ALBERT

Notes: Summary Somatic fusions between an accession of the diploid wild speciesSolanum verrucosum and a dihaploid S.tuberosum genotype were produced in order to incorporate resistance to potato leafroll virus (PLRV). In total 15 somatic hybrids out of 16 regenerants were obtained. Identification of hybrids was based on additive RAPD patterns, general morphological characteristics, chromosome numbers and chloroplast counts in stomata guard cells. A field trial was performed with the hybrids, their two parents and the control cultivar Kennebec to assess field performance and phenotypic variability. Yield parameters varied considerably among somatic hybrids. Some of the hybrids gave significantly higher yields, tuber numbers and tuber weights than both parents. Pollen fertility of hybrids ranged from 19 to 77%. Twelve hybrids were found to be resistant to PLRV.

Notes: Abstract A suitable screening method for heterozygous DNA markers in shiitake,Lentinula edodes (Berk.) Pegler, is reported. Monokaryons were derived from a dikaryon by de-dikaryotization via protoplast formation. Compatibility of the monokaryons was determined by pairwise culture on agar plates. We selected the primers to amplify polymorphic fragments among the original strain (Hokken600∶H600) and two monokaryons (H600PP-39 and H600PP-67) showing compatibility. A total of 135 fragments were selected as specific random amplified polymorphic DNAs (RAPDs) resulting from 56 primers of the 147 primers tested. Furthermore, we tested whether the polymorphic fragments segregated into 2∶2 among four strains isolated from a basidium. Most of the polymorphic fragments (about 97.8%) showed 2∶2 segregation among the four strains. We concluded that the polymorphic fragments were heterozygous if they were detected in either of the monokaryons (H600PP-39 and H600PP-67) and segregated to 2∶2 among four meiotic strains (H600B-1,-2, -3, and -4). A total of 132 heterozygous DNA markers were therefore selected from a dikaryon of shiitake (Hokken600∶H600).

Notes: Abstract This paper describes a novel knowledge-based approachfor helping workflow process designers andparticipants better manage the exceptions (deviationsfrom an ideal collaborative work process caused byerrors, failures, resource or requirements changesetc.) that can occur during the enactment of aworkflow. This approach is based on exploiting ageneric and reusable body of knowledge concerning whatkinds of exceptions can occur in collaborative workprocesses, and how these exceptions can handled(detected, diagnosed and resolved). This work buildsupon previous efforts from the MIT Process Handbookproject and from research on conflict management incollaborative design.

Notes: Abstract This study describes the development of species-specific pairs of PCR primers for the root-knot nematodes Meloidogyne chitwoodi, M. fallax and M. hapla that amplify species-specific RAPD fragments. After sequencing the fragments, longer primers were designed to complement the terminal sequences of the polymorphic DNA fragments. The resulting pairs of primers were used to generate the sequence-characterized amplified regions (SCARs). Using the developed pairs of SCAR primers, SCAR fragments of M. chitwoodi, M. fallax or M. hapla were easily amplified from DNA extracts from juveniles, egg masses, females of the particular nematode species investigated, either present alone, in a mixture with other nematode species or in infested plant material. A specially designed multiplex assay using three pairs of SCAR primers enabled the identification of multiple species in a mixture in a single PCR step. Single juveniles were easily identified by applying this multiplex assay followed by a subsequent multiplex PCR using three pairs of nested primers. The SCAR-PCR-based assays described have potential to be optimized for routine practical diagnostic tests. The usefulness of converting RAPD markers into SCAR markers is discussed.

Notes: Abstract Previously chromosome 3A of wheat (Triticum aestivum L.) was reported to carry genes influencing yield, yield components, plant height, and anthesis date. The objective of current study was to survey various molecular marker systems for their ability to detect polymorphism between wheat cultivars Cheyenne(CNN) and Wichita (WI), particularly for chromosome3A. Seventy-seven `sequence tagged site' (STS), 10simple sequence repeat (SSR), 40 randomly amplified polymorphic DNA (RAPD) markers, and 52 restriction fragment length polymorphism (RFLP) probes for wheat homoeologous group 3 chromosomes, were investigated. Three (3.9%) STS-PCR primer sets amplified polymorphic fragments for the two cultivars, of which one was polymorphic for chromosome 3A. Sixty percent of SSR markers detected polymorphism between CNN and WI of which 50% were polymorphic for chromosome 3A. Twenty percent of RAPD markers detected polymorphism between CNN and WI in general, but none of these detected polymorphism for chromosome 3A. Of the fifty-two RFLP probes, 78.8% detected polymorphism between CNN and WI for group 3 chromosomes with one or more of seven restriction enzymes and 42% of the polymorphic fragements were for chromosome 3A. These high levels of RFLP and SSR polymorphisms between two related wheat cultivars could be used to map and tag genes influencing important agronomic traits. It may also be important to reconsider RFLP as the most suitable marker system at least for anchor maps of closely related wheat cultivars.

Notes: Abstract Thirteen varieties of Brazilian barley selected for malting qualities were analysed by RAPD (Random Amplified Polymorphic DNA). Amplification with 18 random primers generated 221 reproducible bands, of which 206 bands were polymorphic (93%):of this number, 137 fragments (62%) detected diversity among varieties and 56 bands (25.34%) allowed the distinction of varieties or groups of them. Variation was detected in all Brazilian varieties studied. Within-variety similarities estimated by Jaccard Similarity Coefficient ranged from 0.28 to 0.94, with averages ranging from 0.57 to 0.83, and an overall average of 0.72. Nevertheless, in the cluster analysis representatives of the same variety always fell into the same group and only later joined the other varieties. The average intervarietal similarities estimated by Jaccard Similarity Coefficient ranged from 0.45 to 0.62, with an overall average of 0.52. Many bands or combinations of bands which were responsible for the differentiation of all varieties were detected. Nevertheless, the majority of these bands cannot be considered as diagnostic markers because a great number of them did not occur in many representatives of the variety or had low intensity or even because they were not easily identified in the total pattern of bands.

