Method for obtaining dihaploid plants of sweet peppers

A double haploid, sweet pepper technology, applied in the biological field, can solve the problems of restricting the wide application of sweet pepper, waste of haploid plants, unsatisfactory doubling effect, etc., to achieve the inhibition of spindle formation, equivalent doubling effect, and cultivation The effect of improving efficiency

Inactive Publication Date: 2012-05-23
BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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  • Abstract
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  • Application Information

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Problems solved by technology

The doubling effect of the field colchicine doubling method commonly used in the prior art is also unsatisfactory
Therefore, a large number of haploid plants are wasted, which limits the widespread application of sweet pepper haploid breeding technology

Method used

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  • Method for obtaining dihaploid plants of sweet peppers
  • Method for obtaining dihaploid plants of sweet peppers
  • Method for obtaining dihaploid plants of sweet peppers

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Embodiment 1

[0032] The concrete steps of sweet capsicum double haploid cultivation method of the present invention are as follows:

[0033] (1) Select a plant with good growth and no pests and diseases as the donor plant, select flower buds whose microspore development is in the single-nucleate marginal stage to the early binucleate stage, and pretreat the flower buds at a low temperature of 4°C for 1-3 days according to the results of microscopic examination;

[0034] (2) Take flower buds of appropriate size, peel off the calyx, soak them in 75% (v / v) alcohol for 30 seconds, then sterilize them with 8-10% (v / v) sodium hypochlorite for 10-20 minutes, and finally use aseptic Wash with water 3 times, 3-5min each time;

[0035] (3) Peel off the anthers intact on the workbench, and inoculate them on the N4-3 solid-liquid double-layer medium with a density of 12-18 anthers per 60mm diameter petri dish. The formula of the N4-3 medium is:

[0036] Solid layer: NTH medium+3-6% sucrose+0.5% gac+0...

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Abstract

The invention provides a method for obtaining doubled haploid (DH) plants of sweet peppers, which comprises the following steps of: (1) selecting buds of a donor plant, the microspores of which are in a development state between nonokaryotic stage with the nucleus located aside and early dikaryotic stage, and pretreating at 4 DEG C for 1-3 days; (2) stripping the calyces off, soaking in alcohol, disinfecting through oscillation with sodium hypochlorite, and cleaning with sterile water; (3) stripping the anthers, and inoculating the anthers on an N4-3 solid-liquid double-layer culture medium by the density that 12-18 anthers are inoculated to a culture dish with diameter of 60 mm; (4) firstly, culturing the anthers for 1-10 days in the dark at 28-35 DEG C, and then, transferring to the condition of 25-28 DEG C to continue culturing in the dark; and (5) culturing for 4-7 weeks, when a large quantity of embryoids occur, transferring leaf type embryoids to an MS basic culture medium free of hormones, and cultivating the seedlings. According to the cultivating method provided by the invention, the occurrence rate of the embryoids breakthrough the limitations of genotype, the culturing efficiency of sweet peppers is greatly increased, and meanwhile, the ratio of the DH plants is also increased.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a haploid breeding technology. Background technique [0002] Abundant segregation, variation, and recombination are manifested in the present generation through haploid culture, and can be permanently stabilized in homozygous form through chromosome doubling. Compared with conventional breeding technology, haploid breeding technology has the following advantages: first, it can significantly speed up the homozygous speed, and the application of haploid breeding can shorten the crop breeding cycle by 5-6 generations; second, double the recessiveness of haploid The gene can be normally expressed without being covered by the dominant gene, which can significantly improve the selection efficiency and selection accuracy of the genotype, and is beneficial to the selection of multiple gene recombination and recessive genes. In addition, haploid culture has important application and research...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01H4/00
Inventor 张晓芬耿三省陈斌张宇
Owner BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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