栽培荞麦种子醇溶蛋白遗传多样性分析

利用A-PAGE（Acid-polyacrylamide gel electrophoresis）对来源于7个国家的76份栽培荞麦（苦荞54份,甜荞22份）醇溶蛋白遗传多样性进行评价。结果表明,荞麦醇溶蛋白位点存在丰富的等位变异,共分离出18条迁移率不同的谱带,每份材料具有6~12条不等,平均9.5条,多态性带占88.89%。材料间平均遗传相似系数（GS）为0.777,变幅为0.389~1.000。在GS为0.63的水平上供试材料可聚为苦荞和甜荞两大类,绝大部分来自于相同或相似生态地理环境的材料聚成一类,表明荞麦醇溶蛋白所揭示的遗传关系与地理来源有较高的相关性。     

High-performance liquid chromatographic analysis of anthocyanins in Japanese garden iris and its wild forms

Summary Following bud pollination of tartary buckwheat (Fagopyrum tataricum) with pollens from common buckwheat (F. esculentum), cv. Mancan, 31% of the ovaries began to grow but all turned brown and withered after 6 to 14 days. Fluorescence microscopy of the growing ovaries showed the pollen-tube entering the embryo-sac. The ovules from the growing ovaries failed to produce any embryo in the culture medium in which the immature embryos from the self-pollinated (compatible) tartary flowers were able to mature and germinate. No embryo development was observed after cross-pollination. The interspecific incompatibility is attributed to the failure of gametes to fuse.Abbreviations BAP benzylaminopurine - IAA indoleacetic acid     

Selection of wild Cicer accessions for the generation of mapping populations segregating for resistance to ascochyta blight

In order to determine genetically diverse parents for the generation of mapping populations segregating for resistance to ascochyta blight in wild Cicer species, the genetic diversity between a selection of resistant and susceptible accessions was assessed using molecular markers. Twenty Cicer accessions — comprising eight C. reticulatum accessions, six C. echinospermum accessions, five C. bijugum accessions, and one C. arietinum accession — were compared using a combination of seven RAPD primers and seven ISSR primers. A total of 231 polymorphic bands were scored and used to determine the genetic distances between accessions using Jaccard similarity coefficients. The most genetically diverse parents for the generation of intraspecific and interspecific populations segregating for resistance to ascochyta blight are reported. This revised version was published online in July 2006 with corrections to the Cover Date.     

Geographical differentiation and diallel analysis of seed dormancy in barley

Seed dormancy is one of the most important parameters affecting the malting process and pre-harvest sprouting in barley (Hordeum vulgare L.). Variation of seed dormancy in 4365 cultivated and 177 wild barley (ssp. spontaneum) accessions derived from different regions of the world was investigated in Okayama University, Kurashiki, Japan. Seed dormancy of each accession was estimated from their germination percentages at 0, 5, 10 and 15 weeks post-harvest after-ripening periods. All of the wild barley accessions showed less than 10% germination at 0 week after-ripening period. Level of seed dormancy in 4365 cultivated barley accessions showed a clear geographical differentiation. Seventy seven percent of Ethiopian accessions showed high germination percentages, while 86% of Japanese, Turkish and North African accessions showed low germination percentages at 0 week after-ripening period. A half diallel cross using eleven barley accessions with different level of dormancy revealed that seed dormancy was predominately controlled by additive gene effects. These results suggest that large genetic diversity for seed dormancy in barley is explained as different levels of additive accumulation of genetic factors. Barley varieties showing appropriate dormancy could be developed by crossing among barley germplasm accessions used in the present study.     

Evidence of a partial reproductive barrier between wild and cultivated pearl millets (Pennisetum glaucum)

Summary The occurrence of seed malformation in association with reduced thousand grain weight and germination ability has been observed in crosses between cultivated female plants and wild male plants. A survey of 16 cultivated accessions (P. glaucum subsp. glaucum) and 11 wild accessions (P. glaucum subsp. monodii) ranging over the whole species diversity showed this postzygotic incompatibility was general, but its intensity varied greatly with the cultivated female accession used and very little with the wild male parent origin. About 15% of the 123 cultivated x wild crosses observed gave normal seeds. Seed malformation has never been observed in crosses between cultivated accessions and appeared independent of genetic distances between the parents. The reciprocal crosses between wild female plants and cultivated male plants gave normal-looking seeds with good germination but consistently reduced thousand grain weight. Both seed malformation and seed small size are an expression of a genetic imbalance. These slight reproductive barriers seem to have been built during the domestication process.Abbreviation ICRISAT International Crop Research Institute for the Semi-Arid Tropics     

