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A method for in vitro regeneration and genetic transformation of moso bamboo immature embryos

A technology of in vitro regeneration and genetic transformation, applied in the field of in vitro regeneration and genetic transformation of moso bamboo embryos, can solve the problems of imperfect regeneration system of moso bamboo, achieve the effects of shortening the transformation cycle, improving transformation efficiency, and improving proliferation and state

Active Publication Date: 2021-10-15
RES INST OF SUBTROPICAL FORESTRY CHINESE ACAD OF FORESTRY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] The purpose of the present invention is to provide a method for in vitro regeneration and genetic transformation of moso bamboo immature embryos, which solves the problem that the regeneration system of moso bamboo is not perfect, can improve the proliferation and state of resistant callus, shorten the transformation cycle, and improve Conversion efficiency

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  • A method for in vitro regeneration and genetic transformation of moso bamboo immature embryos
  • A method for in vitro regeneration and genetic transformation of moso bamboo immature embryos

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Embodiment 1

[0034] A method for in vitro regeneration and genetic transformation of moso bamboo embryos, the method comprising:

[0035] (1) Induction of callus from immature embryos of Phyllostachys pubescens:

[0036] Surface sterilize the immature moso bamboo seeds with 75% alcohol for 60 seconds, wash them twice with sterile water, soak them in 0.1% mercuric chloride solution for 20 minutes, wash them with sterile water for 5 times, then peel off the embryos on the callus induction medium , 25 ℃, dark culture 21d, to obtain callus.

[0037] The induction medium used is: MS+2.0-6.0mg / L2,4-D+30g / L sucrose+7g / L agar, pH=5.8.

[0038] (2) Proliferation of Phyllostachys pubescens callus:

[0039] The induced callus was transferred to the subculture medium for proliferation, cultured in the dark at 25° C., and subcultured every 21 d (days) to meet the subsequent genetic transformation requirements.

[0040] The adopted subculture medium is: MS+0.1-1.0 mg / L 2,4-D (2,4-dichlorophenoxyaceti...

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Abstract

The invention discloses a method for in vitro regeneration and genetic transformation of immature bamboo embryos of Moso bamboo, the method comprising: induction of callus of immature moso bamboo embryos, proliferation of callus of moso bamboo, construction of transformation vector pER8-WUS-RdCodA, and Agrobacterium Infection, selection of resistant callus of Moso bamboo and differentiation of adventitious buds, regeneration of transgenic plants, transplanting of hardened seedlings and detection of transgenic regenerated plants of Moso bamboo. Wherein, the construction of the transformation vector pER8-WUS-RdCodA includes: carrying out Xho I / Spe I double enzyme digestion of the estradiol-inducible vector pER8; adding vector homologous sequences to both ends of the corn WUS gene, and inserting it into pER8 after digestion In the vector, the complete expression frame of the CodA gene induced by the Rd29A promoter was inserted into the EcoR V site to construct the transformation vector pER8‑WUS‑RdCodA. The method of the invention can shorten the transformation period and improve the transformation efficiency.

Description

technical field [0001] The invention relates to a method for genetic transformation, in particular to a method for in vitro regeneration and genetic transformation of moso bamboo immature embryos. Background technique [0002] Bamboo is one of the important forest resources, which has the characteristics of wide distribution, fast growth, multiple uses, and high ecological and economic value. Bamboo has a long flowering cycle and dies after flowering. Hybrid breeding is very difficult, and there is a lack of excellent new varieties that are urgently needed in production. Since the 1980s, more than 20 genera of Phyllostachys, Dendrocalamus Nees, Dendrocalamus, Sasa Makinoet Shibata and Thysostachys Gamble have been studied. More than 70 bamboo species with different explants and culture conditions were studied in tissue culture, callus or clustered shoots were obtained, and complete regenerated plants were induced on some bamboo species. However, 90% of the current reports ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/84A01H5/00A01H6/46A01H4/00
CPCA01H4/001A01H4/005C12N15/8205
Inventor 乔桂荣卓仁英黄碧芸邱文敏王雨珺
Owner RES INST OF SUBTROPICAL FORESTRY CHINESE ACAD OF FORESTRY
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