RAPD variation in wild, weedy and cultivated azuki beans in Asia

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.     

Genetic variation in wild and cultivated artichoke revealedby RAPD markers

Determination of intraspecific genetic diversity is an important first step for the utilization of genetic resources and canprovide useful information on crop evolution. A living collection of Italian and foreign artichoke varieties and ecotypes is maintained at the Germplasm Institute, Bari, Italy. A total of 32 accessions of cultivated (Cynara cardunculus L. var.scolymus (L.) Fiori), three of wild (Cynara cardunculus var.cardunculus L.) artichoke and two ofcultivated cardoon (Cynara cardunculus var.altilis L.) were analysed in order to studygenetic variation and relationships within the species, using RAPDmarkers. Cultivated accessions were selected according tomorphological variation and geographical distribution available inthe collection. Fifty arbitrary decamer primers were initially tested, 18 ofwhich showed from 3 to 10 unambiguously interpretable fragments. Anintra-accession analysis using 4 varieties and 8 polymorphicprimers revealed that no RAPD variation was detected amongindividuals. Jaccard's similarity index (JSI) was comprisedamong 1 and 0.693 when all accessions were considered, on the otherhand, within the cultivated artichoke, JSI ranged between 1 and0.817. A UPGMA dendrogram based on the similarity matrix showed thatwild artichokes were clearly separated from cultivated accessions.Moreover, within cultivated artichokes, several groups could bedistinguished.     

Polyphenols in wild and weedy Mexican common beans (Phaseolus vulgaris L.)

The polyphenolic composition of 62 wild and weedy Mexican bean collections from diverse origins, grouped by their seed coat color, was assessed. According to spectrophotometric analysis, the range of total phenols, condensed tannins, and total anthocyanins presented wide differences. Furthermore, the phenolic acid, flavonoid, and anthocyanin profiles were analyzed using HPLC. Ferulic was the main phenolic acid. Kaempferol and quercetin were the main flavonoids, and the isoflavones daidzein and coumestrol were found in only low levels in few collections. Delphinidin was the main anthocyanidin found, followed by petunidin, cyanidin, malvidin, pelargonidin, and peonidin. The wide variation observed in polyphenolic contents was more related to their genotype than to the color factor. These results show that some wild and weedy beans are good sources of phenolic compounds for use in breeding programs focused on nutrition and health.     

Ecogeographical distribution of wild, weedy and cultivated Sorghum bicolor (L.) Moench in Kenya: implications for conservation and crop-to-wild gene flow

The potential gene flow between a crop and its wild relatives is largely determined by the overlaps in their ecological and geographical distributions. Ecogeographical databases are therefore indispensable tools for the sustainable management of genetic resources. In order to expand our knowledge of Sorghum bicolor distribution in Kenya, we conducted in situ collections of wild, weedy and cultivated sorghum. Qualitative and quantitative morphological traits were measured for each sampled wild sorghum plant. Farmers’ knowledge relating to the management of sorghum varieties and autecology of wild sorghum was also obtained. Cluster analysis supports the existence of several wild sorghum morphotypes that might correspond to at least three of the five ecotypes recognized in Africa. Intermediate forms between wild and cultivated sorghum belonging to the S. bicolor ssp. drummondii are frequently found in predominantly sorghum growing areas. Crop-wild gene flow in sorghum is likely to occur in many agroecosystems of Kenya.     

Genetic relationships among cultivated types of Perilla frutescens and their weedy types in East Asia revealed by AFLP markers

