Interspecific hybrids between <Emphasis Type="Italic">Chrysanthemum grandiflorum</Emphasis> (Ramat.) Kitamura and <Emphasis Type="Italic">C. indicum</Emphasis> (L.) Des Moul. and their drought tolerance evaluation

Chrysanthemum grandiflorum ‘Yuhuaxingchen’ is an important commercial chrysanthemum cultivar with excellent ornamental quality but low drought tolerance, whereas C. indicum has exceptional drought tolerance. In our earlier study, many hybrid seeds between them were obtained through interspecific hybridization. In the present study, we selected six putative hybrid lines with most drought tolerance from all the hybrid lines by withholding water, indentified their facticity by chromosome counting, and then evaluated their drought tolerance through determining foliar electrolyte leakage (EL), contents of malondialdehyde (MDA) and proline, and plant survival rate after 20% polyethylene glycol 6000 treatment. It was found that 155 out of 282 seeds germinated and only 132 seedlings survived. In addition, chromosome and morphological analysis showed that the six putative hybrids were real hybrids and their morphological features were intermediate between their parents. Furthermore, the density of leaf epidermal hair, proline content, and plant survival rate were the highest in C. indicum and the lowest in C. grandiflorum among the six hybrids and their parents. In contrast, EL value and MDA content were the highest in C. grandiflorum and the lowest in C. indicum. These results suggest that some true hybrids with improved drought tolerance can be obtained through interspecific hybridization in chrysanthemum breeding. Therefore, interspecific hybridization between chrysanthemum cultivars and their wild species may become a promising way to improve their biotic and abiotic resistance in the future breeding.     

Male fertility and occurrence of 2n gametes in East African Highland bananas (Musa spp.)

The quantity and quality of pollen produced by a genotype are important attributes to be considered in the selection of male parents for breeding. The objective of this research was to determine the quantity and quality of pollen in 13 selected East African Highland bananas (EAHB) in relation to six diploids commonly used as male parents. Pollen quantity was scored on a scale of 0–6 while the variation of pollen stainability along the rachis was determined using acetocarmine glycerol jelly. Actual male fertility was determined by using pollen from the EAHB varieties on M. acuminata spp. burmannicoides, ‘Calcutta 4’, as the female. The diameter of 200 viable pollen grains was measured under a microscope with the aid of a graduated eyepiece. Significant differences were obtained for mean pollen stainability and pollen diameter. The pollen stainability for diploids was higher than that for the EAHB. Node numbers contributed significantly to variation of pollen stainability in EAHB (P ≤ 0.01) and ‘Pisang lilin’ (P ≤ 0.001). ‘Pisang lilin’ and the EAHB varieties had about 100 nodes as opposed to about 200 in other diploids. The EAHB varieties produced less pollen than most of the diploids. The actual male fertility of EAHB varieties was low (1 seed/bunch), compared to that of ‘Calcutta 4’ (223.3 seeds/bunch) and ‘Pisang lilin’ (13.7 seeds/bunch). Although the production of 2n pollen in EAHB varieties was low (0–7%), the potential exists for using them in new breeding approaches.     

High regeneration potential in vitro of sunflower (Helianthus annuus L.) lines derived from interspecific hybridization

Successful selection of interspecific hybrid progenies with superior ability to regenerate shoots from apical meristems was performed in sunflower which now allows for the development of lines for improved biotechnological applications. Early generations of interspecific hybrids originating from crosses between the two H. annuus CMS lines ‘HA89’ and ‘Baso’, and 9 wild species were screened for their ability to regenerate in vitro. Evaluation of 36 progenies allowed to identify seven progenies from crosses involving H. mollis, H. giganteus, H. strumosus, and H. decapetalus which showed a significantly higher regeneration potential than the commercial hybrid ‘Albena’ regarding the number of shoots per explant. Among these progenies, 47.2 to 62.4% of explants produced shoots with an average of 2.3 to 3.5 shoots per cultured explant. Regeneration in vitro was significantly determined by the genotype. More than half of the investigated interspecific hybrids performed better than the inbred ‘HA89’ demonstrating that the high regeneration potential available in the wild species can be efficiently transferred to cultivated sunflower. The seven progenies with high regeneration potential in vitro were characterised by agronomic performance in the field. Two of the interspecific hybrids derived from H. strumosus and H. decapetalus not only showed a superior regeneration potential but also proved to be competitive to commercial hybrids with regard to important agronomic traits, e.g. fat content and TGW. This revised version was published online in July 2006 with corrections to the Cover Date.     

