Scientia Agricultura Sinica ›› 2015, Vol. 48 ›› Issue (2): 398-406.doi: 10.3864/j.issn.0578-1752.2015.02.20

• RESEARCH NOTES • Previous Articles    

Rapid Karyotype Analysis of Cucumber Varieties Based on Genomic in situ Hybridization

ZHANG Yun-xia, LOU Qun-feng, LI Zi-ang, WANG Yun-zhu, ZHANG Zhen-tao, LI Ji, CHEN Jin-feng   

  1. College of Horticulture, Nanjing Agricultural University/State Laboratory for Crop Genetics and Germplasm Enhancement, Nanjing 210095
  • Received:2014-06-24 Online:2015-01-16 Published:2015-01-16

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Abstract: 【Objective】Using genomic in situ hybridization (GISH), metaphase chromosome analysis was performed for two variants of cucumber (C. sativus L.，2n = 2x = 14) [cultivated cucumber (C. sativus var. sativus) and wild cucumber (C. sativus var. hardwickii)] to quickly establish the chromosome karyotype of cucumber varieties, which will provide useful information for molecular cytogenetic research of cucumber. 【Method】Genomic DNA from cultivated cucumber ‘9930’ isolated using CTAB method and 45S rDNA were labeled with Dig-dUTP or Bio-dUTP through nick-translation method, and used as probes and hybridized onto the metaphase chromosomes of cultivated cucumber itself and wild cucumber using genomic in situ hybridization technique. Individual chromosome of two cucumber variants was analyzed based on the GISH banding, and 45 rDNA signals as well, and the karyotypes of these two variants were constructed. 【Result】 The GISH results showed that cucumber genomic DNA probes exhibited distinguished signal patterns on the chromosomes of two variants. The unique distribution and intensity of signals were observed on each chromosome, rather than the even distribution of signals across all chromosomes. For C. sativus var. sativus ‘9930’, GISH produced obvious signals at the both ends of chromosomes and pericentromeric heterochromatin regions on all other chromosomes except for chromosome 6 which only gave signals at the end of short arm and pericentromeric heterochromatin region. The signal patterns from each chromosome exhibited an obvious difference. 45S rDNA loci were mapped adjacent to centromeric regions on chromosomes 1, 2, 3, 4 and 7, among which three pairs of strong signals and two pair of weak signals were observed. However, the signal patterns from wild cucumber showed an obvious difference compared with that from ‘9930’. The GISH signals were observed only at one end of chromosomes 1, 2, 4 and 5, and pericentromeric heterochromatin regions of all chromosomes. 45S rDNA signals were observed only on the chromosomes 1, 2 and 3, in which chromosome 1 produced extremely strong signals and chromosomes 2 and 3 gave very weak signals. These results indicated that unique signal patterns for cucumber variants could be produced by GISH using cucumber genomic DNA. Each chromosome was characterized unequivocally according to the signal patterns, and a karyotype for each cucumber variant was constructed. Compared with the distribution of cucumber repetitive sequences from previous reports, the GISH patterns were found to locate mainly in tandem repetitive blocks of cucumber chromosomes. 【Conclusion】Results of this study showed that the cucumber genomic in situ hybridization could quickly reveal the distribution of genomic tandem repetitive sequences at one time, and could be efficiently applied to rapid karyotype analysis of different variants. It was also found that significant differentiation of repetitive sequences happened among cucumber variants.

Key words: cucumber varieties, GISH, chromosome, karyotype analysis, repetitive sequence

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