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Method for quickly introducing embryonic callus through anther to regenerate plant

An embryogenic callus and callus technology, applied in the field of tissue culture, can solve the problems of limited application of grape anther culture, long cultivation period, low embryogenic callus induction efficiency, etc. The effect of short cycle and rich genotype

Inactive Publication Date: 2014-11-12
TOBACCO RES INST HENAN ACADEMY OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

During the induction process, the induction efficiency of embryogenic callus is low, accompanied by the appearance of non-embryogenic callus, and the cultivation period is long, which limits the application of grape anther culture in actual breeding

Method used

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  • Method for quickly introducing embryonic callus through anther to regenerate plant
  • Method for quickly introducing embryonic callus through anther to regenerate plant
  • Method for quickly introducing embryonic callus through anther to regenerate plant

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Embodiment Construction

[0018] 1. Collection and disinfection of anthers:

[0019] (1) Collection of anthers: 12-14 days before the full flowering of grapes, that is, the pollen tetrad to mononuclear stage, pick the flower buds with full anthers in the morning when the weather is sunny, and store them in the dark at 4°C for 3 days.

[0020] (2) Disinfection of anthers: Disinfect the surface of the flower buds, that is, first soak them in 75% (v / v) alcohol for 1 minute, then rinse them with sterile water for 3 times, and then wash them with 2% (v / v) alcohol. Disinfect with sodium hypochlorite for 8 minutes, and finally rinse with sterile water 5 times.

[0021] (3) Inoculation: cancel the poisonous flower buds, remove the petals with a dissecting needle through a microscope under sterile conditions in the ultra-clean table, clamp the filaments with tweezers, take out the anthers and inoculate them directly and evenly on a callus-containing induction medium with a diameter of 9 cm (Callus induction me...

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Abstract

The invention provides a method for quickly introducing embryonic callus through anther to regenerate a plant, and belongs to the field of agricultural science. The method comprises the following steps: getting wine grape Riesling anther as a material; inducing the embryonic callus through the anther; performing differentiation culture on the callus; performing rooting culture on the differentiated seedlings; and acclimatizing and transplanting to accomplish the grape plant regeneration. The culture medium and the cultivation method have good effect, and have the advantages that the inductivity of the embryonic callus is high, the quality is high, and the cultivation cycle is relatively short; simultaneously, subculturing can be performed for a long term, and a constant loose embryonic callus state can be kept, therefore, sufficient materials are prepared for the study on production of secondary metabolite for grape cell culture, and the study on biotechnology. The culture medium and the anther cultivation method applied to grape genetic breeding can shorten the breeding cycle and provide an important technology platform for improving the grape varieties.

Description

technical field [0001] The invention belongs to the field of tissue culture, in particular to a method for inducing grape anthers to form embryogenic callus through tissue culture. Background technique [0002] Establishing a stable and high-frequency grape regeneration system is of great significance for optimizing transgenic conditions, improving transgenic efficiency, and conducting new grape variety selection and genetic improvement. There are many advantages in using callus as recipient material for genetic transformation research. First, the differentiated cells return to the level of dedifferentiated meristematic cells, which are easy to accept exogenous genes, and the transformation efficiency is high; second, the amount of expansion is large, and the transformed callus obtained by subculture expansion culture, More transformed plants can be differentiated; thirdly, callus can be induced in various tissues and organs, and the test materials of explants are extensive...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01H4/00
Inventor 赵凤霞高相彬王正平李耀宇宋鹏飞李海峰宋学立孟智勇
Owner TOBACCO RES INST HENAN ACADEMY OF AGRI SCI
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