Analysis of genetic variation in eggplant and related Solanum species using sequence-related amplified polymorphism markers

Collection and characterization of all sorts of germplasm resources are required for the development of new cultivars. Molecular characterization is more reliable than morphological characterization. Here, we employed sequence-related amplified polymorphism (SRAP) markers to evaluate genetic variation in a diverse collection of 56 Solanum accessions. Fifty-five SRAP primer combinations were used and a total of 635 polymorphic bands were observed. Cluster analysis by the unweighted pair-group method with arithmetic averages based on similarity matrices indicated that there were three clusters: (i) S. melongena; (ii) S. aethiopicum; (iii) S. surattense. The coefficients of genetic similarity among all the accessions ranged from 0.04 to 0.96 with an average of 0.73, and averaged 0.78 among S. melongena accessions originated from China, indicating extensive genetic variation. These results demonstrated that SRAP can be efficiently used to estimate genetic diversity and analyze phylogenetic relationship.     

Genetic structure of Buxus sinica var. parvifolia, a rare and endangered plant

Buxus sinica var. parvifolia, a rare and endangered tree species in some semitropics alpine areas of China, plays an important role in the maintenance of the landscape and ecosystem. In this study, RAPD and ISSR markers were used to investigate the genetic diversity and structure of five natural populations and one tamed population of B. sinica var. parvifolia. 21 RAPD primers amplified 209 bands with 167 (79.90%) polymorphic and 21 ISSR primers amplified 518 bands with 467 (90.15%) polymorphic. The genetic diversity, estimated by Shannon’ index, was 0.4343 (by RAPDs) and 0.3661 (by ISSRs). Both RAPD and ISSR analyses revealed a high level of genetic diversity in natural populations of B. sinica var. parvifolia. Furthermore, analysis of molecular variance (AMOVA) was used to apportion the variation within and between populations. The proportion of variation attributable to within-population differences was very high (69.2% by RAPDs; 84.51% by ISSRs). Moderate differentiation was detected among populations using RAPDs (30.80%), while only a small amount of variation (15.49%) was detected among populations using ISSRs. We suggest that the present genetic structure is due to high levels of environmental variability and gene flow, which still need further study to confirm. Conservation measures are suggested, including in situ and ex situ strategies, based on the observed population genetic information.     

Characterisation of cultivated breeding lines of eggplant (Solanum melongena L.) and related wild Solanum species from India

Immature fruit of cultivated species of eggplant (Solanum melongena L.) are commonly used as a summer vegetable in India. Rich morphological variation exists among the cultivated species of eggplant in different growing regions of the country. We have characterised 24 breeding lines of Solanum spp., including 20 eggplant cultivars and four wild species of eggplant, based on 13 morphological characters using Mahalonabis D² statistics. All 24 breeding lines of Solanum spp., including the 20 eggplant cultivars and four wild species, were grouped into four clusters by agglomerative clustering. Cluster II and Cluster IV contained the most accessions (eight each), while Cluster I and Cluster III had four accessions each. The highest inter-cluster (D²) distance (158.33) was observed between Cluster I and Cluster II, followed by Cluster I and Cluster III (108.48), and Cluster II and Cluster IV (102.96), which indicated that accessions in Cluster I and Cluster II were more divergent than those in the other clusters. The highest intra-cluster distance (5.80) was observed in Cluster IV, with eight genotypes, and the lowest intra-cluster distance (2.21) was observed in Cluster II, also with eight genotypes. The intra-cluster distances in all four clusters were lower than the inter-cluster distances, which indicated that genotypes within the same cluster were closely related. Genotypes in Cluster IV had the maximum number of flowers per cluster (3.63), the highest number of fruit per cluster (3.25), and number of fruit per plant (208.63), which revealed that genotypes could be selected from Cluster IV for these characters. The first three principal components (PC1, PC2, and PC3) accounted 73.99% of the total variation among the 24 genotypes. These phenotypic data increase the feasibility of prioritising breeding lines in a crossing programme based on the uniqueness of their desirable morphological traits.     

