Marker-assisted optimization of an expert-based strategy for the acquisition of modern lettuce varieties to improve a genebank collection

To regularly improve the composition of the lettuce collection of the Centre for Genetic Resources, the Netherlands (CGN) with modern varieties, feedback from crop experts is used to select approximately 10% of the new material for incorporation in the collection. In the present study, assessments of six experts were compared to microsatellite data of 414 new varieties and 1408 existing accessions. Based on the microsatellite data, the extent to which the genetic diversity of the collection would be enriched (added value) was calculated for specific sets of new varieties. When individual assessments of experts were evaluated, the total added value of expert-based selections was not significantly higher compared to randomly chosen groups, except for a single expert. Unfamiliarity with new varieties was shown to be a crucial factor in the assessment of crop experts. According to the current acquisition protocol that seeks for consensus among experts, varieties are selected based on recommendations from at least three experts. This protocol also did not perform better than randomly chosen groups of new varieties. However, significantly better results were obtained with alternative protocols. It was concluded that breeding value was a more decisive criterion in the current acquisition protocol than maximal extension of the genetic diversity within the collection. A modified protocol addressing both commercial and diversity aspects was suggested in order to meet the demands of plant breeders as well as conservationists.     

Analysis of genetic diversity in the endangered tropical tree species <Emphasis Type="Italic">Hagenia abyssinica</Emphasis> using ISSR markers

Genetic diversity within and among 12 populations of the dioecious tropical tree species Hagenia abyssinica (Bruce) J.F. Gmel. in Ethiopia was examined with eight inter simple sequence repeat (ISSR) primers. A total of 104 clearly scorable bands were generated, among which 84 (81%) were polymorphic. Jaccard similarity coefficient was calculated for pairwise comparisons among all 120 individuals and ranged from 0.30 to 0.88 while average within-population similarity ranged from 0.53 to 0.66. Within-population variability was estimated as percentage polymorphic loci (ranging from 52% to 87%), Shannon’s information index (0.30–0.50) and Nei’s genetic diversity (0.21–0.35). The highest variability values were obtained for one recently planted population and for one wild population growing in an undisturbed primary forest area. Significant overall differentiation among populations was detected by both Shannon’s information index (0.26) and G _ST (0.25). Relatedness among samples was estimated with a principal coordinate analysis, and relatedness among populations was estimated with a cluster analysis (UPGMA). A Mantel test indicated a significant association between genetic and geographic distances, and an autocorrelation analysis showed significant evidence of gene flow over distances up to 30 km. This study is the first of its kind for H. abyssinica, which has decreased recently in Ethiopia and now must be regarded as an endangered species. Both within-population and between-population diversity estimates are typical of outcrossing, longlived and late successional species, suggesting that recent anthropogenic disturbances have not yet had much impact on population genetic parameters. DNA marker data can, however, be used to identify the most suitable sites for in situ conservation and for collection of material for establishment of genebanks and plant improvement programs.     

Genetic variation of Ethiopian and Eritrean sorghum (Sorghum bicolor (L.) Moench) germplasm assessed by random amplified polymorphic DNA (RAPD)

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.     

Comparison between the radical scavenging activity and antioxidant activity of six distilled and nondistilled mediterranean herbs and aromatic plants

Thirty-six different extracts of six herbs and aromatic plants (fennel, common melilot, milfoil, lavandin cv. Super, spike lavender, and tarragon) were evaluated for their radical scavenging activity by the DPPH*, NBT/hypoxanthine superoxide, and *OH/luminol chemiluminescence methods, and for their antioxidant activity by the beta-carotene blenching test. The total phenolic content was also determined by the Folin-Ciocalteu method. The plant material included cultivated plants and their wastes after being distilled for essential oils. Both remarkably high phenolic content and radical scavenging activities were found for the ethyl acetate and dichloromethane fractions among the different plant extracts. In general, the distilled plant material was found to exhibit a higher phenolic content as well as antioxidant and radical scavenging activities than the nondistilled material. Ethyl acetate and dichloromethane extracts, and even some crude extract, of both distilled and nondistilled plants exhibited activities comparable to those of commercial extracts/compounds, thus making it possible to consider some of them as a potential source of antioxidants of natural origin.     

