A method for identifying the complete set of chromosomes of cotton d genome and D sub-genome
A chromosome and cotton technology, applied in the field of molecular cytogenetics, can solve problems such as centromere is difficult to distinguish, chromosome shortening degree is different, chromosome size is small, etc.
Inactive Publication Date: 2016-05-04
INST OF COTTON RES CHINESE ACAD OF AGRI SCI
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Problems solved by technology
However, in actual experiments, different types of reagents, different concentrations and differences in treatment time often lead to different degrees of shortening of chromosomes. In addition, the size of cotton chromosomes is small, the difference between chromosomes is not large, and the centromere is not easy. Factors such as discrimination make it difficult to obtain ideal and reliable results by relying on chromosome morphological characteristics to identify chromosomes
Method used
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Embodiment 1
[0088] Example 1 Use the 2kb sequence as a probe to hybridize upland cotton (AD) 1 , Asia Cotton A 2 , Raymond Cotton D 5 Metaphase chromosomes
[0089] 1 Materials and methods
[0090] 1.1 Experimental materials
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Abstract
The invention belongs to the field of molecular cytogenetics and particularly relates to a method for identifying D genome and D sub-genome complete chromosome. The method comprises the following step: by taking a sequence as shown in SEQ ID NO.1 as a probe to perform fluorescence in-situ hybridization on the DNA (deoxyribonucleic acid) of a cotton genome. The sequence is taken as the probe, and chromosomes in the mitosis metaphase of the upland cotton, the asiatic cotton and the Raymond's cotton are taken as target DNA to perform fluorescence in-situ hybridization, and the results indicate that obvious hybrid signals are shown on the D subgenome chromosome of the upland cotton and the D genome chromosome of the Raymond's cotton, the signals are distributed on all chromosomes, and no obvious signals are shown in the A subgenome chromosome of the upland cotton and the A genome chromosome of the asiatic cotton. The sequence adopted by the method disclosed by the invention can be used for quickly identifying the cotton D genome and D sub-genome complete chromosome by virtue of a FISH (fluorescence in situ hybridization) method.
Description
Technical field [0001] The invention belongs to the field of molecular cytogenetics, and specifically relates to a method for identifying a complete set of chromosomes of cotton Dgenome and Dsub-genome. Background technique [0002] Gossypium is classified into 52 cotton species, including A, B, C, D, E, F, G and K 8 chromosomes. Nowadays, the main cultivars of cotton worldwide are the tetraploid cotton species Upland cotton (AD) and Sea island cotton (AD), which account for about 95% and 5% of the cultivated area respectively. At present, the genome sequencing of group A cotton species Asian cotton and group D cotton species Raymond's cotton has been completed (Wangetal., 2013; Lietal., 2014). The comparative study of cotton A and D genomes will be of great help to the evolution of tetraploid cotton and to cotton breeding. Fluorescence in situ hybridization technology was born in the 1960s, and its emergence opened a new era of cell molecular biology. Subsequently, the fluore...
Claims
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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6841C12Q2543/10C12Q2563/107
Inventor 崔兴雷刘方刘玉玲孟菲王春英周忠丽蔡小彦王星星王玉红彭仁海王坤波
Owner INST OF COTTON RES CHINESE ACAD OF AGRI SCI
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