Application of genomic in situ hybridization for phylogenetic study between Mangifera indica L. and eight wild species of Mangifera

The phylogenetic relationship between mango (Mangifera indica L.) and eight wild species of Mangifera were analyzed by comparing signal intensity of genomic in situ hybridization (GISH) on somatic metaphase chromosomes of M. indica, using labeled DNA of eight wild Mangifera species. The eight wild species were divided into four groups based on intensity and number of hybridization signals on chromosomes of M. indica in GISH analyses. The probe of Mangifera sylvatica Roxb. gave the highest intensities on the chromosome of M. indica, indicating a close relationship between M. indica and M. sylvatica. For the other species, classification of GISH was comparable to that of the phylogenetic analysis using AFLP markers, as previously reported ( Eiadthong et al., 2000). This suggested a possibility that GISH analysis can be effectively used in the classification of Mangifera species.     

Assessment of genetic purity of tomato (Lycopersicon esculentum L.) hybrid using molecular markers

Three DNA molecular marker systems, RAPD, ISSR and SSR, were used to test seed genetic purity of two commercial hybrid tomato (Lycopersicon esculentum L.) cultivars ‘Hezuo 903’ and ‘Sufen No. 8’. Genomic DNA from the two F₁ hybrid cultivars and their corresponding parental lines was screened with 218 RAPD decamer primers, 54 ISSR primers and 49 SSR primers. Among the 321 primers, 4 primers for ‘Hezuo 903’ and 3 for ‘Sufen No. 8’, which could produce both female and male parent-specific markers, were selected for testing the genetic purity. A total of 210 hybrid individuals of each cultivar were analyzed using the identified primers. The combined results of the marker analysis showed that eight of the 210 F₁ plants in ‘Hezuo 903’ and 13 of 210 in ‘Sufen No. 8’ were false hybrids, and the overall genetic purity of the two F₁ hybrid seed lots was 96.2 and 93.8%, respectively. This study showed that RAPD and SSR markers could provide a practical and efficient tool in quality control of the tomato commercial hybrid seeds.     

Assessment of genetic diversity and species relationships in eggplant (Solanum melongena L.) using STMS markers

Ninety six accessions including 92 of Solanum melongena and four related non-tuberous species (Solanum insanum, S. incanum, S. integrifolium and S. sysimbriifolium) were taken for the assessment of genetic diversity using 23 STMS primers. Eleven of the 23 primers tested showed polymorphism. The number of alleles per primer ranged from three to six with an average of 4.4. S. melongena had maximum average similarity with its closely related species, S. insanum (0.67) and minimum average similarity with the wild species, S. sysimbriifolium (0.50). The two weedy species S. incanum and S. integrifolium showed more average similarity value of 0.62 and 0.61, respectively with the cultivated S. melongena. S. insanum. S. incanum and S. integrifolium were relatively similar to each other with similarity index value of 0.61 (between S. insanum and S. incanum), 0.63 (between S. insanum and S. integrifolium) and 0.62 (between S. incanum and S. integrifolium). In contrast S. sysimbriifolium was most divergent with the similarity value of 0.49, 0.47 and 0.51 with S. insanum, S. incanum and S. integrifolium, respectively. The closely related species S. insanum and S. incanum, which clustered along with S. melongena accessions, being crossable with cultivated species, constitute important sources of genes that can be introgressed by backcross breeding. Molecular markers can be employed to identify the hybrids and also to monitor introgression of the useful genes.     

Characterization of Tunisian pomegranate (Punica granatum L.) cultivars using amplified fragment length polymorphism analysis

The amplified fragment length polymorphism (AFLP) analysis of DNA was used to characterize 34 pomegranate cultivars. By using a combination of six primers, a total of 327 markers were scored with a mean of 57.5. The high percentage of polymorphic bands (ppb) of 94.7 and the resolving power (R_p) collective rate value of 129.14 were scored. Data proved that the tested primers were informative to discriminate among cultivars and to survey the genetic diversity in this fruit crop. It has been assumed that the local pomegranate germplasm is characterized by a typically continuous genetic diversity. The derived dendrogram proved that cultivars are clustered independently from their geographical origin and their denomination. In addition, AFLP permitted the generation of a nearly unlimited number of molecular markers that are reliable in differentiating the cultivars and/or the polyclonal varieties.     

