Molecular marking method for distinguishing salt-tolerance character of tomato

Abstract

The invention provides a molecular marking method for distinguishing salt-tolerance character of tomato. The method comprises the following steps of: 1) positioning a salt-tolerance major gene of the tomato; 2) artificially designing and synthesizing a pair of nucleotide sequences as primers, wherein the primers are oligomers of specific sequences consisting of basic groups; the forward primer is5'GCCAATCCAAACAAACCA'3; and the reverse primer is 5'CATTGTCTCCTTCGCCTCG'3; 3) extracting the DNA of the genome of the tomato; performing PCR amplification on the DNA of the genome of the tomato without digestion by using specific restrictive endonuclease so as to obtain product bands with difference; and marking the sizes of the segments of the PCR product with the bands so as to distinguish the salt-tolerance material and the salt-sensitive material of the tomato; and 4) performing electrophoresis and ethidium bromide staining on the PCR product on 1.5 percent agarose gel; observing and photographing the product with a gel imaging system so as to ensure that the PCR product of the salt-tolerance material has one piece of band of 1,000bp and the salt-sensitive material has one piece of band of 900bp. The method comprises the following steps of: positioning the salt-tolerance gene; selecting the introgressive segments containing the salt-tolerance genes; extracting the DNA of the seedling; establishing a CAPS reaction system; and performing digestion control on the PCR product by using the restrictive endonuclease and the like.

Description

technical field [0001] The invention relates to the localization of tomato salt-tolerant main effect genes, DNA extraction, PCR amplification, design of PCR primers and restriction endonucleases on the basis of existing RFLP marker information, and convert RFLP markers with complicated operation and high cost into The CAPS marker, which is simple to operate and affordable in cost, can identify the salt-tolerant traits of the species. Background technique [0002] Salinity is one of the main environmental factors that limit plant growth and yield. Soil salinization is a worldwide problem. Due to excessive application of chemical fertilizers, secondary salinization of soil, especially vegetable protection land, is an extremely serious phenomenon. It often hinders the growth and development of vegetable crops, resulting in a substantial reduction in production and a decline in product quality. Breeding of new salt-tolerant varieties of crops has always been the focus of resear...

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