Identification of self-incompatibility alleles in pear cultivars (Pyrus communis L.)

Self and cross-incompatibility determination by means of fruit and seed set experiments or pollen tube growth observations in the style has been frequently reported to be unclear in pear (Pyrus communis L.). Thus,in order to develop a reliable in vivo method to test pollen-pistil incompatibility in pear, pollen tube performance has been studied along the pistil following self and cross-pollinations. Results show that, while pollen tube growth in the style may be an unclear test, ovule observation at the microscope for the presence of pollen tube in the nucellus is a proper method to test incompatibility in this crop. With this analysis we could identify S-alleles of ‘Williams’ (S₁S₂) and ‘Coscia’(S₃S₄), and three of the four possible S-genotypes resulting from the ‘Williams’ × ‘Coscia’ cross, as represented by ‘Butirra Precoz Morettini’ (S₁S₃), ‘Santa Maria Morettini’ (S₂S₃)and ‘Tosca’ (S₁S₄). This result demonstrates that ‘Williams’ and ‘Coscia’ cultivars do not share any allele in common. We also established two new inter-incompatibility groups in pear. Furthermore, the presence of a common allele between ‘Williams’ and ‘Agua de Aranjuez’,and ‘Coscia’ and ‘Agua de Aranjuez’, three apparently unrelated old cultivars, may indicate a narrower genetic base than expected for European pear. This finding together with the fact that 40% of new released cultivars have direct or indirect parental relationship with the cultivars ‘Coscia’ and/or ‘Williams’, anticipates the possibility of new cases of cross-incompatibility for this crop in the future. Both the method described and the determination of the S-genotypes will facilitate the characterisation of self and cross-incompatibility relationships in this species. This revised version was published online in August 2006 with corrections to the Cover Date.     

Creating novel chrysanthemum germplasm via interspecific hybridization and backcrossing

The interspecific cross between Chrysanthemum × grandiflorum (Ramat.) Tzvel. ‘rm20-12’ (R, 2n = 54) and C. makinoi Matsum., and Nakai (M, 2n = 18) was achieved using embryo rescue, and a single backcross progeny using C. × grandiflorum ‘rm20-12’ as paternal parent was obtained. The morphology of the two independent F₁ hybrids (MR1 and MR2) differed from that of both parents. MR1 had a larger inflorescence diameter along with narrow leaves and a reduced number of ray and tubular florets. MR2 was shorter and its inflorescence developed fewer tubular florets than either M or R. The BC₁F₁ hybrid was similar to its maternal plant MR2 in terms of leaf length and width, inflorescence diameter and the number of ray florets, while it produced fewer tubular florets than either MR2 or R. The flower color in both F₁ hybrids was lavender, while the BC₁F₁ plant bore purple flowers. The aphid resistance and heat tolerance of MR1, MR2 and the BC₁F₁ hybrid were both significantly superior to that of C. × grandiflorum ‘rm20-12’. Interspecific hybridization followed by backcrossing shows clear potential for cultivar improvement in chrysanthemum.     