Notes: Abstract Genetic variation and genotype × environment interactions for important agronomic characters were estimated for eleven Norwegian populations of white clover (Trifolium repens L.), originating from between 58°52′N and 69°30′N latitude, and from altitudes of 10 to 450 m a.s.l. The populations were evaluated in monoculture clonal field plots for two years at two locations in Norway. The medium-leaved Danish cultivar Milkanova was also included in the experiment as a standard market cultivar. Considerable variation between populations was observed for winter survival, spring growth, morphological characteristics, dry matter yield, general performance, earliness and seed yield. Winter survival, measured as plant cover after two years, was significantly different at the two locations. All the populations had sufficient winter survival at the southern, continental highland site (61°07′N).At the northern coastal location (69°30′N),where the winter climate is unstable with physical stresses such as frost, ice-encasement and water-logging frequently occurring, only the northernmost, coastal populations showed sufficient winter survival. Highly significant interactions were found between populations and locations, and between populations and years for all characters measured at both locations and in both years, respectively. Interaction effects between populations, locations and years were significant only for leaflet length. The largest leaf lamina's were observed in the most southern populations. However, genotypes possessing relatively long leaflets were also found in the northernmost populations, although at low frequencies. In general, the populations originating from the northernmost locations had lower biomass production, and were smaller-leaved, more prostrate and exhibited shorter internodes and stolons compared to the populations from South and Middle Norway. These populations also initiated inflorescences earliest, and were among the lowest seed yielding populations. Otherwise, no clear clinal variation in growth characteristics was observed which could be related to the latitude or altitude of origin of the populations.

Notes: Abstract Similar to SCAR, an extended random primer amplified region (ERPAR) marker is a PCR amplified genomic DNA fragment at a single genetically defined locus. However, ERPAR uses specific primer pairs derived from RAPD primers by adding bases sequentially to their 3′-ends. As an example, an ERPAR marker was derived from a RAPD marker (OT11900) linked to a dominant male sterility gene in cabbage (Brassica oleracea var. capitata). After two cycles of base adding and primer pair screening, a primer pair (5′-TTCCCCGCGACT-3′and 5′-TTCCCCGCGAGA-3′) amplified a single intense band with the same size as OT11900. The identity of the new marker and OT11900 was verified by segregation analysis. The new marker amplified by this extended primer pair was named as EPT11900. The development of ERPAR exploits the importance of 3′-end bases of primers in PCR ERPAR shares advantages of SCAR, but eliminates the need for cloning and sequencing. It is a fast and universal way of converting RAPD markers into stable markers.

Notes: Abstract Wild emmer wheat, Triticum dicoccoides, grows naturally in several habitats in northern Israel. The assumption that a genotype is better adapted to the `native' soil from which it was collected than to other soils, was tested. Each of nine T. dicoccoides accessions from nine different habitats and three wheat cultivars was sown in soils taken from all habitats, and grown in a greenhouse over 3 years. To enhance the biological nutrition absorption forces from the soil, three common wheat cultivars were added to the wild genotypes. No interaction in grain yield between wild wheat genotype and soil type was found within experiments. Soil type was the main factor that affected development and yield. Seed nutrient ability (SNA) of each soil was defined as the mineral element content in the seeds, averaged over all genotypes. Multiple regression analysis revealed diversity between the SNA related to growth and yield of the genotypes. Total seed yield per plant of each accession was related to several SNAs, and mainly to S and K, R2 = 0.5–0.85. The spikelet number per spike was determined by N and Na in five accessions and by Ca in the other four (R2 = 0.39–0.93).Heading date was affected mainly by the genotype, and the soil effect exhibited Fe and P dependence. A genotype-habitat adaptation exhibited by yield components was related to yield quality rather than to yield quantity. When a mineral nutrient is deficient in a natural soil, natural selection leads to establishment of plants that store a higher concentration of that nutrient in the seed, for the benefit of the succeeding generation.

Notes: Abstract The diversity of 27 superior tea (Camellia sinensis var. sinensis) accessions from Korea, Japan and Taiwan was examined with RAPD-PCR (Random Amplified Polymorphic DNA Polymerase Chain Reaction) markers. Out of the 50 primers screened, 17 primers generated 58 polymorphic and reproducible bands. A minimum of 3 primers was sufficient to distinguish all the 27 accessions studied. The Shannon's index used to partition diversity into inter- and intra-group, revealed that 71 percent of variability resided within groups and 29 percent between groups. Diversity was greatest within the Korean group followed by Taiwan and Japan. The relatively high diversity observed in Korea might reflect the larger genetic base of its plantations while the low diversity in Japan could be explained by the long and intensive tea selection programme in this country. A dendrogram based on the UPGMA-link method using Jaccard's distances and multivariate Factorial correspondence analysis clustered the tea accessions into two main groups, regrouping the Taiwan cultivars on the one side and the Korean and Japanese accessions on the other side. This suggests that the Taiwan tea studied here may have a different origin from that of Korea and Japan.

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

Notes: Abstract A set of 60 random primers was used to analyse 11accessions from six taxa of Mentha developed byCIMAP. These accessions were maintained in the nationalgene bank for medicinal and aromatic plants at CIMAP.A total of 630 bands could be detected as amplifiedproducts upon PCR amplification, out of which 589 werepolymorphic (93.5%). Further analysis of these RAPDprofiles for band similarity indices clearlydifferentiated five of the Mentha arvensis L.accessions from the rest. Among two accessions of Mentha spicata L. CIMAP/C33 could bedistinguished from CIMAP/C32. Mentha × gracilis Sole cv. cardiaca showed a muchhigher similarity with Mentha spicata L. as wellas Mentha arvensis L. which amongst themselvesshowed rather a greater distance indicating that Mentha × gracilis Sole cv. cardiaca might have evolved as a natural hybridbetween M arvensis L. and M. spicataL. In terms of uniqueness of amplified bands fordeveloping RAPD markers, it was observed that at taxalevel 298 bands were unique to one of the six taxa,singly amounting to 47.3% of total amplifiedfragments. Primers MAP 10 and 17 produced polymorphismonly in case of M. spicata L. and Menthaspicata L. cv. viridis while MAP 08 producedpolymorphic bands in all 4 other species than thesetwo. Similarly unique patterns were observeddifferentiating all six species and could be used asRAPD markers for differentiating Mentha species.