AFLP markers were employed to detect genetic diversity in two cultivated Perilla frutescens (i.e. var. frutescens and var. crispa) and their weedy types and to assess their genetic relationships. Analysis of 60 Perilla accessions from China, Korea and Japan by seven AFLP primer combinations identified a total of 125 fragments, of which 80 (64%) were polymorphic at the species level. The phenotypic diversity meassured by Shannon's index of information for the cultivated type of var. frutescens, the weedy type of var. frutescens, the cultivated type of var. crispa and the weedy type of var. crispa were on average 1.07, 2.23, 1.24 and 1.75, respectively. The weedy types exhibited high within-type variation in spite of a small number of samples. In the neighbor joining tree, two major clusters were recognized: (1) cultivated type of var. frutescens, (2) weedy type of var. frutescens, and cultivated and weedy types of var. crispa. The cultivated types of var. frutescens and of var. crispa were sharply separated by AFLPs. However, there remained ambiguities in regard to the intraspecific relaltionships, due to the clustering of the weedy types which occured in each of the clusters of the cultivated types. Two cultivated types of P. frutescens and their weedy types should be taxonomically considered as a P. frutescens complex. The present AFLP data are consistent with the hypothesis that China is the original place of cultivation of var. frutescens and Korea is a secondary center of diffusion of var. frutescens.     

Phytochemical variation in echinacea from roots and flowerheads of wild and cultivated populations

Quantitative phytochemical variation was determined from roots and inflorescences of native plant populations in the genus Echinacea. Specimens were collected in situ throughout the natural range of each putative taxon and transplanted to greenhouse cultivation. Ethanolic extracts from individual plants were separated by reversed-phase HPLC to quantify the alkamides, polyenes/ynes, and phenolics, and then grouped by age and taxonomically, according to a recent morphometric taxonomic revision of the genus. Canonical discriminant analysis revealed that cichoric acid, the diene alkamides 1-3 and 7, and ketoalkene 24 were the best taxonomic markers. Mean content for each of 26 phytochemicals revealed useful agronomic information, such as those varieties and organs with the highest accumulations, as well as the optimal age and growth conditions for each variety. The highest amounts of cichoric acid were measured from the older, wild inflorescences of E. pallida var.sanguinea, whereas the highest quantities of the alkamides 1-3 and 7 were present in roots of wild and transplanted E. purpurea. Baseline phytochemical data and chromatographic profiles for all types of wild Echinacea may be used for protection of wild stands, germplasm identification, and crop improvement.     

Isozyme variation in Passiflora subgenera Tacsonia and Manicata. Relationships between cultivated and wild species

Isozyme variation was studied in 87 plants from 32 cultivated and wild accessions of banana passion fruit (P. tripartita var. mollissima, P. tripartita var. tripartita, P. tarminiana, and P. mixta), rosy passion fruit (P. cumbalensis), tin-tin (P. pinnatistipula), gulián (P. ampullacea), P. antioquensis, P. bracteosa, and P. manicata, from the Andes of Venezuela, Colombia, and Ecuador. Six polymorphic enzyme systems (IDH, PGDH, PGM, DIA, PRX, and ACP) revealed 31 zymotypes characterized by the presence or absence of 31 electromorphs. Cluster analysis separated clearly the accessions of P. tarminiana, P. tripartita, P. mixta, and P. cumbalensis from the less typical species of subgenus Tacsonia, which is consistent with morphological evidence. P. mixta showed the highest intraspecific variation and the closest affinity with P. tripartita. The accessions of these two species formed two clusters, one dominated by Colombian genotypes and the other dominated by Ecuadorian genotypes. One of the P. tripartita var. mollissima accessions clustered close to P. tarminiana accessions. The affinity between these three species is particularly interesting for conservation and use of banana passion fruit genetic resources. All the other species formed monospecific clusters.     

Diversity in Allium ampeloprasum: from small and wild to large and cultivated

Allium ampeloprasum evolved as a complex of different cyto- and morpho-types widely distributed either in the wild or domesticated range of the Mediterranean regions. The assessment of genetic and phylogenetic relationships between Tunisian A. ampeloprasum and specimens from different origins and with variable degree of domestication can promote conservation and breeding. Minisatellite M13, microsatellite (GTG)₅ and nucleotide sequence analysis of the internal transcribed spacer region (ITS) were used to assess DNA polymorphism and genetic diversity. M13 and (GTG)₅ molecular markers efficiently discriminated A. ampeloprasum gene-pool from A. sativum. Geographic genetic patterns of variation of the wild gene-pool were not detected. However, domesticated A. ampeloprasum (great headed garlic, kurrat and leek) clustered consistently within the ampeloprasum group. A. sativum was found to be closer to A. ampeloprasum than A. fistulosum and A. schoenoprasum. A high number of single point mutations (SNPs) was recorded over the ITS1-2 spacer sequence. Most of these SNPs were heterozygous only in great headed garlic. It is inferred that heterozygosity played the major role in promoting great headed garlic domestication. Thus, great headed garlic adaptation to horticultural conditions along with its yield trait sizes are mainly associated to heterozygosity rather than to polyploidy.     