Reproductive barriers in the intergeneric hybridization between <Emphasis Type="Italic">Chrysanthemum grandiflorum</Emphasis> (Ramat.) Kitam. and <Emphasis Type="Italic">Ajania przewalskii</Emphasis> Poljak. (Asteraceae)

In order to reveal the causes of low seed set in the intergeneric cross between Chrysanthemum grandiflorum (Ramat.) Kitam. ‘Zhongshanjingui’ (female parent) and Ajania przewalskii Poljak. (male parent), we systematically investigated megasporogenesis and megagametogenesis of female parents, pollen germinability of male parents, pollen–pistil interaction and hybrid embryo development using paraffin section and light, fluorescence and scanning electron microscopy. The results indicate that development of megagametophyte of female parent belongs to Polygonum type and 84.6% ovules developed to normal 8-nucleate embryo sac at the stage of pollination; the pollen germinability in vitro of male parent was 39.3%; a large number of pollen grains germinated normally and no callose deposited on the female stigmas within 24 h after pollination. Furthermore, about 75% ovules were fertilized, whereas only 17.2% ovules developed to normal maturing embryos at 15 days after pollination due to abortion at various developmental stages. Taken together, these results suggest that sterile ovules of female parent, pollen viability and pollen–pistil interaction have no significant influence on seed set in the cross. A high percentage of embryo abortion or degeneration, i.e. post-fertilization barrier, may be the main factor for low seed yield.     

Characterization of segregation distortion on chromosome 3 induced in wide hybridization between indica and japonica type rice varieties

We previously surveyed chromosomal regions showing segregation distortion of RFLP markers in the F₂ population from the cross between a japonica type variety ‘Nipponbare’ and an indica type variety ‘Milyang23’, and showed that the most skewed segregation appeared on the short arm of chromosome 3. By comparison with the marker loci where distortion factors were previously identified, this region was assumed to be a gametophytic selection-2 (ga2) gene region. To evaluate this region, two near isogenic lines (NILs) were developed. One NIL had the ‘Nipponbare’ segment of this region on the genetic background of ‘Milyang23’ (NIL9-23), and the other NIL had the ‘Milyang23’ segment on the genetic background of ‘Nipponbare’ (NIL33-18). NIL9-23 and ‘Milyang23’, NIL33-18 and ‘Nipponbare’, and ‘Nipponbare’ and ‘Milyang23’ were respectively crossed to produce F₁ and F₂ populations. The F₁ plants of NIL9-23 × ‘Milyang23’ and NIL33-18 × ‘Nipponbare’ showed high seed fertility and the same pollen fertility as their parental cultivars, indicating that ga2 does not reduce seed and pollen fertility. Segregation ratio of a molecular marker on the ga2 region in the three F₂ populations was investigated to clarify whether segregation distortion occurred on the different genetic backgrounds. Segregation distortion of the ga2 region appeared in the both F₂ populations from the NIL9-23 and ‘Milyang23’ cross (background was ‘Milyang23’ homozygote) and the ‘Nipponbare’ and ‘Milyang23’ cross (background was heterozygote), but did notin the F₂ population from the NIL33-18 and ‘Nipponbare’ cross (background was ‘Nipponbare’ homozygote). This result indicates that ga2 interacts with a ‘Milyang23’ allele(s) on the different chromosomal region(s) to cause skewed segregation of the ga2 region. In addition, segregation ratio was the same between the F₂ populations from NIL9-23 × ‘Milyang23’ and ‘Nipponbare’ × ‘Milyang23’ crosses, suggesting that the both genotypes, ‘Milyang23’ homozygote and heterozygote, of gene(s) located on the different chromosomal region(s) have the same effect on the segregation distortion. This revised version was published online in July 2006 with corrections to the Cover Date.     

Evaluation of leaf rust resistance genes Lr1 , Lr9 , Lr24 , Lr47 and their introgression into common wheat cultivars by marker-assisted selection

Epidemiological field controls in different Italian locations and seedling evaluations of the ‘Thatcher’ near-isogenic lines (NILs) carrying the leaf rust resistance genes Lr1, Lr9, Lr24 and Lr47 were conducted during 5 years of testing. These genes confirmed their effectiveness in both field and greenhouse conditions. Moreover a backcross program was carried out by using as recurrent parents the susceptible high-quality common wheat cvs ‘Bolero’, ‘Colfiorito’, ‘Serio’ and ‘Spada’ and the ‘Thatcher’ NILs carrying the above mentioned genes as donor parents. The progenies of different cross combinations were selected by both resistance tests and marker assisted selection using molecular markers (STS, SCAR, CAPS) closely linked to Lr genes: a complete cosegregation was observed between the resistance genes used and the corresponding molecular markers.     

