The Genetic Diversity and Similarities Among <Emphasis Type="Italic">Kengyilia</Emphasis> Species Based on Random Amplified Microsatellite Polymorphism (RAMP)

The genetic diversity and similarities among 32 Kengyilia accessions, distributed to 14 species and one variety were analyzed by using random amplified microsatellite polymorphism (RAMP) markers. Of the 160 RAMP primer combinations tested, 40 (25%) produced polymorphic and clear bands. A total of 264 bands were produced by 40 primer combinations, among which 231 out of 264 bands (87.5%) were polymorphic. Two to 11 polymorphic bands could be amplified from each primer combination, with an average of 5.8 bands. The data of 264 bands were used for RAMP assay. By NTSYS-pc program, genetic similarity coefficients were generated and dendrogram was constructed using UPGMA. The genetic similarity coefficients ranged from 0.477 to 0.965 with the mean of 0.714. The results showed as follows: (1) distinct genetic differences were present among the different species; (2) the different accessions in a species were clustered together, respectively, which had larger genetic similarities and closer relations; (3) the species with similar morphological characters and the species from the same areas or neighboring geographical regions were clustered together; (4) the lowest genetic similarity was found between K. hirsuta (PI531618) and K. laxiflora (PI531631), while the highest genetic similarity was observed between K. hirsuta (Y2364) and K. hirsuta (Y2368); (5) RAMP results are basically comparable with those obtained from studies on morphology and cytology. It is a useful method for analysis of the genetic diversity and similarities in Kengyilia.     

Diversity Analysis of Garlic (Allium sativum L.) Germplasms Available in Nepal Based on Morphological Characters

A total of 179 garlic (Allium sativum L.) accessions were collected from various parts of Nepal in 2000. Each accession was planted in each of a plot of 2.25 m² at the Institute of Agriculture and Animal Science (IAAS) Horticulture farm (225 m asl) and at the Agriculture Research Station, Dailekh (1400 m asl) of Nepal Agriculture Research Council (NARC) in the first week of November 2000. All accessions were characterized for leaf erectness, leaf color, leaf wax, leaf cross section, bulb regularity, bulb skin color, bulb outer scales number, days to emergence, bulbing period, number of green leaves at 135 days after planting, days to maturity, plant height, bulb weight, bulb diameter, number of cloves per bulb, clove diameter and bulb yield. Data were analyzed by using principal component and cluster analysis procedures to reveal three major clusters. Four principal components were identified explaining more than 86% of total variation. Major characters included in the principal components were bulb weight, diameter, yield, number of cloves per bulb, maturity, plant height, number of green leaves at 135 days after planting, and bulbing period. The level of variation found in the collection showed the great potentiality of improving agronomic characters in garlic.     

Genetic Diversity and Phylogenic Relations Among Sahelian Sorghum Accessions

To establish the genetic relationship among Sahelian sorghum [Sorghum bicolor (L.) Moench S.L.] landraces from Burkina Faso were submitted to electrophoretic analysis for 10 enzymatic systems and 18 loci. Four enzymatic systems (ADH, LAP, MDH, PGD) and five loci revealed polymorphism both within and among landraces. Thirty-eight per cent of the landraces were monomorphic in all the 18 loci. The genotypic frequencies in most of the landraces deviated markedly from Hardy–Weinberg proportions due to a major heterozygote deficit, the landrace being homozygous or a mixture of homozygotes. Multivariate analysis yielded three main groups, containing native landraces and five minors, containing introduced cultivars, randomly distributed over the territory. The pattern of allelic occurrence was random and unrelated to external selection pressures. The major diversity among landraces appears to be from genetic shift caused by farmers’ selection of their seeds. It could also be due to the low rates of outcrossing (19%) and migration (0.06) prevailing in the set.     

