Seed storage protein electrophoresis in Arachis pintoi and A. repens (Leguminosae) for evaluating genetic diversity

Seed storage protein profiles were studied to evaluate the genetic diversity of a germplasm collection of the Caulorrhizae section of genus Arachis. Total seed storage protein of 28 accessions of Arachis pintoi and one of A. repens, collected in the Paranã, São Francisco and Jequitinhonha river basins, was compared by native-PAGE and SDS-PAGE. Significant variation was detected among the protein profiles of different accessions of Arachis pintoi, and three major protein profiles A, B, C were revealed in native-PAGE. Protein variation was much higher between accessions than within accessions. Nevertheless an exceptional case of intra-accession variation was observed in accession W 34b. In SDS-PAGE major variation between the accessions was confined to the region of arachin (basic group), which corresponds to region III in native-PAGE. There was no correlation between seed protein profiles and geographic distribution. However, protein polymorphism is predominantly in accessions of the São Francisco (A, B and C phenotypes) and Paranã (A and B phenotypes) basins, with the B phenotype predominating in the São Francisco basin. A specific protein profile to differentiate between the typical forms of A. pintoi and A. repens was not found.     

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.     

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.     

Improved fractionation of glycinin and beta-conglycinin and partitioning of phytochemicals

Glycinin-rich and beta-conglycinin-rich products are prepared by soy protein fractionation. Physicochemical characteristics of these proteins affect their unique, important functionality in food systems and industrial applications. Soybean isoflavones and saponins are phytochemicals with potential health benefits. Objectives of this protein fractionation research were to (1) improve protein and phytochemical extraction from defatted soy flakes and recovery in product fractions and (2) evaluate phytochemical partitioning and profile changes during fractionation. Extraction environments (pH, ethanol concentration, temperature, and water-to-flake ratio) were each varied during bench-scale optimization. Optimized conditions of 45 degrees C and 10:1 water-to-flake ratio were compared with previous conditions of 20 degrees C and 15:1 water-to-flake ratio and a soy protein isolate process at pilot scale. Optimized conditions yielded more beta-conglycinin with higher isoflavone and saponin concentrations, but fraction purity was diminished by glycinin contamination. Bench-scale data demonstrated that increased phytochemical extraction did not translate into increased concentrations in product fractions.     

Genetic Diversity of Japanese Melon Cultivars (<Emphasis Type="Italic">Cucumis melo</Emphasis> L.) as Assessed by Random Amplified Polymorphic DNA and Simple Sequence Repeat Markers

The genetic diversity among 67 melon (C. melo L.) cultivars from five Japanese seed companies was assessed using 25 10-mer RAPD primers (56 bands) and nine SSR (36 alleles) markers. These cultivars belong to three horticultural varieties (synom. Groups) spanning eight melon market classes: Group Cantalupensis (market classes Earl’s, House, Galia, Charentais, and Ogen), Group Inodorus [Honeydew and Casaba melons (market classes Amarillo, Piel de Sapo, Rochet, Negro, Crenshaw, and Tendral)], and Group Conomon (market class Oriental). Genetic variation among these cultivars was compared to variation in a reference array (RA) consisting of 34 selected melon accessions from previous studies. Cluster analysis resulted in 11 of 15 Japanese Oriental accessions forming a group with South African RA accessions. The remaining Group Conomon Japanese accessions grouped either with Casaba or with Honeydew cultivars. Japanese Group Conomon accessions and South African RA accessions formed a genetic group that was distinct from all other accessions studied, and suggests either an Asiatic origin for the South African melon germplasm examined or an independent domestication involving similar ancestors. The majority of Japanese House and Earl market class accessions shared genetic affinities, and were genetically different from the Japanese Group Inodorus accessions examined. These Japanese accessions were most similar to Casaba RA accessions. Japanese Galia accessions were similar to either House and Earl’s market classes or to Galia, Ogen, Casaba, and Honeydew RA accessions. Genetic differences exist between melon types that were domesticated from wild, ‘free-living’ subspecies agrestis and from melo. Mention of trade name, proprietary products, or specific equipment does not costitute a guarantee or warranty by the USDHA and does not imply its approval to the exclusion of other products that may be suitable.     