Notes: Abstract Growth habit, heading date and Vrn genotypewere examined for wheat landraces cultivated in China,Korea and Japan, to study their ecogeographicaldifferentiation in east Asia. Spring type landracesaccounted for 43.6% of the whole, and the frequencyvaried between the localities, being closely relatedto the degree of winter coldness. Spring typelandraces mainly adapted to north and south Chinawhere average January temperature is under –7 °Cand over 4 °C, respectively. On the contrary,winter type adapted to areas of average Januarytemperature from –7 °C to 4 °C. As toheading date, significant difference was not observedbetween spring and winter type landraces but betweenlocalities, and those cultivated in north China weresignificantly later in heading. It is thereforeindicated that spring type mainly adapts to areaswhere wheat is sown in spring to avoid frost injury,and where winter temperature is not low enough tovernalize winter type wheat. Genetic analysis forspring type landraces showed that the relativefrequency of four Vrn genes was different witheach other. Vrn3 was most widely and frequentlyfound among the four genes, followed by Vrn1 andVrn2. Only seven landraces proved to be thecarrier of Vrn4. The frequency was alsodifferent between localities. Genotype with Vrn1plus other dominant gene(s) adapted to spring sowingto avoid severely cold winter in north China, whilegenotype with only Vrn3 adapted to winter sowingin south China and southwest Japan. It is thereforeconcluded that at least three ecotypes, differing ingrowth habit and Vrn genotype, areallopatrically distributed in east Asia, as a resultof adaptation to winter coldness in each locality.

Notes: Abstract We have examined the effectiveness of similar numbers of markers from four molecular marker systems (AFLP, isozymes, ISSR and RAPD) for revealing genetic diversity and discriminating between infraspecific groups of Oryza sativa germplasm. Each marker system classifies the germplasm into three major groups (most effectively with isozymes and AFLPs), but with differences (primarily with ISSR) between the precise classifications generated. However, at the highest levels of genetic similarity there was only partial agreement as to relationships between individual accessions when different markers were used. When variance was partitioned among and within the three subspecific groups, although the differences were not significant, greater variation was found among than within groups using AFLP and isozymes, with the reverse for RAPD and ISSR. Measurement of polymorphism using average heterozygosity and effective number of alleles gave similar results for each marker system. These results are discussed in relation to various genetic resources conservation activities, and the advisability of extrapolating to other sets of germplasm particularly of other crop species.

Notes: Abstract Málaga is a province of Spain where olive-trees are cultivated in a large range of environments, climates and soils. We have developed a reliable and reproducible method to detect RAPD and AP-PCR polymorphisms, using DNA from olive-tree (Olea europaea L.) leaves. Starting from their natural orchards, fifty-six olive-tree cultivars throughout Málaga province, including oil and table olive cultivars, were screened and grouped into 22 varieties. A total of 62 informative polymorphic loci that provide 601 conspicuous bands were enough to differentiate the varieties. Clustering analyses managing 3 different pairwise distances, as well as phylogenetic analyses, led to the same result: olive-trees in Málaga can be divided into three main groups. Group I (90% of certainty) contains wild type and two introduced varieties, group II (83% of certainty) covers some native olive-trees, and group III (58% of certainty) is an heterogeneous cluster that includes varieties originating and cultivated in a number of Andalusian locations. Geographic location seems to be the first responsible of this classification, and morphological traits are needed to justify the group III subclustering. These results are consistent with the hypothesis of autochthonic origin of most olive-tree cultivars, and have been used to support a Label of Origin for the olive oil produced by the varieties included in group II.

Notes: Abstract Marker assisted selection (MAS) may improve the efficiency of breeding downy mildew resistant cucumber cultivars. A study was conducted to identify random amplified polymorphic DNA (RAPD) markers linked to the downy mildew resistance gene (dm) which would be suitable for MAS. A total of 145 F3 families from two populations (55 from the WI 1983G × Straight 8 population and 90 from the Zudm1 × Straight 8 population) were evaluated over five locations in North America and Europe. Resistant and susceptible F3 families were identified and mean family resistance ratings were used to type individual F2 plants. No evidence for race differences in the pathogen (Psuedoperonospora cubensis (Berk. & Curt.) Rostow) between North America and Europe was found. Phenotypic correlations between locations ranged from 0.3 to 0.7. Of the 135 polymorphic RAPD markers identified from 960 primers, five were linked to dm - G14800, X151100, AS5800, BC5191100, and BC5261000. In the WI 1983G × Straight 8 population, G14800 was linked to dm at 16.5 cm, AS5800 at 32.8 cm, BC5191100 at 9.9 cm, and BC5261000 at 19.2 cm. In the Zudm1 ×Straight 8 population, G14800 was linked at 20.9 cm, X151100at 14.8 cm, AS5800 at 24.8 cm, and BC526_1000 at 32.9 cm. MarkersG14800 and BC5191100 were linked in repulsion to the dm allele, and X151100, AS5800, and BC5261000 were linked in coupling phase. These genetic markers may be exploited to develop an efficient MAS strategy for breeding resistant cucumber cultivars.

