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Method for establishing dactylis glomerata tissue culture regeneration system

A technology for tissue culture and establishment of methods, applied in the biological field, can solve problems such as time-consuming experiments, and achieve the effect of shortening the experimental period, overcoming seasonal restrictions, and good quality.

Active Publication Date: 2017-01-11
CHENGDU URBAN MODERN AGRI IND TECH RES INST CO LTD
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Problems solved by technology

Traditional tissue culture methods mostly use the young flower spikes, stem nodes or seedlings of Dactylis chinensis as explants to induce callus, and then obtain complete plants through differentiation. However, in these methods, the selection of explants is also affected by the season. Restrictions cannot provide enough explants for experiments throughout the year, and experiments are relatively time-consuming

Method used

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Embodiment Construction

[0019] The present invention adopts the mature seeds of Dactylis chinensis as explants, washes and sterilizes the mature seeds of Dactylis chinensis, inoculates them in the induction medium and cultivates them for 26-30 days to obtain callus tissue, and the callus tissue is prepared by N6 medium, Vb stock, 30g / L sucrose, The induction medium consisting of 200mg / L acid hydrolyzed complexin, 500mg / L proline, 7.0mg / L 2,4-D, and 4.5g / L agar can greatly increase the induction rate, and then the callus Transfer to callus subculture medium and culture for 5-9 days, consisting of MS medium, 30g / L sucrose, 5.0mg / L 2,4-D, 1.0mg / L 6-BA, 1.0g / L The subculture medium composed of activated carbon and 4.5g / L agar can make the callus proliferate the fastest and have good quality, and then transfer it to the differentiation medium and cultivate it for 18-22d. The MS medium, 30g / L The callus differentiation medium composed of sucrose, 0.2mg / L NAA, 0.5mg / L GA, 1.0mg / L 6-BA, and 4.5g / L agar can g...

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Abstract

The invention discloses a method for establishing a dactylis glomerata tissue culture regeneration system. The season limitation of explant supply can be overcome, and the experimental period can be greatly shortened. Dactylis glomerata mature seeds serve as explants, after the dactylis glomerata mature seeds are cleaned and sterilized, an induction medium is inoculated with the dactylis glomerata mature seeds for culture for 26-30 d, calluses are obtained, the calluses are transferred into a callus subculture medium for culture for 5-9 d, the calluses are transferred into a differential medium for culture for 18-22 d, and when seedlings grow to 2-3 cm, the seedlings are transplanted. The efficient dactylis glomerata tissue culture regeneration system is established through the method, a foundation is laid for establishing a dactylis glomerata tissue efficient genetic transformation system, technical reference is provided for breeding good dactylis glomerata tissue varieties, the season limitation of explant supply is overcome, and the experimental period is greatly shortened. The method is suitable for being applied and popularized in the field of the biotechnology.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for establishing a Dactylis duckweed tissue culture regeneration system. Background technique [0002] The wild species of Dactylis chinensis (also known as Coppergrass and Orchard Grass) are distributed in Xinjiang, the forest edge of Tianshan Mountains, the forest edges of Mount Emei, Erlang Mountain, Qionglai Mountains, Liangshan and Minshan Mountains in Sichuan at an altitude of 1600-3100 meters , shrubs and hillside grasslands; and scattered in the southeastern slopes of Daxing'an Mountains. In addition to domesticating local wild species, the cultivated duckweed is mostly imported from Denmark, the United States, Australia and other countries. At present, it is cultivated in Qinghai, Gansu, Shaanxi, Jilin, Jiangsu, Hubei, Sichuan and Xinjiang and other provinces (regions); Dactylis native to Europe, North Africa and temperate Asia, and later introduced to ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
CPCA01H4/001A01H4/008
Inventor 张新全冯光燕汪霞黄琳凯许蕾黄婷马啸
Owner CHENGDU URBAN MODERN AGRI IND TECH RES INST CO LTD
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