Lagerstroemia plant stem tip chromosome tablet preparation method

Abstract

The invention discloses a lagerstroemia plant stem tip chromosome tablet preparation method which comprises the following steps of: selecting a part with a length of 0.5-1.0 cm at the innermost side of a lagerstroemia plant annual branch stem tip growing point or a newly-grown stem tip growing point after a branch which is pruned or refrigerated in early winter is subjected to water culture, fixing with stationary liquid (Ethanol: glacial acetic acid: chloroform is equal to 5:3:2 according to the to volume ratio), carrying out hypotonic treatment, carrying out hypotonic treatment again after mixed enzyme liquid is subjected to enzymatic hydrolysis and preparing a tablet by adopting a flame drying method. The chromosome tablet preparation method has the beneficial effects that the processes of pretreatment, dissociation and chromosome tabletting are omitted, the tablet preparation efficiency is improved through directly fixing, the toxic effect of pretreatment liquid and dissociation liquid on a chromosome is avoided, the original form of the chromosome is maintained, the better chromosome dispersion effect is obtained, and meanwhile, the sampling range and the time of lagerstroemia plant stem tip chromosome preparation are enlarged. The tablet prepared by the method can be used for a fluorescence in-situ hybridization experiment to lay a good foundation for the further research of a lagerstroemia genome.

Description

technical field [0001] The invention relates to a method for preparing chromosomes of plants, in particular to a method for preparing chromosomes of shoot tips of the genus Lagerstroemia. Background technique [0002] Lagerstroemia belongs to Lythraceae, and its main species, such as Lagerstroemia grandiflorum and Lagerstroemia urophylla, are important ornamental flowers and trees in summer in my country. Compared with other garden flowers, the research on the chromosomes of Lagerstroemia genus lags far behind, and most of the plants of this genus have small and medium chromosomes, which are not easy to dye. It is difficult to identify their chromosomes by classical cytological methods, which limits the in-depth understanding of the molecular cytogenetic mechanism of Lagerstroemia genus. carried out. [0003] Because most plants of the genus Lagerstroemia are aneuploid and have small chromosomes (Graham and Cavalcanti, 2001), the number of chromosomes reported in this genus...

AI Technical Summary

Problems solved by technology

Compared with other garden flowers, the research on the chromosomes of Lagerstroemia genus lags far behind, and most of the plants of this genus have small and medium chromosomes, which are not easy to dye. It is difficult to identify their chromosomes by classical cytological methods, which limits the in-depth understanding of the molecular cytogenetic mechanism of Lagerstroemia genus. carry out

[0003] Because most plants of the genus Lagerstroemia are aneuploid and have small chromosomes (Graham and Cavalcanti, 2001), the number of chromosomes reported in this genus is inconsistent. Bowden believes that the number of chromosomes of Lagerstroemia indica and Lagerstroemia grandiflora is 2n=50 (Bowden, 1945 ); Guha thinks that the number of chromosomes is only 2n=48 (Guha, 1972). The number of chromosomes of Lagerstroemia in several important domestic chromosome maps is also different, mainly concentrated in 2n=50 or 2n=48. Ping (2009), Tong Jun (2009) and others all used the conventional tablet method in the identification of the ploidy of Lagerstroemia indica polyploidy, and no one has systematically studied the sheet-making technology of the Lagerstroemia indica chromosome

[0004] Lagerstroemia genus plants have thick cell walls and small chromosomes. When using the conventional compression method, the chromosomes will be shortened due to the toxic effect of the pretreatment solution and the dissociation solution, and the chromosomes are easy to overlap during the compression process, making it difficult to obtain Clear cleavage phase, it is difficult to meet the chromosome standard required for fluorescence in situ hybridization experiments, which is not conducive to the experiment

Therefore, the chromosome pellets prepared by the conventional pelleting method can only be used for routine observation and counting and preliminary karyotype analysis, and cannot be used for accurate karyotype analysis by fluorescence in situ hybridization. There is an urgent need for a new chromosome pellet production technology. The chip can meet the needs of accurate karyotype analysis by fluorescence in situ hybridization

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