Production of phenolic compounds in hairy root culture of tartary buckwheat (Fagopyrum tataricum Gaertn)

Fagopyrum tataricum Gaertn (tartary buckwheat) is an excellent medicinal and nutrient-rich crop. It has a high content of rutin and other phenolic compounds. An experiment was conducted to investigate in vitro production of phenolic compounds from hairy root culture of tartary buckwheat. Hairy root growth was promoted by increasing culture time in MS medium. The highest hairy root growth reached up to 11.2 g/l dry weight at 18 d after placement. Transformation was confirmed by PCR using rol genes, rol A (304 bp), B (797 bp), C (550 bp), and D (1035 bp) genes which is transferred into hairy roots from the Ri-plasmid in Agrobacterium rhizogenes and is responsible for the induction of hairy root from plant species. Rutin, quercetin, (−) epicatechin, (−) catechin hydrate, gallic acid, ferulic acid, chlorogenic acid, and caffeic acid were identified both in hairy and wild type roots of tartary buckwheat. The main compound found in the both types of root was epicatechin followed by rutin. The concentration of phenolic compounds in the hairy roots of tartary buckwheat was several-fold higher compared with wild type roots of same species. Our results indicate that hairy root culture of F. tataricum is a valuable alternative approach for the production of phenolic compounds.     

Taxonomy of Portuguese Tronchuda cabbage and Galega kale landraces using morphological characters,nuclear RFLPs,and isozyme analysis: A review

Summary Morphological characters, nuclear RFLPs, and isozyme analysis were used to study the similarity between 32 Portuguese Tronchuda cabbage and Galega kale landraces, and some cabbage cultivars traditionally grown in Portugal. Forty-six morphological characters observed in two consecutive years, RFLP data from 55 nuclear probes, detecting 291 polymorphic nuclear DNA restriction fragments, and allelic frequencies in 21 putative loci, generated by nine isozymes, were analyzed by the unweighted pair group method, using arithmetic averages (UPGMA), in order to present the results in the form of a phenogram. The three methods resulted in different clustering patterns of the 32 cole accessions. Morphological characters gave consistent clustering according to the traditional landrace definition and denomination, producing clear separation between Tronchuda cabbages and Galega kales. RFLPs were unable to separate Tronchuda cabbages from Galega kales and defined five landrace groups corresponding to their geographic origins rather than to their morphological similarities. Isozymes showed poor accession discrimination and an intermediate clustering pattern with some accessions being clustered according to their geographic origins and others according to their morphological similarities. Portuguese Tronchuda cabbages and Galega kales constitute a distinct and relatively homogenous group within Brassica oleracea, sharing the same genetic background. It is concluded that Portuguese coles have evolved independently from a common ancestor to the present cultivated forms. Portugal should be considered as an important region of domestication of specialized leafy coles.     

Genomic variation and genetic relationships in Ipomoea spp.

Random amplified polymorphic DNA (RAPD) markers were used to develop genetic fingerprints and analyse genetic relationships among 29 Ipomoea accessions from different geographical locations around the world, including unique wild species, and reproducible profiles were obtained for all accessions using random decamer primers. The primers generated 46 polymorphic markers, one primer alone having 10 products, enabling the discrimination of all 29 accessions. A high level of genetic variability in sweet potato collections was suggested by the degree of polymorphism. Half of the Japanese land races were closely related while accessions from Papua New Guinea and The Philippines were distinct and exhibited the greatest genetic diversity. The wild species Ipomoea gracilis and Ipomoea tiliacea formed a group distinct from the cultivated sweet potato. The wild tetraploid accession K233 and the species Ipomoea trifida were progressively more related genetically to the cultivated sweet potato and are the probable progenitors of Ipomoea batatas, and may be suitable as germplasm for genetic enhancement. RAPDs proved to be useful for sweet potato systematics and should be valuable for germplasm management, gene tagging and efficient choice of parents in breeding programmes.     

