Genetic differentiation of watermelon landrace types in Mali revealed by microsatellite (SSR) markers

This study describes the genetic differentiation of a collection of 134 watermelon landrace accessions from Mali, representing red fleshed dessert and white fleshed seed and cooking type watermelons from five regions, plus three commercial dessert type cultivars with red flesh. The material was represented by in total 397 plants and was analysed using 24 microsatellite primer sets, which differentiated 129 alleles across all loci. Analyses of molecular variance ascribed 51 % of the variation among the landrace accessions, and 14 % of the variation could be ascribed to the regions. Partitioning the accessions into use groups (dessert, cooking, seed processing) explained 25 % of the variation. When categorising the accessions further into 10 landrace types, differentiated on the basis of use groups, local accession name, flesh colour and seed phenotype, these landrace types explained 26 % of the variation. Analysis with the software Structure revealed that the accessions with confidence could be separated into two major genetic groups, related to flesh colour (red and white) of the watermelon fruits. The same analysis further indicated that the material may be differentiated into eight genetic sub-groups. One group included again the red fleshed dessert types with local and commercial origin, while the remaining seven genetic sub-groups comprised the white fleshed landrace types used for seed processing and cooking, as well as white fleshed types of one dessert type. Some of the seed and cooking types were to a large extent different and assigned to one genetic sub-group each. A Mantel test, based on geographical and genetic distance matrices showed a positive correlation, indicating that seed exchange has not overcome local adaptation.     

Genetic Diversity of Traditional Chinese Mustard Crops Brassica juncea as Revealed by Phenotypic differences and RAPD Markers

This investigation was aimed at exploring the genetic diversity among nine typical accessions of Chinese mustard crops using random amplified polymorphic DNA (RAPD) markers and morphological comparison. Totally, 111 reproducible DNA bands were generated by 16 arbitrary primers, of which 91 bands were proved to be polymorphic. Based on pair-wise comparisons of the amplified bands, genetic similarities were obtained using Nei & Li's similarity coefficients and a dendrogram reflecting their relationships was made using the unweighted pair–group method with arithmetic averages (UPGMA). The result of cluster analysis indicated that the nine accessions were capable of being classified into two primary groups, one including accession 2 with expanded root (root mustard), accession 3 with entirely expanded whole stem (long-stem mustard), accession 6 with edible leaves (leaf mustard), accession 8 with edible seed stalk (seed stalk mustard) and another one including accession 4 with expanded basal stem (short-stem mustard), accession 5 with bulgy petiole (leafy bulgy mustard), and accession 9 with mustard-rich seed (seed mustard). Besides, accession 1 with expanded root (root mustard) and accession 7 with edible leaves and seed stalk (seed stalk mustard) were independent of other accessions in the dendrogram. Additionally, by cluster analysis based on highly reproducible RAPD markers, the accessions with similar edible parts of leaves or roots were not actually in the same phylogenetic groups. This implied that they were probably derived from different geographical origins with dissimilar genetic background and possessed higher genetic diversification. Furthermore, the results indicated that the traditional method for classifying Chinese mustard crops was not much reliable as it was largely dependent on phenotypic behaviors. Meanwhile, the phenotypic differences among individuals did not necessarily mean they must have sharp difference in genetic background as they met in the same group. Undoubtedly, these results aforementioned make this crop quite interesting to researchers for further investigating the molecular evolution of this special AABB group.     

Phenotypic and RAPD diversity among Cymbopogon winterianus Jowitt accessions in relation to Cymbopogon nardus Rendle

The species Cymbopogon winterianus Jowitt is believed to have originated from the well-known species Cymbopogon nardus, type Maha Pengiri, referred to as Ceylonese (Sri Lankan) commercial citronella. Cymbopogon winterianus Jowitt was named after Winter, who raised its population as a separate species in the 19th century. C. winterianus was introduced into Indonesia and became commercially known as the Javanese citronella. The Javanese type C. winterianus material was introduced into India for the commercial cultivation of this crop during 1959. Varieties of this species have been developed later by the use of breeding procedures from the same introduced material. A comparative analysis of yields of herb, oil percentage and oil constituents for eight prevalent C. winterianus cultivars comparing them between themselves as well as against an accession of C. nardus has been carried out. All these accessions were analyzed at the molecular level for the similarity and genetic distances through RAPD profiling, using 20 random primers. More than 50% divergence was observed for all the C. winterianus accessions in relation to C. nardus accession CN2. The clustering based on the similarity matrices showed a major cluster of six accessions, consisting of two sub-clusters. The accession C. nardus CN2 got carved out along with two C. winterianus accessions, CW2 and CW6. On the other hand, the accessions CW2 and CW6 demonstrated distinct identities compared to CN2 at the DNA level.     