Overcoming pre-fertilization barriers in the wide cross between Chrysanthemumgrandiflorum (Ramat.) Kitamura and C. nankingense (Nakai) Tzvel. by using special pollination techniques

In our earlier study, pre-fertilization barrier was one of the main factors causing the failure of the wide hybridization between Chrysanthemum grandiflorum ‘Yuhuaxingchen’ and C. nankingense, and seriously restricted the utilization of the excellent chrysanthemum germplasm, C. nankingense. In order to overcome the pre-fertilization barrier and obtain some hybrids, we carried out wide hybridization between them again in this study, and tentatively adopted three pollination techniques including mentor pollen, delayed pollination, and gibberellic acid treatment. The average number of pollen grains germinating on stigma at 24 h after pollination and the percentage of embryo were investigated with a temporary mount method and the technique of fluorescence microscopy. It was found that, compared with the control, delayed pollination, gibberellic acid treatment, and mentor pollen significantly increased the average number of pollen grains germinating on each stigma at 24 h after pollination by approximately 152, 235, and 182%, respectively. In addition, all the three pollination techniques significantly increased the percentage of normal embryos at different periods after artificial pollination in comparison to the control. Furthermore, seed sets of the crosses using the three special pollination techniques were 1.1, 1.4, and 2.0%, respectively, whereas we did not obtained any seeds in the control. Taken together, these results suggest that delayed pollination, gibberellic acid treatment, and mentor pollen are three useful pollination techniques for overcoming pre-fertilization barriers and increasing seed set in the wide hybridization between C. grandiflorum ‘Yuhuaxingchen’ and C. nankingense, and they may be widely applied for increasing breeding efficiency in other chrysanthemum wide hybridization facing similar pre-fertilization barriers.     

Confirmation and characterization of interspecific hybrids of <Emphasis Type="Italic">Passiflora</Emphasis> L. (Passifloraceae) for ornamental use

Three new varieties of Passiflora hybrids were developed from crosses between P. sublanceolata J. M. MacDougal (ex P. palmeri var. sublanceolata Killip) versus P. foetida var. foetida L. Twenty putative hybrids were analyzed. Hybridizations were confirmed by RAPD and SSR markers. The RAPD primer UBC11 (5′-CCGGCCTTAC-3′) generated informative bands. The SSR primer A08FP1 amplified species-specific fragments and heterozygote status was observed with the two parent bands 240 and 280 bp. The molecular markers generated by primers were analyzed in terms of the presence or absence of specific informative bands. The morphological characterization of the hybrids enabled their differentiation into three groups, identified as: (1) Passiflora ‘Alva’, composed of five hybrid plants with white flowers, large corona, light purple filaments at base, white and purple/white banding to apex; (2) P. ‘Aninha’, composed of six hybrid plants with pale pink flowers, corona filaments reddish/purple at base, white, purple/white banding to apex; (3) P. ‘Priscilla’, composed of nine hybrid plants with white flowers, small corona, filaments dark purple at base, white and purple to apex. The genomic homology of parent plants was verified by cytogenetic analysis. Both parents were 2n = 22. Meiosis was regular in genitors and hybrids. Aneuploidy was observed at hybrid groups P. ‘Alva’ and P. ‘Priscilla’ (2n = 20). Other authors had already observed the same number of chromosomes for some P. foetida genotypes. Obtaining valuable interspecific hybrids opens up new perspectives to offer opportunities in agribusiness for producers and to arouse the interest of consumers into using passion flowers in the Brazilian ornamental plant market.     

