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Regeneration culture method of whole cotyledonary joint of hyacinth bean

A technology of regenerative culture and cotyledon nodes, which is applied in the field of modern biology, can solve the problems of large differences between genotypes, low regeneration ability, and small number of adventitious buds, and achieve the effects of short regeneration cycle, fast bud elongation, and fewer capped buds

Inactive Publication Date: 2011-05-18
上海市浦东新区农业技术推广中心
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The existing lentil tissue culture technology has difficulties such as low regeneration ability, small number of adventitious buds, long regeneration cycle and large differences between genotypes. Using biotechnology to improve lentils and establish an efficient lentil regeneration system is a modern biotechnology researcher. The goal of efforts, but there are few research reports on the regeneration culture of lentils

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] The test variety is 'Green Lentil No. 1', 30 capsules.

[0017] Select plump lentil seeds, rinse them with tap water, disinfect with 70% alcohol for one minute, disinfect with saturated sodium hypochlorite solution for 50 minutes, then rinse with sterile water for 4 times, and soak in sterile water for 1 hour. Then umbilical down to the MSB 5 Grow on germination medium for 4 days. For the germinated lentils, remove the seed coat, protoleaf, epicotyl and radicle, keep two cotyledons, leave 3cm of hypocotyl, and make a small wound at the junction of the two cotyledons, and transfer the explants to MSB 5 + 6-BA 1.0 mg / L + IBA 0.2 mg / L adventitious bud induction medium for 14 days to induce buds. After inducing germination, the entire cotyledon and part of the hypocotyl were excised, and the explants were transferred to MSB 5 + 6-BA 0.05 mg / L + IBA 0.1 mg / L in adventitious bud elongation medium for 20 days. When the adventitious buds elongate to 3 cm, cut the buds and t...

Embodiment 2

[0020] The test variety is 'Green Lentil No. 2', 30 capsules.

[0021] Select plump lentil seeds, rinse them with tap water, disinfect with 70% alcohol for one minute, disinfect with saturated sodium hypochlorite solution for 50 minutes, then rinse with sterile water for 4 times, and soak in sterile water for 1 hour. Then umbilical down to the MSB 5 +6-BA 0.5 mg / L germination medium for 5 days. For the germinated lentils, remove the seed coat, protoleaf, epicotyl and radicle, keep two cotyledons, leave 3cm of hypocotyl, and make a small wound at the junction of the two cotyledons, and transfer the explants to MSB 5 + 6-BA 0.6 mg / L + IBA 0.2 mg / L Adventitious bud induction medium cultured for 15 days to induce buds. After inducing germination, the entire cotyledon and part of the hypocotyl were excised and the explants were transferred to MSB 5 + 6-BA 0.1 mg / L + IBA 0.1 mg / L adventitious bud elongation medium for 25 days. When the adventitious buds elongate to 3 cm, cut the...

Embodiment 3

[0024] The test variety is 'Yanhongbian', 30 capsules.

[0025] Select plump lentil seeds, rinse them with tap water, then disinfect them with 70% alcohol for one minute, then disinfect them with saturated sodium hypochlorite solution for 50 minutes, then rinse them with sterile water 4 times, and soak them in sterile water for 1 hour. Then umbilical down to the MSB 5 +6-BA 0.5 mg / L germination medium for 6 days. Remove the seed coat, protoleaf, epicotyl and radicle of the germinated lentils, keep two cotyledons, and leave 3cm for the hypocotyl, and make a small wound at the junction of the two cotyledons, and transfer the explants to MSB 5 + 6-BA 2.0 mg / L + IBA 0.2 mg / L adventitious bud induction medium for 13 days to induce buds. After inducing germination, the entire cotyledon and part of the hypocotyl were excised, and the explants were transferred to MSB 5 + 6-BA 0.1 mg / L + IBA 0.2 mg / L adventitious bud elongation medium for 25 days. When the adventitious buds elongat...

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Abstract

The invention relates to a regeneration culture method of the whole cotyledonary joint of a hyacinth bean, belonging to the field of modern biotechnology. The method is characterized by comprising the following steps of: (1) sterilizing, soaking and inoculating a hyacinth bean seed to 0-0.5 mg / l of MSB5+6-BA as a germination culture medium to be cultured and germinating an aseptic seedling; (2) removing original leaves, an upper hypocotyl and a radicle from the aseptic seedling, keeping two cotyledons and 3cm of lower hypocotyl, making a micro wound at the joint of the two cotyledons and transferring to 0.6-2.0 mg / l of MSB5+6-BA and 0.2 mg / l of IBA as an adventitious bud inducement culture medium to be induced and germinating; (3) transferring an explant obtained in the step (2) to the adventitious bud inducement culture medium to lengthen; and (4) transferring to 0.1-0.2 mg / l of MSB5+IBA as a rooting culture medium to root and then transplanting to obtain a regeneration plant. The invention has the advantages of large generated adventitious bud quantity, less capping buds, high bud growth speed, short regeneration period, vigorous regeneration plant growth, less tissue culture difference among various seeds and good repetitiveness. The invention is suitable for the tissue culture of various hyacinth bean varieties.

Description

technical field [0001] The invention belongs to the field of modern biotechnology, and relates to a method for regenerating and cultivating whole cotyledon nodes of lentils. Background technique [0002] The use of biotechnology to improve crops has become an important way for the development of modern agriculture. Tissue culture is the foundation of biotechnology. The success of tissue culture depends on having a good regenerative culture method. Ma Xiaohong et al. wrote an article "Establishment of the explant regeneration system of the whole cotyledon node of soybean and comparison with the regeneration system of the cotyledon node and embryo tip" in "Soybean Science" in the 27 (3) issue of 2008. In this paper, the whole cotyledon node explant regeneration system of soybean was established, and the regeneration frequency, bud number, bud elongation and regeneration cycle of different soybean explant regeneration systems were compared. The research showed that the soybea...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
Inventor 袁娟周超英汪洁金彩华吴寒冰
Owner 上海市浦东新区农业技术推广中心
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