Genetic resource enhancement by isolation of diversegenotypes from seed progeny in predominantly sterile rose scentedgeranium Pelargonium graveolens

Four new accessions (BSP1-4) of rose scentedgeranium Pelargonium graveolens were derivedfrom the spontaneous half-sib seed progeny of mostly sterilepopulations of the cultivar accession Bipuli. The seed producingplants of Bipuli had been growing among the populations of cultivaraccessions Hemanti and Kunti in the temperate agroclimate ofKodaikanal, Tamilnadu in India. The four new accessions were comparedwith the cultivar accessions Bipuli, Hemanti and Kunti in fieldexperiments under subtropical agroclimate of Lucknow, Uttar Pradeshfor essential oil yield and quality related traits and in terms oftheir RAPD profiles. The crop yield and essential oil parameters andDNA profiles of the four new accessions and three cultivar accessionsallowed the conclusion that the accessions BSP-1, BSP-2 andBSP-3 were Bipuli × Kunti hybrids and BSP-4was a Bipuli × Hemanti hybrid. The accession BSP-4demonstrated hybrid vigour in several of the essential oil yieldrelated traits; on average basis it out yielded other accessions by afactor of about 2.4. The essential oil of BSP-4 had 89%rhodinol content in which citronellol to geraniol ratio was 1:1 andthe contents of isomenthone, menthone, 10-epi--eudesmol, 6,9-guaiadiene, decanoic acid andisodecanoic acid were relatively lower than in the oils of accessionsBipuli and/or Hemanti. The present study has shown a way for thegeneration of new genotypes in rose scented geranium in which thecultivars have been vegetatively maintained for decades and thegenetic resources are scanty.     

Microdetermination of diosgenin from fenugreek (Trigonella foenum-graecum) seeds

Sulfuric acid hydrolysis of steroidal glycosides of Amber fenugreek was studied by capillary gas chromatographic analysis of diosgenin [(25R)-spirost-5-en-3-ol] and isomeric spirostadiene artifacts from 100 mg samples of seed material. Following extraction with 80% ethanol, highest recoveries of diosgenin occurred when hydrolyses were conducted in sulfuric acid, prepared at 1 molar (M) concentration in water containing 60-80% 2-propanol. Compared to a previous method with aqueous hydrochloric acid, the selected conditions of hydrolysis at 100 degrees C for 2 h with sulfuric acid in 70% 2-propanol reduced diene formation but did not completely eliminate these artifacts. Extraction of steroidal saponins with various alcohol/water mixtures prior to sulfuric acid hydrolysis gave similar recoveries of diosgenin. Application of the quantitative method to experimental samples of Amber, Quatro, and ZT-5 fenugreek, using 10 mg subsamples of crushed seed that had been defatted with petroleum ether and dried at 60 degrees C, gave diosgenin levels of 0.55, 0.42, and 0.75%, respectively. Levels of smilagenin and sarsasapogenin were very low in hydrolyzed seed extracts from ZT-5, a Canadian breeder line of fenugreek.     

Comparison of oil and phytosterol levels in germplasm accessions of corn, teosinte, and Job's tears

Seeds of 49 accessions of corn (Zea mays ssp. mays), 9 accessions of teosinte (Zea species that are thought to be ancestors and probable progenitors to corn), and 3 accessions of Job's tears (Coix lacryma), obtained from a germplasm repository, were ground and extracted with hexane. Whole kernel oil yields and levels of four phytonutrients (free phytosterols, fatty acyl phytosterol esters, ferulate phytosterol esters, and gamma-tocopherol) in the oils were measured. Among the seeds tested, oil yields ranged from 2.19 to 4.83 wt %, the levels of ferulate phytosterol esters in the oil ranged from 0.047 to 0.839 wt %, the levels of free phytosterols in the oil ranged from 0.54 to 1.28 wt %, the levels of phytosterol fatty acyl esters in the oil ranged from 0.76 to 3.09 wt %, the levels of total phytosterols in the oil ranged from 1.40 to 4.38 wt %, and the levels of gamma-tocopherol in the oil ranged from 0.023 to 0.127 wt %. In general, higher levels of all three phytosterol classes were observed in seed oils from accessions of Zea mays ssp. mays than in seed oils from accessions of the other taxonomic groups. The highest levels of gamma-tocopherol were observed in teosinte accessions.     