Identification and genetic relationship based on ISSR analysis in a germplasm collection of sea buckthorn (Hippophae L.) from China and other countries

Sea buckthorn (Hippophae L.) is a woody, outcrossing dioecious pioneer plant, being widely planted as a new berry crop with rich nutritional and medicinal compounds, as well as a means of prevention of soil erosion. Inter simple sequence repeat (ISSR) markers were generated in 219 plants of 40 cultivars from China and other countries, representing two species of Hippophae salicifolia and Hippophae rhamnoides that includes two subspecies (ssp. mongolica and ssp. rhamnoides) and intraspecific hybrids between ssp. mongolica and ssp. sinensis. Fifteen selected ISSR primers generated 385 bands with an average of 25.6 bands per primer. Thirty hundred and eighty-five bands were used to generate a Dice's coefficient matrix of pairwise comparisons between individual ISSR profiles. Cluster analysis based on this matrix showed clustering of plants into groups which was in good agreement with their taxonomic classification. The results also revealed the presence of different cultivars with the same name, identical cultivars but with a different name and the genetic relationship of cultivars with unknown parentage to those with known parentage. Our data will be helpful in collection and conservation of sea buckthorn germplasm and potential hybrid strategy for breeding.     

Eggplant relatives as sources of variation for developing new rootstocks: Effects of grafting on eggplant yield and fruit apparent quality and composition

We propose the utilization of eggplant (Solanum melongena L.) interspecific hybrids derived from crosses with closely related species as an approach for developing new improved rootstocks for eggplant. Here we investigate rootstock effects on fruit yield, apparent quality and proximate and mineral composition of S. melongena ‘Black Beauty’ (BB) scions grafted on interspecific hybrid rootstocks developed from crosses of S. melongena with Solanum incanum L. (SI × SM) and Solanum aethiopicum L. (SM × SA). The results are compared with non-grafted (BB control) and self-grafted (BB/BB) controls and with S. melongena ‘Black Beauty’ scions grafted onto Solanum torvum Sw. (STO) and Solanum macrocarpon L. (SMA) rootstocks. All treatments were grown in a soil naturally infested with root-knot nematodes (mostly Meloidogyne incognita (Kofoid and White) Chitwood). SI × SM and SM × SA interspecific hybrids had high germination (≥90%) and total graft success (100%). Contrary to what occurred with all other treatments, no plants from scions grafted onto these hybrid rootstocks died during the experiment. In particular, the SI × SM hybrid rootstock conferred the highest vigour to the scion, which resulted in the highest values for fruit earliness and early and total yield. Little difference was observed among treatments for apparent fruit quality traits, except for a greater fruit calyx length and prickliness of fruit grafted onto SMA rootstocks. A similar result was obtained for fruit composition where phenolics content was higher in fruit from plants grafted onto SMA rootstocks. Grafting eggplant onto interspecific eggplant hybrids, especially on the SI × SM hybrid, has proved advantageous for eggplant production, as the high vigour and good compatibility of the rootstock with scion results in improved early and total yield without negative effects on apparent fruit quality or composition. Interspecific hybrids represent an alternative to the commonly used STO rootstock, which is a wild species with irregular germination.     

