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Annual slide preparation method of plum blossom chromosome

A technology of chromosomes and plum blossoms, which is applied in the preparation of test samples, etc., can solve the problems of short chromosome production cycle and limited time, and achieve the effect of good production effect, long cycle and overcoming time constraints

Active Publication Date: 2014-08-13
BEIJING FORESTRY UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In order to solve the problem of short cycle and limited time for chromosome preparation and material collection in the process of plum blossom chromosome research, the present invention provides a method for annual plum blossom chromosome preparation

Method used

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  • Annual slide preparation method of plum blossom chromosome
  • Annual slide preparation method of plum blossom chromosome
  • Annual slide preparation method of plum blossom chromosome

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1 Chromosomal sheeting of new shoot tips from early spring shoots

[0029] The sprouting leaf buds of plum blossom cultivar Fenpetal were collected from the plum blossom greenhouse of Beijing Jiufeng Forest Park in early March 2012.

[0030] When the leaf buds germinate and elongate, peel off the scales and large leaves, and remove the sprouting shoot tip.

[0031] The obtained stem tips were directly fixed with Carnot's fixative solution at a low temperature of 4°C for more than 20 hours. After fixing, they were rinsed with distilled water and dried with absorbent paper.

[0032] Take 0.20 g of cellulase, 0.20 g of pectinase and 9.60 g of distilled water to prepare an enzyme solution of cellulase and pectinase.

[0033] Put the fixed stem tip into the enzyme solution, and perform enzymatic hydrolysis at 37°C for 150 minutes. After the enzymatic hydrolysis, rinse with distilled water and dry with absorbent paper.

[0034] Cut off the top of the embryo, place i...

Embodiment 2

[0036] Example 2 Immature Embryo Chromosome Preparation

[0037] The immature fruit of the plum blossom cultivar Fenpetal was collected in Qingdao Plum Garden in early May 2011 (25-35 days after pollination).

[0038] Remove the pulp with a stone hammer, open the core, take out the seeds, and take out the immature embryos.

[0039] The obtained immature embryos were directly fixed with Carnot's fixative solution at a low temperature of 4°C for more than 20 hours. After fixing, they were rinsed with distilled water and blotted dry with absorbent paper.

[0040] Take 0.25 g of cellulase, 0.25 g of pectinase and 9.50 g of distilled water to prepare an enzyme solution of cellulase and pectinase.

[0041] The fixed immature embryos were put into the enzyme solution, enzymatically hydrolyzed at 37°C for 90 minutes, rinsed with distilled water, and blotted dry with absorbent paper.

[0042] Cut out the top of the immature embryo, place it on a glass slide, mash it with a dissecting...

Embodiment 3

[0044] Example 3 Preparation of growth point chromosomes obtained from immature embryo tissue culture

[0045] The tissue culture of immature embryos of plum blossom cultivar powder petals was carried out, and the medium formula was MS+6-BA (1 mg / L)+IBA (0.2 mg / L). The cotyledons were cut in half before inoculation. Culture in dark for one week, and when the embryo elongates, cut out the elongated part of 0.5 cm (take the material after one week, the tissue will expand and it is not easy to operate).

[0046] The obtained materials were directly fixed with Carnot’s fixative solution at a low temperature of 4°C for more than 20 h. After fixing, they were rinsed with distilled water and blotted dry with absorbent paper.

[0047] Take 0.25 g of cellulase, 0.25 g of pectinase and 10 g of distilled water to prepare an enzyme solution of cellulase and pectinase.

[0048] Put the fixed material into the enzyme solution, and enzymolyze it at 37°C for 110 minutes. After the enzymolys...

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Abstract

The invention provides an annual slide preparation method of plum blossom chromosome. According to the method, the following meristems are selected as materials for preparing the slide according to time: 1) spring: new stem tips of branches in early spring; 2) early summer: immature embryos of plum blossom and growth points obtained from tissue culture of immature embryos; 3) summer and autumn: top growth points of annual seedlings of plum blossom; and 4) winter: stem tips obtained from hydroponics of dormant branches of plum blossom. The above materials for preparing the slide are treated with chromosome slide preparation, which comprises successive steps of low temperature fixing, enzyme and dyeing tabletting, to obtain a clear chromosome image with good dispersion. The method provided by the invention solves problems of short material collection cycle and time restriction of chromosome slide preparation in the study course of plum blossom chromosome.

Description

technical field [0001] The invention relates to a method for producing plant chromosomes, in particular to a method for producing annual chromosomes of plum blossoms. Background technique [0002] Plum blossom (Prunus mume Sieb.et Zucc.) is an important woody flower, and the study of Prunus mume at the chromosome level is an important prerequisite for its gene mapping and construction of its physical map. However, limited to woody plants, the research on the chromosome level is extremely limited. [0003] According to existing literature records, the number of chromosomes of Mei is 2n=16, 24 as reported by foreign Okabe (1927, 1929), and 2n=16 as published by Oginuma et al. (1987, 1989). In China, mainly Huang Zhe (1989) studied the chromosomes of plum blossoms for the first time; ) studied plum blossom chromosomes, identified the ploidy of plum variety resources, and provided cytological basis for the origin of plum classification and genetic breeding. The experimental m...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N1/28
Inventor 张启翔陈晶鑫杨炜茹孙丽丹
Owner BEIJING FORESTRY UNIVERSITY
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