Flavonoids of zoysiagrass (Zoysia spp.) cultivars varying in fall armyworm (Spodoptera frugiperda) resistance

Flavonoid profiles of 12 zoysiagrass (Zoysia spp.) cultivars sampled six times in 1998 were correlated to fall armyworm (Spodoptera frugiperda J. E. Smith) larval weights and survival on replicated field-grown plant material and analyzed to determine genetic and seasonal variations of flavonoids among zoysiagrass cultivars. From multiple regression analyses and correlations, flavonoid peak 10 (luteolin-glucoside) had the greatest positive association with average fall armyworm weight; however, resistance appeared to be correlated with a number of other flavonoids. The flavonoid profiles of cultivars subjected to clustering procedures showed consistent genetic variability for five of six samplings and was used to genotype 23 cultivars. The dendrogram supported the results of the FASTCLUS procedure in clustering certain genotypes such as fall armyworm-resistant Cavalier and Zeon together, as well as J-36 and Meyer. Flavonoid evaluations measure genetic relatedness among cultivars and could be used for selective breeding of resistance to fall armyworm.     

The RAPD evidence for the phylogenetic relationship of the closely related species of cultivated apple

The phylogenetic relationship of cultivated apple and its closely related species is still not clear in the taxonomy of genus Malus. To try to find new evidence for the origin and evolution of the cultivated apple, random amplified polymorphic DNA (RAPD) markers of 14 taxa of Malus, among which a reference species (M. toringoides) and six presumably ancestral species of cultivated apple in the genus were investigated. The RAPD data obtained were used to construct both unrooted and rooted trees using TREECON software package. The result showed in our rooted tree that M. sieversii from the Xinjiang Autonomous Region of China is the species which is most closely related to the cultivated apple, M. domesticacv. `Golden Delicious'. The phylogenetic relationships among the species studied are discussed.     

Isozyme studies of genetic diversity and evolution in Hippophae

Summary To provide information on the genetic variation, differentiation and evolution in Hippophae seed samples of 25 populations from China, Finland and Russia were electrophoretically analyzed. Of six loci investigated, four were good genetic markers for identifying species and subspecies. The percentages of polymorphic loci per population were 40.3% and 62.5% at 0.95 and 0.99 polymorphic criteria respectively. The mean number of alleles per locus per population was 2.1. Total genetic diversity in the material was 0.4614. Genetic diversity partitioning showed that there was a large amount of diversity residing within geographical populations (0.1354), between subspecies within species (0.1046) and between species (0.2566) but not between geographical populations (0.0114). There were nearly twice as many negative fixation indices as positive ones in Hippophae populations. The phylogenetic tree agreed very well with botanic classifications of the species and subspecies and their geographical distributions, and quantitatively presented the genetic relationships of 25 populations. A detailed view of the evolutionary stages in Hippophae showed clearly a general decline of similarity as evolutionary divergence continued, which further explained the evolution process in Hippophae.     

Evaluation of Genetic Diversity of Wild Rice (Oryza rufipogon Griff.) in Myanmar using Simple Sequence Repeats (SSRs)

As a part of an in situ survey of wild rice (Oryza rufipogon Griff.) in Myanmar (Burma), 16 strains of wild rice were collected, and analyzed for allelic diversity over 74 loci with simple sequence repeat (SSR) markers to obtain a basic information for their conservation. Three each of indica and japonica cultivars were added for a comparison. In the six cultivars and 16 strains of wild rice, three to 15 alleles were detected per locus with an average of 7.9. The wild rice revealed a large number of unique alleles throughout their chromosomes with much wider ranges of variation than those detected in the six cultivars of O. sativa L.. The alleles found in the wild rice were classified into those specific to wild rice, common to wild rice and cultivars, and those similar to indica or japonica cultivars. According to the classification, the genotype of each of the 16 strains of wild rice was schematically depicted. The genetic variation among individual strains within a collection site was larger than the variation among the collection sites.     