Interspecific relationships of Lycoris (Amaryllidaceae) inferred from inter-simple sequence repeat data

Inter-simple sequence repeat (ISSR) markers were used to evaluate interspecific relationships of Lycoris. Twenty-four samples were included in the study representing a total of 20 among species and varieties. Thirteen primers produced 228 discernible DNA fragments, 205, or 89.91%, of which were polymorphic, indicating a high level of interspecific genetic variation in Lycoris. Our UPGMA cluster analysis recognized four major groups of species, which were consistent with morphological and karyotype observations. The first group included species with telocentric and metacentric chromosomes, while the second consisted of species with a haploid genome of 11 subtelocentric chromosomes. The third group contained species with a mixture of subtelocentric, telocentric and metacentric chromosomes. The last group included the Japanese and Korean species. Lycoris anhuiensis was clustered within accessions of Lycoris longituba, suggesting that it could be recognized as a variety of L. longituba. Our ISSR data also suggested that L. straminea may be a hybrid between Lycoris chinensis and Lycoris radiata var. pumila, and that L. caldwellii, an allotriploid, may be of a hybrid origin of L. chinensis and L. sprengeri.     

Variety identification and comparative analysis of genetic diversity in yardlong bean (Vigna unguiculata spp. sesquipedalis) using morphological characters,SSR and ISSR analysis

Genetic diversity and relatedness of 23 yardlong bean (Vigna unguiculata spp. sesquipedalis) accessions and 7 accessions of a hybrid between cowpea (V. unguiculata spp. unguiculata) and yardlong bean (dwarf yardlong bean) in Thailand were estimated using morphological characters, simple sequence repeat (SSR) and inter-simple sequence repeat (ISSR) markers. In addition, two mungbean (Vignaradiata (L.) Wilczek) and two blackgram (Vigna mungo (L.) Hepper) accessions were also used as outgroup species for molecular analysis. Five morphological characters were diverse among most accessions. However, five groups of 2–3 accessions could not be distinguished from one another based on these morphological characters alone. Unweighted pair-group arithmetic average (UPGMA) analysis of these characters separated these 30 accessions into 2 major groups; the yardlong bean group and the dwarf yardlong bean group. Eleven of the sixteen SSR primers yielded clear SSRs, ten of which were polymorphic (90.91% polymorphism), detecting a total of 54 alleles with an average of 4.91 alleles per locus. These 10 polymorphic SSR markers successfully distinguished 28 yardlong bean and dwarf yardlong bean accessions. The polymorphic information content (PIC) among genotypes varied from 0.251 to 0.752 with an average of 0.597. Among the 16 ISSR primers used, a total of 312 ISSR fragments were amplified for these three Vigna species, revealing the polymorphism percentage of 91.03%. The average ISSR PIC value (0.197) with the range of 0.137–0.276 was lower than that of SSR. Nevertheless, the average marker index of this multilocus marker was 3.495, which was higher than that of SSR (0.669), owing to the differences in the effective multiplex ratio. In addition, Mantel test cophenetic correlation coefficient was higher for ISSR (0.566) than that of SSR (0.198). These results indicated higher efficiency of ISSR for estimating the levels of genetic diversity and relationships among yardlong beans and dwarf yardlong beans in this study. Pair-wise coefficients of SSR- and ISSR-based genetic similarity among all yardlong bean and dwarf yardlong bean accessions averaged 0.87 and 0.91, respectively, suggesting a narrow genetic base that emphasizes the need to broaden genetic diversity to ensure continued breeding success. Clustering of genotypes within groups was not similar when SSR and ISSR derived dendrograms from UPGMA analysis were compared. It appeared that ISSR was the most effective marker system in determining the genetic variability and relationships among yardlong bean and dwarf yardlong bean accessions and differentiating three Vigna species. In addition, ISSR was also most useful for variety identification since all 30 yardlong beans and dwarf yardlong bean accessions can be effectively distinguished by only four ISSR primers with the highest PIC values.     

