Comparative cytological study of Solanum aethiopicum Gilo group,Solanum aethiopicum Shum group and Solanum anguivi

Summary Comparative cytological studies were carried out on Gilo and Shum cultivar groups of Solanum aethiopicum (scarlet eggplant) and their ancestor-Solanum anguivi, in order to elucidate the evolutionary relationships between them. Results indicated preferential pairing of chromosomes in S. anguivi and non-preferential pairing of chromosomes in Gilo and Shum cultivar groups of S. aethiopicum during early stages of meiosis. The low degree of pairing in Gilo and Shum cultivar groups of S. aethiopicum and higher percentage of chromosomal abnormalities (12.50 and 10.58% respectively showed cases of translocation heterozygotes and suggest that the two taxa have hybrid origin. Conversely, the high degree of pairing and very low percentage of chromosomal abnormalities (2.50%) tend to suggest that S. anguivi is more ancient than either the Gilo and Shum cultivar groups of S. aethiopicum. It had already been proved that the Gilo and Shum cultivar groups of S. aethiopicum together with the Kumba and Aculeatum groups originated from S. anguivi (Lester & Niakan, 1986). Also studies on external morphology and cytological studies have shown the Gilo cultivar group to be more complex morphologically and having higher percentage of chromosomal aberrations (12.50% as against 10.58% observed in Shum cultivar group). It is suggested that the Gilo cultivar group might have evolved from the Shum cultivar group through hybridization and selection, while the S. anguivi was already stabilized as their ancestor.     

Inheritance in Solanum aethiopicum,the scarlet eggplant

Summary Analyses of F₁ and F₂ hybrid progenies from crosses between the ancestral species Solanum anguivi and cultivars of its domesticated derivative S. aethiopicum, have proved the mode of inheritance of prickles, stellate hairs and some other morphological characters and suggested the mode for several more. Most wild-type traits were dominant: many recessive domesticated traits involved imperfect morphogenesis suggesting loss of genetic control. F₁ hybrids between cultivars show marked heterosis and are recommended for crop production.     

Fertility restoration of hybrids between Solanum melongena L. and S. aethiopicum L. Gilo Group by chromosome doubling and cytoplasmic effect on pollen fertility

Reproductive fertility traits were studied in the reciprocal hybrids of the eggplant(Solanum melongena L.) and S. aethiopicum L. Gilo Group, and in synthetic amphidiploids to discover whether fertility in these reciprocal hybrids was restored by chromosome doubling. Isozyme and RAPD analyses confirmed hybridity of the hybrids and amphidiploids. Analyses of chloroplast and mitochondrial DNAs confirmed that the cytoplasm of each of the hybrids and amphidiploids was from the maternal parent. Pollen sterility of S. melongena × S. aethiopicum Gilo Group [F₁ (Mel × Aet)] was restored by chromosome doubling, while the reciprocal hybrid S. aethiopicum Gilo Group ×S. melongena [F₁ (Aet × Mel)]and its amphidiploid did not produce any pollen grains; their microspores degenerated without being released from tetrads. Hence the cytoplasm of S. aethiopicum Gilo Group seems to beresponsible for their pollen-non-formation type sterility of the hybrid. Both the F₁ hybrids did not set any fruits by either selfing or backcrossing, while their amphidiploids set fruits after pollinating with pollen from the amphidiploid of F₁ (Mel × Aet). Seeds obtained from both the amphidiploids germinated normally. Chromosome doubling has been effective in restoring fertility of the hybrids. This revised version was published online in July 2006 with corrections to the Cover Date.     

Segregation of isozymes in selfed progenies of a synthetic amphidiploid between Solanum integrifolium and S. melongena

Isozyme and cytogenetic analyses were performed on selfed progenies of a synthetic amphidiploid between scarlet eggplant, Solanum integrifolium (= S. aethiopicum),and eggplant, Solanum melongena `DMP', for estimating genetic uniformity. Isozymes in the 379 examined seedlings segregated into five genotypes (phenotypes) each at the four loci examined, Pgd-2 of phosphogluconate dehydrogenase (E.C.1.1.1.43), Idh-2 of isocitrate dehydrogenase (E.C.1.1.1.41), Pgm-2 of phosphoglucomutase (E.C.2.7.5.1)and Skdh-1 of shikimate dehydrogenase (E.C.1.1.1.25), indicating that the selfed seedlings were not genetically uniform. Most of the examined 15 selfed seedlings exhibited a somatic chromosome number of 48, that is the same number of the synthetic amphidiploid, whereas isozyme genotypes among them were variable. It is suggested that the segregation of isozymes was not caused by variation of chromosome number but by genetic segregation of isozyme genes. The genome of the synthetic amphidiploid was indicated to be unstable. This revised version was published online in July 2006 with corrections to the Cover Date.     

