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Method for increasing embryoid induction rate in culture of isolated microspores of hot (sweet) pepper

A technology of microspore culture and embryoid body, applied in the field of cytogenetics theory and breeding, can solve the problem of low induction rate of embryoid body, and achieve the effect of specific steps, wide genotype and good repeatability

Inactive Publication Date: 2010-09-08
HEBEI AGRICULTURAL UNIV.
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Problems solved by technology

[0003] Hot (sweet) pepper free microspore culture technology to obtain embryoid body technology has been successfully reported at home and abroad, Zhang Juping obtained a patent (patent number: ZL 200610042616.0), but the current domestic reports show that hot (sweet) pepper free The embryoid body induction rate of microspore culture is still relatively low, the highest is 1.83 embryoid bodies / anther (Liu Fan, Zhao Hong, Chen Bin, Zhang Yueyun. Embryoid body formation in free microspore cell mass culture of pepper. Acta Molecular Cell Biology , 2007, 40(6): 371-379), subject to certain limitations in practical applications

Method used

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  • Method for increasing embryoid induction rate in culture of isolated microspores of hot (sweet) pepper
  • Method for increasing embryoid induction rate in culture of isolated microspores of hot (sweet) pepper
  • Method for increasing embryoid induction rate in culture of isolated microspores of hot (sweet) pepper

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Embodiment Construction

[0036] According to the technical scheme of the present invention, the specific implementation steps are:

[0037] (1) Determination of the test material: the sweet pepper genotype 08-488 with excellent comprehensive properties was selected as the test material.

[0038] (2) Cultivation of test materials and improvement of flower bud quality: cultivate strong seedlings, plant in time, rationally fertilize and water, pick fruits and flowers that have opened in time, and maintain vigorous growth of plants; It is in an environment with a daytime temperature of 25-30°C and a nighttime temperature of about 20°C.

[0039] (3) Preparation of medium

[0040] The basic medium is the composition of Nitsch and Nitsch (1969) (quoted from J.P. Nitsch, C. Nitsch. Haploid Plants from Pollen Grains. Science, 1969, 1(3): 85-87. See the list for details).

[0041] Solid medium-Nitsch component + 2% maltose + 1.0% activated carbon + 0.6% agar, pH 6.0, sterilized at 121° C., 1.1 Mpa for 18 minu...

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Abstract

The invention discloses a method for increasing the embryoid induction rate in the culture of isolated microspores of hot (sweet) pepper, which belongs to the technical field of the utilization of the theory of cytogenetics and breeding. On the basis of the published culturing method for obtaining embryoids by culturing isolated microspores of hot pepper, the quality of buds for culture is increased, bud sampling, preprocessing, disinfection and anther picking are controlled under lower temperature, anthers are precultured under the temperature of 7 DEG C in the darkness for one week and then cultured under the temperature of 28 DEG C in the darkness, and consequently, up to 8.4 embryoids / anther can be obtained, 1.3 embryoids / anther in average, and are higher than 1 embryoid / anther in the published patent technology and up to 1.83 embryoids / anther in the recently reported domestic research. The invention is characterized in that: the method is practical, the steps are specific, the method is applicable to a wide range of genotypes, the repeatability is good, and the method can ensure that the culture of isolated microspores of hot (sweet) pepper can be widely applied to haploid breeding and the establishment of genetic mapping populations.

Description

technical field [0001] The invention relates to a method for improving the induction rate of embryoid body cultured by free microspores of pepper (sweet) pepper, and belongs to the technical field of using cytogenetics theory and breeding. Background technique [0002] In the hot (sweet) pepper breeding work, the selection of inbred lines is a very important task, but the conventional breeding methods have problems such as long cycle, high cost, and large land occupation. Haploid breeding is an important method to solve this problem. It can greatly improve breeding efficiency, reduce the size of breeding groups, and shorten the breeding period. Free microspore culture is the main way to obtain haploid. The traits of the pollen lines obtained by microspore culture have a large variation range, and the frequency of super-parenting and special good traits is significantly higher than that of conventional breeding; the traits within the strain are neat and stable between genera...

Claims

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Application Information

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IPC IPC(8): A01H4/00C12N5/04
Inventor 申书兴银婷宋彦平顾爱侠陈雪平王彦华轩淑欣李晓峰罗双霞
Owner HEBEI AGRICULTURAL UNIV.
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