Wheatear in-vitro culture solution formula of hybrid induced haploid embryos of wheat and maize and preparation method thereof
An in vitro culture and haploid technology, which is applied in the field of agricultural biochemistry, can solve the problems of inconvenience of planting in the same season and inconvenience in research and improvement, and achieve the effects of high fruiting rate, full development and cost reduction of caryopsis.
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Embodiment 1
[0024] Ear culture solution formula (P) of the present invention: 2,4-D100mg / L+ sucrose 35g / L+ potassium dihydrogen phosphate 2g / L+ sulfurous acid 6ml / L+ sodium tetraborate 0.5g / L+ carbonamide 2g / L+ silver nitrate 8mg / L.
[0025] Control ear culture medium formula (C): 2,4-D 100mg / L+ sucrose 40g / L+ silver nitrate 10mg / L+ sulfurous acid 8ml / L, the formula comes from the journal Euphytica (1998, 104: 17-23).
[0026] Preparation of culture solution: (1) Weigh 100mg of 2,4-dichlorophenoxyacetic acid into a beaker, add 50ml of sodium hydroxide with a concentration of 0.1N, and stir with a glass rod or stirrer to make 2,4-dichlorophenoxyacetic acid Phenoxyacetic acid is completely dissolved; (2) Add 600ml of tap water to a volumetric flask with a capacity of 1000ml, then add 35g of sucrose, 2g of potassium dihydrogen phosphate, 0.5g of sodium tetraborate, 2g of carbonamide, 6ml of sulfurous acid, and 8 mg of silver and dissolved 2,4-dichlorophenoxyacetic acid; (3) add tap water t...
Embodiment 2
[0036] Ear culture solution formula (P): 2,4-D100mg / L+ sucrose 45g / L+ potassium dihydrogen phosphate 4g / L+ sulfurous acid 8ml / L+ sodium tetraborate 1.5g / L+ carbonamide 4g / L+ silver nitrate 10mg / L .
[0037] Control (C) panicle culture solution formula: 2,4-D100mg / L+ sucrose 40g / L+ silver nitrate 10mg / L+ sulfurous acid 8ml / L+ potassium dihydrogen phosphate 3g / L, the formula is quoted from "Journal of Wheat Crops", 2008, 28(1):1-5.
[0038] The preparation method of two culture fluids is the same as embodiment 1.
[0039] In vitro culture of wheat-corn hybrid ears: In a comparative experiment on August 7, 2011, a total of 24 wheat ears of two wheat sterile lines K78S (S1) and K456S (S2) that had been pollinated with corn were cut from the field. Back in the room, 12 wheat ears of each wheat material were divided into two groups, with 6 ears in each group, inserted into the culture solution (P) prepared by this formula and the control culture solution (C) respectively and culti...
Embodiment 3
[0046] Ear culture medium formula (P): 2,4-D100mg / L+ sucrose 40g / L+ potassium dihydrogen phosphate 3g / L+ sulfurous acid 7ml / L, sodium tetraborate 1.0g / L, carbonamide 3g / L, silver nitrate 9mg / L. The preparation method of culture medium is the same as embodiment 1.
[0047] In vitro culture of wheat and corn hybrid wheat ears: On July 22, 2012, a total of 156 wheat ears from 15 wheat sterile line materials (S1-S15) that had been inoculated with corn pollen were cut from the field and inserted into the culture prepared by this formula. Cultured in liquid for 14 days (until August 5, 2012), the ear was taken, the caryopsis (grain) was peeled off, the embryo was peeled off under a dissecting microscope, and the number of haploid embryos of each sterile line material was counted. The test results are as follows:
[0048] S1: 12 spikes, 459 pollinated florets, 453 fruiting caryopsis, 180 haploid embryos, induction rate 39.2%
[0049] S2: 15 panicles, 611 pollinated florets, 574 fr...
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