Discrimination between Oryza malampuzhaensis Krish. et Chand. and Oryza officinalis Wall ex Watt based on RAPD markers and morphological traits

Oryza malampuzhaensis, one of the tetraploid taxa of the genusOryza, is geographically restricted to Western Ghats of South India. Its taxonomic status is not well established and is generally treated as a tetraploid race of O. officinalis. Sixty-three morphological traits and 262 random amplified polymorphic DNA (RAPD) markers generated by 23 random decamer primers were used to assess the genetic relationship between O. malampuzhaensis and O. officinalis. Pair wise comparisons based on both RAPDs and morphological traits revealed 60% genetic distance between the two taxa and was significantly higher (p <0.01) than the corresponding intra-specific distances. Cluster and principal component analysis (PCA) of genetic distance estimations based on RAPDs and morphological traits clearly differentiated the two taxa. High frequency of discrete O. malampuzhaensis specific RAPDs (21%) and the significantly higher (p < 0.05) mean number of amplification products per individual in O. malampuzhaensis observed in the study reflect its allopolyploid nature. Low genetic diversity within O. malampuzhaensis revealed by RAPD analysis indicates the recent origin of this taxa. The RAPD analysis further revealed the possibility that the putative ‘C’ genomeprogenitor of O. malampuzhaensis is a close relative of O. officinalis. In addition, amplification products diagnostic to O. malampuzhaensis were identified. The results of the present study clearly demonstrated that the O. malampuzhaensis is a distinct entity, and support the recent conclusion that O. malampuzhaensis has diverged enough to deserve species status. This revised version was published online in August 2006 with corrections to the Cover Date.     

Induction of novel organelle DNA variation and transfer of resistance to frost and Verticillium wilt in Solanum tuberosum through somatic hybridization with 1EBN S. commersonii

Somatic hybridization can be used to induce genetic variability in plastidial and mitochondrial genomes, and transfer multiple uncloned genes across sexual barriers. Somatic hybrids were produced between a dihaploid clone of the common potato, S. tuberosum subsp. tuberosum, and the wild sexually incongruent diploid species S. commersonii. Fusion products were selected on the basis of callus growth and regeneration in vitro. Genome composition of putative somatic hybrids was determined by flow cytometric analysis of nuclear DNA content, RAPD analysis, and Southern analysis with probes specific to organellar DNA. All regenerated fusion products proved to be hybrids based on RAPD analysis. Seventy per cent of somatic hybrids were (near) tetraploids, 22% (near) hexaploids and 8%(near) octoploids. A high correlation was found between the nuclear DNA content determined by flow cytometry and the number of chloroplasts in stomata guard cell pairs. Somatic hybrids inherited the parental plastids in a random manner. On the contrary, they preferentially inherited the mitochondrial DNA fragments of S. tuberosum. The majority of them had a rearranged mitochondrial genome with fragments from both parents. Hybrids were highly vigorous and morphologically more similar to the cultivated than to the wild parent, produced tubers on long stolons under long photoperiod conditions, showed a high degree of flowering, but did not produce pollen. In addition, somatic hybrids were generally more resistant to frost and Verticillium wilt than the cultivated parent, indicating the introgression of relevant resistance genes from the wild species into the genetic background of S. tuberosum. This revised version was published online in July 2006 with corrections to the Cover Date.     

