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Aseptic grafting method solving difficulty of soybean transformed adventitious buds in rooting

A technology of sprouts and soybeans, which is applied in the field of crop genetics and breeding, can solve the problems of difficult rooting of soybeans transformed into adventitious shoots, low survival rate of transplanting, etc., and achieve the effect of overcoming the low survival rate of grafting, overcoming technical defects, and solving the problem of difficult rooting

Inactive Publication Date: 2013-01-16
SOUTH CHINA AGRI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] Aiming at the disadvantages in the prior art that soybean transformed adventitious shoots are difficult to root and the survival rate of transplanting is low, as well as the defects and deficiencies in the above-mentioned existing soybean grafting technology, the present invention provides a method for aseptic grafting of soybean adventitious shoots, It solves the technical problem in the prior art that it is difficult to obtain a complete plant that can survive cultivation after soybean genetic transformation treatment

Method used

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  • Aseptic grafting method solving difficulty of soybean transformed adventitious buds in rooting
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  • Aseptic grafting method solving difficulty of soybean transformed adventitious buds in rooting

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Embodiment 1

[0032] (a) Use a sharp knife to separate the soybean bud strips that have undergone conventional Agrobacterium-mediated genetic transformation and screening from the parent body by 0.7cm, and then gently scrape off the epidermis at the base of the stem of the bud strips, as a scion, as attached figure 1 right;

[0033] (b) Select clean and plump mature soybean seeds, and inoculate them with hormone-free MSB after routine sterilization 5 cultured in the dark at 25°C, respectively take 5-day-old seedlings, cross-cut at 5 mm below the cotyledons, discard the cotyledons and epicotyls above, and obtain hypocotyls with radicles as rootstocks, such as figure 1 left. The hormone-free MSB 5 The composition of the culture medium is inorganic components of MS formula + organic components of B5 formula + 25 g / L sucrose + 100 mg / L inositol + 300 mg / L peptone + 7 g / L agar, pH 6.0;

[0034] (c) Use a needle of appropriate thickness (toothpicks, bamboo sticks, needles, etc.) to insert v...

Embodiment 2

[0038] (a) Use a sharp knife to separate the soybean bud strips that have undergone conventional Agrobacterium-mediated genetic transformation and screening from the parent body by 0.7 cm, and cut the base of the bud strips into a "V" shape as a scion, as shown in the attached image 3 shown on the right;

[0039] (b) Select clean and plump mature soybean seeds, and inoculate them into hormone-free MSB after routine sterilization 5 On the culture medium, cultivate in the dark at 25°C, take 5-day-old seedlings, cross-cut at 5 mm below the cotyledons, discard the cotyledons and epicotyls above, and obtain hypocotyls with radicles as rootstocks, as attached image 3 Shown left; hormone-free MSB described 5 The composition of the culture medium is inorganic components of MS formula + organic components of B5 formula + 25 g / L sucrose + 100 mg / L inositol + 300 mg / L peptone + 7 g / L agar, pH 6.0;

[0040] (c) Use a scalpel to cut vertically 0.5 cm downward in the middle of the cross...

Embodiment 3

[0047] In order to compare the influence of rootstocks obtained by different germination days on the growth of grafted seedlings, the present embodiment takes 3, 4, 5, and 8-day-old seedlings respectively, cuts them across at 5 mm below the cotyledons, discards the cotyledons and epicotyls above, Obtain the hypocotyl with radicle, as stock, other methods are with embodiment 1. The growth status of the grafted seedlings obtained is shown in Table 2. As can be seen from Table 2, the rootstocks obtained by different germination days are grafted, and the survival rate of grafted seedlings can reach 100%. The rootstocks germinated for 5 days were used for micro-grafting, and the number of roots increased by 68, which was significantly more than 15.67 for 3 days, 26.33 for 4 days, and 4 for 8 days, and the difference was significant; The increase in length reached the maximum on the 5th day, which was 13.77 cm, significantly greater than 0.43 cm on the 8th day; the fresh weight inc...

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Abstract

The invention discloses an aseptic grafting method solving the difficulty of soybean transformed adventitious buds in rooting. Hypocotyls of soybean seedlings aseptically germinated are used as rootstocks. Genetically transformed soybean buds are scions to be grafted to the rootstocks. The buds grafted with the scions are provided with radicles below the hypocotyls with no need of reforming a root system, grafting is performed aseptically with no need of tedious operations of binding and fixing the scions and rootstocks, the buds which are grafted with the scions can heal with the rootstocks to grow into complete plants able to survive in transplanting and can also grow, develop, flower and fruit normally, and survival rate of grafted seedlings is 100%. The technical problem that the soybean transformed adventitious buds root difficulty is solved effectively.

Description

technical field [0001] The invention belongs to the technical field of crop genetics and breeding, and in particular relates to an aseptic grafting method for solving the problem of difficult rooting of transformed adventitious buds of soybeans. Background technique [0002] soybean ( Glycine max L. Merrill) is a diploid (2n=40) plant belonging to the Fabaceae, Papilioninae, Glycine genus. It originated in China, and later spread to Japan, Europe, the United States and other places. It is one of the main sources of vegetable protein for human beings, and it is also an important livestock feed and a raw material for many industries, and it is also of great value in medicine. Therefore, it is a very important crop. When biology entered the era of molecular biology, scientific research on soybeans also entered a period of rapid development, especially the research on genetic engineering of soybeans has brought huge economic benefits to production. [0003] Plant genetic tr...

Claims

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Application Information

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IPC IPC(8): A01G1/06A01H4/00
Inventor 杨跃生刘颖于磊金亚红郭振飞年海
Owner SOUTH CHINA AGRI UNIV
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