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Chinar tissue culture method suitable for Agrobacterium rhizogenes mediation

A technology mediated by Agrobacterium and tissue culture, applied in horticultural methods, botanical equipment and methods, biochemical equipment and methods, etc., can solve the problem of affecting the regeneration rate of adventitious buds in transformed explants, affecting the development of multi-gene transformation work, etc. problem, to achieve the effect of thick leaves and high frequency of adventitious bud regeneration, strong regeneration ability of adventitious buds, and not easy to brown and die

Inactive Publication Date: 2012-04-25
JIANGSU ACAD OF AGRI SCI +1
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Problems solved by technology

[0004] The previous research work all adopted continuous subculture on the same medium. Due to the accumulation of hormones, micropropagated seedlings are prone to vitrification, which in turn affects the amount of transformed explants and the regeneration rate of adventitious buds.
Thus affecting the development of multigene transformation work mediated by Agrobacterium syringae

Method used

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  • Chinar tissue culture method suitable for Agrobacterium rhizogenes mediation
  • Chinar tissue culture method suitable for Agrobacterium rhizogenes mediation
  • Chinar tissue culture method suitable for Agrobacterium rhizogenes mediation

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Experimental program
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Effect test

Embodiment Construction

[0043] a) Materials and methods

[0044] (1) Material

[0045] The stem section of Platanus biscuica, which is strong and straight, is selected as the explant material;

[0046] The strains and plasmids are LBA4404 and pGBI4AB containing the target gene nptII gene (Yu Jianming, Liang Liufang, Zhang Baolong, He Xiaolan, Chen Zhiyi, Ni Wanchao. Agrobacterium-mediated method to obtain bluegrass bluegrass transgenic plants. Jiangsu Agricultural Science Journal, 2005, 21( 2): 102~105)

[0047] (2) culture medium

[0048] Basic medium: WPM or MS, the mass ratio of sucrose is 3%, the mass ratio of agar strip is 0.8%;

[0049] Axillary bud initiation medium: WPM+0.1mg / L zeatin (ZT)+0.1mg / L 6-benzylaminopurine (6-BA)+0.01mg / L indolebutyric acid (IBA)

[0050] Micropropagation medium: MS+2.0mg / L 6-BA+0.5mg / L kinetin (KT)+0.1mg / L IBA

[0051] Strong seedling medium: MS+2.0mg / L ZT+0.5mg / L KT+0.1mg / L IBA

[0052] Transition medium: WPM+0.2mg / L ZT+0.1mg / L indole acetic acid (IAA)

[...

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Abstract

The invention relates to a chinar tissue culture method suitable for Agrobacterium rhizogenes mediation, which belongs to a plant tissue culture method. The method is used in a chinar micropropagation strain, the successive transfer culture is carried out by alternately using a sound seedling culture medium, a transitional culture medium and a micropropagation culture medium, thereby ensuring vigorous growth and basic stability of endogenesis hormone level of the micropropagation seedling. In the adventitious bud induction culture, the leaf blade growing in the transitional culture medium for30 days is used for carrying out the differentiation of the adventitious buds, and the regeneration frequency of the adventitious buds stays above 80%. The alternate successive transfer culture used on the micropropagation seedling ensures the sufficient leaf blade exophyte source and the high regeneration frequency of the adventitious buds of the leaf blade, thereby being more suitable for the recipient material processed by gene transformation by using an Agrobacterium rhizogenes mediation method. When the adventitious bud regeneration system is used for Agrobacterium rhizogenes mediation, the nptII gene is transferred into the chinar strain.

Description

1. Technical field [0001] The invention is a method for plant tissue culture suitable for Agrobacterium-mediated plane tree tissue culture, which belongs to plant tissue culture methods, and is specially used for the cultivation of acceptor materials for gene transformation through the Agrobacterium-mediated method. 2. Background technology [0002] Platanus spp. is one of the four famous street tree species in the world. It has the effects of resisting smoke and dust, cooling and humidifying, and reducing noise. Recent studies have also shown that it has a certain ability to absorb sulfur dioxide, chlorine, fluorine, and lead. Strong resistance to ozone, benzene, hydrogen sulfide, etc. Our country introduces and cultivates one-ball plane tree (American plane tree, P.occidentalis Lirm.), two-ball plane tree (English plane tree, (P.acerifolia (Ait.) Willd.) and three-ball plane tree (French plane tree, P.orientalis Linn.), etc. It has a history of more than 100 years. At pre...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01H4/00C12N15/84
Inventor 何晓兰倪万潮易金鑫隋德宗张大勇教忠义韩杰峰王宝松徐照龙刘桂华
Owner JIANGSU ACAD OF AGRI SCI
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