Diversity,relationships, and genetic fingerprinting of the Listada de Gandía eggplant landrace using genomic SSRs and EST-SSRs

Listada de Gandía is one of the most renowned Spanish eggplant (Solanum melongena L.) landraces. Assessing its genetic diversity and relationships, as well as devising tools for its identification, is of great relevance for the enhancement and protection of this landrace. Forty-two eggplant accessions, which included 25 Striped accessions, of which 19 were of the Listada type (six accessions of Listada de Gandía, eight of Other Spanish Listada, and five of Non-Spanish Listada) and six of the Other Non-Spanish Striped group, and 17 Non-Striped accessions were characterized with 17 genomic SSRs and 32 EST-SSRs. Genomic SSRs had, as a mean, a greater polymorphism and polymorphic information content (PIC) than EST-SSRs. Although Listada de Gandía proved to be genetically diverse, specific and universal alleles for two SSR markers were found for this landrace. All the Listada accessions cluster together in the multivariate PCoA and UPGMA phenograms performed, and are separated from the Other Non-Spanish Striped and Non-Striped accessions. Also, Listada de Gandía accessions were clearly differentiated from the Other Spanish Listada and Non-Spanish Listada accessions in these analyses. SSR markers revealed of great utility to obtain a specific fingerprint for the Listada de Gandía eggplant as well as to establish the uniqueness and distinctness of this landrace. This information will be very helpful for the enhancement and protection from imitation of Listada de Gandía, and contributes to support its potential recognition with a Protected Designation of Origin (PDO) status.     

Breaking the intergeneric hybridization barrier in Carica papaya and Vasconcellea cauliflora

The present investigation was undertaken to develop PRSV (Papaya ringspot virus) resistant hybrids through intergeneric hybridization. Intergeneric hybridization was done involving nine Carica papaya cultivars as female and Vasconcellea cauliflora as male. To break the intergeneric hybridization barrier, various nutrient combinations were used. Among the combinations used, sucrose 5%, sucrose 5% + boron 0.5% and sucrose 5% + CaCl₂ 0.5% improved the fruit set and seed set percentage. A total number of 1197 flowers were pollinated and 308 fruits were obtained. On extraction, 721 seeds were obtained from CO 7, Pusa Nanha and CP 50. Out of 721 F₀ seeds (crossed seeds) sown, 419 seeds germinated and artificial screening for PRSV was carried out 27 days after sap inoculation. Out of 29 F₁ hybrid plants from CO 7 x V. cauliflora cross, only six plants namely CO 7V1 to CO 7V6 were found free from PRSV symptoms. Similarly, out of 55 F₁ hybrids from cross involving Pusa Nanha x V. cauliflora only 23 plants namely PNV1 to PNV23 were found free from the symptoms and 70 plants namely CPV1 to CPV70 out of 335 plants of CP50 x V. cauliflora cross were found free from PRSV symptoms. Among the crosses, Pusa Nanha x V. cauliflora had higher yield under PRSV infected conditions, however, total soluble solids and total sugars were found lesser than the CO 7 x V. cauliflora cross. The hybridity of the progenies were confirmed by using ISSR (Inter Simple Sequence Repeats) primers by the amplification of DNA from progenies and their parents. ISSR primers UBC 856, UBC807 and ISSR primer combinations UBC 856-817, UBC 810-817, UBC 861-817, UBC 856-810, UBC 861-810 and UBC 856-817 clearly amplified specific bands of the male parent, which were present in F₁ progenies, but it was absent in female parents.     

Utility of RAPD,ISSR, IRAP and REMAP markers for the genetic analysis of Citrus spp.

The application and informativeness of RAPD, ISSR, IRAP and REMAP markers to study the genetic diversity and relationship among the Citrus and its relative genotypes were investigated. High levels of polymorphism were observed for all four marker systems. The RAPD technique generated the highest number of polymorphic bands and average number of polymorphic band per assay unit. Average limit of the discriminating power was very close to its actual discriminating power of each marker. The highest and the lowest values of effective number of patterns were obtained from the marker REMAP (5.94) and RAPD (4.48), respectively. Correlation between the genetic similarities matrices were estimated from all four markers using the Mantel matrix correspondence test, and results showed significant correlations among the RAPD, ISSR, IRAP and REMAP similarity matrices. The highest correlations were found comparing RAPD and ISSR markers, whereas RAPD and REMAP (r = 0.143) markers were poorly correlated. To assess the usefulness of the overall information provided by these marker data for establishing phylogenetic relationships and Citrus germplasm classification, cluster analysis was performed. All four techniques, solely or in combination, discriminated the genotypes very efficiently and generated a high similarity in dendrogram topologies, although some differences were observed. The linkage analysis was conducted based on the segregation of 38 RAPD, 13 ISSR, 19 IRAP and 9 REMAP loci in 96 progeny of an intergeneric cross Citrus sinensis and Poncirus trifoliata. Among the 81 studied loci 65 loci distributed on five linkage groups. Comparing the result obtained with RAPD, ISSR, IRAP and REMAP markers in this study, IRAP and REMAP proved to be as a reliable molecular marker for fingerprinting, mapping and diversity study of Citrus and its relatives.     