深圳市土壤微量元素含量成因分析

对深圳地区９种母质和６种土类构成的１６个母质─土类单元共８３个土壤样品中１２种微量元素的含量进行侧定。在此基础上分析了微量元素含量分布的成因。研究结果表明，成土母质是影响土壤微量元素含量和分布的首要因素，相同母质上的不同类型土壤中的微量元素含量大多无明显差异。土壤中微量元素的共生组合关系表现出明显的母质继承性特征，由于微量元素主要以同晶替代方式分散在硅酸盐矿物中，其分布与离子半径关系密切。第四周期过渡元家具有相似的分布格局，而Ｐｂ、Ｚｎ、Ｃｄ等另成一类。     

Evaluation of the discriminance capacity of RAPD, isoenzymes and morphologic markers in apple (Malus x domestica Borkh.) and the congruence among classifications

Twenty-one apple accessions were characterised and classified taxonomically with randomly amplified polymorphic DNA (RAPD). The results were compared with morphological and isoenzymatic classifications of the same material by calculating the congruence between similarity matrices. The results confirmed the greater discrimination capacity of RAPD compared with isoenzymes, although the taxonomic clusters obtained with both types of markers were congruent. The genetic relationships calculated using RAPD and isoenzymes showed little congruence with the morphological relationships among accessions. Isoenzymatic markers are useful in germplasm banks to evaluate the diversity of newly introduced accessions. The use of RAPD markers are especially beneficial to discriminate between material that is genetically similar, to evaluate genetic variability within a collection and to choose the components of the core collection.     

Geographic variation in tetraploid wheat (Triticum turgidum spp. turgidum convar. durum) landraces from two provinces in Ethiopia

There is considerable wealth of genetical and morphological variation in tetraploid wheat collected from Ethiopia by past expeditions. Several authors have speculated on the reasons for this concentration of diversity so far removed from the centre of origin of wheats in West Asia. The present study reports results of evaluation carried out on material collected in the early 1970's from Ethiopia. The landraces from the two provinces Shewa and Tigray were found to be distinctly different. This divergence was attributed to the differences in environmental conditions between them. Wide differentiation among landraces within each province was also present. The Ethiopian farmer's practice of growing composite cultivars in the field has resulted in bulk samples of Ethiopian landraces containing several agrotypes. The proportion of total variance due to differences among agrotypes within landraces was by far the greatest found in this study, followed in most cases by the variation among landrace populations within provinces which exceeded, in turn, the variation between province gene pools. The optimal strategy for further germplasm collections and implications for improvement and conservation in locally-based community gene banks are discussed.     

Evaluation of some phosphoroamides as soil urease inhibitors

Summary The effectiveness of six phosphoroamides for retardation of urea hydrolysis in soils was studied by determining the effects of 10 g g^–1 soil of each compound on the amounts of urea hydrolyzed when soils treated with urea were incubated at 10°, 20°, 30°, and 40°C for 3, 7, and 14 days. The phosphoroamides used wereN-(diaminophosphinyl)-cyclohexylamine,N-benzyl-N-methyl phosphoric triamide, diethyl phosphoric triamide, trichloroethyl phosphorodiamidate, dimethyl phosphoric triamide, andN-butyl phosphorothioic triamide [N-(n-butyl) thiophosphoric triamide]. The soils used were selected to obtain a range in properties, and the effects of the six phosphoroamides studied were compared with those of two compounds known to be among the most effective compounds thus far proposed for retardation of urea hydrolysis in soils (phenylphosphorodiamidate and hydroquinone). The data obtained showed that all six of the phosphoroamides evaluated compared favorably with hydroquinone as soil urease inhibitors and that two of them [N-butyl phosphorothioic triamide andN-(diaminophosphinyl)-cyclohexylamine] were superior to phenylphosphorodiamidate for retardation of urea hydrolysis in soils at 20°, 30°, or 40°C.     

