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Method for breeding somatic embryo of immature soybean cotyledon, preserving succeed generation and revegetating plant

A somatic embryo and immature technology, applied in plant regeneration, botany equipment and methods, horticultural methods, etc., can solve the problems of complicated operation techniques and difficulty in mastering, and achieve low production costs, simple and easy-to-master operation methods, and save The effect of removing the device

Inactive Publication Date: 2003-05-07
MILITARY SUPPLIES UNIV THE CHINESE PLA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This system has greatly improved the efficiency of soybean tissue culture, but liquid suspension culture requires equipment such as a constant temperature shaker, and the operation techniques are complicated and difficult to master. Only certain cultivated soybean varieties such as Fayette can obtain embryogenic cell clusters, and there are only a few laboratories. This system was used (Bailey, 1993), but has not been widely used

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] Somatic Embryogenesis, Globular Embryo Proliferation, Subculture Preservation and Plantlet Regeneration of Hefeng 25 Soybean Variety

[0016] Plant Hefeng 25 in the field, pick soybean pods that have bloomed for 16-20 days, and sterilize with 75% ethanol for 2 minutes, 0.1% HgCl 2 Sterilize for 15 minutes, rinse with sterile water 3 times, peel off the seed coat, select 3-5mm cotyledon to remove the hypocotyl, and inoculate the cotyledon with the adaxial side up in MS medium (additional 20mg / L 2,4-D, pH = 6.0), cultured under dark conditions at 25°C, the somatic embryogenesis rate (number of somatic embryogenic cotyledons / number of inoculated cotyledons × 100%) was 41.36% at 42 days.

[0017] From the somatic embryos, select the yellow-green, small granular, and clustered spherical embryos with a diameter of 0.9-1.1 mm that have not been swallowed by the surrounding callus, and transfer them to MS medium (additional 20mg / L 2, 4-D) Induced proliferation of somatic embry...

Embodiment 2

[0021] Somatic Embryogenesis, Globular Embryo Proliferation, Subculture Preservation and Plantlet Regeneration of Dongnong 40 Soybean Variety

[0022] Plant Dongnong 40 in the field, pick soybean pods that have bloomed for 16-20 days, and sterilize with 75% ethanol for 2 minutes, 0.1% HgCl 2 Sterilize for 12 minutes, rinse with sterile water 3 times, peel off the seed coat, select 3-5mm cotyledons to remove the hypocotyl, and inoculate the cotyledons with the adaxial surface upward in MS medium (additional 40mg / L 2,4-D, pH = 6.0), cultured under dark conditions at 25°C, the somatic embryogenesis rate (number of somatic embryogenic cotyledons / number of inoculated cotyledons × 100%) was 40.34% at 50 days.

[0023] From the somatic embryos, select the yellow-green, small granular, and clustered spherical embryos with a diameter of 0.9-1.1 mm that have not been swallowed by the surrounding callus, and transfer them to MS medium (additional 20mg / L 2, 4-D) Induce the proliferation ...

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PUM

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Abstract

A tissue culture technique for soybean includes using the immature soybean cotyledon as explant to induce the reproduction of somatic embryo in solid culture medium, successive transfer storage in the solid culture medium, and normally germinating to regenerate plant.

Description

Technical field: [0001] The invention relates to the technical field of crop tissue culture methods, and relates to a method for somatic embryo proliferation, subculture preservation and plant regeneration of immature soybean cotyledons. Background technique: [0002] Soybean is an important economic crop and the main source of human edible oil and vegetable protein. In soybean production, the yield and quality of soybean are often seriously affected by the occurrence of diseases, weeds and insects. The conventional breeding methods are used to improve soybean varieties, but the improvement effect is not good due to the lack of resistance sources. [0003] With the development of molecular biology and molecular genetics, it has become one of the important methods of modern molecular breeding to transfer exogenous genes into plants to improve some undesirable traits by using transgenic technology. Tissue culture technology is the basis of transgenic work, although immature ...

Claims

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Application Information

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IPC IPC(8): A01H4/00
Inventor 王罡王萍季静吴颖
Owner MILITARY SUPPLIES UNIV THE CHINESE PLA
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