Pentaploid hybrid seedling of trifolium ambiguum and trifolium repens and tissue culture propagation method of pentaploid hybrid seedling

A technology of tissue culture and hybrids, applied in the field of breeding, can solve the problems of sterility of pentaploid hybrids and inability to obtain offspring plants, etc., achieve sufficient material supply and strong technical support, strong viability and multiplication quantity, shorten The effect of the breeding cycle

Inactive Publication Date: 2018-11-06
GRASSLAND RES INST OF CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The object of the present invention is to provide a method for tissue culture and propagation of pentaploid hybrids of Caucasian clover and white clover, so as to overcome the problem that pentaploid hybrids are infertile and cannot obtain offspring plants

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] This embodiment provides a method for tissue culture and propagation of pentaploid hybrids of Caucasian clover and white clover, which includes:

[0049] a. Cut the stems of pentaploid hybrid aseptic seedlings into small pieces of about 1 cm, inoculate them on the induction medium as selected explants, and inoculate them at a temperature of 26°C, a sunshine time of 12 hours, and a light intensity of 18000 μmol·m -2 ·s -1 The induction culture was carried out under certain environmental conditions, and the induction of adventitious buds was observed every 5 days.

[0050] Among them, the pH of the induction medium is 5.8, which is added 3.0mg / L 6-benzylaminopurine, 0.3mg / L 2,4-dichlorophenoxyacetic acid and 25g / L sucrose, 7g / L of agar formed.

[0051] b. After 30 days, inoculate the induced young adventitious buds on the subculture medium, and inoculate them at a temperature of 26°C, a sunshine time of 12h, and a light intensity of 18000μmol·m -2 ·s -1 The subcultur...

Embodiment 2

[0060] This embodiment provides a method for tissue culture and propagation of pentaploid hybrids of Caucasian clover and white clover, which includes:

[0061] a. Cut the stems of pentaploid hybrid aseptic seedlings into small pieces of about 1 cm, inoculate them on the induction medium as selected explants, and inoculate them at a temperature of 28°C, a sunshine time of 10 h, and a light intensity of 22000 μmol m -2 ·s -1 The induction culture was carried out under the environment of , and the induction of adventitious buds was observed every 7 days.

[0062] Among them, the pH of the induction medium is 6.0, which is added 2.5mg / L of 6-benzylaminopurine, 0.25mg / L of 2,4-dichlorophenoxyacetic acid and 23g / L of sucrose, 7.5 g / L of agar formed.

[0063] b. After 25 days, inoculate the induced young adventitious buds on the subculture medium, and inoculate them at a temperature of 28°C, a sunshine time of 10h, and a light intensity of 22000μmol·m -2 ·s -1 The subculture was...

Embodiment 3

[0072] This embodiment provides a method for tissue culture and propagation of pentaploid hybrids of Caucasian clover and white clover, which includes:

[0073] a. Cut the stems of pentaploid hybrid aseptic seedlings into small sections of about 1 cm, inoculate them on the induction medium as selected explants, and inoculate them at an ambient temperature of 27°C, a sunshine time of 11 hours, and a light intensity of 20,000 μmol m -2 ·s -1 The induction culture was carried out under the environmental conditions, and the induction of adventitious buds was observed every 7 days.

[0074] Among them, the pH of the induction medium is 5.9, which is added 3.5mg / L 6-benzylaminopurine, 0.35mg / L 2,4-dichlorophenoxyacetic acid and 28g / L sucrose, 6g / L of agar formed.

[0075] b. After 35 days, inoculate the induced young adventitious buds on the subculture medium, and inoculate them at a temperature of 27°C, a sunshine time of 11 hours, and a light intensity of 20,000 μmol·m -2 ·s ...

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Abstract

The invention provides a pentaploid hybrid seedling of trifolium ambiguum and trifolium repens and a tissue culture propagation method of the pentaploid hybrid seedling, and belongs to the field of breeding. The culture method of the pentaploid hybrid seedlings comprises the following steps: taking stem segments of the pentaploid hybrid aseptic seedling as an explant, and inoculating into an induction medium for carrying out induction culture of adventitious buds; inoculating the induced young adventitious buds into a subculture medium for subculturing; after subculturing, inoculating the obtained adventitious bud clumps into a differential medium for carrying out differentiation culture; after regeneration seedlings grow to 2.5 to 4.0 cm, separating the regeneration seedlings into singleplants and inoculating the single plants into a rooting medium for carrying out rooting culture. By adopting the issue culture propagation method, massive pentaploid hybrid seedling plants can be successfully obtained in a short period of time, adequate supply of materials in a backcross and fertility recovery test is ensured and a technical support is provided for the cultivation of new trifoliumvarieties which can be planted in drought and cold areas of Inner Mongolia.

Description

technical field [0001] The invention relates to the field of breeding, in particular to a pentaploid hybrid seedling of Caucasian clover and white clover and a tissue culture propagation method thereof. Background technique [0002] Clover is an annual or perennial forage grass belonging to the genus Trifolium of the family Trifolieae of the leguminous family. For the United States, New Zealand and other developed countries, there is almost no developed modern grassland agriculture and animal husbandry without clover. In most areas of Inner Mongolia, due to the double adversity of drought and cold, it is almost difficult for clover that can be directly cultivated or used for grazing to survive. [0003] White clover (T.repens L.) is a perennial herb of the genus Clover, which only grows in some parts of the Daxing'anling forest area in Inner Mongolia. Long-term natural selection has given it the ability to resist low temperatures of -30°C, but it must survive in areas with...

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 黄帆李俊刘磊李志勇李鸿雁师文贵解永凤
Owner GRASSLAND RES INST OF CHINESE ACAD OF AGRI SCI
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