Genetic variation and species relationships in Himalayan buckwheats as revealed by SDS PAGE of endosperm proteins extracted from single seeds and RAPD based DNA fingerprints

SDS PAGE of endosperm proteins and␣RAPD profiles from different accessions of Himalayan buckwheat were studied to determine their genetic variation and phylogenetic relationships. Comparisons based on Jaccard’s coefficient and UPGMA clustering revealed interrelationships broadly in conformity with conventional treatments. Cluster analysis of the endosperm protein profiles of the selected accessions revealed three broad clusters. A moderate level of intraspecific variability was detected in the endosperm protein profile of different accessions of Fagopyrum esculentum, Moench. Three subgroups were detected in cluster 1. Subgroup 1 included varieties designated as Local, Kamroo local, OC-2 and␣VL-7 which were collected from VPKAS, Almora. Local, Kamroo local and OC-2 showed a similarity coefficient of 1.0 inspite of their being identified as different accessions. VL-7 emerged out separately from the rest of the three accessions. Accessions having winged grains and those having striations on the seed coat formed a 2nd and 3rd subgroup, respectively. IC-13145 which been identified as “F. himalianum”, showed 100%␣similarity in endosperm protein profile with IC-13376 (F. esculentum) and 85–90% similarity which other accessions of F. esculentum. Our results indicate that “F. himalianum” belongs to the esculentum group and should not be regarded as a different species. Cluster 2 included all the accessions of F. tataricum (L.) Gaertn. with Sangla 1, 2, 3, 5, 6, 7 and IC-412863 showing 100% similarity. F. cymosum emerged as a separate group distinct from both esculentum and tataricum. Accessions of F. tataricum and F. cymosum did not show significant intraspecific variation in the SDS PAGE profile of endosperm proteins. Out of the 20 primers used, 3 generated robust, easily interpretable amplified products. While a 1490 bp and a 300 bp RAPD was detected only in F. tataricum, a 1154 bp RAPD was detected in all accessions of F. tataricum except in Shimla B-1. This variety is early maturing and has high seed yields.     

Seed protein electrophoretic characterization of cowpea (Vigna unguiculata) germplasm from IITA gene bank

Summary The total seed protein, globulin and albumin fractions of 20 cowpea (V. unguiculata) accessions from IITA gene bank were investigated by SDS-polyacrylamide gel electrophoresis. Total seed protein extracts were prepared from the defatted meal by homogenisation and centrifugation in Tris-glycine buffer. The globulins and albumins were obtained from the total protein extract by exhaustive dialysis against sodium acetate buffer. Two main total seed protein electrophoretic patterns were observed with respect to the 39 and 20 kD subunits, which were present in six out of the twenty accessions analysed. While there was no correlation between seed colour and total seed protein banding pattern, the six insect-resistant cultivars were characterized by the presence of the 39 and 20 kD subunits. The globulins were the predominant class of the total seed proteins and consisted mainly of 64, 58, 56 and 14 kD subunits which make up CP1 and CP2, the major globulins. The albumins in all accessions were a heterogeneous protein fraction consisting of both high and low molecular weight subunits. It was suggested that the insect-resistant cultivars may be genetically related and that the 39 and 20 kD subunits may be involved in the insect resistance mechanism.     

Wet Processing of Barley Grains into Concentrates of Proteins, β‐Glucan,and Starch

An improved wet method was developed to process barley into fractions concentrated in protein, (1‐3)(1‐4)‐β‐d ‐glucan (BG), starch, or other carbohydrates (CHO). Alkaline concentration, solvent to barley flour ratio (SFR), and extraction temperature were evaluated for their effects on concentration and recovery of protein, BG, starch, oil, ash, and other CHO in each fraction type. Results show that the three parameters and their interactions all had significant effects, resulting in varying nutrient concentrations and recovery rates in each type of fractions. For protein fractions, protein content varied from 37.7 to 75.2%, protein recovery from 8.5 to 75.7%, and increasing alkaline concentration and SFR improved nutrient recovery. For BG fractions, BG content ranged from 21.5 to 87.0%, BG recovery from 28.6 to 78.0%, and increasing alkaline concentration decreased BG content but increased its recovery significantly. For starch fractions, starch content varied from 76.9 to 93.9%, starch recovery from 33.6 to 63.9%, and all parameters had little effect on the nutrient concentrations, but alkaline concentration and SFR improved recovery of starch, other CHO, and mass. Overall, the improved wet method was effective in concentrating the major nutrients from barley into their respective fractions, but process optimization through manipulating the three parameters is necessary to achieve a maximum concentration or recovery rate of a nutrient of interest in a specific fraction.     

Protein heterogeneity in European wheat landraces and obsolete cultivars

Identity and present degree of genetic homogeneity and heterogeneity, respectively of 52 European wheat accessions, maintained in the collection of wheat genetic resources, have been characterized using analyses of glutenins by sodiumdodecylsulphate polyacrylamide gel electrophoresis (SDS-PAGE). Six of the analyzed wheat accessions were observed to be homogeneous, while 46 (88.5%) of them were heterogeneous in protein profiles. Heterogeneous accessions possessed 2 to 13 different protein lanes. Together, 17 high molecular weight glutenin subunit (HMW-GS) alleles have been found. The most frequent HMW-GS alleles at the Glu-A1, Glu-B1, and Glu-D1 complex loci were 1, 7+9, and 2+12, respectively. However, also low frequented HMW-GS alleles or allelic combinations, such as 7+15, 13+16, 20, 6, 7, and 9 were observed. Furthermore, another new allele encoding HMW glutenin subunit with relative molecular weight 98.6 kDa has been found in one of the lines of the cultivar Eritrospermum 917. The Glu-score in the examined accessions varied in broad range, some of the lines reached the maximum value 10.     