Solute adjustment to salinity and calcium supply in cultivated and wild barley

Ion relations, water content, leaf water potential, and osmotic adjustment were determined for cultivated barley (cv Harrington) and wild barley grown under mixed sulphate (SO₄) salts with varied calcium (Ca) supply using a hydroponic system. Salinity induced significant increases of leaf, stem and root sodium (Na) concentrations in both species. Salt‐stressed wild barley roots accumulated more Na than shoots, and transport of Na from roots to shoots was low compared to Harrington. Cultivated barley had lower Ca concentrations than wild barley, especially in the low Ca salt treatment. Although potassium (K/Na) and Ca/Na ratios were higher in control wild barley plants than in Harrington, they declined under salt stress, irrespective of Ca supply. Major osmotica in wild barley leaves were K, sugars, organic acids, and quaternary ammonium compounds, while in Harrington they were cations, including Na, K and Mg, and anions such as phosphate (PO₄) and SO₄. Wild barley maintained better water status than Harrington under low Ca salt treatment. Supplemental Ca improved water status more in Harrington than in wild barley. Lack of osmotic adjustment to salinity in wild barley apparently resulted from its ion exclusion. Low Ca salt treatment caused Ca deficiency, Na toxicity, and loss of turgor in Harrington. In the high Ca salt treatment, Harrington had improved water and ion relations, as well as positive turgor.     

Sources of resistance to cyst nematode in cultivated and wild Cicer species

Summary Among the nematodes infesting chickpea (Cicer arietinum L.) plants in Syria, cyst nematode (Heterodera ciceri Vovlas, Greco et Di Vito) is the most important. It is uneconomical to grow chickpea in fields infested with cyst nematode and to control this nematode with nematicide. Therefore, investigations were conducted at ICARDA, Syria from 1987 to 1991 to identify sources of resistance to cyst nematode in 7258 lines of C. arietinum and 102 lines of eight annual Cicer species including C. bijugum K.R. Rech. (13 lines), C. chorassanicum (Bge) M. Pop. (3 lines), C. cuneatum Hochst. ex Rich. (3 lines), C. echinospermum P.H. Davis (8 lines), C. judaicum Boiss. (18 lines), C. pinnatifidum Jaub. & Sp. (18 lines), C. reticulatum Ladiz. (36 lines), and C. yamashitae Kitamura (3 lines). All lines were grown in a greenhouse at 15–25°C in pots containing soil infested with 20 eggs of the nematode g^-1 soil. Nematode infestation was evaluated on a 0 to 5 scale based on number of females and cysts on roots. Resistance was found in one line of C. bijugum, six lines of C. pinnatifidum, and one line of C. reticulatum. No lines of C. arietinum, C. chorassanicum, C. cuneatum, C. echinospermum, C. judaicum, or C. yamashitae was resistant to cyst nematode. Plants with resistance have been recovered in the F₃ generation from crosses between the cultigen and C. reticulatum, indicating the possibility of transfer of gene(s) for resistance to cyst nematode from wild to cultivated Cicer species.Joint contribution from Istituto di Nematologia Agraria, ICARDA and ICRISAT (International Crops Research Institute for the Semi-Arid Tropics), Patancheru P.O., A.P. 502 324, India.     

Seed protein diversity in Solanum melongena L. and its wild and weedy relatives

Seed protein profiles by SDS-PAGE were studied in 72 accessions ofSolanum melongena L. and its related taxa in subgenusLeptostemonum (Dunal) Bitter. Comparisons based onJaccard's similarity and UPGMA clustering revealed interrelationships broadly inconformity with the conventional taxonomic treatments. Thus,S. viarum Dunal (sectionAcanthophora Dunal), S.torvum Swartz (section Torva Nees) andS. sisymbriifolium Lam. (sectionCryptocarpum Dunal) were located on separate branches ofthe dendrogram while all taxa of section Melongena Dunal,except S. macrocarpon Dunal, wereclustered closer together. S.melongena, incanum and insanum,all members of the eggplant complex, were revealed to bear very high similaritywith each other. Most of their accessions had identical band patterns supportingthe contention that these represent an interbreeding complex with limitedgenetic differentiation.     

