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Hotness-resistant Lucerne transgenic culturing method

A technology of alfalfa and breeding method, which is applied in the field of transgenic breeding of heat-resistant alfalfa, can solve problems such as hindering the development of grass-fed livestock and poultry in the south, increasing feeding costs, and difficulty in ensuring quality, so as to increase farmers' income, promote rural economic development, The effect of increasing supply

Inactive Publication Date: 2010-02-10
ZHEJIANG UNIV
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Problems solved by technology

[0003] Due to the high temperature in summer, it is difficult to plant alfalfa in southern my country. The alfalfa hay in the south is basically transported from the north. The transportation cost is extremely high, which not only increases the feeding cost, but also the quality is difficult to guarantee, which greatly hinders the development of grass-fed livestock and poultry in the south.

Method used

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Embodiment 1

[0028] The cotyledons of the alfalfa variety "Victoria" were used as explants and inoculated into the induction medium to induce the formation of callus; the callus that had been induced for 2 weeks was transferred to the subculture medium and subcultured to the callus Enter the rapid proliferation phase; take the fast-proliferating callus and pre-culture the recipient callus for 3 days under dark conditions at 25°C; take the pre-cultured callus (about 3000 culture dishes, containing more than 30000 Injured tissue), soaked in the Agrobacterium suspension for 15 minutes, then discarded the bacterial solution, blotted dry with sterile filter paper, inoculated on the co-culture medium, and co-cultivated in the dark at 23°C for 3 days; the co-cultured callus After the tissues were washed and dried, they were transferred to the screening medium, and cultured in the dark at 25°C for 4 weeks, and the grown resistant calli (about 200 pieces) were collected, transferred to a new screeni...

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Abstract

The invention relates to a hotness-resistant Lucerne transgenic culturing method which is characterized by comprising the following steps of: selecting cotyledons of a main-bred Lucerne variety as explants; inducing calli; subculturing the calli till a fast proliferation period; pre-culturing the calli in a dark condition; carrying out agrobacterium co-culture transformation; carrying out 3 turnsof selection by transferring the calli into a selection culture medium; selecting the resistant calli for induction and differentiation so as to obtain seedlings; carrying out histochemical staining and molecular detection on resistant plants; identifying the differentiated seedlings containing target genes; determining hotness resistance of selected transgenic plants; remaining the transgenic plants which are free from hotness damage and grow well; continuing to grow the transgenic plants; carrying out indoor and outdoor assessment and verification on the expression stability of the hotness resistance; carrying out seed proliferation on stable and fine plants; and culturing the hotness-resistant Lucerne which can normally go through the summer. The hotness-resistant Lucerne is more suitable for the growth in southern China and increases the supply of fine forage grass.

Description

technical field [0001] The invention relates to a method for transgenic breeding of heat-resistant alfalfa. Background technique [0002] Alfalfa is known as the "King of Grass" because of its high yield, high quality, long useful life and well-developed root system, which can improve soil structure and increase soil fertility. In my country, with the adjustment of agricultural industrial structure and animal husbandry structure, grain-saving animal husbandry, especially herbivorous animal breeding, has shown a good development trend, because alfalfa has played a decisive role in increasing the yield of dairy cows and mutton sheep. It has become the first-choice feed crop for my country's "three-way" planting structure adjustment, and the alfalfa industry, as an emerging sunrise industry, has a broad space for development. [0003] Due to the high temperature in summer, it is difficult to plant alfalfa in the south of my country. The alfalfa hay in the south is basically tra...

Claims

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Application Information

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IPC IPC(8): C12N15/82A01H4/00
Inventor 黄新舒小丽蒋永清刘新华叶红霞吴殿星
Owner ZHEJIANG UNIV
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