Cultivation method for Isolated microspore of cucumber

Abstract

The invention provides a method for cultivating isolated microspore of cucumber, belonging to the field of biological technology. The method of the invention comprises the steps as follows: taking buds, identifying the development period of the microspore, pre-disposing, disinfecting the buds, isolating the microspore with an extrusion method by using B<5>-13 extracting solution, filtrating, centrifuging and purifying the microspore by cell screens with the size of 76mum, carrying out static dark cultivation at the temperature of 25 DEG C till the embryoid can be seen by naked eyes, forming cotyledonary embryoid by surging dark cultivation under the frequency of 60r / min<-1> subsequently, forming regenerated plants or rootless seedlings on MS-BA embryoid germination culture mediums, and finally gaining entire regenerated plants by inducing the roots on the rooting culture medium MS-IBA. The method of the invention can gain haploid or double haploid. The double haploid is good seedling raising material and can be used for the selection of new type or improvement of type, thus greatly shortening the seedling raising period and improving the selection efficiency; the double haploid isalso ideal material for carrying out the molecular marking such as AFLP, RFLP, RAPD, and the like, the drawing and researching of genetic maps, the species evolution research, genetic analysis, plantgene cloning selection, etc.

Description

technical field [0001] The invention relates to a method for cultivating cucumber free microspores, which relates to the isolation, purification and suspension preparation of cucumber microspores and a method for cultivating free microspores, and belongs to the field of biotechnology science. Background technique [0002] Haploid refers to an individual with only the number of gamete chromosomes, which is of great significance in crop breeding practice and theoretical research on genetic breeding. Homozygous double haploids can be obtained by chromosome doubling of haploids. In breeding practice, pure lines, that is, double haploid populations, can be obtained by doubling the chromosomes of haploids, but it takes 6 to 7 years or even longer to cultivate inbred lines by conventional breeding methods. At the same time, double haploids are absolutely homozygous, and the genotype and phenotype are completely consistent, while the inbred lines obtained through advanced selfing s...

AI Technical Summary

Problems solved by technology

Among them, the first three methods need to plant larger plant populations for artificial induction, which has a huge workload, long cycle, time-consuming and labor-intensive, high cost and low induction rate, and is difficult to use; while unpollinated ovary and ovule culture, anther culture The embryoid body induction rate and plant regeneration rate are low, and there is a problem of somatic cell interference, which increases the workload of identification of regenerated plants

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