Comparative Analysis of Genetic Diversity in Two Tunisian Collections of Fig Cultivars Based on Random Amplified Polymorphic DNA and Inter Simple Sequence Repeats Fingerprints

This study characterized the genetic diversity of 18 Tunisian fig cultivars using random amplified polymorphic DNA (RAPD) and inter simple sequence repeats (ISSR). Both random and ISSR primers tested generated a total of 116 RAPD and 47 ISSR markers. Considerable genetic variation was observed among fig cultivars sampled from two regional Tunisian collections with an average diversity of 4.57. RAPD and ISSR banding patterns and genetic distances values reflected the high level of diversity among the collections and lower variability between the two collections. The correlation between the RAPD and ISSR similarity matrices computed for the 153 pairwise comparisons among the 18 varieties was lower and significant. An analysis of molecular variance showed that 92% of the total genetic diversity resided within collections, whereas only 8% between collections. The results indicated that in the local fig germ plasm the information provided by RAPD and ISSR is not analogous, most likely as a consequence of the fact that the two classes of markers explore, at least in part, different portions of the genome.     

Polymorphism and Genotype-specific Markers for Dichanthium Identified by Random Amplified Polymorphic DNA

One hundred decamer arbitrary primers were tested for PCR based amplification of seven genotypes (IG2208-S-1, IG2177, IG2180, IG2178, IG2165-S-1-1, IG2165-1 and Local-1) of an apomictic grass, Dichanthium annulatum, with the aim of screening polymorphic primers and genotype-specific markers. Out of 100 decamer primers tested, 42 produced no amplification or smeared non scorable bands, 12 amplified only single band and 46 yielded more than one polymorphic bands. Thirty-two primers out of 46 selected showed high level of polymorphism, producing 3–15 reproducible bands each for the seven Dichanthium genotypes examined. Among the total of 307 amplified fragments 222 were polymorphic, 53 bands were unique to the genotypes and 32 were monomorphic. Thus, with selected primers sufficient polymorphism could be detected to allow identification of individual genotypes. Genetic similarities of RAPD profiles generated were estimated via a coefficient of DICE and then the data were processed by cluster analysis (UPGMA). The maximum similarities between two genotypes (IG2180 and IG2178) was 58% and these two made a cluster with genotype IG2177 having similarity of only 54%. It clearly corroborated existence of high levels of polymorphism in this grass though being apomictic in nature. Primers like OPE-16, OPG-02, OPG-18, OPH-05, OPH-09, OPH-16, OPI-07 and OPF-06 found most informative as they produced specific bands pertaining to five out of seven genotypes. Polymerase chain reaction (PCR) offers a substantially simple, rapid and reliable method for identification of large number of Dichanthium genotypes once enough number of reproducible and suitable primers is screened.     

Genetic diversity in bambara groundnut (Vigna subterranea (L.) Verdc) landraces assessed by Random Amplified Polymorphic DNA (RAPD) markers

Genetic diversity in 12 landraces of bambara groundnut (Vigna subterranea), an indigenous African legume, was evaluated using Random Amplified Polymorphic DNA (RAPD) markers. DNA from individuals of each landrace was also analysed to determine the level of heterogeneity within landraces. RAPDs revealed high levels of polymorphism among landraces. The percentage polymorphism ranged from 63.2% to 88.2% with an average of 73.1% for the 16 RAPD primers evaluated. The construction of genetic relationships using cluster analysis groups the 12 landraces in two clusters. RAPDs are useful for the genetic diversity studies in V. subterranea and can identify variation within landraces.     

Genetic Diversity within and Between Populations of Lathyrus genus (Fabaceae) Revealed by ISSR Markers

In order to evaluate the genetic diversity in Lathyrus genus, the Inter Simple Sequence Repeats method (ISSR) was exploited in five populations. These consisted of two cultivated species belonging to section Lathyrus (L. sativus L. and L. cicera L.) and a wild one belonging to the section Clymenum (L. ochrus DC.). Two 3′anchored ISSR primers and two unanchored ones, generated a total of 60 useful polymorphic DNA bands. Our data provide evidence of high molecular polymorphism at the intra- and the inter-specific levels showing that both wild and cultivated forms constitute an important pool of diversity. Moreover, among the generated DNA bands, a 500 bp band, totally absent in the banding patterns of the section Clymenum, appears to be a molecular marker of section Lathyrus. Results provided for lineage and suggest recent origin of these species that might have evolved from a common ancestor producing both L. ochrus species and the two other species L. sativus and L. cicera. These relationships support previous studies based on morphological variation and molecular analysis.     