Variation in temperature requirements for germination and early seedling root development in Chamaecrista rotundifolia and three allied species

Two experiments are discussed. In the first, rates of root elongation during the first four days of growth at 25 °C were studied in Chamaecrista rotundifolia (34 accessions), C. nictitans (2 accessions), C. pilosa (1 accession) and C. serpens (3 accessions). In the second, a comparison was made of germination characteristics of 14 accessions of C. rotundifolia and one of C. pilosa at nine constant temperatures (11–27 °C). Entries included accessions from a wide latitudinal range. There were statistically significant differences in root length on all four days of measurement. No accession had significantly longer roots than the commercial cultivar cv. Wynn on day 1, but over the following three days, 4, 10 and 13 accessions had significantly longer roots, respectively. Root length on day 4 was positively correlated with latitude of origin. In the second experiment, maximum germination occurred at 25.5–27 °C, with progressively lower percentages at lower temperatures. There were highly significant differences between accessions at each temperature (P<0.001), and a highly significant accession × temperature interaction. No accession had a higher germination percentage than cv. Wynn at any temperature. Percentage mortality differed significantly (P<0.05) at all temperatures except 27 °C. Mortality at low temperature was negatively correlated with latitude of provenance. Increased temperatures up to 22.5 °C reduced the time taken to reach 50% of maximum germination (T₅₀), and thereafter it was reasonably constant. With one exception (22.5 °C), no accession germinated significantly more quickly than cv. Wynn, and at 13 °C cv. Wynn germinated significantly more rapidly than all other entries. Over all temperatures, there was a significant negative correlation between days to 50% germination and latitude of provenance. Estimated base temperature for germination (T _b ) ranged from 10.6 °C to 13.6 °C. T _b was positively correlated with latitude of provenance. It is argued that accessions from higher latitudes have developed mechanisms for survival of soft seed at low temperatures, low base germination temperature, germination rates and root elongation rates that adapt them to an environment with summer-dominant rainfall and winters which are too cool to support growth.     

The genetic diversity associated with seed proteins in a collection of Spanish underground vetches (<Emphasis Type="Italic">Vicia sativa</Emphasis> L. subsp. <Emphasis Type="Italic">amphicarpa</Emphasis> (Dorthes) Asch. et Graebn.)

The characterization of plant genetic resources is the first step towards improving their use. The Spanish Plant Genetic Resources Centre, which belongs to the National Institute for Agricultural and Food Technology Research (CRF-INIA), conserves accessions of wild underground vetches collected in Spain. In the present work, 26 underground vetch accessions were characterized in terms of their seed storage proteins (separated by SDS-PAGE) as a means of assessing the genetic variation of these plants and their agronomic value. Vicia sativa cv. Vereda was used as control. A total of 54 bands were detected, of which 49 were polymorphic. Fifty eight different electrophoretic patterns were observed in total. Protein bands were scored in terms of their presence (1) or absence (0) for all the seeds studied, and two matrices constructed, one with all the bands present in each accession, the other with the different patterns for each accession. Dendrograms based on the Jaccard similarity index and the UPGMA clustering method were produced from these matrices, and the degree of genetic variation between and within accessions was calculated. The groups obtained were compared with the chromosome number for each accession. The results reflect the great diversity of underground vetch seed storage proteins. The aerial and underground seeds of 16 accessions were then analysed separately. In some cases, the aerial and underground flowers of the same plant produced different proteins.     