Evaluation of genetic variation in Lachenalia bulbifera (Hyacinthaceae) using RAPDs

RAPD (randomly amplified polymorphic DNA) analyses were carried out on 21 accessions of Lachenalia bulbifera (Cyrillo) Engl. Five pre-selected primers produced an average of 88% polymorphisms. Fifteen of the 21 accessions could be identified using the five primers. In a pairwise comparison genetic distance values ranging from 0.11 to 1.08 were obtained. These values reveal a high amount of variation within the species. The genetic distance values within the tetraploid and hexaploid groups on the south coast were low, but values were high between the groups on the south coast and those on the west coast. A dendogram was constructed from the RAPD banding profiles, using UPGM cluster analysis. The dendogram clusters certain accessions together. These clusters are supported by their geographical locality and chromosome data. The hexaploid group, tetraploid group and octoploid group on the south coast are respectively clustered together. It is concluded that RAPDs can be used to assess the genetic variation at an intra-specific level in Lachenalia. This revised version was published online in July 2006 with corrections to the Cover Date.     

A biosystematic study of the origin of the cultivated diploid potato,Solanum x Ajanhuiri Juz. et Buk.

Summary The origin and nature of the diploid cultivated potato species S. ajanhuiri Juz. et Buk. was studied. Several lines of evidence indicate that S. ajanhuiri might be derived from natural crosses between primitive cultivars of the diploid species S. stenotomum and the wild species S. megistacrolobum. Morphological comparisons were made between S. stenotomum x S. megistacrolobum F₁ hybrids and naturally occurring S. ajanhuiri to investigate this hypothesis. Comparisons were also made between S. ajanhuiri x S. stenotomum crosses and the F₂ generation of the first-mentioned cross.Crosses between the two major groups of S. ajanhuiri cultivars, Ajawiri and Yari, showed not only genetic breakdown but also a wide range of phenotypic variation similar to those of artificial F₂ families of S. stenotomum x S. megistacrolobum. Furthermore, there was strong evidence showing that the Yari group of S. ajanhuiri could almost certainly be an F₁ S. stenotomum x S. megistacrolobum hybrid, whereas the Ajawiri group could be a backcross of an F₁ hybrid to S. stenotomum. These results added further support to the hypothesis of a hybrid origin of S. ajanhuiri, as well as indicating its putative parents. It is suggested that this hybridogenic taxon be retained at the species level under the name Solanum x ajanhuiri.     

Genetical assessment of diploid progenitors of <Emphasis Type="Italic">S. scabra</Emphasis> by isozyme,RAPD and STS markers: a possible strategy for improvement of drought tolerant allo-tetraploid <Emphasis Type="Italic">S. scabra</Emphasis> species

Assessment of genetic variations of two progenitors of S. scabra was investigated employing isozyme, RAPD and STS markers. Earlier reports have conclusively indicated S. seabrana and S. viscosa as two diploid progenitors of allo-tetraploid S. scabra. A total of 476 RAPD bands generated from 32 primers, 104 STS fragments from 17 primer pairs and 73 isoforms from 14 enzyme systems indicated significant sharing of these bands to S. scabra with that of S. seabrana in comparison to its other progenitor i.e., S. viscosa. High frequencies of isozyme band sharing between S. seabrana and S. scabra (0.125–0.30) as well as high genetic similarities (59%) of it with S. seabrana as reveled from dendrogram analysis indicated impact of close proximity of the occurrence of S. seabrana in the development of S. scabra. The proportion of bands shared by RAPD (40%) and STS (40%) markers were also observed high between S. seabrana and S. scabra. However, the numbers of unique bands were less in all these species. More correspondence and sharing of bands by S. seabrana towards S. scabra suggested using selected lines of S. seabrana the genetic base of allo-tetraploid, drought tolerant S. scabra lines can be broaden.     

Natural hybridization between Solanum acaule Bitt. and S. megistacrolobum Bitt. in the province of jujuy,Argentina