Hybrids between wild and cultivated carrots in Danish carrotfields

It is well known that wild carrots may pollute the seed cropsof cultivated carrots, but whether cultivated carrots can alsodisperse pollen and seed, and thereby introduce genes into wildcarrot populations, is only little described. In Denmark, there is nocommercial seed production of carrots, and as biennials they shouldnormally not flower before harvest of the roots. Still, floweringindividuals can be found in most Danish fields, and sometimes in veryhigh numbers. At least 75% of the flowering plants are malefertile, with ca. 83% of the pollen being viable. More thanhalf of the plants produce seeds. Pollen and seed dispersal fromfields into wild carrot populations is probably rather frequent inDenmark. A closer inspection of the morphology of flowering plantsindicate that some of these (2–60%) arebolters of pure cultivar origin, as indicated primarily by orangeroot colour. The remainder is probably first or advanced generationhybrids between wild and cultivated plants, as indicated by theirwhite roots and combinations of morphological characters from eitherplant type. Some of these hybrids are imported to Denmark togetherwith the sowing seed, as indicated by significantly differentfrequencies of bolters with white roots in different carrot cultivarstested in the field.     

Genetic diversity and relationships among cultivated and wild accessions of tartary buckwheat (Fagopyrum tataricum Gaertn.) as revealed by RAPD markers

The methodology of sampling and the selection of a proper marker systemfor the analysis of accessions are major concerns in the evaluation of gene bank material. In our study the RAPD analysis of bulked DNA samples and single seedsDNA was successfully employed to evaluate intra- and inter-population geneticvariability of cultivated and wild tartary buckwheat accessions. The bulkingapproach enabled the distinction of all 40 analysed accessions and theirseparation into geographically well defined clusters. Three wild populations,two from Sichuan and one from Qinghai, formed a group that was geneticallyrelatively distant from wild populations from Tibet and all cultivatedlandraces which, on the other hand, exhibited very close relationships. Thesingle seed study that was used after bulked DNA analysis provided detailedinformation of the genetic variation present within some accessions of specialinterest. A moderate level of genetic variability was detected betweenaccessions and the variability was partitioned into between- andwithin-population components. On average, most of the detected variation ispresent between F. tataricumpopulations. The genetic and geographic distribution of variability is furtherdiscussed. We demonstrated the usefulness of combining bulking and single seedstudy approaches for the effective evaluation of genetic variability inF. tataricum accessions that couldalso have wider applicability in the management of plant genetic resources andphylogenetic studies.     

Spontaneous gene flow and population structure in wild and cultivated chicory, Cichorium intybus L.

Spontaneous gene flow between wild and cultivated chicory, Cichorium intybus L., may have implications for the genetic structure and evolution of populations and varieties. One aspect of this crop-wild gene flow is the dispersal of transgenes from genetically modified varieties, e.g. gene flow from GM chicory to natural chicory could have unwanted consequences. With the purpose to identify and quantify crop-wild gene flow in chicory, we analysed introgression in 19 wild chicory populations and 16 accessions of chicory varieties and landraces distributed across Northern, Central and Mediterranean Europe. The analysis used 281 AFLP markers and 75 SSAP markers giving a total of 356 polymorphic markers. Results from model based assignments with the program STRUCTURE indicated many incidents of recent gene flow. Gene flow was observed both between cultivars and wild populations, between landraces and wild populations, between different wild populations as well as between cultivars. Population structure visualized by distance-based clustering showed a North–South geographical structuring of the wild populations, and a general grouping of the cultivars corresponding to known origin. The results indicated, however, that the structuring between the two groups of wild and cultivated types was weak. As crop and wild recipients are genetically close and genes are transferred between the two types rather frequently, focus on mitigating crop-wild gene flow should be increased, before transgenic varieties are cultivated openly.     