Patterns of SSR variation in bread wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis> L.) seeds under ex situ genebank storage and accelerated ageing

Maintaining seed viability and germplasm integrity is a challenging task in conservation of plant genetic resources, as seeds under storage will lose viability and genetic changes will occur. Attempt was made to analyze the patterns of genetic changes in wheat germplasm under ex situ genebank storage and accelerated ageing treatments. A set of 16 naturally aged wheat accessions under ex situ genebank storage since 1994 were sampled. Four recently regenerated wheat accessions were selected, four random seed samples were chosen from each accession, and three of them were exposed to three different accelerated ageing treatments. These 32 seed samples in two germplasm sets displayed a range of germination rates from 4 to 98 %. Thirty-seven microsatellite markers representing 21 wheat chromosomes were applied to screen 12 seeds of each sample and 449 SSR alleles were scored. Large SSR variation was found in each germplasm set. There was 73.1 % of the total SSR variation present among the naturally aged samples and 78.2 % present among the accelerated ageing samples. Several analyses for genetic association consistently revealed no clear genetic separations among samples of high or low germination rates in both germplasm sets. Samples under different accelerated ageing treatments did not show much genetic differentiations from the original sample of each accession. Mantel tests revealed non-significant associations between SSR variability and sample germination rates for both germplasm sets. These findings are useful for understanding seed deterioration under different ageing conditions and suggest that genome-wide SSR variability may not provide sensitive markers for the monitoring of wheat seed viability.     

Geographical comparison of genetic diversity in Asian landrace wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis> L.) germplasm based on high-molecular-weight glutenin subunits

Glutenin largely determines wheat bread baking quality. As high-molecular-weight glutenin subunit (HMW-GS), related to Glu-1 loci, determines wheat flour elasticity, it correlates strongly with bread-making quality. This study was aimed at clarifying genetic variations in bread-making characteristics between East and West Asian wheat landrace germplasms, by investigating HMW-GS allelic composition of 1068 wheat accessions. Herein, the accession number having reported HMW-GS pattern in previous studies was 855. However, the accession number with newly detected HMW-GS patterns was 114. These new HMW-GS patterns were classified into 4 types based on similarity. Eight Korean accessions with these four types were identified. Concerning landrace germplasm nature, 99 accessions showed heterogeneous patterns caused by seed mixture. The Glu-1 loci allelic variation analysis, revealed that the percentages of Glu-A1c (73.6%), Glu-B1b (60.2%), and Glu-D1a (68.5%) were highest at Glu-A1, Glu-B1, and Glu-D1 loci, respectively. The incidence of preferable alleles for bread baking was high in Chinese accessions. In bread-making quality evaluation using Glu-1 score, 24 among 35 accessions with full score were from China. The polymorphic information content index of each origin based on HMW glutenin subunit combination showed that West Asian and neighboring-regional landraces, excluding Afghanistan ones, were more diverse than East Asian landraces excluding Chinese ones. Cluster analysis based on Glu-1 allelic combination showed that many Korean, Japanese, and Afghan accessions were in the same group. However, many Chinese and other West Asian accessions were in the other group despite geographical distance.     

Analysis of Genetic Diversity in Colonial Bentgrass (<Emphasis Type="Italic">Agrostis capillaris</Emphasis> L.) using Randomly Amplified Polymorphic DNA (RAPD) Markers

Colonial bentgrass (Agrostis capillaris L.) is a cool-season grass, native to temperate Asia and Europe. It has good tolerance to low temperatures and partial shade and is well suited to golf course fairways and tees. Little information is available regarding levels and patterns of genetic variation among populations of colonial bentgrass, which would be useful for breeding programs. To study the genetic relationships among 27 colonial bentgrass accessions obtained from the US National Plant Germplasm System (NPGS), randomly amplified polymorphic DNA (RAPD) markers were scored and analyzed. Out of 80 primers screened, 16 were selected for further analysis, which yielded a total of 120 polymorphic bands used to differentiate the accessions. Dice's similarity coefficients for pair-wise comparisons ranged from 0.23 to 0.84 based on the RAPD data. Since there was no similarity coefficient value close to 1 between any two accessions, there was no apparent duplication among the sampled accessions. A dendrogram constructed on the basis of the Unweighted Pair Group Method with Arithmetic average (UPGMA) clustering algorithm clearly separated 26 of the accessions into three clusters with one accession distinct from the rest. The least similar pair of accessions was PI 204397 from Turkey and PI 628720 from Bulgaria, and the most similar pair was PI 509437 from Romania and PI 491264 from Finland. Clustering patterns based on principal components analysis (PCA) corresponded well with the dendrogram. A high cophenetic correlation (r = 0.82) was found between the RAPD data matrix and cophenetic matrix. The accession PI 628720, from Bulgaria, did not cluster with any other accessions.     