Population genetic structure of the clonal plant Zingiber zerumbet (L.) Smith (Zingiberaceae), a wild relative of cultivated ginger, and its response to Pythium aphanidermatum

Zingiber zerumbet (L) Smith, a wild clonal species related to the cultivated ginger (Zingiber officinale Roscoe), is a potential resistance donor for soft rot disease in ginger caused by Pythium aphanidermatum (Edson) Fitzp. In this study we evaluated the genetic diversity and P. aphanidermatum resistance of 74 Z. zerumbet accessions belonging to 15 populations from eight districts in Kerala state, India. The disease index (DI) of the accessions varied from 0% to 72.24% and the accessions could be separated into six frequency classes according to their DI values. More than 65% of the accessions had a DI < 20%. Eight accessions were found to be immune to the infection. The relative frequency of resistant accessions was higher in the central and northern regions of Kerala. Amplified fragment length polymorphism (AFLP) analysis of Z. zerumbet accessions using five primer combinations yielded 215 bands in total, of which 175 (81.4%) were polymorphic. Nei’s genetic diversity (h) of 0.2738 and Shannon information index (I) of 0.4012 revealed a high genetic diversity in Z. zerumbet unexpected for a clonal species. In the UPGMA dendrogram, accessions were clustered mostly according to their geographical origin and no clear correspondence was observed between the clustering pattern of accessions and their responses to Pythium aphanidermatum. The study revealed high genetic diversity and variability for pathogen resistance among Z. zerumbet accessions and confirmed the value of Z. zerumbet as a potential donor for soft rot resistance for the genetic improvement of ginger.     

Genetic diversity in tef [Eragrostis tef (Zucc) Trotter] and its relatives as revealed by Random Amplified Polymorphic DNAs

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. This revised version was published online in July 2006 with corrections to the Cover Date.     

Genetic diversity and phylogenetic relationships among the annual Cicer species as revealed by isozyme polymorphism

Summary There are few estimates of genetic variability within and among populations of the nine annual Cicer species and for the wild species this information is based on few accessions. The present study was undertaken to examine genetic variation within and between annual Cicer species. One hundred and thirty-nine accessions of nine annual Cicer species were used for electrophoretic analysis at ICARDA. High levels of polymorphism in all eight wild annual Cicer species was found. This is in contrast to earlier research which had shown high polymorphism only in C. reticulatum. Cicer reticulatum had the highest proportion of polymorphic loci. However, for the cultigen, among 14 loci assayed, only two were polymorphic, ADH and PGD2. The nine species formed four phylogenetic groups based on the neighbor-joining method. The first group comprised C. arietinum, C. Reticulatum and C. echinospermum, the second C. bijugum, C. judaicum and C. pinnatifidum, the third C. chorassanicum and C. yamashitae; and the fourth group consisted of one species, C. cuneatum. The phylogenetic tree developed from the neighbor-joining technique illustrated that C. reticulatum is the probable progenitor of C. arietinum and that C. echinospermum split off from a common ancestor at an earlier stage in the evolutionary history of Cicer. Genetic diversity data showed that the greatest diversity was within C. reticulatum and the lowest with the cultigen, C. arientinum. With the exception of C. reticulatum, genetic diversity increased with genetic distance from the cultigen. Little geographic variation in genetic diversity was found.     

裸燕麦品质资源AFLP标记遗传多样性分析

用20对AFLP引物组合对281份栽培裸燕麦(Avena nuda)进行遗传多样性分析,共得到1 137条带,其中260条为多态性带,引物的平均多态性百分率为22.96%,平均多样性信息指数(PIC)为0.0326。以地理来源分组,不同来源的组群Simpson指数在1.235~1.495之间,Shannon指数范围为0.1558~0.4437,组群内变异贡献率为83.45%,组群间变异占16.55%。组群大小与多态性位点数、组群内变异贡献率、Simpson指数及Shannon指数显著相关。内蒙古和山西资源多样性丰富,东北地区资源独特,西部地区资源遗传结构单一,东欧组群与内蒙古组群遗传关系最近。国内组群的遗传多样性水平高于国外组群。地方品种与育成品种相比,组群内变异贡献率较高。建议在遗传多样性丰富地区进一步收集裸燕麦资源,并加强对材料少、代表性较差的地区,如西北和西南地区的裸燕麦地方品种的收集,以丰富我国的裸燕麦基因源。     

