Hairy roots were induced by infecting leaf explants of Scrophularia buergeriana with Agrobacterium rhizo- genes strains A4 and R1601. Hairy roots could grow rapidly and autonomously in liquid or solid plant-growth regulator-free MS medium. The transformation was confirmed by PCR amplification of the rolB gene of the Ri plasmid from S. buergeriana hairy roots. Hairy roots could form green callus in MS medium with 0.5 mg·L^-1 6-BA and 0.1 mg·L^-1 NAA, then regenerated plantlets were induced from the callus. A whole transgenic plantlet was obtained. Adventitious buds could be directly induced from hairy roots in MS medium with 0.5 mg·L^-1 6-BA and 0.02 mg·L^-1 NAA after about 15-20 d, with the induction rate of buds up to 85%.

Key words: Scrophularia buergeriana, Agrobacterium rhizogenes, hairy root, plant regeneration

Ren Ruyi, Xue Jukun, Guo Huiyan, Wei Jicheng. Induction of Hairy Roots of Scrophularia buergeriana and Its Plant Regeneration[J]. Chinese Bulletin of Botany, 2017, 52(6): 783-787.

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URL: https://www.chinbullbotany.com/EN/10.11983/CBB16246

https://www.chinbullbotany.com/EN/Y2017/V52/I6/783

Figures/Tables 3

Figure 1 Induction, in vitro culture and plants regeneration of hairy root in Scrophularia buergeriana(A) Hairy root formation from leaf explants 16 days after infection with the strain of Agrobacterium rhizogenes A4; (B) Green callus induction on hairy roots in medium MS+0.2 mg·L-1 6-BA+0.1 mg·L-1 NAA; (C) Green callus and adventitious buds in- duction on medium MS+0.5 mg·L-1 6-BA+0.1 mg·L-1 NAA; (D) Adventitious shoots formation on medium MS+0.5 mg·L-1 6- BA+0.02 mg·L-1 NAA; (E) Adventitious buds; (F) Transplanted plant

Figure 2 Gel electrophoresis analysis of PCR fragments of rolB gene amplified from the genome DNA of hairy roots (A4)1-4: Fragments amplified from genome DNA of hairy roots; 5, 6: Amplification products from genome DNA of untransformed roots; 7: Fragments amplified from the colony of Agrobacte- rium rhizogenes; M: DL2000 marker

Table 1 Effect of 6-BA and NAA on callus induction and adventitious shoots formation of hairy roots in Scrophularia buergeriana

Med- ium	6-BA (mg·L^-1)	NAA (mg·L^-1)	Change of hairy roots
1	0.1	0.02	Few adventitious buds induced
2	0.1	0.1	Formation of light yellow callus
3	0.2	0.02	Few adventitious buds induced
4	0.2	0.1	Formation of green callus
5	0.5	0.02	More adventitious buds developed
6	0.5	0.1	Firstly formed green callus, then developed more adventitious buds

References 17

[1]	付春祥, 金治平, 杨睿, 吴风燕, 赵德修 (2004). 新疆雪莲毛状根的诱导及其植株再生体系的建立. 生物工程学报 20, 366-371. DOI URL
[2]	刘莉莉, 卢淑波, 徐佳萍, 张庆田, 李昌禹 (2015). 以黄花乌头发根为外植体的再生培养体系建立. 植物学报 50, 623-627. DOI URL
[3]	吕政, 张淑丽, 路放, 杨世海 (2014). 关苍术毛状根培养体系建立及其多糖含量测定. 中国药学杂志 49, 1386-1392. DOI URL
[4]	施和平, 梁朋, 权宏 (2003). 商陆毛状根的诱导、培养及其皂甙的产生. 生物工程学报 19, 46-49. DOI URL
[5]	施和平, 王蓓, 杨树楠, 郭亚鹏 (2016). 五寸石竹毛状根诱导及其植株再生. 植物学报 51, 363-368. DOI URL
[6]	闻玉莉, 杨世海 (2010). 罗勒毛状根的诱导及培养. 安徽农业科学 38, 1727-1730. DOI URL
[7]	吴喜民, 张刘强, 陈小冲, 冯丽, 邢旺兴, 李医明 (2014). 北玄参根中的一个新环烯醚萜衍生物. 药学学报 49, 1019-1021.
[8]	张继栋, 杨雪清, 乔爱民, 孙敏, 何生根, 雷栀, 尹彩霞 (2008). 木本曼陀罗毛状根植株再生体系的建立. 热带亚热带植物学报 16, 480-485. DOI URL
[9]	张雯洁, 刘玉青, 李兴从, 浦湘渝, 金永清, 杨崇仁 (1994). 中药玄参的化学成分. 云南植物研究 16, 407-412.
[10]	Bercetche J, Chriqui D, Adam S, David C (1987). Mor- phogenetic and cellular reorientations induced by Agrobacterium rhizogenes(strains 1855, 2659 and 8196) on carrot, pea and tobacco. Plant Sci 52, 195-210. DOI URL
[11]	Habibi P, de Sa MFG, da Silva ALL, Makhzoum A, da Luz Costa J, Borghetti IA, Soccol CR (2016). Efficient genetic transformation and regeneration system from hairy root of Origanum vulgare. Physiol Mol Biol Plants 22, 271-277. DOI URL PMID
[12]	Lioshina LG, Bulko OV (2014). Plant regeneration from hairy roots and calluses of periwinkle Vinca minor L. and foxglove purple Digitalis purpurea L. Cytol Gen 48, 302-307.
[13]	Park SU, Chae YA, Facchini PJ (2003). Genetic transfor- mation of the figwort, Scrophularia buergeriana Miq., an oriental medicinal plant. Plant Cell Rep 21, 1194-1198.
[14]	Qian JF, Hunkler D, Rimpler H (1992). Iridoid-related aglycone and its glycosides from Scrophularia ningpoensis. Phytochemistry 31, 905-911.
[15]	Rogers SO, Bendich AJ (1985). Extraction of DNA from milligram amounts of fresh, herbarium and mummified plant tissues.Plant Mol Biol 5, 69-76. DOI URL
[16]	Sharafi A, Sohi HH, Azadi P, Sharafi AA (2014). Hairy root induction and plant regeneration of medicinal plant Dra- cocephalum kotschyi. Physiol Mol Biol Plants 20, 257-262. DOI URL PMID
[17]	Wu J, Wang Y, Zhang LX, Zhang XZ, Kong J, Lu J, Han ZH (2012). High-efficiency regeneration of Agrobacterium rhizogenes-induced hairy root in apple rootstock Malus baccata (L.) Borkh. Plant Cell Tissue Organ Cult 111, 183-189.