Summary Evidence is provided that hybridization of the tetraploid (2n=4x=48), self-fertile tuber-bearing species Solanum acaule Bitt. with the diploid (2n=2x=24), self-incompatible, tuber-bearing S. megistacrolobum Bitt. takes place in several localities of the province of Jujuy in the high, cold plateaux of the Argentine Puna. The triploid hybrids (2n=3x=36) closely resemble S. acaule in growth habit, leaf morphology and floral structures and for this reason they can be easily overlooked for that species in the field. Experimental data show that S. acaule can be crossed with S. megistacrolobum though the crossability is rather low and variable according to the particular cross considered. The artificial hybrid obtained compares well with the natural hybrid in morphology and chromosome number. The hybrids, though almost completely male sterile, are successful colonizers of disturbed areas around farmers' dwellings, cattle enclosures and other areas where the soil is rich in organic matter.There is some evidence that the Tilcara material of S. acaule subsp. aemulans and the hybrids of S. acaule x S. megistacrolobum have some characters in common, which can be interpreted as having a similar origin.It is postulated that S. acaule subsp. aemulans, in Jujuy at any rate, is not a primitive form of S. acaule as thought by Hawkes and Hjerting, but rather a fertile hybrid derivative of S. acaule x S. megistacrolobum through the functioning of 2n gametes.We also provide evidence that S. bruecheri Correll should not be considered a hybrid of S. acaulle x S. megistacrolobum but a synonym for S. gourlayi Hawkes. The new name, S. x indunii Okada et Clausen, is proposed to designate this hybrid.     

Identification of somatic hybrids of dihaploid Solanum tuberosum lines and S. brevidens by species specific RAPD patterns and assessment of disease resistance of the hybrids

Summary Symmetric somatic hybrids were produced by electrofusion of protoplasts of two dihaploid tuber-bearing potato (Solanum tuberosum L.) lines and Solanum brevidens Phil., a diploid non-tuber-bearing wild potato species. A total of 985 plants was obtained. Verification of nuclear hybridity of putative hybrids was based on additive RAPD patterns, general morphological characteristics and chromosome counts. 53 (90%) calli regenerated into plants which were identified as somatic hybrids. Most of the hybrids were aneuploids at the tetraploid (4×) or hexaploid (6×) level. The 20 hybrids tested expressed a high level of resistance to potato virus Y (PVY ^N ) characteristic of the S. brevidens parent. Resistance to late blight (Phytophthora infestans (Mont.) de Bary) varied between hybrids, but was on average better than that of the fusion parents. Resistance of hybrids to bacterial stem rot (Erwinia carotovora subsp. atroseptica (van Hall) Dye) was not superior to that of commercial potato cultivars.     

Genetic diversity in tef [Eragrostis tef (Zucc) Trotter] and its relatives as revealed by Random Amplified Polymorphic DNAs

Tef is one of the staple cereal crops in Ethiopia. To evaluate genetic diversity of tef and its relatives, 47 accessions of tef, three accessions of E. pilosa, and six accessions of E. curvulawere analyzed using random amplified polymorphic DNA (RAPD) markers. The level of polymorphism among the wild species was extremely high, while low polymorphism was detected among tef accessions. All cultivars and wild species under study could be distinguished with the help of different primers, thereby indicating the potential of RAPD in the genetic fingerprinting of tef. Accessions from E. curvula and E. pilosa can be differentiated by a single selected primer. In spite of low polymorphism within tef, accessions under study could be distinguished by a combination of selected primers. Cluster analysis indicated that tef is a very closely related species to E. pilosa with 45%similarity, supporting the hypothesis that tef originated from E. pilosa based on morphological data. Given that RAPD are relatively quick, simple to use, and are not subjected to environmental influences, they provide a valuable new approach for the genetic fingerprinting and study of genetic diversity in tef. This revised version was published online in July 2006 with corrections to the Cover Date.     

Phylogenetic relationships as in Ocimum revealed by RAPD markers

Genetic relationships were examined among thirty germplasm accessions belonging to five Ocimum species using RAPD markers. A very high degree of polymorphism (98.20%) was observed. UPGMA cluster analysis of genetic similarity indices grouped all the accessions into two major clusters corresponding to previously reported botanical sections. Intra-clustering within the two clusters precisely grouped the accessions belonging to one species in one sub-cluster as expected from their genetic background. Our results show that RAPD technique is a sensitive, precise and efficient tool for genomic analysis in Ocimum species, that may be useful in future studies, by assigning new unclassified germplasm accessions to specific taxonomic groups and reclassifying previously classified accessions of other Ocimum species by traditional criteria on a more objective basis. This revised version was published online in August 2006 with corrections to the Cover Date.     