RAPD and ISSR fingerprinting in cultivated chickpea (<Emphasis Type="Italic">Cicer arietinum</Emphasis> L.) and its wild progenitor <Emphasis Type="Italic">Cicer reticulatum</Emphasis> Ladizinsky

Detection of genetic relationships between 19 chickpea cultivars and five accessions of its wild progenitor Cicer reticulatum Ladizinsky were investigated by using RAPD and ISSR markers. On an average, six bands per primer were observed in RAPD analysis and 11 bands per primer in ISSR analysis. In RAPD, the wild accessions shared 77.8% polymorphic bands with chickpea cultivars, whereas they shared 79.6% polymorphic bands in ISSR analysis. In RAPD analysis 51.7% and 50.5% polymorphic bands were observed among wild accessions and chickpea cultivars, respectively. Similarly, 65.63% and 56.25% polymorphic bands were found in ISSR analysis. The dendrogram developed by pooling the data of RAPD and ISSR analysis revealed that the wild accessions and the ICCV lines showed similar pattern with the dendrogram of RAPD analysis. The ISSR analysis clearly indicated that even with six polymorphic primers, reliable estimation of genetic diversity could be obtained, while nearly 30 primers are required for RAPD. Moreover, RAPD can cause genotyping errors due to competition in the amplification of all RAPD fragments. The markers generated by ISSR and RAPD assays can provide practical information for the management of genetic resources. For the selection of good parental material in breeding programs the genetic data produced through ISSR can be used to correlate with the relationship measures based on pedigree data and morphological traits to minimize the individual inaccuracies in chickpea.     

Phenotypic diversity for qualitative and phenologic characters in germplasm collections of tef (Eragrostis tef)

A total of 3600 entries of tef [Eragrostis tef (Zucc.) Trotter] germplasm collections representing 36 populations from six regions of Ethiopia were evaluated for phenotypic diversity in six qualitative and phenologic characters. High values of Shannon–Weaver diversity index ( ) were recorded for most characters in each population. Monomorphism was high for anther color (purple) and panicle form (loose). Larger variances for were obtained among regions than among populations within a region from hierarchical analysis of variance. Significant (P<0.05) to highly significant (P<0.01) t-values were observed among values of some regions for seed color, panicle form and days to maturity. A substantial level of variance (P<0.05) was obtained for populations within altitudinal zones only for days to maturity. Altitudinal zones exhibited similar levels of diversity across characters for these collections. Generally, considerable variations important for tef improvement work have been observed and regions with highest diversity for some traits have been suggested for future in situ or ex situ germplasm conservation works. Although similar mean diversity indices were obtained among regions, they were non overlapping.     

Isozyme diversity and analysis of the mating system of the wild rice Oryza latifolia Desv. in Costa Rica

Oryza latifolia is the most abundant and widespread wild relative of rice in the lowlands of Costa Rica. It also has the most diverse morphology. However, little is known about the genetic diversity and mating system of this allotetraploid species. Genetic analyses of nine populations from different life zones from the Pacific and Atlantic slopes were performed using six isozyme loci. Differences in genetic variability were observed among populations, of which Palo Verde and Santa Rosa were the most diverse. Most Atlantic populations clustered together, and a similar result was obtained with the Pacific populations. High levels of interpopulation diversity were observed while most populations were monomorphic for at least one genotype, suggesting little genetic flow within populations. Even in polymorphic populations no variation was observed within progeny groups, combined with Hardy-Weinberg disequilibria in most populations, commonly observed in autogamous species or in species with clonal reproduction. However, the high frequency of heterozygous-like patterns may suggest that the reproductive system of O. latifolia might be more complex. Future research may explain the genetic patterns as well as the reproductive biology of this species. The knowledge of the genetic diversity and mating system of O. latifolia could contribute to the implementation of genetic resource conservation strategies and gene flow analyses, as well as of breeding programs for rice improvement.     

Molecular characterization of mulberry genetic resources indigenous to India

Mulberry (Morus L.) is essential for the sericulture industry as the primary feed for the silkworm Bombyx mori L. in India, with its long tradition of practicing sericulture, has a large number of indigenous cultivars. Since knowledge on the genetic divergence of these cultivars/varieties is essential for proper conservation and utilization, Inter Simple Sequence Repeat (ISSR) profiling was employed to assess genetic relationships among 34 mulberry accessions, collected from different regions of India. By using 12 ISSR primers, which produced 72 markers displaying a high degree of polymorphism (94.4%), genetic dissimilarity coefficients were calculated for each pair of the accessions and clustering of the accessions with Unweighted Pair Group Method using Arithmetic average (UPGMA) analysis was done to unravel the genetic diversity among the accessions. The dissimilarity coefficients varied from 0.111 to 0.692. UPGMA analysis generated a dendrogram with six groups and five isolates. Clustering of the accessions did not correspond with the information on the geographic origin of many of the accessions. Two-dimensional representations of the relative positions of the accessions with regards to divergence using the first two canonical variates as co-ordinate axes revealed considerable variability among the cases in each group. Further, Discriminant Function Analysis (DFA) showed changes in the geographic origin of 11 accessions and species status of 20 accessions.     

