Polymorphisms Among Chloroplast and Mitochondrial Genomes of <Emphasis Type="Italic">Citrullus</Emphasis> Species and Subspecies

Twelve and six DNA clones representing various parts of chloroplast and mitochondrial genomes, respectively, were used to detect polymorphism among five watermelon cultivars and 21 U.S. Plant Introductions (PIs) collected from diverse geographical locations and representing major groups of Citrullus species. Cluster analysis based on 20 chloroplast DNA (cpDNA) and 10 mitochondrial DNA (mtDNA) restriction fragment length polymorphism (RFLP) markers differentiated the accessions into three major phenetic groups: PIs and watermelon cultivars of Citrullus lanatus subsp. vulgaris (Schrad. ex Eckl. et Zeyh.) Fursa (also designated as C. lanatus var. lanatus) (group I), PIs of C. lanatus var. citroides (of C. lanatus subsp. lanatus Schrad. ex Eckl. et Zeyh.)(group II), and C. colocynthis (L.) Schrad. PIs (group III). The chloroplast and mitochondrial genomes of watermelon cultivars are distinct, but closely related to those of the C. lanatus var. lanatus PIs. On the other hand, the chloroplast and mitochondrial genomes of the wild species C. colocynthis are more similar to those of C. lanatus var. citroides. Polymorphic cpDNA and mtDNA markers identified in this study can complement isozyme and nuclear DNA data used in earlier phylogenetic and phenetic classifications of Citrullus PIs. These cpDNA and mtDNA markers are being used in experiments designed to enhance watermelon cultivars by replacing the chloroplast and mitochondrial genome of cultivated watermelon with those of the wild species C. colocynthis.     

Isozyme relationships between cultivated artichoke and the wild relatives

Summary The genetic affinities between the cultivated artichoke Cynara cardunculus L. var. scolym (L.) Fiori [= C. scolymus L.] and its wild relatives were assessed by tests of 20 enzyme systems (28 loci). Six representative cultivars and the following wild taxa were examined: (i) wild cardoon C.cardunculus L. var. sylvestris (Lamk) Fiori, (ii) C. syriaca Boiss., (iii) C. cornigera (Lindley) [=C. sibthorpiana Boiss.], (iv) C. algarbiensis Cosson, (v) C. baetica (Spreng.) Pau [=C. alba Boiss.], and (vi) C. humilis L. Twenty one out of the 28 loci tested were polymorphic (mainly between species). The genetic identity between the cultivars and the wild cardoon forms ranged from 0.92 to 0.96; while that between these two taxa and the five other wild Cynara species ranged between 0.67 and 0.79. This implicates wild C. cardunculus in the wild ancestry of the cultivated artichoke.     

The wild ancestry of the cultivated artichoke

Summary The genetic affinities between the cultivated artichoke Cynara cardunculus L. var. scolymus (L.) Flori (= C. scolymus L.) and its wild relatives were tested by means of a crossing programme. The following wild taxa were involved: (i) wild cardoon C. carcundulus L. var. sylvestris (Lamk) Fiori, (ii) C. syriaca Boiss., (iii) C. cornigera Lindley, (iv) C. algarbiensis Cosson, (v) C. baetica (Spreng.) Pau (= C. alba Boiss.), and (vi) C. humilis L. Only the wild cardoon was found to be fully cross-compatible and fully infertile with the crop. In contrast, all other five wild Cynara species turned to be almost fully-or fully-cross-incompatible with the crop, and the few interspecific F₁ hybrids recovered were partly or almost fully sterile. These finds establish the wild cardoon as the wild ancestor of the cultivated vegetable.     

The Corsican citron melon (<Emphasis Type="Italic">Citrullus lanatus</Emphasis> (Thunb.) Matsum. et Nakai subsp. <Emphasis Type="Italic">lanatus</Emphasis> var. <Emphasis Type="Italic">citroides</Emphasis> (Bailey) Mansf. ex Greb.) a traditional and neglected crop

During a collecting mission in Corsica (France) three landraces of citron melon (Citrullus lanatus (Thunb.) Matsum. et Nakai subsp. lanatus var. citroides (Bailey) Mansf. ex Greb.) were collected in the northern areas. Today in Corsica this old and neglected crop is in decline and risks extinction. A strategy for its characterization and safeguarding is in progress at the genebank of IGV of Bari (Italy).     

