Method for culturing wheat haploid embryo test-tube seedlings

A culturing method and a test-tube seedling technology are applied in the field of culturing wheat haploid embryonic test-tube seedlings, which can solve the problems affecting the transplanting survival and doubling efficiency of haploid seedlings, affecting the application of haploid technology breeding, stopping growth and tillering. Death and other problems, to achieve the effect of alleviating the decline of summer, improving the survival rate, and increasing the success rate

Inactive Publication Date: 2017-05-31
襄阳市农业科学院
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Problems solved by technology

For the cultivation of indoor haploid test-tube plantlets, when the test-tube plantlets germinated from wheat haploid embryos produced by wheat-corn hybridization grow to 2-3 leaves and 3-5 cm high, the temperature, light intensity, and light duration can be set. The stable and adjusted strong seedlings are cultivated in an incubator; when they are cultivated to 2-3 tillers, they can be transferred to other incubators where the temperature, light intensity, and light duration can be set and adjusted for 3-4 months or even longer Oversummer cultivation over time; when the field temperature drops below 15°C, the test-tube seedlings cultivated oversummer will be tempered before transplanting and then transplanted to open field cultivation management. It takes 3-4 months or even longer to cultivate in summer until transplanted to the field. During this period, it is easy to cause yellow leaves, pollution, stop growing, tillering and even death of test tube seedlings due to improper operation, which will affect the transplanting survival and survival of haploid seedlings. Doubling efficiency, affecting breeding applications of haploid technology

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  • Method for culturing wheat haploid embryo test-tube seedlings

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Embodiment 1

[0030] The configuration of No. 1 nutrient supplement solution, No. 2 nutrient supplement solution and strong seedling medium

[0031] 1. The preparation method of No. 1 nutrient supplement solution is: dissolve ammonium nitrate, potassium nitrate, and potassium dihydrogen phosphate in water, and add paclobutrazol dissolved in 95% ethanol, among which ammonium nitrate is 1650 mg / L and potassium nitrate is 1900 mg / L , Potassium dihydrogen phosphate 170 mg / L, paclobutrazol 0.5 mg / L, adjust the pH value to 5.6-5.8, sterilize, and store at 4°C for 2 weeks;

[0032] 2. The preparation method of No. 2 nutrient supplement solution is: dissolve ammonium nitrate, potassium nitrate, and potassium dihydrogen phosphate in water, among which, ammonium nitrate is 1650 mg / L, potassium nitrate is 1900 mg / L, and potassium dihydrogen phosphate is 170 mg / L , adjust the pH value to 5.6-5.8, sterilize, and store at 4°C for a period of 2 weeks;

[0033] 3. Preparation of strong seedling medium:

...

Embodiment 2

[0038] This example is used as a comparative example, without using No. 1 nutrient supplement solution and No. 2 nutrient supplement solution prepared in Example 1; the strong seedling medium adopts MS basic medium, that is, except that paclobutrazol is not added, other components and contents are the same as those in the example 1 prepared strong seedling medium.

[0039] 1. Preparation: preparation of wheat haploid embryonic test-tube plantlets

[0040] (1) Carry out the inoculation test of wheat distant heterozygous haploid immature embryos, which lasted for 1 month, and rescued a total of 1200 immature embryos under sterile conditions;

[0041] (2) Place the inoculated immature embryos on the culture rack in the tissue culture room, and cultivate them under the conditions of 20-22°C, 12 hours of 5000 lx light and 12 hours of darkness; check bottle by bottle every 2-3 days, and pick out the ones that have germinated. Wheat haploid embryonic test-tube plantlets are placed i...

Embodiment 3

[0051] In this example, No. 1 nutrient supplement solution, No. 2 nutrient supplement solution and seedling growth medium prepared in Example 1 were used.

[0052] 1. Preparation: preparation of wheat haploid embryonic test-tube plantlets

[0053] (1) The inoculation test of wheat distant heterozygous haploid immature embryos was carried out, and a total of 1600 immature embryos were rescued under sterile conditions;

[0054] (3) Place the inoculated immature embryos on the culture rack in the tissue culture room, and cultivate them under the conditions of 20-22°C, 12 hours of 5000 lx light and 12 hours of darkness; check bottle by bottle every 2-3 days, and pick out the ones that have germinated. Wheat haploid embryonic test-tube plantlets are placed in the seedling area of ​​the culture rack.

[0055] 2. a kind of culture method of wheat haploid embryogenic test-tube plantlet, concrete method is as follows:

[0056] (1) From May 4th to June 24th, 2014, according to the dif...

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Abstract

The invention discloses a method for culturing wheat haploid embryo test-tube seedlings. The method comprises the following steps that when each of the wheat haploid embryo test-tube seedlings grows to have 2-3 leaves, the seedlings are transferred to a strong seedling culture box for strong seedling culture; when 50% of strong seedlings in the strong seedling culture box reach a three-leaf stage, strong seedling culture is performed; after the strong seedling culture is performed for 2-3 weeks, nutritional supplementing liquid is added; when the test-tube seedlings are cultured to produce 2-3 tillers, the seedlings are transferred to a culture box for over-summering, over-summering culture is performed, and nutritional supplementing liquid is added; from the last ten days of October to the first ten days of November, the culture conditions of the strong seedling culture box are recovered, the seedlings are transplanted and hardened when the field temperature is reduced to 15 DEG C or below, and the seedlings are transplanted in open ground in 2-3 days after seedling hardening. The nutritional supplementing liquid is added at different stages, the wheat haploid embryo test-tube seedlings can grow into one-step strong seedlings through one step, the over-summering decline and fall of the test-tube seedlings are simply and effectively relieved, the survival rate of the test-tube seedlings and survival rate of bottling out and transplantation are improved, and the doubling efficiency of the haploid seedlings is improved indirectly.

Description

technical field [0001] The invention belongs to the technical field of wheat haploid breeding, and in particular relates to a method for cultivating wheat haploid embryogenic test-tube plantlets. Background technique [0002] The technology of wheat haploid induced by hybridization of wheat and corn has become a way to produce wheat haploid with high efficiency through more than 20 years of domestic research. Haploid technology breeding can shorten the breeding period and improve breeding efficiency. Wheat haploid has important application value in screening gene mutants and gene function research, etc. Doubled haploid has important application value in gene mapping, construction of linkage genetic map, fixation of heterosis, and acceleration of wheat breeding process. [0003] Wheat haploid test-tube seedlings produced by hybridization of wheat and corn were formed in the mature stage of field wheat (middle and late May), and then the outside temperature became higher an...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01G1/00A01G31/00A01H4/00
CPCA01G22/00A01G24/00A01G31/00A01H4/001A01H4/008
Inventor 赵翠荣刘莹卢阳王立峰唐清陈科海彭萍
Owner 襄阳市农业科学院
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