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Establishment of a Multi‐color Genomic in situ Hybridization Technique to Simultaneously Discriminate the Three Interspecific Hybrid Genomes in Gossypium
Author(s) -
Guan Bing,
Wang Kai,
Zhou BaoLiang,
Guo WangZhen,
Zhang TianZhen
Publication year - 2008
Publication title -
journal of integrative plant biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.734
H-Index - 83
eISSN - 1744-7909
pISSN - 1672-9072
DOI - 10.1111/j.1744-7909.2007.00623.x
Subject(s) - genome , biology , genetics , somatic cell , chromosome , gossypium hirsutum , botany , gene
To identify alien chromosomes in recipient progenies and to analyze genome components in polyploidy, a genomic in situ hybridization (GISH) technique that is suitable for cotton was developed using increased stringency conditions. The increased stringency conditions were a combination of the four factors in the following optimized state: 100:1 ratio of blocking DNA to probe, 60% formamide wash solution, 43 °C temperature wash and a 13 min wash. Under these specific conditions using gDNA from Gossypium sturtianum (C 1 C 1 ) as a probe, strong hybridization signals were only observed on chromosomes from the C 1 genome in somatic cells of the hybrid F 1 ( G. hirsutum × G . sturtianum ) (A t D t C 1 ). Therefore, GISH was able to discriminate parental chromosomes in the hybrid. Further, we developed a multi‐color GISH to simultaneously discriminate the three genomes of the above hybrid. The results repeatedly displayed the three genomes, A t , D t , and C 1 , and each set of chromosomes with a unique color, making them easy to identify. The power of the multi‐color GISH was proven by analysis of the hexaploid hybrid F 1 ( G . hirsutum × G . australe ) (A t A t D t D t G 2 G 2 ). We believe that the powerful multi‐color GISH technique could be applied extensively to analyze the genome component in polyploidy and to identify alien chromosomes in the recipient progenies.