Phylogenetic relationships in the North African genus <Emphasis Type="Italic">Hedysarum</Emphasis> as inferred from ITS sequences of nuclear ribosomal DNA

Sequences of the internal transcribed spacers (ITS) of nuclear ribosomal DNA were analysed to precise their length (637–643 bp) and resolve phylogenetic relationships among eight Mediterranean species of the genus Hedysarum (Fabaceae). The infra-specific variability levels of the ITS sequences of spontaneous population of H. coronarium proved a lack of polymorphism both in the length and in the sequences examined in this species. Hence, a consensus ITS sequence characterising each Hedysarum species has been investigated for analysis of inter-specific polymorphisms. The level of variation of ITS sequence was high enough to make the ITS1 and ITS2 a useful tool for phylogenetic reconstruction. However, ITS2 seems to be relatively more polymorphic and more informative than ITS1 regarding length or GC percent. The phylogenic relationships in the genus Hedysarum based on ITS1 and ITS2 sequences taken independently or together, are discussed in the context of current work in molecular biosystematics. Results exhibited the distinctiveness of the two H. spinosissimum subspecies (i.e. H. spinosissimum ssp. capitatum and H. spinosissimum ssp. spinosissimum). In addition, the great similarity of the ITS sequences between H. coronarium (the only cultivated species of the genus) and H. carnosum suggests the usefulness of the latter in selection programmes to improve pastoral production in semi-arid areas.     

Diversity and origin of cultivated and citron type watermelon (<Emphasis Type="Italic">Citrullus lanatus)</Emphasis>

Cultivated (Citrullus lanatus var. lanatus) and citron type (var. citroides) watermelon collected from different areas on the African continent are remarkably diverse in fruit and seed morphology. Chloroplast DNA investigations using PCR-RFLP and sequencing analysis of several non-coding regions were conducted to infer their biogeographic and evolutionary relationships, origin and domestication history. Variability within C. lanatus was observed at regions of high A + T content, resulting in indels and transversions mainly. Distinct chlorotype lineages were identified separating the cultivated and egusi-type watermelon from var. citroides accessions. This suggests an ancient split from a common ancestor and haplotype fixation. Three haplotypes as a result of relatively recent indel events were detected within var. citroides. The geographical range of two of the main citroides haplotypes is relatively similar across southern Africa. Accessions with the most ancient citroides haplotype originated in Swaziland and South Africa resulting in colonization routes from this area all over the world. Chloroplast divergence is not associated with morphological divergence. The cultivated and wild watermelon appear to have diverged independently from a common ancestor, possibly C. ecirrhosus from Namibia.     

The genetic characterization of Turkish watermelon (<Emphasis Type="Italic">Citrullus lanatus</Emphasis>) accessions using RAPD markers

Genetic diversity of the Turkish watermelon genetic resources was evaluated using different Citrullus species, wild relatives, foreign landraces, open pollinated (OP) and commercial hybrid cultivars by RAPD markers. The germplasm was consisted of 303 accessions collected from various geographical regions. Twenty-two of 35 RAPD primers generated a total of 241 reproducible bands, 146 (60.6%) of which were polymorphic. Based on the RAPD data the genetic similarity coefficients were calculated and the dendrogram was constructed using UPGMA (Unweighted pair-group method with arithmetic average). Cluster analysis of the 303 accessions employing RAPD data resulted in a multi-branched dendrogram indicating that most of the Turkish accessions belonging to var. lanatus of Citrullus lanatus (Thunb.) Matsum et Nakai were grouped together. Accessions of different Citrullus species and Praecitrullus fistulosus (Stocks) Pangalo formed distant clusters from C. lanatus var. lanatus. Among 303 accessions, a subset of 56 accessions was selected representing different groups and a second dendrogram was constructed. The genetic similarity coefficients (GS) within the Turkish accessions were ranged from 0.76 to 1.00 with 0.94 average indicating that they are closely related. Taken together, our results indicated that low genetic variability exist among the watermelon genetic resources collected from Turkey contrary to their remarkable phenotypic diversity.     