Genetic diversity of wheat wild relatives in the Near East detected by AFLP

In order to reveal the molecular genetic diversity of wheat wild relatives, an AFLP analysis was conducted with 16 accessions of five Triticum andAegilops species originating from the Near East. Variation within population was studied with at least seven individuals per accession. Four primer combinations were used for selective amplification. Based on the scored bands, we estimated percentage of polymorphic bands, 1 – proportion of shared bands (1-psb) and nucleotide diversity (π). Of the five species used in this study, Ae. speltoides had the highest level of `within population' variation. This species had also the highest value of the variation among populations. As for Triticum species, the level of variation within population was low in diploid species (T. urartu and T. boeoticum),whereas two tetraploid species (T. dicoccoides and T. araraticum) had relatively high levels of variation within population. While the two diploid Triticum indicated a clear interspecific divergence, the two tetraploid wild wheats were not clearly divergent in this study. The variance portioning analysis indicated that the variation detected for diploid Triticum species was mainly composed of `between species' variation, on the other hand that for tetraploid Triticum was mostly composed of `within population' variation. In conclusion, AFLP analysis reveals molecular variation in all accessions used in this study, suggesting a potential genetic diversity of the wheat wild relatives in natural populations. These results have implications for the design of strategies to maintain genetic diversity within genebank collections. This revised version was published online in August 2006 with corrections to the Cover Date.     

Genetic diversity of wild coffee (Coffea arabica L.) using molecular markers

Genetic diversity was studied using RAPD markers among119 coffee (Coffea arabica L.) individuals representing 88 accessions derived from spontaneous and subspontaneous trees in Ethiopia, the primary centre of species diversity, six cultivars grown locally in Ethiopia, and two accessions derived from the genetic populations Typica and Bourbon, spread in the 18^th century, which gave rise to the most currently grown cultivars. Twenty-nine polymorphic fragments were used to calculate a similarity index and construct dendrograms. The Ethiopian material was separated from the Typica- and Bourbon-derived accessions and classified in four groups: one with most of the collected material from southwestern Ethiopia and three from southern and southeastern Ethiopia. Almost all detected diversity was found in the southwestern group while the southern and southeastern groups presented only 59% of identified markers. The genetic distances were low between the southwestern group and the southern and southeastern groups, and between the southwestern group and the Typica- and Bourbon-derived accessions. The cultivated coffee derived from the genetic populations Typica and Bourbon appeared little differentiated from wild coffee growing in the southwest. The results supported the hypothesis that southwestern Ethiopian coffee trees could have been introduced recently in the south and southeast. A separate analysis of the 80accessions classified in the southwestern group allowed identifying particular spontaneous- and subspontaneous-derived accessions and redundancies in the collected material from southwestern Ethiopia. RAPD markers did not detect any within-collection polymorphism except for two trees that were identified as off-types in the CATIE field genebank. This revised version was published online in July 2006 with corrections to the Cover Date.     

Genetic diversity and taxonomic relationships within the genus Lens as revealed by allozyme polymorphism

Summary A survey of allozyme polymorphism at 11 loci was carried out on 439 accessions from the genus Lens. This comprised 153 Lens culinaris subsp. orientalis, 35 L. odemensis, 117 L. ervoides, 32 L. nigricans, 2 of a differentiated cytotype of L. nigricans and 100 landrace accessions of the cultivated lentil (L. culinaris subsp. culinaris), from 10 different countries. The aim of the survey was to determine intra-specific genetic diversity and species relationships, based on phylogenetic and phenetic analyses, particularly regarding the position of L. odemensis and the differentiated cytotype of L. nigricans. Diversity was described by three statistics. The level of diversity in the cultivated taxon was lower than in any of the wild species according to two of these statistics, the percentage of polymorphic loci and mean number of alleles per locus. For the third measure (Nei's mean genetic diversity) it was only greater than L. ervoides. Genetic diversity statistics of the wild species indicated differences in the nature of between-population genetic diversity within the different taxa. Phylogenetic analysis suggests that L. odemensis and L. ervoides evolved from a common ancestor, and L. culinaris subsp. orientalis subsequently evolved from L. odemensis. Phenetic analysis, however, places L. odemensis closer to L. culinaris subsp. orientalis than to L. ervoides. Nei's mean genetic distance of L. odemensis from both L. culinaris subsp. culinaris (0.204) and L. culinaris subsp. orientalis (0.110) was greater than the distance between them (0.062). This evidence is not conclusive in determining whether L. odemensis should retain its specific status. Further crossability studies should be carried out on a range of genotypes to assess the potential for gene flow. The evidence presented shows the differentiated cytotype of L. nigricans to be quite distinct from other L. nigricans accessions, both phenetically and phylogenetically. This indicates that the differentiated cytotype of L. nigricans may constitute a new taxon. Discriminant function analysis reveals that isozymes may be useful in validating species classification.     