Notes: Abstract Random amplified polymorphic DNA (RAPD) and simple sequence repeat (SSR) markers were used to characterize genetic relationships among 46 accessions in two C. melo L. subsp. melo (Cantalupensis, Inodorus) and subsp.agrestis (Conomon, and Flexuosus) groups. Genetic distance (GD) estimates were made among and between accessions in four melon market classes [Galia, Ogen, Charentais, and Shipper (European and U.S. types)] of Cantalupensis, one market class of Inodorus (Cassaba and Honey Dew), one accession of Conomon, and one accession of Flexuosus by employing three GD estimators; simple matching coefficient, Jaccard's coefficient, and Nei's distance-D. Differences detected among 135 RAPD bands and 54 SSR bands (products of 17 SSR primers) were used to calculate GD. Band polymorphisms observed with 21 RAPD primers and 7 SSR primers were important (p =0.01) in the detection of genetic differences. Estimators of GD were highly correlated (p 0.0001; rs = 0.64 to0.99) when comparisons were made between estimation methods within a particular marker system. Lower correlations (rs = 0.17 to 0.40) were detected (P 〉 0.001) between marker systems using any one estimator. The GD of the Conomon and Flexuosus accessions was significantly different (p〉 0.001)from the mean GD of all the market classes examined. The mean GD (Jaccard's coefficient) among accessions of Ogen, Galia, Cassaba, Charentais, European shipper, and U.S. shipper groups was 0.11 ± 0.04, 0.33± 0.09, 0.21 ± 0.04, 0.26 ± 0.10, 0.17± 0.05 and 0.22 ± 0.08, respectively. Market classes were distinct (p 〉 0.001), such that GDs between Galia and other accessions were the largest(mean GD 0.34 to 0.35), and GDs between Ogen and other accessions were the smallest (mean GD 0.29 to 0.30). Contrasts between the U.S. shipper cultivar Top Mark and accessions within any market class was relatively large (mean GD = 0.42 ± 0.06). Empirical estimations of variances associated with each marker type in the accessions examined indicated that, per band, lower coefficients of variation can be attained in the estimation of GD when using RAPDs compared to SSRs. Nevertheless, the genetic relationships identified using these markers were generally similar. The disparity between the analyses of the two markers made may be related to the amount of genome coverage which is characteristic of a particular marker system and/or its efficiency in sampling variation in a population. Results of RAPD marker analysis suggest that 80 marker bands were adequate for assessing the genetic variation present in the accessions examined.

Notes: Abstract Wild Arachis germplasm includes potential forage species, such as the rhizomatous Arachis glabrata and the stoloniferous A. pinto and A. repens. Commercial cultivars of A. pintoi have already been released in Australia and in several Latin American countries, and most of these cultivars were derived from a single accession of A. pintoi (GK 12787). Arachis repens is less productive as a forage plant than is A. pintoi. However, it can be crossed with A. pintoi, and thus has good potential as germplasm for the improvement of A. pintoi. Arachis repens is also used as an ornamental plant and ground cover. Many new accessions of these two stoloniferous species are now available, and they harbor significant genetic variability beyond that available in the few older accessions, previously available. Therefore, these new accessions need to be conserved, documented and considered in terms of their potential for crop improvement and direct commercial use. Sixty-four accessions of this new germplasm were analyzed using RAPD analysis. Most of the accessions of A. repens grouped together into a clearly distinct group. In general, the accessions from the distinct valleys of the Jequitinhonha, São Francisco and Paranã rivers did not group together, suggesting there is not a tight relation between dispersion by rivers and the geographic distribution of genetic variation in these species.

Notes: Abstract The development of a core collection, one which represents the genetic diversity of a crop with minimal redundancy and increases utility of the collection as a whole, is especially important as the funding for germplasm collections decreases. With limited resources, it is difficult to manage large germplasm collections and disperse genetically diverse germplasm to plant breeders. An algorithm was developed to assist in selection of core collections based on estimates of genetic distance. The criteria for selection of the maximum genetically diverse set were based on rankings of genetic distance between an accession with respect to all other accessions. Depending on the size core which a user wished, a zone around each selected accession was determined and no other accession within these limits was selected. The premise for the algorithm was that the genetic variability represented in the core must be representative of the distribution of genetic distances within the population of interest. In the present study, the algorithm was used with RAPD-marker-based estimates of genetic distance for 270 Theobroma cacao L. accessions and 134 Capsicum accessions that chose a set representing 18.5% of the population and representing the breadth of RAPD-based variation.

Notes: Abstract RAPD (Random Amplified Polymorphic DNA) variation was assessed in 42 accessions of azuki bean (Vigna angularis) including wild, weedy and cultivated races and in three accessions of two related species used as outgroups. A much lower level of genetic variation was observed in cultivated and weedy azuki beans compared to wild azuki bean. Wild azuki bean (V. angularis var. nipponensis) has relatively high genetic variation in subtropical highlands of Asia compared to the Far East. Although cultivated azuki bean has low RAPD variation, accessions from subtropical highlands and Southeast Asia showed different RAPD features compared to those of the Far East. It is hypothesized that the cultivated azuki bean has been derived from wild azuki bean in the Far East; the high variation in wild azuki bean has been created through its natural dissemination; and the relatively low variation in cultivated azuki bean has come about through human dissemination after genetic bottleneck reduced by domestication. In addition, high genetic diversity in wild azuki bean in subtropical highlands of Asia is regarded as an important genetic resource in azuki improvement.

Notes: Abstract Following a legume collection mission to south-west Turkey in 1996, five species of Trifolium were analysed for genetic variation within and between species in eleven morphological and flowering characters. The five species included two outcrossing species, T. michelianum and T. resupinatum, and three inbreeding species, T. clypeatum, T. glomeratum and T. tomentosum. The genetic diversity found was related to climate and edaphic factors. All five species showed significant amounts of genetic differentiation between sites and the species could be separated morphologically by principal components analysis and cluster analysis. The most significant source of genetic variation was found to be related to geographical distribution with those species which were widely distributed across south-west Turkey exhibiting much greater amounts of genetic variation between sites, than those which had a narrow distribution. The breeding system was found to be less important, but only the morphology of the outbreeding species showed any environmental clines in relation to climate. A multiple regression analysis was computed to estimate the effect of growing season on the days to flowering of each of the species.