Morphological and reproductive characterization of guar (<Emphasis Type="Italic">Cyamopsis tetragonoloba</Emphasis>) genetic resources regenerated in Georgia,USA

Guar, Cyamopsis tetragonoloba is used worldwide for food stabilization, fiber source, food, and industry. The United States Department of Agriculture, Agricultural Research Service, Plant Genetic Resources Conservation Unit (USDA, ARS, PGRCU) conserves 1,298 accessions originating from India, Pakistan, and breeding lines from the USA. My objectives were to determine: (1) if these guar accessions can successfully be regenerated in Georgia, USA, (2) which traits contribute the most variation, and (3) to provide estimates for genetic relatedness among these accessions. Guar accessions were directly seeded to the field in Griffin, GA between early May and mid June 2006–2008. At 50% maturity, 73 accessions were characterized for morphological, phenological, and reproductive traits during the regeneration cycles. High quality plants regenerated from most of the accessions and produced 80 to more than 9,300 seeds per accession. Guar can be successfully grown and regenerated in Griffin, GA. Coefficients of variation and principal component analysis revealed variability among accessions for these traits evaluated. Cluster analysis separated guar accessions into three groups (clusters) based on low, intermediate or high seed numbers. Guar accessions clustered in groups 2 and 3 appear to be more genetically related than those in group 1.     

Variation of glucosinolates in wild radish (Raphanus raphanistrum) accessions

Glucosinolate composition was determined in wild radish accessions from eight states in the northeastern and southern United States to determine the variability of production among accessions. Glucosinolates were evaluated from roots, leaves, flowers, primary, and secondary branches. Seventeen glucosinolates were identified, with glucoerucin, glucoraphenin, glucobrassicin, and gluconasturtiin contributing 90% to 100% of the total glucosinolates. Flowers contained the highest glucosinolate concentrations, 12.07 to 55.36 μmol/g, but flowers contributed only 5.3 to 21.3% to the total glucosinolates. Of the eight accessions, the Mississippi accession produced significantly higher levels of total glucosinolates and glucosinolates which can be degraded to isothiocyanates per plant, totals of 618.97 and 563.53 μmol/plant, respectively. Total plant biomass did not differ between accessions indicating a difference in the ability of the Mississippi accession to produce glucosinolates. Further studies are needed to determine if this accession would consistently produce higher glucosinolate levels under different environmental conditions.     

Characterization of a USDA Kentucky Bluegrass (Poa pratensis L.) Core Collection for Reproductive Mode and DNA Content by Flow Cytometry

Kentucky bluegrass (Poa pratensis L.) is an important turf and forage grass species with a facultative apomictic breeding behavior. In this study, mature seed and leaf tissue from 38 accessions of a USDA core collection of Kentucky bluegrass were analyzed with flow cytometry to characterize the reproductive mode and DNA content for each accession. Major reproductive pathways for each accession were determined based upon the presence and the position of the peaks observed and the known methods of reproduction for Kentucky bluegrass. While the majority of the accessions exhibited facultative apomictic reproductive behavior with a combination of reduced, zygotic and unreduced, parthenogenic embryo production, obligate sexual or obligate apomictic accessions were also found to be present in this core collection. In addition, reduced, parthenogenic and unreduced zygotic embryos were also detected in several accessions. Flow cytometric analysis of somatic tissue revealed a large range of DNA variation within this core collection. We also examined the sensitivity of flow cytometry in analyzing bulked samples containing a large number of plants with varied DNA content and determined that flow cytometry can effectively detect a plant having a different DNA content within a 15-plant bulk sample. Overall the combination of mature seed and somatic tissue analysis generated important information for the Kentucky bluegrass core collection and can be an effective and affordable tool to characterize even greater numbers of Kentucky bluegrass accessions.     

