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Efficient induction method of plum blossom blade callus

A technology of callus induction and leaves, which is applied in the field of plant tissue culture, can solve the problems of high pollution rate and difficulty in callus induction, and achieve the effect of solving the source and realizing long-term preservation

Active Publication Date: 2012-07-11
BEIJING FORESTRY UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the contamination rate of woody plant explants is relatively high, and the induction of woody plant callus has certain difficulties.

Method used

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  • Efficient induction method of plum blossom blade callus
  • Efficient induction method of plum blossom blade callus
  • Efficient induction method of plum blossom blade callus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] The collection and disinfection of embodiment 1 plum blossom explant

[0021] In spring, collect the fruits of the real plum variety 'Danhanhong' and wash them with clean water. Prepare a 1% potassium permanganate solution and place it in a 2L beaker, soak and clean the exocarp for 1 hour, rinse with running water for 1 hour until the water is clear. Lightly smash with a stone mortar, remove the pulp, lightly smash the endocarp, take the complete and plump seeds, soak in clean water to remove the seed coat, and cultivate the embryos for later use. Disinfect during inoculation, soak the seeds in 70% alcohol for 30s, soak in 1% mercuric chloride for 10min, rinse with sterile water 3 times, and inoculate for later use.

Embodiment 2

[0022] Embryo culture of embodiment 2 plum blossom seeds

[0023] Put the sterilized seeds of Example 1 on sterile filter paper, blot the water on the surface of the seeds; separate the cotyledon with tweezers, cut the lower half of the cotyledon with embryo with a blade, and remove one cotyledon and the other cotyledon 1 / 2 of the explants, and then inoculate the explants in 100ml Erlenmeyer flasks containing 40ml medium, and inoculate 3 explants in each bottle. The medium formula is MS+0.5-1.5mg / L 6-BA+0.1- 0.3mg / L IBA (see Table 1), which added 30g / L sucrose and 7g / L agar; put the culture bottle under dark conditions, culture at 25±3℃ for two weeks, and then switch to light culture , cultivated for two weeks. The light culture condition is light intensity 2000~3000Lx, light time 14h / d. Sterile vaccines can be obtained after inoculation for about one month. The results of hormone concentration and germination rate are shown in Table 1.

[0024] Table 1 Effects of differen...

Embodiment 3

[0027] Induction of embodiment 3 callus

[0028] When the aseptic seedling in embodiment 2 grows to 6-8 sheet leaves, get the 3-4th sheet blade that counts from top to bottom, cut 2-3 cuts along midrib 1 / 3, keep leaf edge to connect, Inoculate to MS + sucrose 30g / L + agar 7g / L + 2,4-D 0~3.0mg / L, pH=5.85 (see Table 2); place at a temperature of 25±3°C and culture in the dark for two weeks .

[0029] Table 2 Phytohormone induced callus

[0030]

[0031] It can be seen from Table 3 that the concentration of 2,4-D 0.25~3.0mg / L can induce callus tissue, but when the concentration of 2,4-D exceeds 1.00mg / L, callus tissue is more likely to occur Browning, necrosis and other phenomena, when the concentration of 2,4-D exceeds 2.50mg / L, the callus induction rate decreases. Considering comprehensively, when the concentration of 2,4-D is 0.5-1.50mg / L, the induction effect is better, and the optimal concentration is 1.0mg / L.

[0032] With the callus induction method established abov...

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Abstract

The invention provides an efficient induction method of a plum blossom blade callus. A tender blade of a seedling obtained by embryo culture of a pure plum blossom variety serves as an explant, the tender blade with the upward paraxial surface enters into a callus induction culture medium, dark culture is performed over 2 weeks, and the callus induction culture medium is a solid culture medium with MS+20-40g / L of sucrose+0.25-2.00mg / L of 2,4-D and pH=5.8-6.0. An efficient induction culture system of the plum blossom blade callus is built. The building of the system enables the callus to be efficiently induced, the inductivity of the blade callus can be over 90%, and the proliferation volume of the callus is large.

Description

technical field [0001] The invention belongs to the field of plant tissue culture, and in particular relates to a method for efficiently inducing plum blossom callus. Background technique [0002] Plum blossom (Prunus mume Sieb. Et Zucc.) is a traditional famous flower in China, with various varieties and abundant germplasm resources. It is native to Southwest my country, the Yangtze River Basin, and the Pearl River Basin, centered on Sichuan, Yunnan, and Tibet. It has a history of application of more than 7,000 years and a history of cultivation of more than 3,000 years in my country. In the process of introduction and cultivation, many excellent new horticultural varieties have been bred, which has greatly enriched the garden landscape of our country. [0003] It only takes about two or three months for plum blossoms to mature from pollination, and the fruit development time is short. When the fruit reaches maturity, the embryo is not yet mature. The seeds have been stor...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
Inventor 张启翔石文芳杨炜茹陈晶鑫
Owner BEIJING FORESTRY UNIVERSITY
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