The breeding of the cultivated potato species solanum x juzepczukii and S. x curtilobum II. The resynthesis of S. x juzepczukii and S. x curtilobum

Summary The possibility of combining anew the genomes of wild and cultivated progenitors of triploid S. x juzepczukii and pentaploid S. x curtilobum by following the known evolutionary pathway of these species was investigated.The resynthesis of S. x juzepczukii was easy, and a wide range of synthetic forms was bred. Among these were forms with higher frost resistance (-5°C) than has been found in natural S. x juzepczukii. The total tuber glycoalkaloid content of several synthetic hybrids was lower than or as low as that of natural clones. Most synthetic hybrids were more vigorous than natural S. x juzepczukii and produced about the same types of tubers as are found in the natural range of variation. The best diploid parents were found in the species S. goniocalyx.The attempt to resynthesize pentaploid S. x curtilobum has not been successful but tetraploid plants were obtained in the process. An explanation for the occurrence of tetraploids resulting from triploid x tetraploid and/or diploid crosses is offered.The newly bred tetraploids contain at least one genome from S. acaule (possibly two) and hybridize easily with ssp. andigena. They thus provide a means for the transfer of S. acaule germ plasm into the tetraploid cultivated gene pool which would profit from the frost resistance of S. acaule.     

Crossability and cytological studies in Solanum macrocarpon and Solanum linnaeanum (Solanaceae).

Summary Crossing experiments involving the various groups of Solanum macrocarpon L. complex, including wild (dasyphyllum), semi-wild and cultivars produced fully fertile F1 and F2 hybrids. This confirmed earlier findings that these groups belong to the same biological species. S. macrocarpon complex and S. linnaeanum Hepper ex Jaeger are isolated by reproductive barriers. F1 and F2 hybrids between various groups of S. macrocarpon showed heterosis. F2 superior hybrids were isolated as candidates for the future breeding programmes. Most wild-type traits like prickliness, hairiness were dominant to those possessed by cultivars. Relatively resistant hybrids to pest attack were noted. S. macrocarpon complex and S. linnaeanum are diploids (2n=24). The overall nature of the karyotype suggests that the two are related. The F1 hybrids between the cultivars of S. macrocarpon and the wild group (S. dasyphyllum) showed normal meiosis. A lack of isolating barriers confirmed that S. macrocarpon complex constitute a single biological species.     

Assessment of genetic relatedness of the genera Ananas and Pseudananas confirmed by RAPD markers

Genetic relationships among 18 accessions, including 16 of Ananas and two of Pseudananas, were investigated using RAPD molecular markers. The procedure for DNA extraction was adapted from the method of Dellaporta et al. (1983) where an incubation in proteinase K and a purification step were included. From the total of 148 markers scored,132 (89.2%) were polymorphic. The similarity matrix was used for cluster analysis. The phenogram developed from the RAPD bands showed that for most of the cases, the accessions within a species grouped together. Nevertheless, a moderate infraspecific genetic variation was observed. For example, DNA data grouped all A. comosus accessions with a mean similarity coefficient of 0.85. Comparable results were obtained with all other species investigated. The highest genetic divergence was found withinA. lucidus where the mean similarity coefficient among accessions was0.75. A similar level of genetic polymorphism was observed among species,therefore, a definition about which species were involved in the constitution of A. comosus genotypes was not possible. These results agree with the breeders standpoint suggesting that all Ananas species belong to the primary gene pool of pineapple. This revised version was published online in July 2006 with corrections to the Cover Date.     

Genetic diversity within cultivated teas based on nucleotide sequence comparison of ribosomal RNA maturase in chloroplast DNA