Chromosomal regions associated with green plant regeneration in wheat (Triticum aestivum L.) anther culture

Genetic capacity for green plant regeneration in anther culture were mapped in a population comprising 50 doubled haploid lines from a cross between two wheat varieties ‘Ciano’ and ‘Walter’ with widely different capacity for green plant regeneration. Bulked segregant analysis with AFLP markers and composite interval mapping detected four QTLs for green plant percentage on chromosomes 2AL (QGpp.kvl-2A), 2BL (QGpp.kvl-2B.1 and QGpp.kvl-2B.2) and 5BL (QGpp.kvl-5B).The three QTLs detected on chromosome 2AL and 2BL all derived their alleles favouring green plant formation from the responsive parent ‘Ciano’.The remaining QTL on chromosome 5BL had the allele favouring green plants from the low responding parent ‘Walter’. In a multiple regression analysis the four QTLs could explain a total of 80% of the genotypic variation for green plant percentage. None of the chromosomal regions with QTLs for green plant percentage showed significant influence on either embryo formation or regeneration frequencies from the anther culture. The three major QTLs located on group two chromosomes were fixed in a second DH population derived from two parents ‘Ciano’ and ‘Benoist’,both with high capacity to produce green plants. A QTL explaining31.5% of the genetic variation for green plant formation were detected on chromosome 5BL in this cross as well. This revised version was published online in July 2006 with corrections to the Cover Date.     

Effect of maternal genotype on development ofPhaseolus vulgaris L. ×P. lunatus L. interspecific hybrid embryos

Summary CertainPhaseolus vulgaris L. ×P. lunatus L. crosses were performed to study the effect of maternal heterozygosity on development and growth of the interspecific hybrid embryos. Interspecific embryos had a much slower growth rate in vitro compared with embryos derived from self-pollination ofP. vulgaris parents. Thus, interspecific embryos could be identified by growth rate in vitro. TheP. vulgaris maternal genotype affected both the number and size of 15-day-old interspecific embryos. Specifically, 76 Spartan Arrow produced significantly more interspecific embryos than did Great Northern as the seed parent, while maternal intraspecific hybrids produced smaller embryos than did maternal pure lines. There were no reciprocal differences between hybrid maternal parents for embryo number or size. Embryo size at excision and final size after culturing were closely correlated (r²=+0.93). The crossP. vulgaris 76 Spartan Arrow ×P. lunatus P.I. 214170 produced both the largest mean size at excision and the fastest growth in culture, indicating that specific combining ability affected both characteristics.Journal Paper No. J-12208 of the Iowa Agriculture and Home Economics Experiment Station, Ames, Iowa. Project No. 2495     

Somatic hybrids between navel orange (Citrus sinensis) and grapefruit (C. paradisi) for seedless triploid breeding

Protoplasts from cell suspension cultures of ‘Bonnaza’ navel orange (Citrus sinensis (L.) Osbeck) were electrically fused with mesophyll protoplasts isolated from seedless ‘Red Blush’ grapefruit (Citrusparadisi). After 6 months of culture, a total of 20 plants were regenerated. Root tip chromosome counting revealed that 4 of them were tetraploids (2n = 4x = 36)and the rest were diploids (2n = 2x = 18) morphologically resembling the mesophyll parent. After 6 months of transplantation into the greenhouse, 4 of the diploidmesophyll regenerants unexpectedly flowered, but this phenomenon disappeared in the next year. This is the first report of precocious flowering in citrus via protoplast fusion. RAPD analysis further confirmed that the tetraploid regenerants were somatic hybrids while the diploid regenerants were mesophyll parent type. This somatic hybrid will be utilized as a possible pollen parent for improving the seedy pummelo cultivars in China by producing triploid seedless pummelo hybrid. The mechanism of early flowering was also discussed. This revised version was published online in July 2006 with corrections to the Cover Date.     

Zingiberene-mediated resistance to the South American tomato pinworm derived from Lycopersicon hirsutum var. hirsutum