Geographic Pattern of Genetic Diversity Among 43 Ethiopian Mustard(Brassica carinata A. Braun) Accessions as Revealed by RAPD Analysis

As an oilseed crop, the cultivation of Ethiopian mustard (Brassica carinata) is restricted only to Ethiopia. Even though geographic diversity is a potent source of allelic diversity, the extent of genetic diversity among germplasm material of Ethiopian mustard from different countries has not been assessed. Forty-three accessions, comprising 29 accessions from eight different geographic regions of Ethiopia and 14 exotic accessions from Australia, Pakistan, Spain, and Zambia were analysed for their genetic diversity using random amplified polymorphic DNA (RAPD) technique. A set of 50 primers yielded a total of 275 polymorphic bands allowing an unequivocal separation of every Ethiopian mustard accession. The usefulness of the 50 RAPD primers in measuring heterozygousity and distinguishing accessions was variable such that polymorphic information content (PIC) varied from 0.05 to 0.40, band informativeness (BI) from 0.05 to 0.65 and primer resolving power (RP) from 0.15 to 6.83. Jaccard's similarity coefficients ranged from 0.44 to 0.87 indicating the presence of a high level of genetic diversity. On the average, Australian and Ethiopian accessions were the most similar while, Spanish and Zambian accessions were the most distant ones. Cluster analysis grouped the 43 accessions into four groups, which has quite a high fit (r = 0.80) to the original similarity matrix. With no prior molecular information, the RAPD technique detected large genetic diversity among the 43 accessions from five different countries and their grouping by dendrogram and principal coordinate analysis (PCoA) was inclined towards geographic differentiation of RAPD markers. Conversely, RAPD differentiation along geographic origin was not apparent within the Ethiopian accessions.     

Genetic Diversity Among Traditional Ethiopian Highland Maize Accessions Assessed by Simple Sequence Repeat (SSR) Markers

Over the past three centuries, maize has become adapted to complex environmental conditions in the highlands of Ethiopia. We analyzed 62 traditional Ethiopian highland maize accessions, using 20 simple sequence repeat (SSR) markers and 15 morphological traits, to assess genetic diversity and relationships among these accessions and to assess the level of correlation between phenotypic and genetic distances. The accessions varied significantly for all of the measured morphological traits. The average number of alleles per locus was 4.9. Pair-wise genetic dissimilarity coefficients ranged from 0.27 to 0.63 with a mean of 0.49. Ward minimum variance cluster analysis showed that accessions collected from the Northern agroecology were distinct from the Western and Southern agroecologies. However, there was no differentiation between the Western and Southern accessions. This suggested gene flow between these regions. The relationship between morphological and SSR-based distances was significant and positive (r = 0.43, p = 0.001). The high genetic diversity observed among these set of accessions, suggests ample opportunity for the development of improved varieties for different agroecologies of Ethiopia. From conservation perspective, sampling many accessions from all agroecologies would be an effective way of capturing genetic variation for future collections and conservation.     

Genetic Relationships and Variation among Biotypes of Dallisgrass (Paspalum dilatatum Poir.) and Related Species Using Random Amplified Polymorphic DNA Markers

The genus Paspalum L. consists of more than 400 species. Around twenty-five informal groups of species are recognized in Paspalum and the Dilatata group is of special interest because its members are excellent potential forage grasses. Seventy-five germplasm accessions, representing 15 taxa, were analyzed using randomly amplified polymorphic DNA (RAPD). Polymorphisms were observed with twenty-two primers in the Dilatata group and 16 of those were analyzed. Four hundred and four different RAPD fragments were generated, resulting in an average of 25.2 bands per primer. Among the 404 markers analyzed, 48 (11.88%) were exclusive for the P. dilatatum Poir. biotypes, 31 (7.67%) were exclusive to taxa belonging to other groups included in this study, 28 markers (6.93%) were diagnosed for other species of the Dilatata group and 16 (3.96%), for natural hybrids. Extensive RAPD variation was found among the species studied. Inter- and intra-taxonomic polymorphisms were detected. A dendrogram based on the RAPD data shows some clusters corresponding to the same taxa. However, the biotypes of P. dilatatum do not form a cluster. The present work confirms that the RAPD technique can be used to determine genetic relationships between the taxa belonging to the Dilatata group.     