Notes: Abstract The extent and patterns of distribution of genetic variation among 80 sorghum (Sorghum bicolor (L.) Moench) germplasm accessions from Ethiopia and Eritrea were investigated using RAPD with 20 oligonucleotide primers. The primers generated a total of 147 polymorphic bands across the 80 accessions with a mean of 7.35 bands per primer. Estimation of the extent of variation by the Shannon-Weaver diversity index revealed an intermediate level of overall variation (H = 53), although the levels varied among regions of origin of the accessions. Partitioning of the total variation revealed considerable variation (77%) within the region of origin of the accessions and the remainder (23%) among regions of origin. Similarly, a large portion (94%) of the total variation was found within the adaptation zones compared to among the adaptation zones (6%). The results suggest a weak differentiation of the sorghum material both on regional and agro-ecological bases, which could be ascribed to the high rate of outcrossing in cultivated sorghum and its free natural hybridization with its wild and weedy relatives, as well as to seed movement by humans. The average genetic dissimilarity was found to be 36% among the 80 accessions and 13% among the 15 regions of origin. Cluster analysis failed to group accessions of the same region or the same adaptation zone, which further confirmed the weak differentiation of the material studied. The clustering pattern of the regions of origin was broadly concordant with previous clustering patterns obtained using morphological characters, in which regions with broad agro-climatic conditions were grouped together.

Notes: Abstract Genetic characterization of germplasm is important for setting objective guidelines for conservation. One common problem found in genebanks is determining the value of populations with insufficient or unreliable data regarding their geographic origin. In this study, a genetic analysis based on RAPD markers was conducted to characterize a `mystery' population of Solanum sucrense, a polysomic tetraploid potato (2n=4x=48), for which adequate documentation was lacking. The comparative analysis of genetic similarities between this mystery population and each one of 30 other S. sucrense populations in the genebank revealed that all populations within this species, including the mystery population, are significantly different from being duplicates, and are therefore worthy of separate conservation. RAPD markers also distinguished the mystery population from closely related tetraploid species S. oplocense, S. gourlayi and S. tuberosum ssp. andigena, suggesting that it is also not a duplicate of a population of these species. If RAPDs can clearly differentiate populations within highly heterogeneous tetraploids like S. sucrense, they should be generally useful for determining germplasm organization within potato species.

Notes: Abstract DNA from twenty-three late maturing cultivars of Guinea yams (D. cayenensis/D. rotundata complex) from the Benin Republic that could not be separated using isozyme markers, were examined using randomly amplified polymorphic DNA (RAPD) markers with decamer primers of arbitrary sequence. All the twelve primers tested were informative and yielded 63 amplified DNA bands from which 47 (75%) were polymorphic. Although no single primer produced polymorphic bands in all cultivars, the great majority of the cultivars were separated with the combinations of polymorphic bands generated by various primers. Putative duplicates and cultivar misclassifications were identified. Many morphologically distinct cultivars were close. The dwarf cultivar Tam-Sam considered as derived from Tabane, appeared more distant from the latter than was believed. RAPD analysis was found as a practical tool for the identification of duplicates toward establishment of an accurate core collection of Guinea yams in Benin Republic and in the other countries of the African yam belt.

Notes: Abstract Variation for 24 morphological traits measured in a greenhouse environment and 36 randomly amplified polymorphic DNA (RAPD) markers was assessed among 318 wild octoploid strawberry (Fragariaspp.) genotypes from diverse habitats across the northern USA. RAPD marker frequencies and certain leaf and flower morphology traits (petiole color, leaf mass/area ratio, leaflet length and width, flower and receptacle diameter, petal width, flowers/inflorescence) were significantly different between the F. chiloensis-platypetala and F. virginiana-glauca species complexes. The proportion of variation accounted for by provenance effects was lower for the RAPD markers than for most morphological traits, especially in the F. virginiana-glauca species complex. Morphological traits of potential adaptive importance group the collection into provenances within each species-complex, and reflect the significant habitat and geographic differences across the region from which the germplasm was collected. Variation among populations within provenances was low for the molecular and most morphological traits, with a much larger amount of variability among plants within populations. Most of the variation for the presumably more selectively-neutral RAPD data was among plants within populations and populations within provenances rather than among the provenances that were recognized based on morphological traits, especially in the F. virginiana-glauca complex. Patterns of diversity for morphological traits must be considered, along with more selectively-neutral molecular characters such as RAPDs, to formulate effective sampling strategies and to properly estimate the quantity and apportionment of diversity within this germplasm.

Notes: Abstract Twenty-five Pyrus communis L. cultivars including eight traditional Portuguese pears, and four commercial Pyrus pyrifolia (Burm.) Nak. (Japanese pear or `nashi') cultivars were analysed by RAPD and AFLP techniques focusing on their molecular discrimination and the assessment of their genetic relatedness. Twenty-five primers generated 324 RAPD markers, among which 271 (84%) were polymorphic. The AFLP technique, using seven primer combinations, revealed a similar level of molecular polymorphisms (87%), representing 418 polymorphic bands among a total of 478 scored in autoradiographs. The high reproducibility of RAPD and AFLP techniques was confirmed comparing DNA samples from different extractions and different digestions of DNA from the same plant. Three genetic similarity matrices and respective dendrograms were elaborated on using RAPD, AFLP or joint RAPD and AFLP data. Both molecular marker techniques proved their reliability to assess genetic relationships among pear cultivars. P. pyrifolia cultivars exhibit a closer genetic relatedness, clustering apart from P. communis cultivars. Within P. communis, `William's', as well as `Doyenne du Comice', cluster close to their hybrids. Most of the Portuguese cultivars tend to cluster together, indicating to constitute a relatively independent genetic pool, which can be of interest in pear breeding programs.