The Lycopersicon hirsutum var. hirsutum accession PI 127826 is recognized as a good source of resistance to arthropod pests due to the action of the allelochemical zimgiberene, a sesquiterpene present in its glandular trichomes. Five genotypes were selected from the F₂ generation of the interspecific cross Lycopersicon esculentum ‘TOM-556’ × Lycopersicon hirsutum var. hirsutum ‘PI 127826’, based on their low levels (BPX-368-clone#56) or high levels(BPX-368-clone#92, BPX-368-clone#105,BPX-368-clone#179, BPX-368-clone#250) of zingiberene. The five F₂ genotypes were tested for resistance to the South American tomato pinworm Tuta absolutaalong with accession L. esculentum ‘TOM-556’ (pinworm susceptible), and the accessions L. hirsutum var. hirsutum ‘PI 127826’ and L. pennellii ‘LA716’ (resistant). The F₂ clones selected for high foliar zingiberene levels showed lower scores for leaflet lesion type(LLT), percent leaflets attacked (PLA) and overall plant damage (OPD) than the low zingiberene genotypes. The results indicated that zingiberene mediates resistance to the South American pinworm, based on feeding and on ovipositing deterrence, in populations derived from the interspecific cross between Lycopersicon. esculentum and Lycopersicon hirsutum var. hirsutum. Indirect selection for high foliar zingiberene content is suggested as an efficient technique for breeding tomatoes for resistance to the South American tomato pinworm. This revised version was published online in July 2006 with corrections to the Cover Date.     

Analysis of introgression of the Tulipa fosteriana genome into Tulipa gesneriana using GISH and FISH

Southern hybridization and genomic in situ hybridization (GISH) have demonstrated that ‘Purissima’ (2n = 2x = 24) is an interspecific hybrid comprised of one genome of Tulipa (T.) gesneriana and one genome of T. fosteriana. Backcrossing T. gesneriana with ‘Purissima’ was partially successful. Simultaneous GISH and fluorescence in situ hybridization (FISH) distinguished chromosomes from both parent genomes, as well as recombinant chromosomes, in interspecific hybrids and their progeny. Chromosome recombination was observed in all cultivars except ‘Purissima’ and ‘Kouki’ (2n = 3x = 36). ‘Kouki’ (2n = 3x = 36) had two genomes of the T. gesneriana and a single genome of the T. fosteriana. The number of nonrecombinant T. fosteriana chromosomes in ‘Judith Leyster’ (2n = 4x = 48) and ‘Purissima’ progeny varied from two in ‘Hatsuzakura’ to six in ‘Kikomachi’ and ‘Momotaro’. The number and type of recombinant chromosomes also differed among cultivars. The total number of translocations ranged from one in ‘Kikomachi’ to six in ‘Hatsuzakura’. Each was a combination of a single T. fosteriana fragment and a single T. gesneriana fragment, indicating that they resulted from a single crossover event. Sequential GISH and FISH analysis with rDNA probes yielded chromosome-specific markers that were used to identify most of the chromosomes in ‘Purissima’ progeny. This is the first report of introgression of T. fosteriana chromatin into the T. gesneriana genome.     

Vitamin C,flower color and ploidy variation of hybrids from a ploidy-unbalanced <Emphasis Type="Italic">Actinidia</Emphasis> interspecific cross and SSR characterization

Seedlings derived from an Actinidia interspecific cross between the hexaploid Actinidia chinensis var. deliciosa ‘Jinkui’ and the diploid male A. eriantha × A. chinensis var. chinensis ‘Chaohong’ hybrid were analyzed using flow cytometry, SSR markers and phenotypic observations. The results show that the leaf vitamin C content of this hybrid population has a mid-parent heterosis. Separation of flower color in the progeny was also observed, progeny with red flowers lighter than ‘Chaohong’, white flowers as in ‘Jinkui’ and intermediate types with a red base to the petals and white margins were all present. Flow cytometry analysis confirmed that most of the progeny were tetraploids, and molecular marker data showed that most of these tetraploid progeny had three alleles from the hexaploid parent and one allele from the diploid parent. UPGMA analysis based on the SSR markers showed that the diploid parent was completely separated from the hexaploid parent and all the progeny.     