Assessing Genetic Diversity of Chinese Cultivated Barley by STS Markers

To assess the genetic diversity among China’s cultivated barley, sequence tagged site (STS) marker analysis was carried out to characterize 109 morphologically distinctive accessions originating from five Chinese eco-geographical zones. Fourteen polymorphic STS markers representing at least one in each chromosome were chosen for the analysis. The 14 STS markers revealed a total of 47 alleles, with an average of 3.36 alleles per locus (range 2–8). The proportion of polymorphic loci per population averaged 0.84 (range 0.71–1.00); the mean gene diversity averaged 0.39 (range 0.28–0.49). The means of P and H_e were highest in the Yangtze reaches and Southern zone (P = 1.00; H_e = 0.46) and lowest (P = 0.71 He = 0.28) in the Yellow river reaches zone. The STS diversity in different zones is quite different from the morphology diversity. The STS variation was partitioned into 17% among the zone and 83% within the zone. Both cluster and principal coordinate analyses clearly separated the accessions into a dispersed group (mostly two-rowed barley with a lower mean GS value) and a concentrated group (mostly six-rowed barley with a higher mean GS value) according to the spike characteristic with only a few exceptions. The accessions from the Qinghai-Tibet plateau formed a distinctive subgroup and can be distinguished from the concentrated group. The role of Tibet in the origin and evolution of cultivated barley has been discussed.     

Genetic Diversity of Asian Cotton (Gossypium arboreum L.) in China Evaluated by Microsatellite Analysis

Asian cotton (Gossypium arboreum L.) was once widely cultivated in China. It has also been a valuable source of genetic variation in modern cotton improvement. In this study, the genetic diversity of selected G. arboreum accessions collected from different regions of China was evaluated by microsatellite (simple sequence repeats, SSRs) analysis. Of the 358 microsatellite markers analyzed, 74 primer pairs detected 165 polymorphic DNA fragments among 39 G. arboreum accessions examined. Twelve accessions could be fingerprinted with one or more SSR markers. With the exception of two accessions, DaZiJie and DaZiMian, genetic similarity coefficients among all accessions ranged from 0.58 to 0.87 suggesting high level of genetic variation in the G. arboreum collections. The UPGMA dendrogram constructed from genetic similarity coefficients revealed positive correlation between cluster groupings and geographic distances. In addition, comparison of the microsatellite amplification profiles of the diploid G. arboreum and tetraploid Gossypium hirsutum L. found that size distribution of amplified products in G. arboreum was dispersive and that of G. hirsutum was relatively concentrated. The information on the genetic diversity and SSR fingerprinting from this study is useful for developing mapping populations for constructing diploid cotton genetic linkage map and tagging economically important traits.Diqiu Liu, Xiaoping Guo: These two authors contributed equally to this work.     

Genetic Diversity in Tef [<Emphasis Type="Italic">Eragrostis tef</Emphasis> (Zucc.) Trotter] Germplasm

One hundred and forty-four heterogeneous tef germplasm accessions collected from 10 major tef growing areas in Ethiopia were evaluated for 18 quantitative traits in a simple lattice design, at Holetta and Ginchi, in 2001 main growing season to study the genetic diversity in tef. The combined analysis of variance revealed the presence of significant variation among germplasm accessions for all traits studied. Cluster analysis revealed the overlapping of the germplasm accessions from different origin and the accessions were grouped into eight distinct clusters of 1–78. Generalized distance square confirmed the presence of significant genetic distance between clusters. In principal component (Prin) analysis, the first four principal components with latent root values above one accounted for about 80.6% of the variability existing among the germplasm accessions. Prin1 constituted about 55% of the variability mainly from almost equal contribution of 10 quantitative traits, indicating that most of the traits are equally important in tef diversity. In this study, the regions and altitudes of origin were not found to have a substantial effect on the genetic diversity in tef germplasm. Diversity within the regions was found to be significant and, hence, an opportunity for exploitation in tef improvement.     

