Method for inducing wild cabbage type cole in vitro microspores and screening mutant using ultraviolet

Abstract

The invention belongs to the technical field of plant cell engineering and in particular relates to a method for screening brassica napus in vitro microspores ultraviolet mutation and mutants, which is characterized in that the method comprises: using aqueous sucrose solution to separate brassica napus microspores, suspending the microspores on NLN culture medium which is modified through adding colchicine to be irradiated with ultraviolet and to be doubled with chromosome, transferring doubled microspores on the NLN culture medium which is modified to induce embryoid, then, transferring the embryoid on B5 culture medium to be induced into double haploid plants, transplanting the plants into filed to indentify, and screening out the mutants. The morphological character mutant rate is 10.53-21.57%, and the variant rates of sulfuric glucoside, the oil content and the erucic acid content are respectively 25.53-78.7%, 22.22-59.09% and 38.89-46.97%. The method of the invention has the advantages that the operation procedure is convenient, the time is saved, the cost is low, and the like. The method is suitable for screening the microspores ultraviolet mutation and mutants of cruciferaecrops.

Description

technical field The invention belongs to the technical field of plant cell engineering, and in particular relates to a screening method for in vitro microspores and mutants of Brassica napus induced by ultraviolet rays Background technique Using physical radiation or chemical mutagens to mutate rape seeds or tissues, organs and single cells is one of the important methods to create new germplasm and improved varieties and to establish mutant libraries. However, the mutants produced by mutagenizing seeds, tissues and organs and multicellular callus are chimeras of mutant and non-mutated genes, and recessive gene mutations do not appear in contemporary times and cannot be identified. Therefore, the discovery and selection of stable mutants must be done through selfing, especially for traits controlled by recessive genes and polygenes, multiple generations of selfing are required to obtain homozygous, genetically stable mutants. The period of obtaining mutants is long and the ...

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Problems solved by technology

However, the UV lamp power and irradiation dose rate used by various researchers are different, and the distance and irradiation time of the isolated microspores from the UV lamp are different. The measured microspore half-lethal dose (LD) 50 ) exposure times vary greatly

Moreover, the correlation between irradiation time and dose rate and variation has not been studied, and it is not clear how much dose rate and how long irradiation time can be used to obtain more variation. So far, there has been no research on the theory and method of ultraviolet mutagenesis of isolated microspores. Systematic research, the technology is not yet mature, let alone programmed

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