Notes: Abstract The present study, using RAPD analysis, was undertaken to characterize genetic variation in three forms of V. angularis, cultivated, wild and weedy forms, and their relationships. The materials used consisted of 171 individuals (plants) or cultivars from 23 populations including 5 wild populations, 6 weedy populations, 6 cultivated populations and 6 populations with plants having wild and weedy or intermediate morphology, denoted here as complex populations. The materials used were collected on Honshu Island, Japan and seeds collected directly from the field were germinated for DNA extraction. In addition, 6 landrace accessions of V. angularis from the genebank were also analyzed. Genetic variation was highest in the wild form (Hg= 0.132; GD = 0.388), followed by the weedy form (Hg= 0.124; GD = 0.341) and the least in the cultivated form (Hg= 0.079; GD = 0.274). Intra-population genetic variation was high in the weedy and in the wild populations. However, inter-population was greater than intra-population genetic variation for all groups of populations studied in the V. angularis complex. 93% of the total diversity in the present study was exhibited by plants from complex populations and specific RAPD bands were found in these populations. Our results provide evidence that complex populations would be a logical focus for efforts to conserve the V. angularis complex in situ. Our results suggest that weedy populations are usually an ecotype of the wild form adapted to a different habitat.

Notes: Abstract To assess the generic delimitation and the interspecific relationships between Hystrix and Elymus, three Hystrix and 10 Elymus species were used for random amplified polymorphic DNA(RAPD) assay. Of the 54 primers tested, 26 (48%) produced polymorphic products. A total of 167 products amplified from 16 primers were selected for RAPD analysis, among which 156 (93.4%) amplified products were found to be polymorphic among the 13 species. The polymorphism produced by each primer ranged from 4 to 13, with an average of 9.8. Data were used to generate Jaccard's similarity coefficients and to construct a dendrogram using UPGMA in the NTSYS computer programs. It is concluded from this study that: (1) there were clear differences between Hystrix and Elymus, which possibly suggest that Hystrix is a valid genus; (2) great diversity existed among the species of Hystrix and Elymus; (3) the species similar to each other in morphological characters were grouped together; (4) the species from neighboring geographical regions were clustered; (5) the species with the same genomes and polyploidy level were clustered together; (6) RAPD results are comparable with those obtained from studies on morphology and cytology. It is a useful additional method for assessing the relationships among genera and species in Triticeae.

Notes: Abstract The internal transcribed spacers ITS1 and ITS2 in the18S-5.8S-26S rDNA repeat units were amplified and cloned from Angelica gigas Nakai, Angelica acutiloba (Siebold & Zucc.) Kitagawa, A. dahurica Maxim, Angelica decursiva (Miq.) Franch. & Savat, Bupleurum falcatum L. and Peucedanum japonicum Thunb. Sequence analyses showed that ITS1 is approx. 215 bp, the 5.8S gene is 162 bp and the ITS2 approx. 221 bp in all six species. The sequences are deposited at the EMBL Nucleotide Database. By including these new sequences in the Apiaceae phylogenetic tree, a third branch consisting of P. japonicum, A. gigas, P. decursivum and A. decursiva is added to theAngelica clade. Peucedanum does not forma distinct branch. The sequence obtained from Angelica dahurica collected in S. Korea is identical to that reported for the same species originating from China. A Bupleurum clade of three species was added to the tree showing closer relationship to theDaucus Laserpitium clade than the Angelica clade. RAPD analysis of all six species showed that the 10-base primer OPC-17 only, out of the20 Kit-C primers from Operon gave polymorphic banding patterns.

Notes: Abstract RAPD and PCR-RFLP analyses were conducted to establish the phylogenetic relationships among collected accessions of shallot and Allium × wakegi, and to assess the origin of A. × wakegi. Twenty out of 100 primers were amplified with 112 scorable bands for cluster analysis. Two main cluster groups consisting of one group for shallot and another group for A. × wakegi were clearly separated. The sub-groups of clusters reflected the phenotype differentiation in shallots and regional specificity in some A. × wakegiaccessions. The present results were also in agreement with previous systematics by isozymes of which the highest genetic variation of A. × wakegi in Indonesia was found in West Java suggesting the possibility that this place might be one of the germplasm centers. From RFLP analysis of amplified matK gene of cpDNA it was demonstrated that A. × wakegi originated from shallot as a maternal plant × Welsh onion as a paternal plant as well as from reciprocal crosses.

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

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

Notes: Abstract It is indicated that up to the year 2030, the annual average temperatures in China will increase by 0.88 to 1.2°C, with increments in the south less than in the north. Annual average precipitation would raise slightly, but the increment could be 4% in northeastern China. The increment of annual mean runoff could rise over 6% in the northeastern area, and decrease in the other regions 1.4 to 10.5%. The increased water shortage due to climate change could achieve 160 to 5090 million m3 in some areas of China. Financial loss due to the lack of water could reach 1300 million yuan, and up to 4400 million yuan in serious drought years in the Beijing-Tianjin-Tangshan area.

Notes: Abstract Adaptation in response to anthropogenic climate change seeks to maintain viability by maximising benefits and minimising losses. It is necessary because some climatic change is now inevitable, despite the international focus on mitigation measures. Indeed, the measures agreed at Kyoto would by themselves result in only a small reduction in the climate changes to be expected over the next century. Discussion of the expected changes and possible impacts leads to the following conclusions regarding climate change scenarios in relation to impacts and adaptation: • Climate change in the foreseeable future will not be some new stable "equilibrium" climate, but rather an ongoing "transient" process; • Climate change predictions relevant to impacts on most sectors and ecosystems are still highly uncertain; • There is a need for a greater focus on developing countries and tropical regions, and on relevant key variables, including the magnitude and frequency of extreme events; • The focus should shift from single predictions, or extreme ranges of uncertainty, to risk assessment; • Thresholds critical to impacted sectors and ecosystems should be identified, and expressed as functions of climatic variables; • Planned adaptations will be necessary to cope with multiple stresses, including those due to non-climatic changes; • A major task of adaptation science is to identify the limits of adaptation, i.e., to identify "dangerous levels of greenhouse gases" beyond which adaptation becomes impractical or prohibitively expensive.