Introgression of unique characteristics of floral initiation under 24 hour day-length of Fragaria chiloensis ‘CHI-24-1’ into F. × ananassa

A unique flower initiation of the wild Junebearing strawberry strain ‘CHI-24-1’ in Fragaria chiloensis occurs under 24 h DL and high temperature conditions. To introduce the floral initiation characteristics of ‘CHI-24-1’ into cultivated strawberries of F. × ananassa, a cross pollination was conducted between ‘CHI-24-1’ and the Japanese short-day type strawberry cultivar ‘Nyoho’. The floral initiation of ‘CHI-24-1’ was induced in both parent and daughter plants linked with runners under a 24 h DL and 23/20 ^∘C, but not 8 and 16 h DLs at the same temperatures. Of the 21 F1 hybrids grown under the 24 h DL, 12 showed flower truss production in the parent and/or daughter plants linked with runners. Among 64 F1 hybrids, 26 exposed to 8, 16 and 24 h DLs for 30 days produced no flower trusses. However, 32, one and five F1 hybrids produced flower trusses under the 24 h DL alone, 8 h DL alone and both the 8 and 24 h DLs, respectively. The results of the experiments indicated that none of the F1 hybrids were day-neutral plants, but approximately 60% had the characteristics of floral initiation under 24 h DL, which was inherited from the pollen parent of ‘CHI-24-1’. The importance of the unique floral initiation characteristics under 24 h DL with high temperature for strawberry breeding was discussed.     

Occurrence of viable unreduced pollen in a <Emphasis Type="Italic">Begonia</Emphasis> collection

Polyploidy is widespread in plants and has played a major role in the evolution and diversification of the plant kingdom. Unreduced (2n) gametes are an interesting tool for polyploidisation and the creation of genetic variation in plant breeding. Especially in ornamentals, polyploidisation can broaden attractive features within a species. A Begonia collection was screened on the occurrence of 2n pollen with the aid of four different techniques: pollen size measurement, flow cytometric analysis of nuclei isolated from germinated and non germinated pollen, investigation of the microsporogenesis and analysis of progeny. In ten of the 70 screened genotypes (B. dregei, B. pearcei, B. ‘Anna Christina’, B. ‘Bubbles’, B. ‘Florence Rita’, B. ‘Orococo’, B. ‘Rubaiyat’, B. ‘Spatflacier’, B. ‘Tamo’ and B276), large pollen were observed with a rather spherical than normal ellipsoidal shape. Flow cytometric data proved that these aberrantly shaped pollen were associated with 2n ploidy levels, although they were not always viable. Meiotic aberrations in these large pollen producers resulted mainly in dyads although also monads, triads and polyads were observed. Successful crosses were obtained with B. dregei, B. ‘Orococo’, and B276 as pollinators; DNA content had increased in all or a part of the progeny. The results show that the occurrence of 2n pollen is not a rare phenomenon in Begonia.     

Interspecific hybridization between urdbean (Vigna mungo (L.) Hepper) and adzukibean (Vigna angularis (Willd.) Ohwi and Ohashi)

Summary To introduce resistance to fungal and viral diseases into urdbean (Vigna mungo), it was crossed with adzukibean (Vigna angularis) to achieve an interspecific hybrid between the two species by using different non-conventional breeding methods. Partial success was obtained with the use of UV irradiated pollen, intraspecific hybrids as parents and spray of immuno-suppressants on the female parent under glass house conditions and to the extent of callusing in aseptic culture.     

Intergeneric hybridization and relationship of genera within the tribe Anthemideae Cass. (I. <Emphasis Type="Italic">Dendranthema crassum</Emphasis> (kitam.) kitam. × <Emphasis Type="Italic">Crossostephium chinense</Emphasis> (L.) Makino)

An intergeneric cross has been made between Dendranthema crassum (kitam.) kitam. (2n = 90; ♀) and Crossostephium chinense (L.) Makino (2n = 18; ♂). Most of the hybrid embryos aborted at an early developmental stage. Using ovule rescue, it was possible to establish a single intergeneric hybrid plant showing 2n = 54 chromosomes. The leaf length, leaf width and epidermal hair density of the hybrid were all intermediate between those of the parents. However the flower diameter, number of tubular florets, epidermal hair height and epidermal hair length exceeded those of both parents. A genomic in situ hybridization approach was able to distinguish between the parental genomes in the hybrid plant.