Culturing method of eurytropic bolting-resisting spring Chinese cabbage free microspores

A technology of microspore culture and culture method, which is applied in the field of culture of free microspores of spring cabbage with broad adaptability and resistance to bolting, can solve problems such as gaps in spring cabbage breeding practices, and achieve the purpose of broadening the selection range of donor plants, improving breeding efficiency, Effects that increase the chance of obtaining

Inactive Publication Date: 2013-09-18
VEGETABLE RES INST OF SHANDONG ACADEMY OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Generally speaking, the free microspore culture system of bolting-resistant spring cabbage is far from

Method used

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  • Culturing method of eurytropic bolting-resisting spring Chinese cabbage free microspores
  • Culturing method of eurytropic bolting-resisting spring Chinese cabbage free microspores
  • Culturing method of eurytropic bolting-resisting spring Chinese cabbage free microspores

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1 Preparation of hybrid combination of different spring cabbage materials and high-generation inbred line materials

[0024] 1.1 Selection and cultivation of test materials

[0025] The test materials included four commercially available spring cabbage varieties and four high-generation inbred line materials. The spring cabbage varieties were one Japanese spring cabbage variety Jujin and three copies (Korea spring cabbage varieties Yangchun, Sambo, Strong, Gao The generation inbred line materials were summer cabbage material 94610, early autumn material J3 and 06-21-1, and spring cabbage material 06-101-4.

[0026] According to the forecast of bolting resistance, the materials and varieties will be bred in 3 batches from late November to mid-December. They will be planted in 50-mesh insect-proof net sheds on March 10 of the following year. The ground will be covered with mulch, and they will bloom in late March. The hybrid combination was prepared by manual hybridizat...

Embodiment 2

[0029] Example 2 Free microspore culture of different spring cabbage materials, high-generation inbred line materials and their cross combinations

[0030] 2.1 Planting of various materials (see 1.1 in Example 1)

[0031] 2.2 Preparation of various media

[0032] Microspore extract and microspore lotion B5W: B5 liquid medium with 13% sucrose, pH 5.8, autoclaved.

[0033] Microspore heat shock treatment culture broth NLN-13: NLN liquid medium plus 13% sucrose, pH 5.8, 0.22μm microporous membrane filter sterilization.

[0034] Microspore dark culture medium NLN-10: NLN liquid medium with 10% sucrose, pH5.8, 0.22μm microporous membrane filter sterilization.

[0035] Microspore embryo growth medium B5G: B5 solid medium supplemented with 6-BA0.2mg / mL+NAA0.02mg / mL+3% sucrose, pH5.8, autoclaved.

[0036] Rooting medium for regenerated plants MS0: MS solid medium, without any hormone added, pH 5.8, autoclaved.

[0037] Please refer to [Guan Chunyun, Li Wei, Li Wenbin, Biotechnology of Brassica Pla...

Embodiment 3

[0048] Example 3 Identification of bolting resistance of regenerated plants cultured with free microspores of 07-ZY4

[0049] 3.1 Domestication, transplantation and selfing of regenerated plants

[0050] After the regenerated plants take root, the medium from which the roots are washed is transferred to a vermiculite: peat = 1:1 matrix to moisturize for 3-5 days for domestication. After the domestication is completed, the plants are transplanted into insect-proof nets for normal management, and transferred to the solar greenhouse at the end of November. After flowering, a single plant with normal pollen is selected to pick buds for self-pollination. Plants that can harvest seeds normally are DH regenerated plants. Seven of the nine regenerated plants of 07-ZY4 harvested selfed seeds, that is, seven double haploid lines (numbered from 09-552 to 09-558).

[0051] 3.2 Identification of bolting resistance of regenerated plants

[0052] In autumn, the double haploid strains and the contro...

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Abstract

The invention discloses a culturing method of eurytropic bolting-resisting spring Chinese cabbage free microspores. The culturing method comprises the following steps of: (1) taking a high-generation selfing line material which can easily carry out free microspore cultivation and carrying out hybridization on bolting-resisting spring Chinese cabbage materials of the microspores to be cultured to obtain hybridized seeds; and (2) sowing the hybridized seeds into a nutrition pot and naturally vernalizing in a greenhouse; from the end of March to the last ten days of April of the next year, taking flower buds at a bolting and flowering period to carry out the free microspore cultivation according to a conventional method to obtain a regenerated plant. According to the culturing method disclosed by the invention, a simple hybridization method is adopted to hybridize spring Chinese cabbages with different backgrounds in the market and the selfing line material which can easily carry out the free microspore cultivation, so that a donor plant gene type state is effectively improved, and a donor plant selection range of the cultivation of the bolting-resisting spring Chinese cabbage free microspores is obviously expanded; and the culturing method is beneficial for improving the obtaining possibility of double haploid materials of the spring Chinese cabbages with the different backgrounds, and the breeding efficiency of the spring Chinese cabbages is greatly improved.

Description

Technical field [0001] The invention relates to a method for culturing free microspores of spring cabbage with wide adaptability and resistance to bolting, and belongs to the field of cell engineering breeding. Background technique [0002] For breeders, rapid purification of breeding materials is the main means to improve breeding efficiency. The conventional method of purifying materials is continuous selfing, and it usually takes 6 to 8 generations to develop parental materials that can be used for cross breeding, which often takes 6 to 8 years. It only takes 1 to 2 years to obtain homozygotes of multiple genotypes through free microspore culture. Therefore, free microspore culture is an ideal method to create stable and variable materials, which can significantly shorten the breeding process and improve breeding efficiency. The culture of free microspores of Chinese cabbage is very obviously affected by the genotype of the donor plant [Cao Mingqing, Li Yan, Liu Fan. The infl...

Claims

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Application Information

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IPC IPC(8): A01H1/02A01H4/00
Inventor 赵智中张志刚刘栓桃李巧云王淑芬卢金东徐文玲刘贤娴付卫民
Owner VEGETABLE RES INST OF SHANDONG ACADEMY OF AGRI SCI
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