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Free small spore culturing technology of non heading cabbage

A technology of microspore culture and head cabbage, which is applied to the method of using spores, plant cells, microorganisms, etc., can solve the problems of low cultivation efficiency and achieve the effect of shortening the breeding cycle

Pending Publication Date: 2006-11-29
NANJING AGRICULTURAL UNIVERSITY
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  • Abstract
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Technical problem: The object of the present invention is to solve the problem of low culture efficiency of non-heading Chinese cabbage free microspores in the above-mentioned prior art, to provide a non-heading Chinese cabbage free microspore culture technology, and to obtain a double haploid pure line

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Embodiment 1 Non-heading Chinese cabbage free microspore culture technology, including:

[0019] 1. Genotypes: Wutaicai (N32), Shulu (N20), and Huangbang (N30), respectively from Hubei, Jiangsu, and Liaoning. Among them, Wutaicai is a conventional species, and Shulu and Huangbang are hybrids. Available in the market.

[0020] 2. Material collection: Flower buds with a length of 2-3 mm were selected twice on April 5 and 20, 2004, and the flower buds were kept in a refrigerator at 4°C with moisturizing.

[0021] 3. Disinfection: Disinfect with 75% alcohol for 30 seconds, then disinfect with 0.1% mercury chloride for 5 minutes, and finally rinse with sterile water for 3 times before use.

[0022] 4. Inoculation: put the sterilized flower buds in a 10ml centrifuge tube, add B 5 Squeeze 5ml of the liquid medium gently with a glass rod to free the microspores, filter with a 400-mesh filter, then centrifuge at 800rpm for 5min, discard the supernatant, and repeat 3 times. Fi...

Embodiment 2

[0033] Example 2 Non-heading cabbage free microspore culture technology, genotype, material collection, disinfection, inoculation, high temperature induction, dark culture, plant regeneration and subculture, transplanting are the same as in Example 1, and the mass ratio concentration is 0.1%. The colchicine soaking root irrigation treatment for 8 hours doubled the chromosomes. As can be seen from Table 3, the doubling rates of different genotype haploid test-tube plantlets were also different, and the doubling rates of N20 and N32 were not much different, being 64.4% and 59.5 respectively, while The doubling rate of N30 is 32.9%, which is significantly different from the former two.

[0034] Genotype code

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PUM

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Abstract

The invention relates to nonconiferous cabbage nomadic small spore culture technique, and it belongs to biotechnology field. It is specially used in nonconiferous cabbage nomadic small spore culturing. It includes material getting and disinfect. The density of inoculate small spore is 1*105-6 / ml, it is high temperature induced at 35 degrees centigrade heat induce 24~48 hours, dark development 15-20d is cultured under condition of 25 degrees centigrade. Then plant rebirth, subculture and transplantation are done. Chromosome is doubled quantitatively by adding 0.1 weight percent colchicines for treatment or using 0.1 weight percent colchicines dipping radicle in transplantation. The successful ratio of inducing nonconiferous cabbage gene type can reach 68.5 percent by using this invention, each flower bud introduce 4-10 in average, and even more than 92 at most.

Description

1. Technical field [0001] The invention relates to a technique for cultivating free microspores of non-heading Chinese cabbage, which belongs to the field of biotechnology and is specially used for culturing free microspores of non-heading Chinese cabbage. 2. Technical background [0002] Non-heading cabbage (Brassica campestris ssp.chinensis Makino), commonly known as green vegetables, Chinese cabbage, rape (northern), is native to my country, accounting for 30-40% of the total multi-planting area of ​​vegetables in large and medium-sized cities in the middle and lower reaches of the Yangtze River, and has It has the characteristics of rich nutrition, fast growth, wide adaptability and low cost. In recent years, the planting area in the northern region has grown rapidly, and Japan, the United States and some European countries have also widely introduced and cultivated it. The proportion of the hybrid generation in cultivars is increasing year by year, and the free microspo...

Claims

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Application Information

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IPC IPC(8): C12N3/00C12N5/04A01H4/00
Inventor 侯喜林耿建峰
Owner NANJING AGRICULTURAL UNIVERSITY
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