Method for cutting wheat chromosomes by SLmuCUT (molecular machine industry, MMI) system

Abstract

The invention relates to a method for cutting wheat chromosomes by a SLmuCUT (molecular machine industry, MMI) system, comprising the steps of taking root tip cell of common wheat (Yannong 15 and Chinese spring), Trititrigia alien substitution line Shannong 0095, and Thinopyrum intermedium as materials; performing microdissection for wheat root tip cell chromosomes on the basis of using the SLmuCUT system to primarily cut the wheat root tip cell chromosomes. The method is simple, quick and convenient to produce slice, fast in slice production speed, and uniform in chromosomes dispersion; on the basis of using the SLmuCUT system to primarily cut the wheat root tip cell chromosomes, main technical parameters for the cutting system for cutting the wheat root tip cell chromosomes is optimized; the microdissection for wheat chromosomes segment, single chromosome, two / three chromosomes is performed successfully; and therefore, the reference is provided for subsequent research such as microdissection and microcloning of specific wheat chromosomes and wheat non-homologous chromosomes, and cloning of exogenous chromosome carrying excellent gene.

Description

technical field [0001] The invention relates to the establishment of a technical system for cutting wheat chromosomes by using the SLμCUT system, which belongs to the field of molecular cytogenetics. Background technique [0002] Chromosome micro-dissection and micro-cloning (chromosome micro-dissection and micro-cloning) technology was proposed by Scalenghe et al. Melanogaster polytene chromosome [J]. Chromosoma, 1981, 82: 205-216) is a new technology of molecular cytogenetics. This technique first cuts the Drosophila chromosome, and successfully obtained 80 clones, which were then used in mice and humans (Rohme Dan, Howard Fox, Bernhard Herrmann, et al. Molecular clones of the mouse complex derived from microdissected metaphase chromosomes[J ]. Cell, 1984, 36:783-788.). With the development of PCR technology, microdissection and microcloning techniques have been greatly developed (Ludecke HJ, Gabriele Senger, Use Claussen, et al. Cloning defined regions of human genome b...

AI Technical Summary

Problems solved by technology

[0003] (1) The existence of plant cell walls and the difficulty of synchronization hinder the preparation of well-dispersed chromosome specimens, and the preparation of chromosome specimens is one of the most basic techniques affecting chromosome cutting; the background is not clean, which affects the cutting efficiency

[0004] (2) The ploidy of the plant genome is diverse, the chromosome structure varies greatly, and the sequence of genes is not conservative enough; the DNA content of plant chromosomes is high and the protein content is low, and most chromosomes cannot display G, R or Q bands like vertebrate chromosomes. Affects DNA sequences (Fukui k., M. Minezawa, Y. Kamisugi, et al. Microdissection of plant chromosome by argon-ion laser beam [J]. Theor Appl Genet., 1992, 84: 787-791); Bands, too much damage to chromosomal DNA, are not conducive to the construction of a complete DNA library; while some plant chromosomes are not only small, but also have no obvious size difference, similar band patterns, and it is difficult to accurately identify chromosomes and their fine structures, all of which increase the analysis. complexity of

If each insert is 1kb, and each chromosome-specific DNA library contains more than 8×105 clones, the workload of library screening is huge

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