Chromosome and karyotype analysis method taking pitaya stem tip as material
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A technology for dragon fruit stem and karyotype analysis, which is applied in the research field of plant cytology and cytogenetics, and can solve problems such as troublesome root tip sampling, unstable growth of dragon fruit, and difficulty in obtaining materials
Pending Publication Date: 2022-04-05
HAINAN UNIVERSITY
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Problems solved by technology
Regarding the karyotype analysis and research methods of dragon fruit, the conventional root tip pressing method or the pitaya aerial root pressing method are basically used. Compared with the root tip pressing method, the pitaya aerial root pressing method is more convenient, but pitaya The growth of fruit aerial roots is not stable, the material is difficult to obtain, and the root tip is relatively troublesome, and the success rate of the experiment is not high
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Embodiment 1
[0044] Embodiment 1 chooses the pitaya stem tip as the experimental material, and the specific operation steps are as follows:
[0045] Step 1: Obtaining materials and preprocessing
[0046] Choose healthy, young and tender dragon fruit stem tips, and dissect it to obtain the dragon fruit plant central stem tip.
[0047] Step 2: Concentrate
[0048] Put the dissected dragon fruit stem tip into a mixture of 0.2% colchicine and 0.002M 8-hydroxyquinoline, and place it at room temperature for 2 hours.
[0049] Step 3: Pre-hypotonic
[0050] The concentrated stem tip is soaked at normal temperature with 0.075M KCl.
[0051] Step 4: Front Fixation
[0052] Put the shoot tip into Carnot's fixative solution (absolute ethanol: glacial acetic acid = 3:1), and place it in a refrigerator at 4°C for more than 4 hours.
[0053] Step 5: Dissociation
[0054] Put the shoot tips after washing off Carnoy's fixative into 1M HCl for dissociation.
[0055] Step 6: Enzymatic Digestion
[00...
Embodiment 2
[0090] Example 2 Select 'Bairou' and 'Jindu No. 1' as the experimental materials of this experimental example, and the specific operations are as follows: Step 1: Take materials, pretreatment
[0091] Choose healthy, young and tender dragon fruit stem tips, and dissect it to obtain the dragon fruit plant central stem tip.
[0092] Step 2: Concentrate
[0093] Put the dissected dragon fruit stem tip into a mixture of 0.2% colchicine and 0.002M 8-hydroxyquinoline, and place it at room temperature for 2 hours.
[0094] Step 3: Pre-hypotonic
[0095] The concentrated shoot tips were soaked with 0.075M KCl at room temperature for 120min.
[0096] Step 4: Front Fixation
[0097] Put the shoot tip into Carnot's fixative solution (absolute ethanol: glacial acetic acid = 3:1), and place it in a refrigerator at 4°C for more than 4 hours.
[0098] Step 5: Dissociation
[0099] Put the shoot tip after washing off Carnot's fixative into 1M HCl, and dissociate at room temperature for 9...
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Abstract
The invention relates to a chromosome and karyotype analysis method taking pitaya stem tips as materials. Specifically, the young and tender stem tips of the pitaya are used as experimental materials, and chromosome morphology observation and research are carried out by utilizing the characteristic that the young and tender stem tips have meristematic region tissue cells with exuberant division. The method comprises the following steps that healthy and tender pitaya stem tips are cut off and then dissected; putting the dissected stem tips into a mixed solution of colchicine and 8-hydroxyquinoline, and carrying out chromosome concentration; soaking the stem tips with KCl; the stem tips are refrigerated and fixed with stationary liquid; cleaning the fixed root tips, and dissociating the root tips in HCl; the dissociated stem tips are cleaned and put into a mixed solution of cellulase and pectinase to be subjected to enzymolysis; cleaning the stem tips subjected to enzymolysis, and putting the cleaned stem tips into double distilled water for post-hypotonic treatment; putting the post-hypotonic stem tips into a stationary liquid for refrigeration treatment; and placing the stem tip on a glass slide, cutting, dyeing, and preparing a sheet for microscopic examination. The problems that the aerial roots of the pitaya are difficult to draw, the root tips of the pitaya are difficult to draw, the plants are damaged, and the experimental effect is unstable are solved. According to the method, the pitaya stem tips serve as materials for the first time, karyotype analysis of the pitaya chromosomes is achieved, and more convenient and stable technical support is provided for correlational researches such as karyotype research of the pitaya chromosomes and ploidy identification of germplasm resources.
Description
technical field [0001] The invention relates to a chromosome and karyotype analysis method using the pitaya stem tip as a material, and belongs to the research fields of plant cytology and cytogenetics. Background technique [0002] Plant chromosomes are aggregates containing important genetic information in plant cells, which are easily stained by basic dyes during cell division and can be observed under a microscope to appear rod-shaped or cylindrical. Chromosomal karyotype analysis is a method of analyzing chromosome information by observing and studying chromosome characteristic information such as chromosome number, morphological characteristics, centromere position, and presence or absence of telomeres. [0003] Dragon fruit is a new tropical characteristic fruit of the genus Hylocereus or Seleniereus of the family Cactaceae, which originated in the Central American tropical rainforest area. The chromosomal basal number of the plants of the genus Liangtianchi is 11, a...
Claims
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