Soybean in-situ fasciculated bud tissue cultivation method

A tissue culture and meristem technology, applied in the fields of biotechnology and modern agriculture, can solve the problems of low reproduction multiples, few adventitious buds, lack and other problems, and achieve the effect of shortened culture time, robust plant growth and small difference in regeneration.

Inactive Publication Date: 2004-11-17
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Soybean tissue culture is difficult due to the lack of an efficient soybean plant regeneration system
It has few adventitious buds, low multiplicity of reproduction and slow growth, and large differences in genotypes during cultivation, which has affected the research on soybean improvement by biotechnology, thus becoming a difficult point in tissue culture at present.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0011] The test variety is Hefeng 35, 50 capsules;

[0012] Wash the seeds with water and put them in 70% alcohol for 25 seconds, take them out and put them in 10% sodium hypochlorite solution for 15 minutes, wash them twice with sterile water, put the sterilized seeds in MSB+6-BA0.1mg / L, pH5.8, and germinate Germinate on the base at 23°C for 5 days; take out the top growth point of the germinated soybeans, keep two cotyledons, put them on MSB+6-BA1mg / L, pH5.8, keep at 23°C for 30 days on the bud growth medium, and cultivate to a height of 3cm; The adventitious buds were cut and placed in MSB+IBA0.1mg / L, pH5.8, rooting medium 23°C to induce rooting for 15 days, transplanted into large pots to obtain regenerated seedlings; 156 regenerated plants of Hefeng 35 were obtained. The regeneration rate is increased by 60% compared with the conventional cotyledon node explant method.

Embodiment 2

[0014] The test variety is Hefeng No. 39, 50 capsules;

[0015] Wash the seeds with water and put them in 70% alcohol for 30 seconds, take them out and put them in 10% sodium hypochlorite solution for 20 minutes, wash them with sterile water for 3 times, put the sterilized seeds in MSB+6-BA0.5mg / L, pH5.8, germination medium Germinate at 24°C for 4 days; take out the top growth point of germinated soybeans, keep two cotyledons, put them on MSB+6-BA2mg / L, pH5.8, keep 24°C for 25 days on the bud growth medium, and cultivate to a height of 3.5cm Cut off the adventitious buds and place them on MSB+IBA0.3mg / L, pH5.8, induce rooting for 20 days at 23°C on the rooting medium, and transplant them into large pots to obtain regenerated seedlings; 189 regenerated plants of Hefeng 39 were obtained. The regeneration rate is increased by 65% ​​compared with the conventional cotyledon node explant method.

Embodiment 3

[0017] The test variety is Dongnong 43, 50 grains;

[0018] Wash the seeds with water and put them in 70% alcohol for 35 seconds, take them out and put them in 10% sodium hyposulfite for 25 minutes, wash them with sterile water for 4 times, put the sterilized seeds in MSB+6-BA0.8mg / L, pH5.8, germination culture Germinate at 25°C on the base for 3 days; take out the top growth point of the germinated soybeans, keep two cotyledons, put them on MSB+6-BA4mg / L, pH5.8, keep on the medium for 26°C for 20 days on the bud growth medium, and cultivate until 4cm high; cut off the adventitious buds and put them on MSB+IBA0.5mg / L, pH5.8, induced rooting for 25 days on the rooting medium at 25°C, transplanted into large pots to obtain regenerated seedlings; obtained 173 regenerated plants of Dongnong 43. The regeneration rate is increased by 50% compared with the conventional cotyledon node explant method.

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PUM

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Abstract

The present invention relates to a soybean in-situ tufted bud tissue culture method, and said method includes the following steps: placing the disinfected soybean seed on the germination culture medium to germinate soybean aseptic seedling, removing shoot tip growing point and remaining two cotyledons, placing explant on the budding culture medium, cutting off grown-up adventitious bud and placing it on rooting culture medium, after its root system is grown up, obtaining regenerated seedling, then transplanting it into large pot. Said invention can quickly obtain soybean group-cultured seedlings.

Description

technical field [0001] The invention relates to a method for culturing soybean tissue, in particular to a method for culturing soybean in situ clustered bud tissue, and belongs to the fields of biotechnology and modern agricultural technology. Background technique [0002] Using biotechnology to improve soybean has become an important technical approach of modern agriculture. Tissue culture is the basis of biotechnology. The success of soybean tissue culture depends on having a good regeneration system. Through literature search, it was found that Cheng Linmei et al. wrote the article "Study on Plant Regeneration of Different Soybean Explants" in "Journal of Chinese Oil Crops", 1998, 20(2): 6-8. The research on the induction rate of different soybean varieties in this article showed that the following Hypocotyl > epicotyl > small true leaf > immature embryo, the regeneration frequency was 34%, the highest was 50%. The report mentions that the frequency of soybean...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01H4/00
Inventor 武天龙马晓红邱承祥
Owner SHANGHAI JIAO TONG UNIV
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