Tissue culture method of haricot in situ cluster seedling

A technology of tissue culture and lentils, applied in horticultural methods, botany equipment and methods, plant regeneration, etc., can solve the problems of large differences between genotypes, few adventitious buds of lentils, and low reproduction number, etc., to achieve small regeneration differences and cultivate Shorter time and better repeatability

Inactive Publication Date: 2005-04-13
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Aiming at the deficiencies and defects of the background technology, the present invention overcomes the problem of low regeneration ability in lentil tissue culture, and provides a method for tissue culture of lentil buds in situ, and solves the problem of indeterminate buds of lentils in tissue c

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0013] The test variety is Jiaoxuan Lentil No. 1, 50 grains;

[0014] Wash the seeds with water and put them in 70% alcohol for 25 seconds, remove them in 10% sodium hyposulfite for 15 minutes, wash them twice with sterile water, put the sterilized seeds in MSB+6-BA 0.1mg / L, pH 5.8, and culture for germination Germinate at 23°C for 5 days; use a scalpel to remove the hypocotyl, the growth points of the terminal buds and lateral buds between the two cotyledons of the germinated lentils under a dissecting microscope, keep the two cotyledons, and put them in MSB+6-BA 2mg / L, pH 5.8, germination at 23°C for 30 days on the bud growth medium, cultured to a height of 2 cm; cut off adventitious buds and put them in MSB+IBA 0.1mg / L, pH 5.8, rooting medium at 23°C to induce rooting for 25 days, Transplant to large pots to obtain regenerated seedlings; obtain 356 regenerated plants of cross-selected lentil No. 1. There are 7-10 clusters of adventitious buds per tree, and the test-tube s...

Embodiment 2

[0016] The test variety is Nicheng No. 1, 50 grains;

[0017] Wash the seeds with water and put them in 70% alcohol for 30 seconds, take them out and put them in 10% sodium hyposulfite for 20 minutes, wash them 3 times with sterile water, put the sterilized seeds in MSB+6-BA 0.5mg / L, pH 5.8, germination medium Germinate at 24°C for 4 days; use a scalpel under a dissecting microscope to remove the hypocotyl, the growth points of the terminal buds between the two cotyledons and the lateral buds, keep the two cotyledons, and put them in MSB+6-BA 3mg / L , pH 5.8, germination at 25°C for 28 days on the bud growth medium, and cultured to a height of 3 cm; cut off adventitious buds and place them in MSB+IBA 0.3mg / L, pH 5.8, induce rooting at 23°C on the rooting medium for 24 days, Transplant to large pots to obtain regenerated seedlings; Obtain Nicheng No. 1, 389 regenerated plants. Produce 7-10 clusters of adventitious buds per tree, cultured for 56 days in test tube seedlings, buds...

Embodiment 3

[0019] The test variety is Nicheng No. 2, 50 grains;

[0020] 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 3 times, put the sterilized seeds in MSB+6-BA 1mg / L, pH 5.8, germination medium Germinate at 25°C for 3 days; use a scalpel to remove the hypocotyl, the growth point between the two cotyledons and the growth points of the lateral buds of the germinated lentils under a dissecting microscope, keep the two cotyledons, and put them in MSB+6-BA 4mg / L , pH 5.8, germination at 27°C for 26 days on the long-bud medium, cultured to a height of 4 cm; excised adventitious buds and placed in MSB+IBA 0.5mg / L, pH 5.8, induced rooting at 23°C on the rooting medium for 23 days, transplanted Planted in large pots to obtain regenerated seedlings; obtained 373 regenerated plants of Nicheng No. 2. Produce 7-10 clusters of adventitious buds per plant, cultured for 52 ...

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Abstract

The present invention is in-situ haricot cespitose bud tissue culturing process in modern agricultural technology. The process adopts regenerated haricot axillary bud meristem and includes the following steps: sterilizing haricot seed, growing bacteria-free haricot seeding in germination culture medium, eliminating the lower plumular axis and top growing points while maintaining two cotyledons, setting the explant in germination culture medium, cutting grown adventitious bud for culturing in rooting culture medium to obtain regenerated seedling with root system, transplanting to big pot and culturing to obtain the regenerated seedling. The present invention can promote the generation of cespitose adventitious bud, and said process is suitable for tissue culture of various haricot varieties to obtain tissue culture seedling fast.

Description

technical field [0001] The invention relates to a method for tissue culture of lentils, in particular to a method for tissue culture of lentil cluster buds in situ, which is used in the field of modern agricultural technology. Background technique [0002] The use of biotechnology to improve crops has become an important technical approach in modern agriculture. Tissue culture is the basis of biotechnology. The success of tissue culture depends on having a good regenerative system. There are few studies on lentil tissue culture. [0003] Find through the retrieval of prior art document, Cheng Linmei etc. are in " Chinese Journal of Oil Crops ", 1998 20 (2): 6~8, author " the research of soybean different explant plant regeneration " this article is to the different soybean varieties. The study of different tissue induction rates showed that hypocotyl > epicotyl > small true leaf > immature embryo, the average regeneration frequency was 34%, the highest was 50%. ...

Claims

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Application Information

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IPC IPC(8): A01G7/00A01H4/00
Inventor 武天龙袁娟马明马晓红
Owner SHANGHAI JIAO TONG UNIV
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