Assessment of Genetic Diversity in Oil Palm (Elaeis guineensis Jacq.) using Restriction Fragment Length Polymorphism (RFLP)

A total of 359 accessions of oil palm (Elaeis guineensis Jacq.) originating from 11 African countries (Nigeria, Cameroon, Congo DR, Tanzania, Angola, Senegal, Sierra Leone, Guinea, Ghana, Madagascar and Gambia) were characterized using the RFLP method using the standard Deli dura as the check. Genomic DNA from each sample was digested using five restriction enzymes and hybridized with four oil palm cDNA probes. Data were analyzed using Biosys-1 computer software to calculate the genetic variability parameters. In general, all the collections exhibited higher levels of diversity than the standard variety, Deli dura. The standard variety, Deli dura, lost 36 alleles as compared to the natural populations indicating a reduction in genetic variability. Material from Nigeria showed the highest mean number of alleles per locus (1.9) and percentage of polymorphic loci (67.2%). These findings, combined with others, suggest that Nigeria may be the center of diversity of wild oil palm. It further suggests that oil palm natural populations maybe possessing adequate genetic variability that are potentially useful for improvement programs.     

秦巴山区黄牛群体的微卫星DNA遗传多样性

为分析秦巴山区西镇牛、赤崖牛、陨巴牛和宣汉牛4个黄牛(Bos taurus)品种的遗传多样性,本研究以蜀宣花牛和郏县红牛作对照,利用12对微卫星引物对6个黄牛品种的289头黄牛个体进行了遗传多样性检测,统计了各品种的等位基因及频率、有效等位基因数、遗传杂合度、多态信息含量及各群体间遗传距离,并对其进行聚类分析.结果表明,6个黄牛品种在12个微卫星位点共发现110个等位基因,等位基因频率为0.001 6～0.517 3,总群体各位点平均有效等位基因数为2.787 7～7.132 6;各位点多态信息含量为0.5192～0.895 3;平均杂合度为0.667 2～0.724 1.群体间发现特有等位基因11个,基因频率为0.008 1～0.381 6;优势等位基因(P＞0.4) 19个,基因频率为0.403 2～0.820 0;共有等位基因41个,占全部等位基因的37.27％,仅有13个共有等位基因为优势等位基因(P＞0.4),占37.71％.群体间Nei's遗传一致度为0.596 5～0.840 8,标准遗传距离(Ds)为0.173 5～0.524 7.聚类分析结果显示,6个黄牛品种聚为3类,陨巴牛与宣汉牛首先聚在一起,然后同西镇牛聚在一类;赤崖牛与郏县红牛聚为一类;蜀宣花牛自成一类.研究结果表明,12对微卫星标记可用于秦巴山区黄牛遗传多样性的分析,秦巴山区各黄牛品种遗传多样性丰富,选育程度不同,4个黄牛品种虽然地理分布格局相近,自然环境相似,但亲缘关系并非相近,应为来源不同的品种.因此,西镇牛、赤崖牛、陨巴牛和宣汉牛不宜合并为1个品种.本研究所揭示的秦巴山区黄牛品种的遗传分化特点及亲缘关系,为研究品种的遗传共适应特点,预测杂交优势,制定育种战略等提供了科学依据.     

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.     

Genetic Diversity for Morphological and Quality Traits in Quinoa(<Emphasis Type="Italic">Chenopodium quinoa</Emphasis> Willd.) Germplasm

Twenty nine germplasm lines of Chenopodium quinoa and two of Chenopodium berlandieri subsp. nuttalliae were evaluated for 12 morphological and 7 quality traits for two test seasons. The 19 traits were analyzed for cluster and principal component analysis. The first four PCs contributed 78.70 % of the variability among the germplasm lines. The first PC accounted for 39.5% of the variation and had inflorescence/plant, plant height and stem diameter as the traits with largest coefficients, all with positive sign. The characters with greatest positive weight on PC₂ were days to maturity (0.309), inflorescence length (0.260) and branches/plant. All the germplasm lines were grouped into six clusters based on average linkage method. Cluster III had high values for seed yield and most of the quality traits but showed a small seed size. The dendrogram separated the two lines of C. berlandieri subsp. nuttalliae from the quinoa lines.     