Exploration of <Emphasis Type="Italic">Vernonia galamensis</Emphasis> in Ethiopia,and Variation in Fatty Acid Composition of Seed Oil

Vernonia galamensis is a new potential industrial crop with very high content of vernolic acid in the seed oil. The species is known to naturally grow as a weed in fields or in woodlands under a wide range of agroecological conditions of Africa. In order to study the existing variability in Ethiopia, germplasm collection was carried out. Vernonia grows wild in various ecosystems. Ten regions were explored from North, South, East, Southeast, Southwest and Central Ethiopia. A diverse range of habitats having different altitudes and ecological conditions was explored. Altitude of collecting sites varied between 1250 and 2050 m, and soil pH from 5.1 to 8.5. The most common soil type was sandy loam, and the organic matter content varied from 0.2% to 12.9%. At 80 sites, about 480 accessions were collected including different maturity time, plant type, flower color, and branching patterns as well as fatty acid composition. The mean vernolic acid content of the seed oil of the accessions was 74%, and ranged from 34% to 87%. A wide variability in composition of other fatty acids was observed. It was not possible to find Vernonia in some locations that were earlier indicated by herbarium specimens collected since 1840. This could be a sign of change in land use system and environmental degradation and, hence, loss of genetic resources of the species.     

Vigna vexillata (L.) A. Rich. seed proteins: heterogeneity in subunits of globulin fraction

Forty-six accessions of Vigna vexillata from Africa, America and Australia were screened for variability in globulin protein fraction by using SDS-PAGE under reducing and non-reducing conditions. The globulin fraction was composed of several bands clustering in two size fractions, both of which were polymorphic. The high molecular weight fraction was constituted by one or two subunits, while the fraction of lower molecular weight exhibited from two to four bands. Two single types were found within the collection. Due to the observed polymorphism only the subunit at 52 kD was species-specific. Southeastern Africa proved to be the region with the highest diversity for Vigna vexillata. Different frequencies of the globulin patterns were observed between the African and American groups of accessions. No significant differences were found comparing the distribution of the globulin types between the varieties vexillata and angustifolia.     

Collections of short-day onion germplasm in West Africa: a survey

Results are presented of a 1994 postal survey of holdings of local onion (Allium cepa L.) germplasm by institutions in West Africa. Data obtained from respondents included the number of accessions of onion or shallot held and the conditions in which onion seed was stored. Twenty-four replies were received and eighteen sites where collections were held were identified. The numbers of local A. cepa accessions in local collections varied in number from 38 down to nil, but many local collections were of less than five accessions. The largest number of national onion accessions was held at CERRA, Maradi, in Niger (16), and the largest regional collection was at the Station de Farako-bâ, Burkina Faso (38). Two sites held substantial shallot collections: CRA Bareng in Guinée and IDESSA, Bouaké, Côte d'Ivoire. Conditions under which onion seed was stored varied from low temperature, controlled storage in moisture-proof packs with desiccants, to storage in plastic envelopes under ambient laboratory conditions.     

Nutrient Content in Buckwheat Milling Fractions

Buckwheat seeds (Fagopyrum esculentum Moench) were milled into 23 fractions: seven fine flours, three coarse flours, four small semolina, two big semolina, six bran, and one husk fraction. A considerable variation in gross chemical composition was found among the milling fractions. The protein content varied from 4.4 to 11.9% (db) in flours and from 19.2 to 31.3% in bran fractions; starch varied from 91.7 to 70.4% in flours and from 42.6 to 20.3 in bran. The percentage of soluble dietary fiber contained in total dietary fiber was higher in flours than in semolina and bran fractions. Ash, Fe, P, tannin, phytate content, and color were also investigated. A unique distribution of phytate was found in starch. Correlation is significantly positive in husk, bran, and semolina fractions, while correlation is significantly negative in flour fractions. Depending on technological or nutritional demands, appropriate fractions may be chosen to achieve the desired end‐use product.     

