Method for quickly establishing metaphase chromosome karyotype of cucumber through genomic in-situ hybridization

Abstract

The invention discloses a method for quickly establishing a metaphase chromosome karyotype of a cucumber through a genomic in-situ hybridization technology. The method includes the steps that genome DNA of the cucumber is extracted and marked as a probe; enzymolysis is conducted on an extracted root tip of the cucumber, a sheet is prepared through a flame drying method, and then a metaphase chromosome sheet with the good dispersity is acquired; the genome DNA serving as the probe is degenerated and then hybridized to the degenerated metaphase chromosome sheet, under the hybridization condition with the high preciseness, clear and differentiable fluorescence signals are produced through a chromosome tandem repeated sequences, and then the metaphase chromosome karyotype of the cucumber is established. The genome DNA of the cucumber serves as the probe, the metaphase chromosome karyotype of the cucumber is quickly established through one-time GISH, and then testing efficiency is improved. Through the method, based on the different signal distribution modes of the genome DNA probe of the cucumber on chromosomes, the metaphase chromosome karyotype of the cucumber can be quickly established, and a novel method and reference are provided for analytical investigation of karyotypes of species.

Description

1. Technical field [0001] The invention discloses a method for rapidly constructing a cucumber mitotic metaphase chromosome karyotype by utilizing the in situ hybridization technology of the cucumber's own genome, which is beneficial to carrying out karyotype analysis and research on cucumbers and other species, and belongs to the field of plant biotechnology. 2. Technical background [0002] Cucumber (Cucumis sativus L., 2n=2x=14) is one of the most important vegetable crops in the world. Cucumber has some special traits, such as diversification of sex phenotype, paternal inheritance of mitochondria, etc.; at the same time, the genome of cucumber is relatively small (about 370Mb), and the number of chromosomes is only 14, which makes cucumber a molecular cytogenetic trait in recent years. One of the model species for scientific research. However, the chromosomes in the cucumber metaphase are small, the dyeing ability is weak, and the morphology of the chromosomes is very s...

AI Technical Summary

Problems solved by technology

[0007] The purpose of the present invention is to use its own genome in situ hybridization to quickly construct the method of cucumber metaphase chromosome karyotype. It is to solve the shortcomings of previous cucumber mitotic metaphase chromosome karyotype construction methods, such as tediousness, and to invent a kind of primary genome that only uses the cucumber's own genome as a probe. In situ hybridization (GISH) can quickly construct the method of cucumber metaphase chromosome karyotype, which has accelerated the progress of cucumber cytogenetics research and provided new methods and ideas for the karyotype analysis of other species

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