Genetic and Phenotypic Diversity in Downy-mildew-resistant Sorghum (Sorghum bicolor (L.) Moench) Germplasm

Genetic and phenotypic diversity among randomly selected 36 downy-mildew-resistant sorghum accessions were assessed, the former using 10 simple sequence repeat (SSR) marker loci and the latter using 20 phenotypic traits. The number of alleles (a _j ) at individual loci varied from five to 14 with an average of 8.8 alleles per locus. Nei's gene diversity (H _j ) varied from 0.59 to 0.92 with an average of 0.81 per locus. High gene diversity and allelic richness were observed in races durra caudatum (H _j = 0.76, a _j = 4.3) and guinea caudatum (H _j = 0.76, a _j = 3.8) and in east Africa (H _j = 0.78, a _j = 7.2). The regions were genetically more differentiated than the races as indicated by Wright's F _st. The pattern of SSR-based clustering of accessions was more in accordance with their geographic proximity than with their racial likeness. This clustering pattern matched little with that obtained from phenotypic traits. The inter-accession genetic distance varied from 0.30 to 1.00 with an average of 0.78. Inter-accession phenotypic distance varied from 0.01 to 0.55 with an average of 0.33. Eleven accession-pairs had phenotypic distance of more than 0.50 and genetic distance of more than 0.70. These could be used as potential parents in a sorghum downy mildew resistance-breeding program.     

Genetic Diversity Among Brazilian Cultivars and Landraces of Tomato Lycopersicon Esculentum Mill. Revealed by RAPD Markers

Random amplified polymorphic DNA markers (RAPD) were used to estimate the variability of 35 tomato accessions (Lycopersicon esculentum Mill.). A total of 257 reproducibly scorable bands were obtained from 20 primers, 78.6% of which were polymorphic. The percentage distribution of RAPD markers shows a bimodal distribution, and the frequency of rare alleles is similar in commercial and landrace accessions. Genetic distances among accessions were calculated and a dendrogram showing the genetic relationships among them was constructed allowing for the separation of four groups. Twenty out of 23 Brazilian landraces fell within one group, whereas commercial cultivars were distributed in the four groups. AMOVA analysis of RAPD data showed that, despite the high within Brazilian landraces and commercial cultivars variation, these two groups are significantly different, indicating that landraces can be a source of variation for breeding programs.     

Assessment of Genetic Relationships between Wild and Cultivated Mulberry (Morus) Species using PCR Based Markers

In order to formulate appropriate strategies for the conservation and utilization of the wild mulberry genetic resources available in India, a study was undertaken with 20 mulberry genotypes from the four different species. Seventeen intersimple sequence repeat primers were used to generate a total of 114 markers, of which 98 (85.96%) were polymorphic. Seven unique bands for Morus serrata Roxb. and one for both M. serrata Roxb. and Morus macroura Miq. were identified, of which one fragment has been sequenced and deposited in the EMBL-GeneBank (AJ-585512). The genetic dissimilarity coefficients varied from 0.078 to 0.530 among these genotypes and from 0.168 to 0.465 among the species. The dendrograms realized from these markers clustered the genotypes into three groups. The outermost group was M. serrata Roxb., which was followed by the group of M. macroura Miq. and the innermost group contained genotypes of Morus indica L. and Morus alba L. This intermixing of genotypes of M. indica and M. alba supports the view that M. indica is merely a synonym of M. alba. Distribution of the genotypes on a two-dimensional figure upon multidimensional scaling with ALSCAL program, further, confirmed the genetic divergence between the cultivated and wild mulberry groups. On the basis of the results a few potential wild mulberry genotypes were identified for its conservation and utilization in breeding programs to confer the stress tolerance to the cultivated varieties of mulberry.