A bac-fish method for simultaneous identification of complete sets of chromosomes in each subgroup in the background of ad or ade in cotton

Abstract

The invention belongs to the field of molecular cytogenetics and particularly relates to a BAC-FISH method for simultaneously identifying whole sets of chromosomes of subgroups under a gossypium AD / ADE background. The method comprises the following steps: carrying out BAC-FISH on different cotton seeds by using BAC cloned 57I23 originated from an Africanum BAC library, and simultaneously identifying chromosomes of an AD genome and AD sub genome of cotton obviously; by using BAC-DNA as a probe, covering most regions out of ends of all chromosomes of the subgroup A in five gossypium tetraploids as well as a small region near the middle part of the all chromosomes of the subgroup D by a hybrid signal; and in allohexaploid hybrids AADDEE hybridized by upland cotton and G.stocksii (E1) in the group E, as the hybrid signal is similar to the tetraploid, obviously differentiating the hybrid signal from the subgroup E (without a signal) so as to simultaneously identify three sets of chromosomes of subgroups (A,D,E). By adopting the BAC-FISH method through BAC cloning, the whole sets of chromosomes of subgroups under the gossypium AD / ADE background can be simultaneously identified.

Description

technical field [0001] The invention belongs to the field of molecular cytogenetics, and specifically relates to a BAC-FISH method for simultaneously identifying a complete set of chromosomes of each subgroup under the background of AD or ADE of cotton. Background technique [0002] Fluorescence in situ hybridization (FISH) technology can locate DNA sequence on chromatin, chromosome or DNA fiber, and is an effective and precise molecular cytogenetics method. Plant genome sequencing is required for functional genome research, and the divergence between genetic distance and physical distance limits the application of linkage maps in guiding genome sequence assembly and map-based cloning. The cytogenetics map represents the actual position of the DNA sequence on the chromosome, and is one of the types of genome physical maps, so it is also the basis for genome sequencing and sequence assembly. Direct positioning of DNA clones on chromosomes by BAC-FISH is currently one of the ...

AI Technical Summary

Problems solved by technology

Especially for allopolyploids with ADE background, involving 3 genome sources, 2-3 gDNA double-color or multi-color probes are needed to realize the identification of subgroup chromosomes, and the workload, technical requirements, and reagent costs increase accordingly

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