Method for interspecific hybrid genome in-situ hybridization of crambe cordifolia

Abstract

The invention belongs to the technical field of crop molecular breeding, and in particular relates to a method for interspecific hybrid genome in-situ hybridization of crambe cordifolia. The method comprises the following steps: preparing a probe marker, preparing a slide, performing in-situ hybridization, performing signal detection and the like. In the slide preparation process, a chromosome slide which is good in morphology and good in dispersity in the meiosis period of crambe cordifolia can be obtained under appropriate enzymolysis conditions by using an acetic acid extension flame drying method. When probes and chromosomes are modified, appropriate temperature and time can be selected, appropriate blocking DNA and a probe concentration ratio can be confirmed, fluorescent in-situ hybridization images with clear signals can be obtained, and different interspecific chromosomes of hybrid crambe cordifolia species can be clearly identified, so that genome in-situ hybridization can be well applied to cytological detection of crambe cordifolia species with characteristics of a large number of chromosomes, small morphology and the like, and novel techniques and methods are provided for identification and structural research on crambe cordifolia chromosomes, evolution research on crambe cordifolia, and the like.

Description

technical field [0001] The invention belongs to the technical field of crop molecular breeding, and in particular relates to a method for in situ hybridization of the interspecific hybrid genome of Crabapple, which is related to the field of crop breeding. Background technique [0002] Genomic in situ hybridization (GISH) is a molecular cytogenetic technique developed in the late 1980s. It uses total genomic DNA from one species as a labeled probe, blocks with total genomic DNA from another species at an appropriate concentration, and performs in situ hybridization on the target chromosome. Between the blocked DNA and the labeled DNA probe, the blocked DNA preferentially hybridizes to the general sequence, and the remaining specific sequences are mainly hybridized by the labeled probe. [0003] Relevant documents have been retrieved in terms of biological in situ hybridization technology, such as document-1, the publication number is CN104099416A discloses "a fluorescent in...

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Problems solved by technology

[0003] Relevant documents have been retrieved in terms of biological in situ hybridization technology, such as document-1, the publication number is CN104099416A discloses "a fluorescent in situ hybridization method for sesame chromosomes", the main point is to use smear technology to make sesame metaphase chromosomes Specimens, using fluorescein-labeled BAC specimens for in situ hybridization of sesame BAC chromosomes, after the probe hybridizes to the chromosomes, the fluorescent signal is detected to determine the copy number and position of the target DNA fragment on the chromosome. The disadvantages are: this patent application is different from this The inventions do not belong to the same reaction system, and the genotypes of the species are also different

[0008] So far, there has been no application or report of the in situ hybridization method for interspecific hybrid genomes of Crabapple

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