xBrassicoraphanus, a new synthetic intergeneric hybrid between Brassica rapa L. ssp. pekinensis and Raphanus sativus L., also locally known as ‘Baemoochae’, is an interesting subject for studying polyploidy and genome plasticity in the family Brassicaceae, but very few genomic and cytogenetic information. Here, we analysed the chromosome complements and pairing of the most fertile lines, BB1 and BB5, using dual-color fluorescence in situ hybridization (FISH) and genomic in situ hybridization (GISH) to check their chromosomal segregation stability. The somatic chromosome complement of B. rapa was confirmed to be 2n=20 (2.8~4.8μm), of R.sativus, 2n=18 (2.0~3.3μm), and of xBrassicoraphanus, 2n=38 (2.2~5.0μm). There were eight, eight, and seventeen metacentric pairs and two, one, and two submetacentric pairs in B. rapa, R. sativus, and xBrassicoraphanus, respectively. Additionally, three, two, and five pairs of 5S rDNA and five, three, and eight pairs of 45S rDNA were observed in B. rapa, R. sativus, and xBrassicoraphanus, respectively. This suggests that both B. rapa (AA) and R. sativus (RR) genomes, particularly the rDNA arrays, co-exist in xBrassicoraphanus (AARR) genome. In meiosis I, nineteen bivalents were most frequent, and GISH analysis showed ten bivalents from the A genome. This study would provide a useful information for further genomic study of xBrassicoraphanus and its improvement as a new promising breeding variety.
Radish, Raphanus sativus L., is an annual vegetable of the family Cruciferae. Radish has RR genome with 18 somatic chromosome numbers (2n=2x=18). Until now, detailed karyotypic analysis is not only constructed only by conventional staining techniques but also other method. Fluorescence in situ hybridization (FISH) is a powerful molecular cytogenetic technique using chromosomal markers that reveal the positions of specific genes, such as ribosomal DNAs, thereby making it easy to identify individual chromosomes. We have constructed detailed karyotypes of four different local and wild varieties of radish, based on the chromosome arm length and fluorescence in situ hybridization (FISH) with the 45S rDNA and 5S rDNA as probes. As for the karyotype of radish, 9 pairs of chromosomes were extremely small in size with about 1 to 3 um in length at mitotic metaphase having metacentrics or submetacentrics. Three pairs of 45S rDNA signals and two pairs of 5S rDNA signals were observed in four radish species. One pair of 45S rDNA signal was located on terminal region of short arm chromosome, while two pairs were in interstitial region. Two pairs of 5S rDNA signals were located on interstitial region of chromosome. In conclusion, it was feasible to identify the radish by karyotype and physical mapping analyzed using ribosomal DNA.
헐떡이풀은 다년생 초본으로 중국, 일본, 대만 그리고 한국에 분포한다. 특히 우리나라에서는 울릉도에서만 분포하는데, 천식 치료, 타박상 그리고 청각장애의 치료에 사용된다. 약용작물로써의 높은 가치에도 불구하고 염색체 수를 제외한 다른 세포유전학적인 연구가 거의 이루어지지 않았다. 따라서 핵형분석 뿐만 아니라 bicolor FISH를 통한 5S 와 45S rDNA의 물리적 지도작성에 관한 연구가 수행되었다. 체세포 염색체 수는 2n=2x=14로 염색체의 길이는 1.66~3.50μm 이다. 또한 염색체의 구성은 4쌍의 차중부 염색체(염색체 1, 2, 3, 6)와 2쌍의 차단부 염색체(염색체 5, 7)그리고 1쌍의 단부 염색체(염색체 4)로 확인되었다. 또한 4번 염색체가 부수체 염색체로 관찰되었다. Bicolor-FISH를 통해 각각 1쌍의 5S와 45S rDNA 위치를 확인하였는데, 5S rDNA의 경우 염색체 3번의 동원체 부위에서 확인되었고, 45S rDNA는 염색체 4번의 단완 말단 부위에서 관찰되었다. Bicolor-FISH는 헐떡이풀 염색체상에 rDNA 유전자의 위치 확인에 매우 유용한 정보를 제공하는 기술로 사용되었다.