Notes: Abstract If current trends continue, human activities will drastically alter most of the planet's remaining natural ecosystems and their composite biota within a few decades. Compounding the impacts on biodiversity from deleterious management practices is climate variability and change. The Intergovernmental Panel on Climate Change (IPCC) recently concluded that there is ample evidence to suggest climate change is likely to result in significant impacts on biological diversity. These impacts are likely to be exacerbated by the secondary effects of climate change such as changes in the occurrence of wildfire, insect outbreaks and similar disturbances. Current changes in climate are very different from those of the past due to their rate and magnitude, the direct effects of increased atmospheric CO2 concentrations and because highly modified landscapes and an array of threatening processes limit the ability of terrestrial ecosystems and species to respond to changed conditions. One of the primary human adaptation option for conserving biodiversity is considered to be changes in management. The complex and overarching nature of climate change issues emphasises the need for greatly enhanced cooperation between scientists, policy makers, industry and the community to better understand key interactions and identify options for adaptation. A key challenge is to identify opportunities that facilitate sustainable development by making use of existing technologies and developing policies that enhance the resilience of climate-sensitive sectors. Measures to enhance the resilience of biodiversity must be considered in all of these activities if many ecosystem services essential to humanity are to be sustained. New institutional arrangements appear necessary at the regional and national level to ensure that policy initiatives and research directed at assessing and mitigating the vulnerability of biodiversity to climate change are complementary and undertaken strategically and cost-effectively. Policy implementation at the national level to meet responsibilities arising from the UNFCCC (e.g., the Kyoto Protocol) and the UN Convention on Biological Diversity require greater coordination and integration between economic sectors, since many primary drivers of biodiversity loss and vulnerability are influenced at this level. A case study from the Australian continent is used to illustrate several key issues and discuss a basis for reform, including recommendations for facilitating adaptation to climate variability and change.

Notes: Abstract Global climate change is likely to have a range of consequences for human health as a result of disturbance or weakening of the biosphere's natural or human-managed life support systems. The full range of potential human health impacts of global climate change is diverse and would be distributed differentially spatially and over time. Changes in the mortality toll of heatwaves and changes in the distribution of vector-borne infectious diseases may occur early. The public health consequences of sea level rise and of regional changes in agricultural productivity may not occur (or become apparent) for several decades. Vulnerability is a measure of both sensitivity to climate change and the ability to adapt in anticipation of, or in response to, its impacts. The basic modes of adaptation to climate-induced health hazards are biological, behavioural and social. Adaptation can be undertaken at the individual, community and whole-population levels. Adaptive strategies should not introduce new health hazards. Enhancement of the acknowledged public health infrastructure and intervention programmes is essential to reduce vulnerability to the health impacts of climate change. In the longer-term, fundamental improvements in the social and material conditions of life and in the reduction of inequalities within and between populations are required for sustained reduction in vulnerability to environmental health hazards.

Notes: Abstract A mechanism has been established to improve integration of international climate-related programmes. Known as the Climate Agenda it outlines a programme, that in a cost-effective way, responds to national obligations to respond to international agreements as well as their national needs for social and economic development. The paper briefly describes the Climate Agenda and the incorporation within it of studies of climate impact assessments and response strategies to reduce vulnerability. The need for increased emphasis on climate impact assessment and for the development of effective adaptation measures is emphasised following the elaboration of a Kyoto Protocol to the United Nations Framework Convention on Climate Change (UNFCCC). UNEP's efforts to identify national programmes contributing to the Climate Agenda are described. The response from developing countries has, however, been very disappointing, even from countries where we know work is ongoing through funding by GEF or US Country Studies Program and other bilateral programmes. Initial compilation of information available so far shows that many developed countries are putting a lot into the area of impact assessment of not just climate change, but also climate variability. There remain some research gaps, especially in the area of assessment of climate impacts on ecosystems, hydrological systems, etc. Considerable efforts are presently being directed at reduction of emissions of greenhouse gases and in the case of developing countries, most efforts are being directed towards completing national communications and providing baseline data for future studies. The paper refers to early activities by UNEP in cooperation with other international organizations to undertake integrated assessments of the impacts of climate change on important socio-economic sectors and the later incorporation of lessons learned into the IPCC Guidelines for Assessing Impacts of Climate Change. Later sections outline the development of a handbook on methods for climate change impact assessment and adaptation strategies as a practical approach to national assessments and the development of appropriate and cost-effective response to climate change. The initiation of a GEF-funded project to apply the methods contained in the handbook and improve the results based on national studies is also described for both developed and developing countries. Working in collaboration with a team of international experts under the coordination of the Institute of Environmental Studies at Vrije University (Amsterdam), the goal of this ongoing project is to develop a valuable methodological tool that Parties to the UNFCCC may apply to develop national climate change impact and adaptation assessments. Development of these guidelines was linked to a series of country studies in Antigua and Barbuda, Estonia, Cameroon and Pakistan funded under a UNEP/GEF project. The application of the first version of the UNEP Handbook by national study teams in these four countries is making valuable technical and practical contributions and will ensure that the next version of the Handbook will be a more useful tool for experts in developing countries undertaking similar studies in the future. The methods contained in the Handbook are also the basis for similar assessments funded under bilateral development programmes in other countries. These and similar studies elsewhere are coordinated with the UNEP programme and will eventually aim to create reliable and comparable assessments, a compatible set of tools for such purpose and the identification of realistic adaptation options for incorporation into national planning for adapting to climate change. The paper also addresses how climate impact assessment and response strategies are undertaken as part of national enabling activities carried out in co-operation with UNEP.