Assessment of genetic diversity, and phylogenetic relationships based on ribosomal DNA repeat unit length variation and Internal Transcribed Spacer (ITS) sequences in chickpea (Cicer arietinum) cultivars and its wild species

Repeat unit length variation and internal transcribed spacer (ITS) sequences of nuclear ribosomal DNA were used to assess genetic diversity, and phylogenetic relationships in chickpea (C. arietinum) cultivars, and its related wild species. Total genomic DNAs of 76 accessions of 10 Cicer species, belonging to three sections of the genus, were restricted with seven enzymes and the restriction fragments were hybridized to heterologous ribosomal clones of wheat pTa71 and Vicia faba probes Ver 6-5 and Ver18-6. A single repeat unit length class of 11.4 kb or 10.5 kb was recognized across Cicer accessions with pTa71. The intraspecific variation was negligible in those species where more than one accession was studied, except the four C. judaicum accessions, which were different from the rest. EcoRI and DraI digests gave two and one-two fragments, respectively. All the accessions produced three and three-five bands with BamHI and SacI, respectively. Both the accessions of C. yamashitae differed in their rDNA repeat unit length as well as restriction site variation. Maximum likelihood tree with rDNA RFLP recognized five clades which were more or less congruent with the previous data. Length of ITS-1 region was more variable (235–239 bp) than the ITS-2 region (212–213 bp). Cladistic analysis of ITS data revealed two major clades, clade I consisting of C. arietinum, C. reticulatum and C. echinospermum, and clade II comprised of C. judaicum, C. chorassanicum, C. bijugum and C. cuneatum. C. microphyllum grouped with the above four species. C. pinnatifidum was present as a separate branch. C. yamashitae emerged as the most distinct species.     

Agricultural potential of Mediterranean grain and forage legumes: Key differences between and within Vicia species in terms of phenology, yield, and agronomy give insight into plant adaptation to semi-arid environments

In order to identify key adaptive traits which affect productivity in Mediterranean grain and forage legumes and simultaneously determine the agricultural potential of a wide range of Vicia species, germplasm collected from the wild throughout the eastern Mediterranean was grown under semi-arid conditions in Tel Hadya, Syria (313 mm growing season rainfall). These included species currently in use in Mediterranean agriculture, such as V. sativa L., as well as those more widely used in the past-such as V. ervilia L., but also a broad selection from Section Narbonensis (B. Fedtsch. ex Radzhi) Maxted, including V. narbonensis L, V. johannis Tamamsch., V. hyaeniscyamus Mout., V. serratifolia Jacq., and V. kalakhensis Khattab et al. V. faba, a near relative of the taxa in Section Narbonensis, was included as a domesticated control. Where applicable, a representative range of subspecies was used. Accessions were chosen from a wide range of habitats in terms of latitude (31.02–40.72 decimal^o), longitude (27.1–43.17 decimal^o), altitude (20–1510 m), rainfall (180–1700 mm/yr) and soil depth (5–50 cm) in order to maximise diversity within species. Agricultural potential was determined by measuring seed, hay and biological yield, as well as agronomic traits such as harvest index, standing crop height, and seed size. The comparative influence of phenology and key agronomic traits such as plant habit and seed size on productivity varied tremendously between species, depending on their reproductive strategies. In V. sativa and V. ervilia, the smaller seed species which rely on long vegetative phases and growing seasons to accumulate sufficient biomass to set seed, and in which there was comparatively little agronomic variation, phenology had a large impact on yield. In early emerging taxa such as V. ervilia and V. s. subsp. sativa, with built-in long vegetative phases and growing seasons, seed yield was negatively correlated with flower ing (r = –0.86 to –0.88), whereas the opposite was the case for later emerging taxa such as V. s. subsp. nigra (L.) Ehrh. (r = 0.95). Within V. narbonensis and relatives, the larger seeded Vicia species which rely on more conservative reproductive strategies where high seedling vigour associated with large seeds enables the species to enter reproductive phases relatively early, phenology had a much smaller impact on yield than did variation of key traits such as seed weight, plant habit and pod shattering. Among the undomesticated germplasm harvest indices ranged from 0.09–0.31, hay yields from 0.1–3.4 t/ha, seed yield from 0–2.0 t/ha, and dry matter at maturity from 1.6–6.5 t/ha. Sub-specific taxonomy was crucial in determining agronomic potential. V. narbonensis var. aegyptiaca Kornhuber ex Asch. et Schweinf. showed the most potential, combining an upright habit, large seeds (212 mg) and tendency to retain intact pods after maturity, with the highest yield, harvest index and crop height of all the wild Vicia species. V. sativa subsp. sativa, V. ervilia and V. narbonensis var. narbonensis were less productive, but still showed agricultural potential. The smaller seeded V. narbonensis, var. affinis, var. jordanica H. Schäf. and var. salmonea (Mout.) H. Schäf., and their close relatives V. johannis, V. hyaeniscyamus, V. serratifolia and V. kalakhensis have little to offer Mediterranean agriculture on the basis of poor agronomy.     