New sources of resistance to race Ro1 of the Golden nematode (Globodera rostochiensis Woll.) among reputed duplicate germplasm accessions of Solanum tuberosum L. subsp. andigena (Juz. et Buk.) Hawkes in the VIR (Russian) and US Potato Genebanks

Cultivated Solanum tuberosum L. subsp. andigena is well known as a rich source of valuable traits for potato breeding, especially for resistance to diseases and pests. The golden potato cyst nematode, Globodera rostochiensis Woll., is considered to be one of todays most serious hindrances to potato production in Europe and North America. Thus, the breeding of new cultivars that have resistance to PCN is of great importance. The USPG (USA) and VIR (Russian) potato genebanks, as well as others, maintain many samples of primitive cultivated and wild potato species originating from Latin America. Many of these samples are assumed to be genetically duplicated because the material in both genebanks came from the same original source. A joint investigation of new genotypes of subsp. andigena forms resistant to potato cyst nematode (PCN) was carried out on samples of subsp. andigena at VIR with reputed duplicate samples at USPG. After careful screening, 14 samples which possessed resistance to PCN were identified. A high level of this resistance was transmitted to sexual progeny at a high frequency for all of the selections. Eleven of the accessions found to be resistant have reputed duplicates in USPG that were not previously known to be resistant. Thus, this research not only broadens the choice of parents available for resistance breeding, but identifies model materials for future research to test the parity of PCN resistance among reputed duplicate samples in the two genebanks.     

Microsatellite variability within and among local landrace populations of tea, Camellia sinensis (L.) O. Kuntze, in Kyoto, Japan

Genetic variability within and among eight landrace populations of tea (Camellia sinensis (L.) O. Kuntze) located in southern Kyoto, Japan, was surveyed with six microsatellite markers. The average number of alleles per locus was 3.83 to 4.67 for landrace populations, whereas the corresponding value among modern cultivars and breeding lines was 6.63. Expected heterozygosity values averaged over loci within landrace populations ranged from 0.498 to 0.723. A similar level of variation, 0.682, was observed for cultivars and breeding lines. High fixation index values (0.177–0.417) for each population are consistent with biparental inbreeding within the population. Genetic differentiation among local populations was extremely low with F _ST = 0.062, although AMOVA revealed significant differentiation among landrace populations. We propose that these populations share a common ancestral gene pool and that some degree of artificial selection within each population has been performed by local farmers. Neighbor-joining analysis revealed that genetic relationships among populations reflect geographical location of populations. This might result from more frequent genetic exchange by nearby farmers.     

Relationship of European persimmon (Diospyros kaki Thunb.) cultivars to Asian cultivars, characterized using AFLPs

Sixty one persimmon (Diospyros kaki Thunb.) selections, including 17 Italian, 11 Spanish, 13 Japanese, six Korean, five Chinese, one Israeli, and eight of unknown origin, were evaluated for genetic differences by AFLP analysis. Relationships among cultivars were evaluated by UPGMA clustering, Neighbor Joining, and MultiDimensional Scaling. While similarities among groups were generally less than 0.60, both UPGMA and Neighbor Joining separated European and Asian cultivars. Spanish and Italian cultivars were not separated by any of the analyses, suggesting that they share a common gene pool, while Japanese, Chinese and Korean cultivars formed distinct clusters. Diversity within groups was greater than diversity between groups. Most cultivars were quite polymorphic (only 0.60–0.80 similarity between cultivars). In addition, the presence of several Japanese cultivars in the European group and a group of European cultivars nested between Chinese and Korean groups suggest that similar, but different progenitors were used in the development of the present European cultivars. ‘Kaki Tipo’ selections from different sources were clearly different by AFLP analysis, indicating that they are separate cultivars.