Microscale ecological stress causes RAPD molecular selection in wild barley, Neve Yaar microsite, Israel

Genetic micro-differentiation was studied in wild barley at the Neve Yaar microsite, Israel, an area of 3182 m², which includes a mosaic of sun-rock, sun-soil, shade-rock, and shade-soil microniches. Fifty-six genotypes were tested for ecological-genetic microniche correlates. Analysis of 75 RAPD loci revealed a significantly higher polymorphism and gene diversity (P _0.05 = 0.920; He = 0.411) on the more stressful sun-rock microniche, compared to the least stressful shade-soil microniche (P _0.05 = 0.653; He = 0.188). Fifty-six (74.7%) out of the 75 loci varied significantly in allele frequencies. On the average, of the 75 loci, 75.7% of the total genetic diversity exists within the four microniches, while 24.3% exists between the microniches. The highest genetic distance was between shade-soil and sun-rock (D = 0.220), which confirmed their sharp microecological contrasts. Gametic phase disequilibria (LD) differed significantly between sunny and shady microniches. The levels of correct classification of individual genotypes into their respective microniche, based on the presence/absence of bands of the 2, 3, 4, 5, and 7 best differentiating loci, obtained by stepwise discriminant analysis, were 79, 84, 89, 95, and 100%, respectively. We conclude that ecological contrasts at the Neve Yaar microsite cause molecular diversifying selection (presumably at both coding and noncoding genomic regions of wild barley) overriding gene flow.     

Intraspecific and interspecific variation in Setaria revealed by RAPD analysis

Random amplified polymorphic DNA (RAPD) analysis was applied to foxtail millet (Setaria italica) and other Setaria species in order to assess the degree of polymorphism among and within the species and to investigate their genetic relationships. Twenty accessions of foxtail millet, five of green foxtail (S. viridis), five of yellow foxtail (S. glauca), one each of S. germanica, S. verticilliformis, S. pallidifusca, S. macrostachya, S. sphacelata and S. faberi, and one unidentified, were evaluated for variability using a set of 19 random 10-mer primers. A total of 148 markers were scored for all the accessions. Wide variations in banding profiles among species were observed with nearly every primer used. The results showed that foxtail millet and S. glauca had a considerable degree of variation. The geographic differentiation of RAPD patterns in foxtail millet accessions examined seemed to exist. The present study confirmed that foxtail millet is more closely related to green foxtail than to other species in Setaria, providing support for the origin of foxtail millet from S. viridis and the conspecificity of these two species. The results confirmed that S. germanica and S. italica are conspecific and the former can be replaced by the latter. Setaria glauca and S. sphacelata are distinct from foxtail millet, implying that elite traits found in these two species are relatively difficult to be used in foxtail millet improvement. This study demonstrates that RAPD analysis can be used in assessing intraspecific and interspecific variation in foxtail millet and its wild relatives.     

Evaluation of cultivated and wild barley for resistance to pathotypes ofPuccinia hordei with wide virulence

An isolate ofPuccinia hordei (ND89-3) originally collected in Morocco is virulent on most barley genotypes reported to possess resistance, except cultivar Estate (CI 3410), which possesses theRph3 gene and exhibits a low to intermediate level of resistance (infection type 12). Isolate ND89-3 possesses one of the widest virulence spectrums reported forP. hordei. Accessions ofHordeum vulgare (1,997 in total) andH. spontaneum (885 in total), mostly originating from the Mediterranean region and parts of North Africa, were evaluated with isolate ND89-3 at the seedling stage to identify new sources of leaf rust resistance. Fifty-eight accessions ofH. vulgare, and 222 accessions ofH. spontaneum exhibited low infection types to this isolate. Further evaluations of these resistant accessions with isolates ofP. hordei virulent forRph3,Rph7, andRph12 suggested that most of the resistantH. vulgare accessions possess theRph3 gene. Data suggested additional sources of effective resistance inH. vulgare are rather limited. FiveH. vulgare accessions and 167H. spontaneum accessions were identified as possible sources of new genes for leaf rust resistance. These accessions likely possess resistance genes that are different fromRph1 toRph12, or gene combinations thereof based on their reaction to four leaf rust isolates. Utilization of these accessions in barley breeding will broaden the germplasm resources available for genetic control ofP. hordei. North Dakota Agricultural Experiment Station Journal Series No. 2123. The U.S. Government right to retain a non-exclusive, royalty free licence in and to any copyright is acknowledged.     