DNA markers and pollen morphology reveal that <Emphasis Type="Italic">Praecitrullus fistulosus</Emphasis> is more closely related to <Emphasis Type="Italic">Benincasa hispida</Emphasis> than to <Emphasis Type="Italic">Citrullus</Emphasis> spp.

The round melon Praecitrullus fistulosus (Stocks) Pangalo has been cultivated in Asia since ancient times and has been considered an underexploited crop in the western world. In the USA, there is an increased interest in using P. fistulosus as a commercial vegetable, and possibly as a rootstock for grafting watermelon, melon, or cucumber. However, the taxonomic classification of P. fistulosus is incomplete and for many years it has been considered a close relative of watermelon [Citrullus lanatus subsp. vulgaris (Schrad. ex Eckl. et Zeyh.) Fursa] and was previously classified as Citrullus lanatus subsp. fistulosus (Stocks) Duthie et J.B. Fuller. Here, we used two sets of DNA markers to assess the genetic similarity of P. fistulosus in relation to Citrullus spp. {including Citrullus lanatus subsp. vulgaris, C. lanatus subsp. lanatus, Citroides group [also known as C. lanatus (Thunb.) Matsum. et Nakai subsp. lanatus var. citroides (Bailey) Mansf. ex Greb.], and C. colocynthis (L.) Schrad.}, Cucumis spp. (including C. melo, C. sativus, C. anguria, C. meeusei, C. zeyheri), Benincasa hispida (Thunb.) Cogn., Lagenaria siceraria (Mol.) Standl. and Cucurbita spp. (including C. moschata Duchesne and the winter squash C. maxima Duchesne). The first marker set comprised 501 markers that were produced by 38 primer pairs derived from watermelon expressed sequenced tags (ESTs) containing simple sequence repeat (SSR) motifs (designated as EST-SSR primers; produced 311 markers), and by 18 primer pairs derived from ESTs that do not contain SSR motives (designated here as EST-PCR primers; produced 190 markers). The second marker set comprised 628 markers that were produced by 18 sequence related amplified polymorphism (SRAP) primer pairs. The phylogenetic data indicated that among these cucurbit species, the wax gourd B. hispida is the closest to the P. fistulosus. Pollen observations, using light microscopy, indicated that each of the cucurbit genera examined here has unique pollen morphology. The Cucurbita spp. have globular pollen grains with a stigmatic surface. The L. siceraria has polygonal pollen grains with symmetrical boundaries, while the Citrullus spp. and Cucumis spp. have ovular (conical) and triangular shaped pollen grains (respectively). The B. hispida and P. fistulosus have spherical or semispherical pollen grains. These pollen features appear to be in agreement with the phylogenetic relationships of these two species based on DNA markers. Analysis with 12 SRAP primer pairs revealed low genetic diversity among 18 United States Plant Introductions (PIs) of P. fistulosus, indicating the need to expand the germplasm collection of this cucurbit crop.     

Agricultural potential of Mediterranean grain and forage legumes: 2) Anti-nutritional factor concentrations in the genus Vicia