Species relationships in the subgenus Ceratotropis (genus Vigna) as revealed by RAPD analysis

Summary The genetic variation among 23 accessions of 5 species in the subgenus Ceratotropis, genus Vigna, were investigated by random amplified polymorphic DNA (RAPD) analysis. A total of 404 fragments amplified with 24 primers were scored and analyzed by cluster analysis. The accessions used were separated into two main groups with an average of 70% differences. Within the main groups, five subgroups were recognized, which are in complete agreement with taxonomic species. Wild forms were always grouped with their most closely related cultivated forms and they showed variation in each species. The largest intraspecific variation was found in V. radiata (mungbean), in which wild forms (V. radiata var. sublobata) were highly different from each other and from cultivated forms. V. angularis (adzuki bean) showed the least variation and thus, was probably differentiated in relatively recent times.     

Isozyme diversity within African Manihot germplasm

Summary Isozyme diversity is described among a collection of 365 Manihot esculenta cultivars plus 109 accessions from wild relatives (M. glaziovii and spontaneous hybrids) from Africa. The study is based on 17 polymorphic loci. A natural hybrid swarm is detected between the two species. Although they were recently introduced, M. esculenta and M. glaziovii show high levels of polymorphism: heterozygosity estimates are 0.225 and 0.252 respectively. For the wild species, diversity is structured at the unilocus level, and the multilocus approach reveals a geographical pattern. The organization of the diversity is not so clear for the cultivated cassava, but a multilocus approach, based on both common and rare alleles, led us to identify different groups of clones with many intermediate genotypes between them. Elements of the secondary diversification process of Manihot in Ivory Coast are discussed.     

Molecular Marker based Genetic Diversity Analysis in Rice (Oryza sativa L.) using RAPD and SSR markers

The availability of an array of molecular marker systems allowed comparing the efficiency of two of these marker systems to estimate the relationships among various taxa. The objective of this study was to assess the genetic diversity among 40 cultivated varieties and five wild relatives of rice, Oryza sativa L. involving simple sequence repeat (SSR) randomly amplified polymorphic DNA (RAPD) markers. The accessions were evaluated for polymorphisms after amplification with 36 decamer primers and 38 SSR primer pairs. A total of 499 RAPD markers were produced among the 40 cultivated varieties and five wild relatives with a polymorphism percentage of 90.0. Out of 38 SSR primer pairs used, only one locus viz., RM115 was monomorphic. The average Polymorphism Information Content (PIC) value was 0.578 and it ranged from a low of zero (RM 115) to a high of 0.890 (RM 202). The Mantel matrix correspondence test was used to compare the similarity matrices and the correlation coefficient was 0. 582. The test indicated that clusters produced based on RAPD and SSR markers were not conserved since matrix correlation value was 0.582 as against the minimum required value of 0.800. The two marker systems contrasted most notably in pair-by-pair comparisons of relationships. SSR analysis resulted in a more definitive separation of clusters of genotypes indicating a higher level of efficiency of SSR markers for the accurate determination of relationships between accessions that are too close to be accurately differentiated by RAPD markers. This revised version was published online in July 2006 with corrections to the Cover Date.     

AFLP分析小豆种(Vigna angularis)内遗传多样性

利用12对AFLP引物,对来自于世界6个国家的146份小豆(Vigna angularis)栽培变种(var. angularis)和野生变种(var. nipponensis)种质的基因组DNA进行扩增,得到580条清晰的显带,其中313(53.93%)条呈多态性,平均每对AFLP引物得到26.08条多态性带;平均遗传距离0.35,变异幅度为0.00～0.87.利用AFLP多态性数据进行的Jaccard''s遗传距