A comparative analysis of the genetic diversity between inbred lines of Zinnia elegans using morphological traits and RAPD and ISSR markers

Genetic diversity within Zinnia elegans is key to the genetic improvement of this important ornamental species. Here, morphological traits and RAPD and ISSR molecular markers were used to assess levels of polymorphism across 20 inbred lines. Thirty-four morphological traits were scored and also 147 RAPD marker-fragments, as amplified by 12 arbitrary primers, and 128 ISSR marker-fragments as generated by 9 primers. The number of polymorphic loci, the percentage of polymorphic loci, Shannon's Information index (I) and the effective number of alleles (Ne) were calculated from the RAPD data as 100, 68.03%, 0.3559 and 1.4169, respectively. From the ISSR data, these respective statistics were calculated as 97, 76.38%, 0.4013 and 1.4728. Thus, ISSR markers were considered slightly superior to RAPD markers for assessing genetic diversity between the accessions; however, Mantel's test indicated significant correlation (R = 0.733) of the RAPD and ISSR results. By contrast, the morphological matrix showed low correlation with both RAPD and ISSR data matrices (R = 0.3814 and 0.3765, respectively). Cluster analysis showed that groupings of the accessions according to all three methods correlated well with their geographic region of origin, but flower color was not strongly associated with the genetic classification of these inbred lines of Z. elegans.     

Genetic diversity of ash gourd [Benincasa hispida (Thunb.) Cogn.] inbred lines based on RAPD and ISSR markers and their hybrid performance

Ten inbred lines of ash gourd [Benincasa hispida (Thunb.) Cogn.] were crossed to produce 45 F₁ hybrids (without reciprocal) which were evaluated along with the parents for 20 growth- and yield-related traits, in a replicated field trial. High level of heterosis was observed among the hybrids for most of the traits examined, including yield. These inbred lines were analysed by using 42 RAPD primers those produced 282 DNA marker bands. A total of 130 RAPD markers were obtained with a mean of 3.1 per primer, which in combination discriminated all the inbreds from each other. Pair-wise genetic distance measurements ranged from 0.07 to 0.31, suggesting a wide genetic diversity for these inbreds. These inberds were also analysed with five ISSR primers of which four were informative. Twenty-six ISSR marker bands were generated of which 11 were polymorphic with an average of 2.80 per primers. The percentage of polymorphic bands produced were higher in ISSR markers (>80%) than generated through RAPD markers (46%). Although the results indicated significant positive correlations of genetic distance with hybrid performance and heterosis, the RAPD based genetic distance measures and use of limited ISSR markers in this present study could not effectively predict hybrid performance in this crop. The genetic variation among ash gourd inbred lines examined, herein, defined a marker array (combined ISSR and RAPD) for the development of a standard reference for further genetic analyses, and the selection of potential parents for predicting hybrid performance and heterosis.     