As part of an investigation into the agricultural potential of under-exploited Mediterranean Viceae, anti-nutritional factor concentrations were measured in the seed of 7 species. These included: V. sativa(subsp. amphicarpa(L.) Batt., subsp. macrocarpa(Moris) Arcang, subsp. nigra(L.) Ehrh., subsp. sativa), V. ervilia L., V. faba L. (var. faba, var. minor Peterm.), V. narbonensis L. (var. aegyptiaca Kornhuber ex Asch. et Schweinf., var. affinis Kornhuber ex Asch. et Schweinf., var. jordanica H. Schaef., var. narbonensis, var. salmonea(Mout.) H. Schaef.), and relatives, including V. johannis Tamamsch., (var. johannis, var. procumbens H. Schaef.), V. hyaeniscyamus Mout., and V. serratifolia Jacq. Both quantitative and qualitative differences in seed chemistry were observed. Condensed tannins, phenolics and proteinase inhibitors were found in varying concentrations in all taxa. The highest concentrations of phenolic secondary metabolites (0.8–1.4% leucocyanidin equivalents, 9.5–16.4 mg/g phenolics) were found in V. faba, the V. narbonensis relatives and V. sativa subsp. macrocarpa. These taxa also tended to contain low levels of proteinase inhibitors (0.25-0.55 mg trypsin/mg protein), whereas the opposite was the case for V. ervilia(2.1 mg t/mg p). V. narbonensis was intermediate both in terms of tannin and proteinase inhibitor levels. In contrast, the non-protein amino acids tended to be more specific in their distribution. Concentrations of canavanine varied from 0.04-0.11% in the seed of V. ervilia, and may constrain the end-use of the grain, given that pigs are sensitive to concentrations as low as 0.08% in their diet. In V. sativa variation of -glutamyl--cyanoalanine within and between subspecies (0.41–1.36%) is not sufficient to make the grain palatable to monogastrics, since > 0.075% inclusion in the diet causes mortality in chicks. In V. narbonensis, there was no significant variation between most varieties for -glutamyl-S-ethenyl cysteine (GEC), a non-protein amino acid peptide which reduces grain palatability. However, lower concentrations were found in V. n. var. jordanica and its near relatives, V. johannis, V. hyaeniscyamus and V. serratifolia(1.3-1.6%, compared with 1.9+/–0.02% in all remaining V. narbonesis sub-species). Anti-nutritional factor concentrations were used to visualize taxonomic relationships using hierarchical cluster analysis. The V. narbonensis varieties were closely aligned with their near relatives, V. johannis, V. hyaeniscyamus and V. serratifolia. In contrast V. faba was positioned closer to the V. sativa subspecies than either to V. narbonesis or its near relatives. V. ervilia was the most distantly related species. Across all taxa proteinase inhibitors and condensed tannins or total phenols were negatively correlated (r=– 0.72, P < 0.009). This may be a mechanism which minimizes wasteful resource allocation to chemical defense, because the 2 anti-nutritional factors are functionally substitutable, since they both reduce the availability of protein in plant material. Thus taxa investing in high concentrations of proteinase inhibitors do not duplicate chemical defense mechanisms by simultaneously producing large amounts of tannins and phenolics.     

Analysis of population substructure,genetic differentiation and phylogenetic relationships among selected Asiatic <Emphasis Type="Italic">Vigna</Emphasis> Savi species

Random amplified polymorphic DNA markers were used to study sub-structure and genetic differentiation amongst 31 populations (seven cultivated and 24 wild populations) belonging to 14 Asiatic Vigna species. Ten pre-selected RAPD primers generated 152 polymorphic amplification products. Estimates of polymorphism indices were higher for the wild taxa in comparison to the cultivated forms. F_ST values between populations ranged from 0.111 to 0.801 and Nei’s genetic diversity values between and within species varied from 0.26 to 0.70 and 0.04 to 0.56 respectively. The high F_ST and F_CT values indicated strong subdivision of populations and high differentiation among species. Analysis of molecular variance was performed by grouping the populations conforming to specific species. AMOVA was also performed separately to better resolve the differentiation of species within mungo–radiata complex. Molecular phylogenetic relationships amongst the species of radiata–mungo complex; namely, black gram (V. mungo (L.) Hepper), green gram (V. radiata (L.) Wilczek), V. radiata var. sublobata, V. radiata var. setulosa, V. mungo var. silvestris and V. hainiana, were studied through cluster analyses. Two distinct groups were recognized within the complex, with population samples of V. hainiana forming one cluster. Further, V. hainiana appeared to be equidistant to both V. radiata and V. mungo.     

Genetic characterisation of olive trees from Madeira Archipelago using flow cytometry and microsatellite markers

In this study, native olive plants from Olea maderensis (≡ O. europaea ssp. cerasiformis) and O. cerasiformis (≡ O. europaea ssp. guanchica), wild olives (O. europaea ssp. europaea var. sylvestris) and cultivated olives (O. europaea ssp. europaea var. europaea) were analysed with respect to genome size and microsatellite markers. The mean nuclear DNA content of O. maderensis was estimated as 5.97 ± 0.191 pg/2C, while the remaining studied taxa presented mean genome sizes ranging from 2.99 to 3.18 pg/2C. These data and the obtained simple sequence repeats (SSR) profiles, i.e., with 2–4 alleles in O. maderensis and a maximum of two alleles in the other taxa, enabled the identification of a new ploidy level, tetraploidy, for a species belonging to the Olea genus. Cluster analysis of the microsatellite data revealed a clear separation of each species in different clusters and a high genetic dissimilarity could be observed among genotypes belonging to different species. This work contributed to a better characterization of olive species and the obtained data can be helpful to support taxonomic studies, and to develop germplasm preservation strategies in endangered populations of O. maderensis from Madeira Archipelago.     