The wild tuber-bearing Solanumspecies represent a genetic pool of enormous diversity, embracing high variability for many agronomic traits. S. kurtzianum (ktz), S. chacoense(chc) and S. ruiz-lealii (rzl) are wild diploid self-incompatible relatives of the common potato, Solanum tuberosumssp. tuberosum, with resistance to adverse biotic and abiotic factors. Pollen-pistil/style compatibility relations and seed set in intra- and interspecific crosses among ktz, chc and rzl were investigated. Pre- and post-zygotic hybridization barriers were detected in the three species, with variable incidence because the percentage of genotypic combinations that produced seeds was15–30% and 7–28% in intra and interspecific crosses, respectively. In pollen-pistil/style incompatible combinations, either pollen grains did not germinate or pollen tube growth was arrested at different sites of the style, but mainly in its upper third. Unilateral incompatibility was detected in both intra-and interspecific crosses, as well as bilateral incompatibility in certain combinations. The incompatibility relations observed could be explained by a genetic model with dominant genes in pistils that interact with corresponding dominant genes in pollen, that are independent of the S-locus. The complex arrangement of compatibility relationships found could not be directly related to taxonomic hypothesis or geographic distribution. These results suggest that divergence between the three species has occurred recently, and that internal barriers of reproductive isolation have not been an important factor in their evolution. This revised version was published online in July 2006 with corrections to the Cover Date.     

Failure in an interspecific cross between Solanum khasianum Clarke and Solanum mammosum L.

Summary Attempts to obtain a successful interspecific cross between Solanum khasianum Clarke and Solanum mammosum L. were unsuccessful, and the probable reasons for this failure were investigated. A reduced number of viable pollen grains and low receptivity of the stigmas of S. mammosum L. and heterostyly in both species are the probable reasons for the failure of the attempted interspecific crosses between S. khasianum Clarke and S. mammosum L.     

Molecular characterization of the progeny of <Emphasis Type="Italic">Solanum etuberosum</Emphasis> identifies a genomic region associated with resistance to potato leafroll virus

Potato leafroll virus (PLRV; Genus Polerovirus; Family Luteoviridae) is one of the most important virus pathogens of potato worldwide and breeders are looking for new sources of resistance. Solanum etuberosum Lindl., a wild potato species native to Chile, was identified as having resistances to PLRV, potato virus Y, potato virus X, and green peach aphid. Barriers to sexual hybridization between S. etuberosum and cultivated potato were overcome through somatic hybridization. Resistance to PLRV has been identified in the BC₁, BC₂ and BC₃ progeny of the somatic hybrids of S. etuberosum (+) S. tuberosum haploid × S. berthaultii Hawkes. In this study, RFLP markers previously mapped in potato, tomato or populations derived from S. palustre (syn S. brevidens) × S. etuberosum and simple sequence repeat (SSR) markers developed from tomato and potato EST sequences were used to characterize S. etuberosum genomic regions associated with resistance to PLRV. The RFLP marker TG443 from tomato linkage group 4 was found to segregate with PLRV resistance. This chromosome region has not previously been associated with PLRV resistance and therefore suggests a unique source of resistance. Synteny groups of molecular markers were constructed using information from published genetic linkage maps of potato, tomato and S. palustre (syn. S. brevidens) × S. etuberosum. Analysis of synteny group transmission over generations confirmed the sequential loss of S. etuberosum chromosomes with each backcross to potato. Marker analyses provided evidence of recombination between the potato and S. etuberosum genomes and/or fragmentation of the S. etuberosum chromosomes.     

Genetic diversity of Tainan-white maize inbred lines and prediction of single cross hybrid performance using RAPD markers

To evaluate the genetic diversity of 13 maize inbred lines, and to determine the correlation between genetic distance and single cross hybrid performance, we employed the randomly amplified polymorphic DNA(RAPD)- a PCR-based technique. Six of these lines came from the Taichung population, and others derived from seven different sites. Forty different primers were used to give a total of 646 reproducible amplification products, 547 (84.7%) of them being polymorphic. Genetic divergence was determined using Jaccard's similarity coefficient, and a final dendrogram was constructed by UPGMA (unweighted pair-group method with arithmetical averages) cluster analysis in the CLUSTER procedure of the SAS system. The RAPD analysis was a useful tool in determining the extent of genetic diversity among Tainan-white maize inbred lines in the present case. Cluster analysis showed that the 13 inbred lines could be classified into distinct heterotic groups. There was no significant linear regression of grain dry weight heterosis value and mean performance of hybrids on genetic distance. And their coefficients of determination(R²) are small, so that predictive value is limited. The present results showed that the Jaccard's similarity coefficients based on RAPD data cannot be used to precisely predict the F₁ hybrids yield performance and heterosis value. This revised version was published online in August 2006 with corrections to the Cover Date.