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Method for establishing a Kangdi No.6 oil sunflower regeneration system

An oil sunflower and system technology, applied in plant regeneration, horticultural methods, botanical equipment and methods, etc., can solve problems such as buds are not easy to grow into plants, affect the cultivation effect, and the rooting rate is low, so as to save rooting steps and operations Simple, fast, and powerful effects

Inactive Publication Date: 2012-01-04
新疆康地种业科技股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In general, the frequency of induced buds is related to specific genotypes, and the buds are not easy to grow into plants due to various factors, such as: early maturity, early flowering, severe vitrification and browning, low rooting rate, etc., which all affect its cultivation. Effect
[0006] To sum up, the existing technology has done a lot of work on the tissue culture of sunflower, but there are still some problems. So far, establishing a mature regeneration system is still a research focus of sunflower tissue culture.

Method used

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  • Method for establishing a Kangdi No.6 oil sunflower regeneration system
  • Method for establishing a Kangdi No.6 oil sunflower regeneration system
  • Method for establishing a Kangdi No.6 oil sunflower regeneration system

Examples

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Effect test

Embodiment 1

[0030] (1) Obtaining sterile oil sunflower seedlings: take Kangdi No. 6 oil sunflower seeds, soak them in 70% alcohol solution for 10 minutes, rinse them twice with sterile water, sterilize them with 10% hydrogen peroxide solution for 30 minutes, and then rinse them with sterile water. Rinse twice, and inoculate the embryos on hormone-free 1 / 2 MS medium with the end facing up. The temperature of the culture room is 26°C-28°C, the light time is 16h, and the light intensity is 2000LX, and grown for 3 days. See attached figure 1

[0031] (2) Induction of oil sunflower callus: select the aseptic seedlings with strong growth and cut the cotyledon near the growth point 1-2mm, insert the cotyledon cut into about 5mm depth in the culture medium, induce 2-3 weeks, and induce A large number of calli, 90% of the explants can obtain callus; the induction medium formula is: MS medium + IAA1.2mg / L + 6-BA 4.5mg / L; see attached figure 2

[0032] (3) Adventitious bud differentiation proces...

Embodiment 2

[0036] The detailed regeneration system establishment process is as follows: Aseptic oil sunflower seedlings are obtained: take Kangdi No. 6 oil sunflower seeds, soak them in 70% alcohol solution for 10 minutes, rinse them twice with sterile water, and sterilize them with 0.1% mercuric chloride solution for 5 minutes. Finally, rinse twice with sterile water, and inoculate the embryo on hormone-free 1 / 2MS medium with the end facing up. The temperature of the culture room is 26°C-28°C, the light time is 16h, and the light intensity is 2000LX. It grows for about 3 days, and the growth The sterile seedlings are relatively weak, and some seeds cannot germinate. Induction of oil sunflower callus: select germinated aseptic seedlings and cut cotyledon at 1-2mm near the growth point, insert the cotyledon incision into the induction medium to a depth of about 5mm, induce for 2-3 weeks, and induce the callus; Probably due to the killing of the material by mercury chloride, the amount of ...

Embodiment 3

[0038] The invention includes the acquisition of sterile oil sunflower seedlings, induction of oil sunflower callus, differentiation of adventitious buds, rooting of differentiated seedlings and survival of grafting. The detailed regeneration system establishment process is as follows: Aseptic oil sunflower seedlings are obtained: take Kangdi No. 6 oil sunflower seeds, soak them in 70% alcohol solution for 10 minutes, rinse them twice with sterile water, and then sterilize them with 10% hydrogen peroxide solution After 30 minutes, rinse twice with sterile water, inoculate the embryos on hormone-free 1 / 2 MS medium with the embryo end up, the temperature of the culture room is 26°C-28°C, the light time is 16h, the light intensity is 2000LX, and the growth is about 3 days. The growth of sterile seedlings is stronger. Induction of oil sunflower callus: select the vigorous sterile seedlings and cut the cotyledon near the growth point 1-2mm, insert the cotyledon incision into the in...

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Abstract

The invention discloses a method for establishing a sunflower Kangdi No.6 regeneration system. The method comprises the following steps of: inoculating Kangdi No.6 self-bred line cotyledons into an induction culture medium with different hormone combinations and inducing embryonic calluses, re-inoculating the induced embryonic calluses into a differentiation culture medium and differentiating adventitious buds at high frequency, transferring the adventitious buds into a growth culture medium after the adventitious buds grow to a certain extent and further growing, and finally grafting regenerated plants and transplanting, wherein the induction culture medium comprises a Murashige and Skoog (MS) culture medium, indole acetic acid (IAA) at the concentration of 1.2mg / L and 6-benzylaminopurine (BA) at the concentration of 4.5 mg / L according to the formula; the differentiation culture medium comprises a MS culture medium, IAA at the concentration of 0.01mg / L and 6-BA at the concentration of 0.04 mg / L according to the formula; and the growth culture medium comprises a two fifth MS culture medium according to the formula. The method has the characteristics of extremely high regeneration frequency, shorter period and the like on a sunflower Kangdi No.6 self-bred line, is easy to operate, and has important application value.

Description

field of invention [0001] The invention relates to the field of biotechnology, in particular, the invention specifically relates to the technical field of establishing an oil sunflower regeneration system. Background technique [0002] Sunflower (Helianthus annuus L.) is one of the four major oil crops in the world, and it is also the main oil crop in northern my country. As an oil or protein resource, people pay more and more attention to it. Tissue culture can not only produce a large number of excellent clones, but also break the boundaries between species, overcome distant hybridization and incompatibility obstacles in the development and application of genetic engineering technology in the cultivation of new varieties of sunflower and the improvement of species has huge potential. Although sunflower plant regeneration methods have been greatly developed and various techniques have been reported, since sunflower is a difficult-to-cultivate plant, there are many factors ...

Claims

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Application Information

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IPC IPC(8): A01H4/00A01G1/06
Inventor 王沛政林霞夏春兰王志安李新鹏
Owner 新疆康地种业科技股份有限公司
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