Cytomolecular characterization of rDNA distribution in various Citrullus species using fluorescent in situ hybridization

The 18S–28S and 5S rDNA sites are useful chromosome landmarks and provide valuable evidence about genome organization and evolution. This investigation was the first attempt to study the dynamics, distribution and directionality of rDNA gains and losses, as well as to understand the contribution of site number variation in the speciation of the genus Citrullus. In this study, we employed fluorescent in situ hybridization (FISH), using the18S–28S and 5S rDNA gene loci, to evaluate the differences between the (1) cultivated type watermelon C. lanatus var. lanatus (sweet watermelon), (2) the “bitter” desert watermelon C. colocynthis (colocynth) that is indigenous to the deserts of northern Africa, the Middle East and Asia, (3) the C. lanatus var. citroides (citron) “Tsamma” or “cow watermelon” that is known as and is indigenous to southern Africa, (4) and C. rehmii that thrive in the Namibian Desert. The FISH analyses showed that the sweet watermelon and colocynth have similar rDNA configuration. The sweet watermelon and colocynth genomes contain two 18S–28S rDNA gene loci, each located on a different chromosome, and one 5S rDNA locus which is co-localized with one of the 18S–28S rDNA gene loci. On the other hand, the C. rehmii has one 18S–28S rDNA locus and one 5S rDNA locus positioned on different chromosomes, while the citron has one18S–28S rDNA and two 5S rDNA loci, each located on a different chromosome. A FISH analysis of F₁ (citron × sweet watermelon) chromosome spreads revealed uniparental homeologous rDNA gene copies pertaining to the sweet watermelon versus the citron chromosomes, with the sweet watermelon chromosome containing the 18S–28S and 5S rDNA locus versus the citron homologue chromosome that has the 5S rDNA locus, but not the 18S–28S rDNA locus. Genomic in situ hybridization (GISH) analysis, using the entire citron genome as a probe to be differentially hybridized on sweet watermelon chromosome spreads, revealed that the citron genomic probes mainly hybridize to subtelomeric and pericentromeric regions of the sweet watermelon chromosomes, suggesting extensive divergence between the citron and sweet watermelon genomes. The FISH and GISH cytogenetic analysis here indicate major differences in genome organization between the cultivated watermelon type sweet watermelon and its counterpart citron that thrive in southern Africa and considered a useful germplasm source for enhancing disease and pest resistance in watermelon cultivars.     

Genetic diversity in cucumber (Cucumis sativus L.): I. A reevaluation of the U.S. germplasm collection

Genetic variation within the U.S. cucumber collection (Cucumis sativus var. sativus L. and var. hardwickii (Royle) Alef.) was assessed by examing the variation at 21 polymorphic isozyme loci and comparing the results of this investigation with a similar previous analysis of 14 loci. About 29% (15 of 51) of the enzyme systems examined in an initial survey were polymorphic. Seven loci (Ak-2, Ak-3, Fdp-1, Fdp-2, Mpi-1, Pep-gl and Skdh) which were not previously used to estimate genetic diversity, were assessed. On average, 1.4 loci were polymorphic per enzyme system and 2.2 alleles were present per polymorphic locus. The frequency of polymorphisms was relatively low for Fdp-1(2) (0.01), Mpi-1(1) (0.03), and Skdh(1) (0.02). Principal component and cluster analyses of allelic variation at polymorphic loci separated a diverse array of 757 cucumber accessions from the U.S. National Plant Germplasm Systesm's (NPGS) collection into distinct groups by country (45 nations examined). All accessions of C. s. var. sativus were isozymically distinct from C. s. var. hardwickii, which were themselves dissimilar from each other. Data suggest that C. s. var. hardwickii is not a feral derivative of extant C. s. var. sativus populations. The allelic profile of C. s. var. sativus accessions originating from Burma, Thailand, Indonesia, Hong Kong, Zimbabwe and Ethiopia were distinct from the other accessions examined. Allelic fixation has occurred at Pgd-2 in accessions from Burma, and at Ak-2 in accessions from Zimbabwe and Ethiopia. Some of the countries examined that were in close geographic proximity (e.g., Thailand, Indonesia and Hong Kong) contained accessions with similar isozyme profiles. Accessions are fixed for certain alleles [e.g., Gr(1) (100%), Fdp-1(1) (100%) and Mpi-2(2) (50%) for accessions from Thailand, Indonesia and Hong Kong]. Grouping countries by continent or sub-continent (i.e., North and South American, China, Eastern Europe, Western Europe) and by numbers of accessions examined (i.e., India/Burma, Iran, Japan, Turkey, and remaining accessions) was used to identify accessions with unique allozymic profiles [PIs 209064 (USA), 257486 (China), 188749 (Egypt), 285607 (Poland), 369717 (Yugoslavia), 357844 (Poland), 255936 (Netherlands), 183127 (India), 200818 (Burma), 200815 (Burma), 137836 (Iran), 227013 (Iran), 227235 (Iran), 451976 (Japan), 181752 (Syria), 181874 (Syria), 169383 (Turkey), 171613 (Turkey)].     