Notes: Abstract Many developing countries, especially in Africa, contribute only very small amounts to the world total of greenhouse gas emissions. For them, the reduction of such emissions is not a priority, and the more important issue is to find ways to reduce their vulnerability to the projected climate change which is being imposed upon them largely as a result of emissions from developed countries. This priority does not accord with the ultimate objective of the United Nations Framework Convention on Climate Change, which is to achieve stabilization of greenhouse gas emissions. This paper reports upon studies in Uganda designed to help in the development of a national adaptation strategy, and addresses the need to reconcile such a strategy with the global priority accorded to mitigation and with national economic development priorities. Some features of a national climate change adaptation strategy are identified and questions are raised about the need for an international regime to facilitate and support adaptation.

Notes: Abstract Climate change impacts on African human settlements arise from a number of climate change-related causes, notably sea level changes, impacts on water resources, extreme weather events, food security, increased health risks from vector home diseases, and temperature-related morbidity in urban environments. Some coastlines and river deltas of Africa have densely populated low-lying areas, which would be affected by a rise in sea level. Other coastal settlements will be subjected to increased coastal erosion. Recent flooding in East Africa highlighted the vulnerability of flood plain settlements and the need to develop adaptive strategies for extreme weather events management and mitigation. In the semi arid and arid zones many settlements are associated with inland drainage water sources. Increases in drought will enhance water supply related vulnerabilities. Inter-basin and international water transfers raise the need for adequate legal frameworks that ensure equity among participating nations. Similarly, water supply and irrigation reservoirs in seasonal river catchments might fail, leading to poor sanitation in urban areas as well as food shortage. Hydroelectric power generation could be restricted in drought periods, and where it is a major contributor to the energy budget, reduced power generation could lead to a multiplicity of other impacts. States are advised to develop other sources of renewable energy. Temperature changes will lead to altered distribution of disease vectors such as mosquitoes, making settlements currently free of vector borne diseases vulnerable. Rapid breeding of the housefly could create a menace associated with enteric disorders, especially in conditions of poor sanitation. The dry savannahs of Africa are projected as possible future food deficit areas. Recurrent crop failures would lead to transmigration into urban areas. Pastoralists are likely to undertake more trans-boundary migrations and probably come into conflict with settled communities. Adaptive measures will involve methods of coastal defences (where applicable), a critical review of the energy sector, both regionally and nationally, a rigorous adherence to city hygiene procedures, an informed agricultural industry that is capable of adapting to changing climate in terms of cropping strategies, and innovations in environment design to maximise human comfort at minimum energy expenditure. In the savannah and arid areas water resource management systems will be needed to optimise water resource use and interstate co-operation where such resources are shared. Climate change issues discussed here raise the need for state support for more research and education in impacts of climate change on human settlements in Africa.

Notes: Abstract Unique DNA bands from strains representative of two groups of Pseudomonas corrugata, as shown by amplification of their genomic DNA by polymerase chain reaction using short random sequence oligonucleotide primers (RAPD-PCR), were isolated, cloned and sequenced. Two pairs of specific primer sequences, based on the ends of the cloned unique DNA bands from strains IPVCT10.3 and IPVCT8.1, were used in multiplex PCR with a range of P. corrugata strains. All strains produced one of the two specific bands, 1100 bp (from the IPVCT10.3-based primers) and 600 bp (from the IPVCT8.1-based primers), representing groups designated I and II, respectively. The primers were also tested on a wider range of Pseudomonas species, including the closely-related fluorescent Pseudomonas genomospecies FP1, FP2 and FP3: none of these bacteria produced any bands following amplification by PCR with these primers. The primer sets detected P. corrugata in tomato pith necrosis-infected plants providing a useful tool for rapid identification and epidemiological studies.

Notes: Abstract The relationships between strains of Pseudomonas savastanoi pv. phaseolicola (P. sav. phaseolicola), P. syringae pv. tabaci (P. syr. tabaci) and P. syr. syringae which all cause disease on bean; the related species P. sav. glycinea and P. syr. actinidiae, and reference bacteria, were evaluated by studying the phenotypic and genetic diversity of a collection of 62 strains. All the P. sav. phaseolicola strains tested produced characteristic watersoaked lesions on bean pods. Other pathovars produced varying combinations of symptoms including necrotic lesions, with or without watersoaked centres and sunken tissue collapse of the lesion (P. syr. tabaci) and necrotic lesions with or without sunken collapse (P. syr. syringae). At the genomospecies level, all the strains of P. sav. phaseolicola, P. sav. glycinea and P. syr. tabaci, belonging to genomospecies 2, could be separated from P. syr. syringae strains (genomospecies 1) and P. syr. actinidiae strains (unknown genomospecies) by BOX-PCR and DNA/DNA hybridisation. To distinguish P. sav. phaseolicola, within genomospecies 2, from P. sav. glycinea and P. syr. tabaci, it was necessary to perform nutritional characterisations myo-inositol negative and p-hydroxy benzoate positive for P. sav. phaseolicola strains), PCR with specific primers designed from the tox region (positive for all of the P. sav. phaseolicola strains) and serotyping, as 71% of the P. sav. phaseolicola strains reacted as O-serogroup PHA1. Important intrapathovar variation was seen by genomic fingerprinting with REP and ERIC primers, as well as with RAPD primers (AE7 and AE10) and esterase profilings. While RAPD fingerprinting detected variability correlated with two race-associated evolutionary lines, REP, ERIC and esterase profiles revealed intrapathovar variation linked to some host origins, that separated the kudzu isolates, and the mungbean isolates, from the other P. sav. phaseolicola strains.