Hypericum perforatum L. Seed Germination: Interpopulation Variationand Effect of Light, Temperature, Presowing Treatments and Seed Desiccation

Germplasm conservation of medicinal plants is of increasing interest and, when possible, seed banking is the most efficient system for ex situ conservation of these plant genetic resources. Hypericum perforatum L. (St. John's wort, Guttiferae) is a medicinal plant with evidence of efficacy as an anti-depressant. The aim of this work was to increase knowledge of its seed germination behaviour by studying 68 wild populations. Seed germination tests were carried out at 25/15 °C under a photoperiod of 16-h light/8-h darkness. Final germination percentages were highly variable depending on the accession, ranging from 6 to 98%. Similarly, germination rate (as expressed by T ₅₀ values) varied significantly from 6.1 to 23.0 days. The effect of seed desiccation with silica gel on subsequent germination was also studied. The effect of two other incubation temperatures (15 and 25 °C) and light (photoperiod or darkness) on seed germination was studied in several accessions. Temperature had no significant effect on final germination percentages. However, light significantly increased the germination of most but not all accessions assayed. Seeds from four accessions with low germination percentages were subjected to different presowing treatments that could increase germination: dry heat, hot water and gibberellic acid. Germination was promoted significantly by gibberellic acid in two of the four accessions assayed, but the thermal treatments did not enhance significantly the germination percentages. This study reveals that conclusions based on one population of Hypericum perforatum cannot characterize the germination behaviour for the entire species.     

Oil and fatty acid contents in seed of Citrullus lanatus Schrad

Intact seed of 475 genebank accessions of Citrullus ( C. lanatus var. lanatus and C. lanatus var. citroides) were analyzed for percent oil content using TD-NMR. Extracts from whole seed of 96 accessions of C. lanatus (30 var. citroides, 33 var. lanatus, and 33 egusi), C. colocynthis (n = 3), C. ecirrhosus (n = 1), C. rehmii (n = 1), and Benincasa fistulosa (n = 3) were also analyzed for their fatty acids content. Among the materials analyzed, seed oil content varied from 14.8 to 43.5%. Mean seed oil content in egusi types of C. lanatus was significantly higher (mean = 35.6%) than that of either var. lanatus (mean = 23.2%) or var. citroides (mean = 22.6%). Egusi types of C. lanatus had a significantly lower hull/kernel ratio when compared to other C. lanatus var. lanatus or C. lanatus var. citroides. The principal fatty acid in all C. lanatus materials examined was linoleic acid (43.6-73%). High levels of linoleic acid were also present in the materials of C. colocynthis (71%), C. ecirrhosus (62.7%), C. rehmii (75.8%), and B. fistulosa (73.2%), which were included for comparative purposes. Most all samples contained traces (<0.5%) of arachidonic acid. The data presented provide novel information on the range in oil content and variability in the concentrations of individual fatty acids present in a diverse array of C. lanatus, and its related species, germplasm.     

Distinguishing among Pisum accessions using a hypervariable intron within Mendel’s green/yellow cotyledon gene

Distinguishing among commercial varieties or accessions in a germplasm repository presents considerable challenges when dealing with common crops or large collections. In the genus Pisum L. the third intron of the gene STAYGREEN (SGR), responsible for the green/yellow cotyledon variation, contains a complex of repeat sequences that display considerable indel and single nucleotide (SNP) polymorphism. In a sample of 137 Pisum L. accessions obtained from various germplasm collections 76 alleles were identified, ranging from about 600 to 1500 nucleotides in length. In most cases alleles differed by insertion/deletion (indel) polymorphisms in addition to single base changes. The amplicon was particularly useful for distinguishing wild accessions (those with dehiscent pods and dormant seeds). Among the 51 P. sativum L. accessions examined with a definite wild phenotype there were 44 SGR alleles. When this allelic variation was combined with location data for the wild accessions, each allele could be assigned a unique geographical location. Similarly, of the 10 P. fulvum Sibth. & Sm. accessions analyzed, only a group of three had the same allele, two of which are known to be replicates. We were able to confirm the identities of several P. sativum accessions from different germplasm collections, as well as identifying two cases where supposedly synonymous accessions gave different intron sequences. In addition, a group of wild pea accessions from Greece, Sicily, and Portugal exhibited a unique repeat sequence not found in any other wild accession, suggesting that these accessions constitute a distinct lineage. The rate of nucleotide site variation in this fragment of the SGR locus was found to be about 2.22 × 10^–8 substitutions per site per year. This value is similar to those found for many other sequences in herbaceous papilionoid tribes, although it is at the faster end of the distribution of herbaceous legume substitution rates. We propose that both a fast substitution rate as well as the formation and rearrangement of indels have contributed to the high level of variation we detected at SGR. This PCR product should prove valuable in the identification of Pisum accessions, for quality control operations in germplasm repositories, and for exploring the evolutionary relationships within the genus.