Genetic diversity among watermelon (Citrullus lanatus and Citrullus colocynthis) accessions

Genetic diversity was estimated among 42 U.S. PlantIntroduction (PI) accessions of the genusCitrullus (of these, 34 PIs are reported tohave disease resistance), and 5 watermelon cultivars, using 30RAPD primers. These primers produced 662 RAPD markers that could berated with high confidence. Based on these markers, geneticsimilarity coefficients were calculated and a dendrogram wasconstructed using the unweighted pair-group method witharithmetic average (UPGMA). The analysis delineated threemajor clusters. The first cluster consisted of a group of fivewatermelon cultivars, a group of C.lanatus var. lanatusaccessions, and a group of C.lanatus var. lanatusaccessions that contained some C.lanatus var. citroidesgenes. The second cluster consisted of the C.lanatus var. citroidesaccessions, while the third cluster consisted of theC. colocynthis accessions.The two C. lanatus clustersdifferentiated from each other and from the C.colocynthis cluster at the level of 58.8%and 38.9% genetic similarity, respectively. Assessment ofgenetic diversity among accessions that have been reported to havedisease resistance indicated that resistance to either anthracnose,downy mildew, powdery mildew, or watermelon mosaic virus is foundamong all major groups of Citrullus PIs.Additionally, resistance to gummy stem blight or Fusarium wilt mayexist among C. lanatus var.citroides PIs. This study demonstrates thatmolecular markers can be useful in assessing genetic diversity, andin sorting Citrullus PIs into phylogeneticgroups prior to their evaluation for disease or pestresistance.     

Molecular differentiation in Indian Citrus L. (Rutaceae) inferred from nrDNA ITS sequence analysis

Molecular differentiation in 24 accessions representing 19 taxa of Indian Citrus has been examined through sequence analysis of Internal Transcribed Spacer (ITS) region of nrDNA. Sequence length in the 24 accessions of Citrus taxa ranged from 512 to 665 bp (ITS1 & ITS2 partial and 5.8S complete sequence). The ITS sequences were very rich in G+C content ranging from 61.40 to 66.60% with an average of 64.2%. Genetic distance within Citrus group ranged from 0 to 13.4% with an average of 4.6%, showing moderate rate of nucleotide divergence. The phylogeny was inferred using the Maximum parsimony (MP) and Neighbor-Joining (NJ) methods. Both MP and NJ trees separated all the 24 accessions of Citrus into six distinct clusters. The disposition of all the accessions of Citrus in separate clusters in ITS-derived dendrograms was partly in accordance with the morpho-taxonomic affinities of the target taxa. This study supports the concept of Citrus medica (citron), C. reticulata (mandarin), and C. maxima (pummelo) as the basic species of the genus. However, ITS marker could not find any clear cut differentiation between subgenera Citrus and Papeda as proposed in Swingle’s Citrus classification system. The present study also supports the distinctiveness of C. indica (Indian wild orange), C. latipes (Khasi papeda) and C. hystrix (Melanesian papeda) as true species, besides elucidating the probable hybrid origin and relationships among the cultivated species/biotypes, such as Citrus ×aurantiifolia (sour lime) C. ×limon (lemon), C. ×taitensis (Indian rough lemon), C. limettioides (sweet lime), C. ×aurantium (including sour and sweet oranges and grapefruit), and other indigenous varieties of Indian origin: C. megaloxycarpa (sour pummelo), C. karna (karna orange), C. pseudolimon (Hill lemon), ‘Memang athur’, ‘Pummelo-lemon’ and ‘Kathairi nimbu’.     