Phylogenetic study and molecular identification of 31 Dendrobium species using inter-simple sequence repeat (ISSR) markers

The genetic diversity of the genus Dendrobium is not well known and the phylogenetic relationship of Dendrobium species are mainly determined by studies of the comparative vegetative anatomy and plant systematics. In the present study, we used the technique of inter-simple sequence repeats (ISSRs) to evaluate genetic diversity and phylogenetic relationship among 31 Dendrobium species from Yunnan region of China. In total, 2368 bands were amplified by 17 ISSR primers, resulting from 278 ISSR loci with 100% polymorphism at genus level. Thirty-one species were unequivocally distinguished based on ISSR fingerprinting. Species-specific ISSR markers were identified in nine of 31 tested Dendrobium species. Unweighted pair-group mean analysis (UPGMA) showed that 31 Dendrobium species were grouped into six clusters, indicating the genus was polyphyletic with several well-supported lineages. The high polymorphism and reliable amplification across species demonstrated the utility of ISSR marker for species diagnosis and genetic diversity study of the genus Dendrobium.     

Comparative analysis of genetic diversity in Indian bitter gourd (Momordica charantia L.) using RAPD and ISSR markers for developing crop improvement strategies

A genetic analysis of 38 diverse Indian bitter gourd (Momordicacharantia var. charantia, and var. muricata) accessions was performed using 29 RAPD and 15 ISSR markers. RAPD primers yielded 208 amplicons of which 76 (36.5%) were polymorphic providing an average of 2.6 amplicons per primer. RAPD amplicons per primer ranged from 3 (OPE-19, OPW-09) to 15 (OPW-05), and varied in size from 200 bp to 3000 bp. Fifteen ISSR primers provided a total of 125 bands of which 94 (74.7%) were polymorphic. Polymorphic ISSR markers ranged from 0 (UBC-841) to 12 (UBC-890) providing a mean of 6.3 amplicons per primer that ranged in size from 150 bp to 2700 bp. Nevertheless, the concordance among bitter gourd accession groupings after cluster analysis was relatively high (r = 0.77), indicating that RAPD- and ISSR-based diversity assessments in this germplasm array were generally consistent. The M.charantia var. charantia (domesticated) and var. muricata (wild, free-living) accessions examined were genetically distinct, and these differences provided for the development of strategies for genetic analyses and crop improvement in this species.     

Genetic diversity of Indonesian cauliflower cultivars and their relationships with hybrid cultivars grown in Australia

The objectives of this study were to investigate genetic variation and relationships between Indonesia-, Australian- and European-based cultivars and to evaluate variation within Indonesia cultivars as all cultivars are open-pollinated. Eight cauliflower cultivars collected from three production regions in Indonesia and four F₁ hybrids cultivars grown in Australia were evaluated using RAPD and ISSR markers. DNA polymorphisms generated from 10 polymorphic RAPD primers were used to construct a dendogram using the unweighted pair-group method with arithmetic averages (UPGMA) and to generate a fingerprinting key. ISSR marker analysis using 14 primers were attempted but DNA polymorphisms could not be clearly identified. The RAPD technique indicated that variation occurred both within and between Indonesian cultivars. Comparison between Indonesian-, Australian- and European-based cultivars showed that Indonesian cultivars have unique genotypes and would be good sources of genes for future crop improvement.     

Development of novel chloroplast microsatellite markers for Dendrobium officinale, and cross-amplification in other Dendrobium species (Orchidaceae)

Nine pairs of polymorphic chloroplast microsatellite primers were developed for Dendrobium officinale Kimura et Migo, an endangered herb. Levels of polymorphism were tested across a total of 55 individuals from four natural populations (12–15 individuals per population). Allele numbers varied from two to four per locus, while the number of haplotypes ranged from four to six per population. Transferability of the nine polymorphic chloroplast microsatellite primers was checked on an additional set of 51 Dendrobium individuals (belonging to 17 different species). Three markers could be transferable to all the species tested, while the remaining six markers successfully cross-amplified in most species tested. Moreover, polymorphism of the nine chloroplast microsatellite primers was tested across Dendrobium moniliforme (L.) Sw. and Dendrobium loddigesii Rolfe. All of them were polymorphic in D. moniliforme, while seven of which were polymorphic in D. loddigesii. These polymorphic chloroplast microsatellite primers developed for D. officinale will be a useful tool for the study of genetic diversity, population genetic structure, evolution of D. officinale and establishment of effective conservation strategies.