     

Reactions of lentil accessions from 25 different countries to Australian isolates of Ascochyta lentis

488 lentil (Lens culinaris) accessions from 25 different countries were evaluated in glasshouse tests for resistance to three Australian isolates of Ascochyta lentis. Disease symptoms on the different accessions ranged from small flecks (resistant) to extensive lesions on both leaves and stems with death of some plants (highly susceptible). No accession was found with complete resistance to ascochyta blight. The 488 accessions showed differential resistance to all three isolates of A. lentis and could be divided into five resistance groups. 26 accessions originating from Greece (1), Pakistan (23) and Turkey (2) were resistant to all three isolates, while 142 accessions showed variable reactions and 320 accessions were susceptible to all three isolates. These results demonstrate there is considerable variation in lentil germ plasm for resistance to A. lentis.     

Molecular evidence of genetic diversity changes in pea (<Emphasis Type="Italic">Pisum sativum</Emphasis> L.) germplasm after long-term maintenance

Maintaining germplasm genetic integrity is a key objective of long-term ex situ conservation. Periodic regeneration, performed on limited plots with small number of individuals, increases the risk of genetic drift and genetic diversity changes. In this study, six accessions of white flowered, dry seed pea varieties (Pisum sativum L. subsp. sativum var. sativum): Bohatyr, Klatovsky zeleny, Hanák, Moravsky hrotovicky krajovy, Raman and Viktoria-75 and four accessions of colour flowered, fodder pea (P. sativum subsp. sativum var. arvense (L.) Poiret: Arvika, Česky banán, Moravská krajová and Niké, representing Czechoslovak varieties and landraces, bred over the last 40–80 years, were analyzed using ten microsatellite locus specific markers. Each accession was represented by 20 individual seeds of two temporally different samples, spanning the period of 20 or 40 years. Together with intra-accession variation (except of cv. Hanák), evidence of genetic changes, e.g. differences in allele frequencies as well as genetic composition of sample, was detected in six out of ten accessions (Arvika, Bohatyr, Česky banán, Moravsky hrotovicky krajovy, Moravská krajová and Raman). Evidence of genetic erosion was found in three accessions (Česky banán, Moravsky hrotovicky krajovy and Raman), while in another three (Arvika, Bohatyr and Moravská krajová) the level of diversity was found to have increased. Moreover in three samples of Bohatyr (2004) and Klatovsky zeleny (1963 and 2004), low levels of heterozygosity was detected. These results demonstrate that in pea, a self-pollinating and highly homozygous plant, the danger of the loss of genetic integrity exists. These findings are significant for long-term ex situ germplasm management.     

Conservation of the genetic variation of Triticum tauschii (Coss.) Schmalh. (Aegilops squarrosa auct. non L.) in synthetic hexaploid wheats (T. turgidum L. s.lat. x T. tauschii; 2n=6x=42, AABBDD) and its potential utilization for wheat improvement

Triticum tauschii (Coss.) Schmalh. (Aegilops squarrosa auct. non L., 2n=2x=14, DD genome) with its diverse range of accessions and distribution provides a unique opportunity for exploiting novel genetic variability for wheat (T. aestivum L.) improvement associated with biotic/abiotic stress factors. From our working collection of 490 T. tauschii accessions we have so far produced 430 different synthetic hexaploids (2n=6x=42, AABBDD) resulting from the chromosome doubling of Triticum turgidum L. s. lat. x T. tauschii F₁ hybrids (each synthetic involving a different T. tauschii accession). We present here our results on hybrid production, plantlet regeneration, cytology, colchicine induced doubling of the 2n=3x=21 chromosome F1 hybrids, seed increase of the doubled progeny and screening for a biotic stress; Cochliobolus sativus Ito and Kuribay (syn. Helminthosporium sativum Pamm. King and Bakke); of 250 of these synthetic hexaploid (2n=6x=42) amphiploids. Application of the direct crossing methodology involving susceptible T. aestivum cultivars with resistant T. tauschii accessions is also alluded to.