Description of Iranian Diploid Wheat Resources

Iran is one of the primary centers of diversity for wheat and its relatives. Wild wheats, in particular diploid species, are extensively distributed in various parts of Iran. Of these, the existence of two “A" genome donor species, i.e., T. baeoticum and T. monococcum, has already been reported in Iranian floras. However, the existence of the other important “A" genome donor, i.e., T. urartu, has not been clearly reported. In a germplasm collection survey in several regions of Iran, T. urartu was identified, collected, and described, and is reported here for the first time. In a detailed and extensive taxonomic program, the collected germplasms of the above three species were studied. Morphological characters and species-specific traits of the three species were evaluated and compared. Using the monographs by Gandilyan (1980) and Dorofeev et al. (1979) for the collected germplasms, 14 varieties were identified of T. boeoticum, 3 varieties of T. urartu, 2 forms of T. monococcum. Of these 5 varieties of T. baeoticum ssp. baeoticum (var. albinigrum, var. garniense, var. zuccarinii, var. baeoticum, var. pseudobaeoticum), 3 varieties of T. baeoticum ssp. thaoudar (var. abovjanii, var. albinigrescens, var. ebrahimzadehae, var. viridinigrireuteri) and 1 variety of T. monococcum (var. mansfeldii) are new to Iran. In addition, 1 variety of T. baeoticum ssp. baeoticum (var. khorramabadicum), 3 varieties of T. baeoticum ssp. thaoudar (var. ebrehimeadena var. taleghanicum, var. ilamicum) and 2 varieties of T. urartu (var. iranicum, var. sardashticum) are new to the wheat botany.     

Genetic diversity and differentiation of <Emphasis Type="Italic">Camellia sinensis</Emphasis> L. (cultivated tea) and its wild relatives in Yunnan province of China,revealed by morphology,biochemistry and allozyme studies

We evaluated morphological, isozyme and biochemical diversity of a total of 87 accessions in the genus Camellia [Camellia sinensis var. sinensis (10), C. talinensis (7), C. sinensis var. dehungensis (3), C. crassicolumna (3) and C. sinensis var. assamica (64)]. Great variation of morphological characters was apparent within each taxa. Across the five taxa, all leaf and most flower quantitative characters showed significant differences while all fruit quantitative characters measured did not differ significantly, and, seven (i.e., life form, bud color, petal texture, pubescence on ovary, style number, stamen location and locule per fruit) of the 33 qualitative characters yield significant differences. As a whole, caffeine content had the highest variation with CV of 22.7%, water extract solid showed the least variation (13.4%) and content of polyphenols (20.0%) and free amino acids (18.8%) showed intermediate variations. Camellia taliensis and C. sinensis var. assamica had significantly higher content of polyphenols and water extract solid than in the other three taxa, while no significant differences were detected for the content of caffeine and free amino acids. For allozyme study, 14 loci presented good resolution, among which, nine loci (64%) were polymorphic in each taxon (AAT-3, FUM-1, 6PDG-1, G6PDH-1, G3PDH-1, ME-1, PGM-1, PGM-2 and SKD-1). The percentage of polymorphic loci (P) for each taxon was 21.4–50.0%. Mean heterozygosity per locus (H _e ) varied 0.114–0.218. F _ST value indicated that only 4.6% of the variations could be ascribable to genetic differences among taxa. Genetic relationships among the five taxa revealed by allozymes, were also exposed by the result of clustering of the morphological and biochemical characters.     

ITS sequence-based identification and utilization evaluation of <Emphasis Type="Italic">“</Emphasis>Nanjiang” <Emphasis Type="Italic">(Lonicera similis</Emphasis> Hemsl.), a local cultivar in Sichuan,China

The internal transcribed spacer (ITS) region was employed to analyze phylogeny of 20 populations, including the local cultivar “Nanjiang”, belonging to 7 species of Lonicera L. 20 ITS sequences that we obtained range from 614 to 618 bp, and the G+C content was higher than 60% with the average value 63.5%. The sequences of 5.8S nrDNA was highly conservative in both length and component, while ITS2 had more variation than ITS1 in sequence component and length. For the ITS complete sequence, the mutation and information sites were 139 and 117 with the content of 22.38 and 18.84%, respectively. The dendrogram analysed by neighbor-joining method showed that “Nanjiang” and Lonicera similis Hemsl. comprised a same cluster with a higher bootstrap support, indicating “Nanjiang” would be a variation or subspecies of L. similis Hemsl. The results of antibacterial activity test and active compounds identification showed that “Nanjiang” had the best antibacterial activity and the highest yield of chlorogenic acid, the active component of Flos Lonicerae, in the six species of Lonicera L.