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Universal induction method for loose type embryonic calluses of beans

A technology of embryogenic callus and callus, applied in the application field of biotechnology in vegetable breeding, can solve the problems of aging, browning, etc., and achieve the effect of solving the effect of low induction efficiency

Active Publication Date: 2016-08-24
TIANJIN RES INST OF VEGETABLE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The purpose of the present invention is to break through the bottleneck of suberization of hard callus internal cells in the bean tissue culture technology, to make the internal and external cells of the induced callus uniform and stable, and to have the ability to continue to grow seedlings; to solve the callus induction process Browning, aging and other problems in

Method used

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  • Universal induction method for loose type embryonic calluses of beans
  • Universal induction method for loose type embryonic calluses of beans
  • Universal induction method for loose type embryonic calluses of beans

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Method for Inducing Loose Embryogenic Callus of Kidney Bean "Shuangfeng No. 1"

[0033] (1) Sterile seedlings obtained by sowing: kidney bean seeds were rinsed with 20% (w / w) sodium hypochlorite for 20 minutes, rinsed with sterile water 3 times, each time for 2-3 minutes, and directly sowed in MS medium, 5 per bottle -6 capsules, get sterile vaccine after 7 days.

[0034] (2) Cut the cotyledons, leaves, and stems between the two leaves of sterile seedlings as explants, and inoculate them on No. 1 medium (MS medium + 0.5mg / mL 2,4-D) for callus induction;

[0035] (3) After 15 days, the induced callus was inoculated on No. 2 medium (MS medium + 0.5 mg / mL 6-BA + 1 mg / mL 2,4-D + 1 mg / mL AgNO 3 ) to induce embryogenic callus. During the induction process from callus to embryogenic callus, subculture once every 10 days. During the subculture process, select smooth surface callus blocks for transfer;

[0036] (4) After subculturing for 4-5 times, the soft callus with a large...

Embodiment 2

[0038] (1) Sterile seedlings obtained by sowing: kidney bean seeds were rinsed with 40% (w / w) sodium hypochlorite for 20 minutes, rinsed with sterile water 3 times, each time for 2-3 minutes, and directly sowed in MS medium, 5 per bottle -6 capsules, get sterile vaccine after 7 days.

[0039] (2) Cut the cotyledons, leaves, and stems between the two leaves of sterile seedlings as explants, and inoculate them on No. 1 medium (MS medium + 0.5mg / mL 2,4-D) for callus induction;

[0040] (3) After 20 days, the induced callus was inoculated on No. 2 medium (MS medium + 0.5 mg / mL 6-BA + 1 mg / mL 2,4-D + 1 mg / mL AgNO 3 ) to induce embryogenic callus. During the induction process from callus to embryogenic callus, subculture once every 15 days. During the subculture process, select smooth surface callus blocks for transfer;

[0041] (4) After subculturing for 4-5 times, the soft callus with a large water content began to transform into a slightly harder granular loose callus, and its...

Embodiment 3

[0043]

[0044] in conclusion:

[0045] (1) The establishment of a general induction method for bean loose embryogenic callus can obtain spherical embryogenic cells with uniform and stable internal and external cells and the ability to continue to grow seedlings.

[0046] (2) More than 90% of the cells constituting the loose embryogenic callus mass can grow into seedlings, which improves the induction efficiency.

[0047] (3) Construct a common experimental system platform for loose embryogenic callus of beans, laying the foundation for scientific research.

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Abstract

The invention discloses a universal induction method for loose type embryonic calluses of beans. The method comprises the steps of carrying out sowing so as to obtain aseptic seedlings, shearing seed leaves, leaves and stem sections between every two leaves of the aseptic seedlings as explants, inoculating the explants to a 1# culture medium for callus induction, 15 to 20 days later, inoculating induced calluses to a 2# culture medium for embryonic callus induction, carrying out subculture once every 10 to 15 days during the induction of embryonic calluses from the calluses, selecting and transferring surface-layer callus blocks with smooth surfaces during subculture, completing the induction of the embryonic calluses of the beans after 4 to 5 times of subculture, wherein the method is a standard induction system for the induction of the loose type embryonic calluses of the beans. The method can be used for solving the problem during the induction of the calluses of the beans that the efficiency of induction is low due to the fact that overall structures and cellular forms of the calluses are nonuniform.

Description

technical field [0001] The invention belongs to the technical field of application of biotechnology in vegetable breeding, and relates to a general method for inducing loose embryogenic callus by tissue culture technology. Background technique [0002] Beans are cultivated in a large area in my country, and the cultivation range is very wide. In recent years, my country's bean production has developed rapidly. According to the FAO report, the planting area of ​​pod-eating cowpea and kidney bean in my country was about 70 million mu in 2013, ranking first in the world. [0003] Callus can be divided into two categories according to histological observation, appearance characteristics, regeneration, and regeneration methods: embryogenic callus (EC) and non-embryonic callus (NEC). . From the perspective of cytology, embryogenic callus is a cell mass composed of embryogenic cells (embryonic cells), which has the characteristics of small cells and equal diameters, thick protopl...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
CPCA01H4/00
Inventor 古瑜刘艳军韩启厚于海龙
Owner TIANJIN RES INST OF VEGETABLE