Identification of chromosome and caryotype analysis in citrus by GISH method and in situ PCR method.

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Although citrus chromosomes, 2n=18, are classified to 5 types (A〜E) by the banding patterns of CMA staining, it is difficult to identify a chromosome in the same type. In this research, CMA staining and GISH method were performed for an identification of chromosome in the same type, and a caryotype analysis of hybrid seedlings from 'Tosa-Buntan' x 'Suisho-Buntan' pummelo was performed to clarify the chromosome paring set of 'Tosa-Buntan'1. Thirteen kinds of chromosome composition were manifested by 38 diploid seedlings from 'Tosa-Buntan' x 'Suisho-Buntan'. The caryotype analysis of the diploid seedlings indicated that the chromosome parting set of 'Tosa-Buntan' would by AB+CC+CC+CD+DE+EE+EE+EE+EE or AB+CC+CC+DD+CE+EE+EE+EE+EE. The caryotype analysis of the tri-and tetraploid seedlings showed that the triploid and tetraploid seedlings from diploid crossing also originated from an unreduced male gamete as well as from an unreduced male gamete.2. By using the relative size of CMA(+) regions as indices, type-C chromosomes were able to classify into two groups in 'Tanikawa-Buntan' and sour orange, and type-D chromosomes were able to classify into three groups in 'Yoshida-Ponkan' and sour orange, and into two groups in 'Mukaku-Kishu'.3. In GISH method, 9 chromosomes of 'Tosa-Buntan' in type-A to E were able to classify into 8 groups depended on coloration and size of the signals by using satsuma mandarin DNA as a probe. Nine chromosomes of 'Suisho-Buntan' in type-A to E were able to classify into 7 groups depended on coloration and size of the signals by using 'Dancan' grapefruit DNA as a probe.4. The GISH for 'Tosa-Buntan' chromosome using several kinds of probe DNA showed that 'Tosa-Buntan' is very closed to 'Suisho-Buntan', relative close to citron and relative far from mandarin and orange. The reciprocal GISH between 'Tosa-Buntan' and navel orange suggested that the same type chromosome between these species may not be homoeologous chromosome.