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Tabletting method of plum blossom shoot tip chromosome

A technology of chromosome production and plum blossom, which is applied in the preparation of test samples and other directions, can solve the problems of limited access to materials, uncertain processing time, time-consuming production technology, etc., to avoid damage, improve production efficiency, and improve production efficiency. The effect of lengthening chromosomes

Active Publication Date: 2012-07-11
BEIJING FORESTRY UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

They mainly use the smear method and the tablet method, and there are some problems in the technical system: the uncertain processing time of each, time-consuming production technology, etc.; at the same time, the materials are limited to the early spring before the leaves of the plum blossoms, and the time for obtaining materials is short , time limited
[0004] In addition, chromosome specimens made by staining and pressing technology, especially Carbo staining, on the one hand, due to the staining, destroyed the chromosome structure, and could not carry out higher-level fluorescence in situ hybridization experiments; During the flipping process, a large amount of split phases will be lost, which is not conducive to the experiment
Therefore, it can only be used for routine observation and preliminary karyotype analysis, and cannot be used for accurate karyotype analysis by fluorescence in situ hybridization

Method used

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  • Tabletting method of plum blossom shoot tip chromosome
  • Tabletting method of plum blossom shoot tip chromosome
  • Tabletting method of plum blossom shoot tip chromosome

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] For the plum blossom variety ('yellow-green calyx'), at 9:00 in the morning, the top growth point of the annual plant was taken. Directly use Carnot’s fixative, fix at low temperature 4°C for 20 hours; after fixing, rinse with distilled water, blot dry with absorbent paper, transfer to 0.075mol / L KCl solution, and perform pre-hypotonic 30min; Rinse with distilled water, blot dry with absorbent paper, and then use the enzyme solution of cellulase and pectinase (0.25g of cellulase, 0.25g of pectinase, 10g of distilled water) for enzymolysis for 1h50min; Rinse, blot dry with absorbent paper, use ddH 2 After O, the hypotonicity was performed, and the hypotonic time was 30 minutes; the base of the stem tip was cut out, smashed with a dissecting needle, and spread flat on a glass slide, and the impurities were removed with tweezers, and the fixative was dripped, and the slices were baked on an alcohol lamp. Then observed under an optical microscope.

[0020] Microscopic exa...

Embodiment 2

[0022] Prunus cultivar ('Button Yudie'), at 11:00 in the morning, get the top growth point of the new shoot after pruning. Directly use Carnot’s fixative, fix at low temperature 4°C for 20 hours; after fixing, rinse with distilled water, blot dry with absorbent paper, transfer to 0.075mol / L KCl solution, and perform pre-hypotonic 30min; Rinse with distilled water, blot dry with absorbent paper, and then use the enzyme solution of cellulase and pectinase (0.25g of cellulase, 0.25g of pectinase, 10g of distilled water) for enzymolysis for 1h50min; Rinse, blot dry with absorbent paper, use ddH 2 After O, the hypotonicity was performed, and the hypotonic time was 30 minutes; the base of the stem tip was cut out, smashed with a dissecting needle, and spread flat on a glass slide, and the impurities were removed with tweezers, and the fixative was dripped, and the slices were baked on an alcohol lamp. Then observed under an optical microscope.

[0023] Microscopic examination effe...

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Abstract

The invention discloses a tabletting method of a plum blossom shoot tip chromosome. The method comprises the following steps: successively carrying out a Kano stationary liquid fixation, front low permeability treatment, cellulase and pectinase enzymolysis and back low permeability treatment on a part of 0.5-1.0cm at the innermost side of a top growth point of an annual branch of plum blossom or the top growth point germinated newly after pruning; and carrying out flame dry-method tabletting on the treated top growth point. According to the invention, tabletting is carried by adopting a flame method, and pretreatment, dissociation and staining and tabletting processes are omitted to directly carry out fixation, thereby improving tabletting efficiency and elongating the length of the chromosome. In addition, more importantly, the obtained tablet can be used for a fluorescence in-situ hybridization experiment, and simultaneously, the problem that sampling is limited by early spring is solved.

Description

technical field [0001] The invention relates to a plant chromosome preparation method, in particular to a plum blossom stem tip chromosome preparation method. Background technique [0002] Plum blossom (Prunus mume Sieb.et Zucc.) is an important ornamental flower and tree, and the research on its genetic breeding develops rapidly. However, due to the small size of plum blossom chromosomes, studies at the chromosome level are extremely limited. [0003] According to the existing literature records, the number of chromosomes of plum is 2n=16, 24 as reported by Okabe (1927, 1929) abroad. Oginuma et al. (1987, 1989) reported that 2n=16. In China, it was mainly Huang Zhe (1989) who first studied the chromosomes of plum blossoms; Huang Yanwen, Bao Manzhu, etc. (1995) studied the number and shape of chromosomes in wild plums, fruit plums and flower plums; Lin Shenghua, Chu Mengyuan (1999) ) studied plum blossom chromosomes, identified the ploidy of plum variety resources, and pr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/28
Inventor 张启翔陈晶鑫罗乐杨冰洁
Owner BEIJING FORESTRY UNIVERSITY
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