Short-period tissue culture method of peanuts
A tissue culture, short-cycle technology, applied in horticultural methods, botanical equipment and methods, plant regeneration, etc., can solve the problems of difficulty in growing plants, large demand for seeds, low repeatability, etc., and shorten the cycle required for tissue culture. , speed up the breeding speed, good repeatability
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Embodiment 1
[0039] The screening of embodiment 1 cluster bud induction medium
[0040] The present embodiment takes kind sesame oil 1-1 as material.
[0041] The basic medium used in this example is MSB 5 Medium, including the large amount, trace amount and iron salt components in MS medium and B 5 For the culture medium of organic components in the culture medium, its mother liquor configuration is shown in Table 1-Table 5:
[0042] Table 1MS a large number of mother liquor (20 times) configuration
[0043]
[0044] Table 2 MS trace mother solution (200 times) configuration
[0045]
[0046] Table 3 MS iron salt mother liquor (200 times) configuration
[0047]
[0048] Table 4B 5 Organic element mother solution (1000 times) configuration
[0049]
[0050]
[0051] Table 5 Inositol mother liquor (100 times) configuration
[0052]
[0053] MSB per liter 5 The medium contained 30 g of sucrose, 7 g of agar, and had a pH of 5.8.
[0054] to the above MSB 5 The medium...
Embodiment 2
[0065] The screening of embodiment 2 elongation medium
[0066] This embodiment uses the MSB in Embodiment 1 5 The medium is the basic medium, which is formulated according to the combination in Table 8 and added with 6-benzylpurine (6-BA), α-naphthaleneacetic acid (NAA), gibberellin (GA 3 ), to obtain the elongation medium to be screened, with a pH value of 5.8.
[0067] Table 8 The hormone type and concentration combination of the elongation medium to be selected
[0068]
[0069] In this example, the clustered buds or clustered bud tissue pieces induced in Example 1 were cut, transferred to 7 elongation mediums to be tested, and placed in a light incubator for cultivation, with light intensity of 1500-2000lx, time of 16h / d, and temperature 26±0.5°C, subculture once every three weeks, the subcultured seedlings are as follows: image 3 As shown, there are both seedlings and newly differentiated cluster buds on the side, which are used to elongate and proliferate the clu...
Embodiment 3
[0075] Embodiment 3 Establishment and verification of peanut short-cycle tissue culture method of the present invention
[0076] According to the result of embodiment 1-2, establish peanut short cycle tissue culture method of the present invention, comprise the steps:
[0077] 1) Surface disinfection and soaking of peanut seeds;
[0078] 2) Taking the embryonic leaflets of the peanut seeds described in step 1), inoculating them on the cluster bud induction medium for culture, and inducing the embryonic leaflets to produce cluster buds or cluster bud tissue blocks;
[0079] The cluster bud induction medium is MSB 5 +0.2mg / L NAA+5~6mg / L6-BA, solid medium with pH=5.8;
[0080] 3) Cut out the clustered buds or clustered bud tissue pieces described in step 2), inoculate them on the No. 1 elongation medium for culture, and subculture once every three weeks; cut newly differentiated clustered buds or clustered bud tissue pieces for each subculture and go to step 2) Cultivate on th...
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