Nitrogen Fixation and N-Balance Studies on Bambara Groundnut (Vigna subterranea L. Verdc) Landraces Grown on Tropical Acidic Soils of Malaysia

Bambara groundnut has great potential as an alternative crop for improving food security in its production regions and beyond. A field experiment was conducted at the Field Research Centre of the Crops for the Future to obtain information on the nitrogen (N) fixation and N balance of Bambara groundnut landraces on tropical acidic soils of Malaysia. Treatments consisted of three Bambara groundnut landraces (Ex-Sokoto, Kaaro, and NN-1) laid out in a randomized complete block design replicated three times. Results obtained revealed that Ex-Sokoto landrace was greater in yield and N fixation, whereas N balance (-haulm) was greater in NN-1 landrace. The results revealed grain yield of 703–2256 kg ha^?1 and N fixation from 32–81 kg ha^?1 and suggest that Bambara groundnut could be integrated into a cereal-based cropping system. Ex-Sokoto landrace appeared to be the most promising for yield and N fixation under Malaysian acidic soil conditions.     

Genotypic diversity and patterns of variation in a germplasm material of Ethiopian mustard (Brassica carinata A. Braun)

The amphidiploid species Brassica carinata A. Braun is believed to have originated in the plateaus of Ethiopia and has been cultivated there as an oilseed crop since antiquity. The species possesses agronomically important genes of rare occurrence. Although there is a large number of collections in Ethiopia, information on the extent of their genetic diversity is very limited. Thirty-six accessions of ecologically diverse regions were tested at three locations and multivariate analyses on 13 morphological and seed characters were performed. There was generally a large amount of divergence in all characters. Geographic isolation of genes, however, was not observed. Both principal component and cluster analyses disclosed complex relationships among the accessions and characters. Accessions with potential genes of interest to improve earliness, yield components and oil and protein contents have been identified. Length of growing period and yield components contributed most for divergence and clustering pattern. Oil, glucosinolate and protein contents also varied among the accessions but not so much between clusters. Increasing protein in high-glucosinolate genotypes reduced oil content and should be more efficient in genotypes of low-glucosinolate genetic background.     

Evaluation of Antioxidant,Lipid, and Protein Fractions of Accessions of Oryza Species

The genus Oryza has given rise to rice (Oryza sativa L.), a major source of food for much of the human population. The Oryza genus is small, including only 23 species, but it is remarkably diverse in terms of its ecological adaptation. This diversity may not be only restricted to ecological characteristics but also to kernel end‐use quality characteristics. This study was undertaken to evaluate Oryza species as a gene pool for improving the properties of rice bran for human consumption. Several accessions of 13 Oryza species were grown in a greenhouse, along with eight rice (Oryza sativa L.) accessions displaying low and high bran oil content, as well as low and high palmitic acid content. The total lipid content of the Oryza species was within the range found for Oryza sativa accessions. However, the level of palmitic acid in the O. species was as high as that reported for soybean high‐palmitic acid mutants. Oryza species also contained higher levels of the γ‐oryzanol and phenolic fractions compared to cultivated rice. Low or not significant phenotypic correlations between lipid, palmitic acid, and γ‐oryzanol suggest that these fractions in cultivated rice could be increased simultaneously using several of the Oryza species accessions identified in this study. A cultivar with enhanced levels of these fractions would be suitable for use in the production of high γ‐oryzanol margarine, shortening, and frying oils.     

RAPD markers in the analysis of genetic diversity among common bean germplasm from Central Himalaya

A set of 99 common bean germplasm collected from central Himalaya was investigated for their genetic variability using random amplified polymorphic DNA (RAPD) markers. Ten oligonucleotide primers, selected from 60 initially screened, generated 123 amplicon products. Of these, two amplicons were shared by all the accessions whereas 112 were polymorphic at least in two pair wise comparison. Nine unique bands identified were as low as 0.32 kb M.W. to as high as 3.5 kb and were confined to eight collections. All primers produced polymorphic amplicons though the extent of polymorphism varied with each primer. The primer OPF-17 was found to be most powerful and efficient as it generated a total of 17 bands of which 15 were polymorphic. RAPD markers data were analysed statistically using NTSYSpc.2.02e software and a dendrogram was generated using Jaccard’s similarity coefficient. The similarity coefficient values varied from 0.19 to 0.91. Grouping analysis revealed the categorization of 99 germplasm into 12 major branches with different level of similarity. Three branches namely branch 2, 3 and 5 out of 12 had only one accession. Branch 1 which consisted of three accessions was the most divergent as revealed by Jaccard’s similarity coefficient. Branching pattern of the accessions did not show any correlation with morphological data or altitudinal alignment of the accessions.     

黄淮麦区部分小麦种质资源的遗传差异分析

利用79对SSR引物对黄淮麦区129份种质资源进行了分析。结果表明,79对引物共检测到335个等位变异,每个引物检测到的等位变异数目2～8个,平均4.240个,变异最大的位点主要位于B组染色体上。79个SSR位点的遗传多态性信./010.015～0.820之间,平均0.498。种质资源间的遗传相似系数在0.253～0.909之间,平均0.492。聚类分析把这些种质45674大类和7个亚类。聚类结果与地域并不吻合,但能较好地反映出亲本的特性和其间的亲缘关系。