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      Inter-sectional hybrids obtained from reciprocal crosses between Begonia semperflorens (section Begonia) and B. ‘Orange Rubra’ (section Gaerdita × section Pritzelia)

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          Abstract

          Inter-sectional hybrids were successfully obtained by the reciprocal crosses between 11 cultivars (including 6 diploids and 5 tetraploids) of Begonia semperflorens (SS & SSSS genomes) and B. ‘Orange Rubra’ (RR genome) with the aid of in vitro culture of mature or immature seeds on MS medium containing 0.1 mg l −1 α-naphthylacetic acid, 0.1 mg l −1 6-benzyladenine, 10 mg l −1 gibberellic acid, 30 g l −1 sucrose and 2.5 g l −1 gellan gum. Embryo rescue as ovary culture with immature seeds 12 th–16 th day after pollination (DAP) generally gave higher efficiency of plantlet formation, but in some cross combinations, culture of mature seeds (30 DAP) resulted in higher yield of plantlets. Flow cytometric analysis revealed that they were consisted of the plants with various genomic combinations (RS, RR, RSS, RRS, RRSS and RRRRSS) as estimated by the DNA contents of both parents. Hybridity of these plants with various genomic combinations including RR was confirmed by random amplified polymorphic DNA analysis. These results suggested that unreduced gamete formation and spontaneous chromosome doubling were involved in the hybrid formation of various ploidy levels and genomic combinations. These hybrids showed various levels of intermediate traits between both parents according to the genomic compositions, and some of them had desirable characters of both parents.

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          Rapid isolation of high molecular weight plant DNA.

          A method is presented for the rapid isolation of high molecular weight plant DNA (50,000 base pairs or more in length) which is free of contaminants which interfere with complete digestion by restriction endonucleases. The procedure yields total cellular DNA (i.e. nuclear, chloroplast, and mitochondrial DNA). The technique is ideal for the rapid isolation of small amounts of DNA from many different species and is also useful for large scale isolations.
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            Unilateral compatibility and genotypic difference in crossability in interspecific hybridization between Dianthus caryophyllus L. and Dianthus japonicus Thunb.

            Reciprocal interspecific crosses were carried out between six lines of Dianthus caryophyllus L. and one line of Dianthus japonicus Thunb. Although no seed was set when D. japonicus was used as the seed parent, six seedlings were successfully obtained from 2,380 immature ovules by applying the embryo-rescue technique. However, they showed seed parent-like morphology and no evidence for the hybridity by flow cytometry and RAPD analyses. When six lines of D. caryophyllus were used as seed parents, a total of 192 seedlings were successfully obtained without using the embryo-rescue technique. Among these seedlings, 12 out of 25 progenies obtained from the carnation line '98sp1651' were confirmed to be the hybrids. The remaining 13 progenies of this line, and the total 167 progenies obtained from the other carnation lines, had carnation-like morphology without any evidence of hybridity by flow cytometry and RAPD analyses. The progenies confirmed as hybrids had intermediate characters of the parents with respect to leaf width and flower size, but they had a uniform flower color, reddish purple, which was different from that of either parent. Since the hybrids obtained in the present study have some profitable characters such as vigorous growth in summer time, upright robust stem, broad leaves and early flowering, they are expected to be used for the breeding of carnation which is suitable for growing under the Japanese climate.
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              Meiotic aberrations during 2n pollen formation in Begonia.

              Unreduced gametes are the driving force for the polyploidization of plants in nature, and are also an important tool for ploidy breeding. The final heterozygosity of a 2n pollen grain depends on the cytological mechanism behind 2n pollen formation. In this study, chromosome pairing and chromosome segregation during the microsporogenesis of seven Begonia genotypes were analysed using fluorescent chromosome staining on (squashed) pollen mother cells. Among the seven genotypes, five genotypes produce 2n pollen (B. 'Bubbles', B. 'Florence Rita', B. 'Orococo', B. 'Tamo' and B276) and two genotypes produce only normal n pollen (B. fischeri and B243). All 2n pollen producers showed a mechanism equivalent to first division restitution (FDR), in which chromosomes did not segregate during meiosis I but only during meiosis II. This FDR was the result of (a) an irregular chromosome pairing in B. 'Tamo', (b) stickiness of chromosomes associated with numerous chromosome bridges in B. 'Florence Rita' and B276, and (c) a combination of irregular chromosome pairing and stickiness of chromosomes in B. 'Bubbles'. The exact mechanism of the nuclear restitution in B. 'Orococo' could not be determined. Other mechanisms, such as early asymmetric cytokinesis, omission of meiosis II, parallel or tripolar spindle formation, were rather uncommon. Unpaired chromosomes (univalents) were observed in all genotypes, but they had moved to one of the poles by the end of anaphase I or II. Only B. 'Tamo' formed a high number of micronuclei. Consequently, this genotype formed a large number of malformed pollen. Obviously, chromosome behaviour during meiosis in Begonia is very dynamic, which may have important consequences for chromosome evolution and biodiversity within the genus.
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                Author and article information

                Journal
                Breed Sci
                Breed. Sci
                BS
                Breeding Science
                Japanese Society of Breeding
                1344-7610
                1347-3735
                19 June 2012
                June 2012
                : 62
                : 2
                : 113-123
                Affiliations
                Laboratory of Plant Cell Technology, Faculty of Horticulture, Chiba University, 648 Matsudo, Matsudo, Chiba 271-8510, Japan
                Author notes
                [* ]Corresponding author (e-mail: miim@ 123456faculty.chiba-u.jp )

                Communicated by T. Sato

                Article
                bs-62-113
                10.1270/jsbbs.62.113
                3405972
                23136522
                208dc2a4-ec07-4ff5-984e-ac1c263b97ae
                Copyright © 2012 by JAPANESE SOCIETY OF BREEDING

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

                History
                : 06 December 2011
                : 02 February 2012
                Categories
                Research Papers

                Animal agriculture
                embryo rescue,intersectional hybrid,unreduced gamete,begonia semperflorens,amphipolyploid,begonia ‘orange rubra’

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