浙江省耕地多功能时空格局变化、影响因素及空间溢出效应

探讨耕地多功能的时空变化规律、影响因素及其空间上的溢出效应或交互作用，对深化耕地多功能认知、提升耕地资源可持续利用能力具有重要意义。该研究选取浙江省为研究区域，基于耕地利用、气象、土壤、社会经济等数据，综合运用GIS制图、空间自相关分析和基于可拓展随机性环境影响评估（Stochastic Impacts by Regression on Population，Affluence and Technology，STIRPAT）的面板空间计量模型探讨了浙江省2000-2015年耕地多功能的时空变化特征、影响因素及空间溢出效应。结果表明：1）2000-2015年浙江省各区县耕地功能综合水平呈现"先减少，后增长"的变化趋势，耕地物质产出功能、净固碳功能和景观美学功能均呈上升趋势，水源涵养功能保持平稳，劳动力承载功能逐步降低；2）耕地多功能水平的提升对邻近地区有较强的辐射影响作用，本县域耕地多功能水平每增长1%将导致邻近县域耕地多功能水平增长0.224%。农业劳动力、农村居民可支配收入和农业机械水平对耕地多功能具有显著的正向效应，第二产业占比和城镇化水平对耕地多功能具有显著的负向效应；3）第二产业占比、农业现代化水平、农业劳动力投入对耕地多功能有直接效应，城乡居民收入、城镇化水平对耕地多功能水平有溢出效应。研究建议重视区县内部以及区县与邻近地区之间的空间交互作用，加强区域间土地整治、生态保护和农业技术升级等多方面合作。应充分考虑各县域资源禀赋的特殊性以及社会经济发展阶段的异质性，制定针对性的耕地保护和农业扶持适应策略。     

Ecological correlates of RAPD DNA diversity of wild barley, Hordeum spontaneum, in the Fertile Crescent

Genetic variability in RAPDs (Randomly Amplified Polymorphic DNA) was studied in 104 genotypes of wild barley, Hordeum spontaneum from 21 populations sampled in Israel, Turkey and Iran, seven population from each country. The band (= loci) frequencies were calculated for each population and correlated with ecogeographical variables. In general, high RAPD genetic diversity indices were associated with stressful environments, either with hot or cold steppes and deserts. Interpopulational genetic distances showed no association with the geographic distance between the populations' provenance. Significant Spearman rank correlations between RAPD band frequencies and ecogeographical parameters of provenance occured. Frequencies of RAPD bands were significantly correlated with the principal component factors of allozymes. The correlation data indirectly suggest that natural selection appears to be the major determinant of both RAPD and allozyme diversities both being correlated with environmental stress.     

Ecogeographic adaptation and genetic variation in wild barley, Application of molecular markers targeted to environmentally regulated genes

Long primer PCR (LP-PCR) markers specifically targeted to sequences involved in the response to abiotic stress were utilised to analyse genetic variation within and among wild barley populations from Israel.Populations were sampled in locations reflecting a wide range of habitats and the potential correlation between genetic variation and ecogeographic parameters was examined. A high level of polymorphism was observed. Most of the variability was due to differences within populations, however a substantial portion of genetic variation (36%) could be attributed to differences among populations, indicating local adaptation of genotypes to microclimatic conditions. Moreover, clustering and ordination techniques showed that genotypes grouped together according to their area of origin and were separated on thebasis of ecogeography, chiefly by a combination of temperature, altitude and rainfall variables. Finally, 28% of the polymorphic PCR fragments were significantly correlated with ecogeographic parameters. Our results demonstrate the effectiveness of PCR-based molecular markers targeted to environmentally regulated genes in detecting useful variation andthus in monitoring the impact exerted by adaptation to the environment ongenetic differentiation.