Gliadin Electrophoretic Analysis of the Genetic Integrity of Wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis> L.) Accessions After Frequent Seed Reproductions

A standard electrophoretic method for wheat cultivar identification was used on single seeds to analyse the genetic integrity of 11 wheat (Triticum aestivum L.) accessions after up to 24 seed reproductions in the Gatersleben genebank. It was clearly demonstrated that the gliadin pattern of single seeds can be used to analyse the genotype composition of wheat accessions. Stability of electrophoretic banding patterns was detected in eight accessions. Very week genetic drift was observed in three accessions. Our investigations confirm experiences of the successful utilisation of protein markers for cultivar verification and genetic integrity testing and demonstrate the high standard of wheat accessions maintenance in the Gatersleben genebank.     

HMW-glutenin and gliadin variations in Tibetan weedrace, Xinjiang rice wheat and Yunnan hulled wheat

Nine Tibetan weedrace, 9 Xingjiang rice wheat and 14 Yunnan hulled wheat accessions were evaluated for the variability of HMW-glutenins and gliadins. Higher variability was observed for both HMW-glutenins and gliadins in Tibetan weedrace and Xingjiang rice wheat, while lower variability was observed in Yunan hulled wheat. There were 4 HMW-glutenin and 9 gliadin patterns in 9 Tibetan weedrace accessions, 5 HMW-glutenin and 8 gliadin patterns in 9 Xingjiang rice wheat accessions, and 3 HMW-glutenin and 8 gliadin patterns in 14 Yunnan hulled wheat accessions. In Xinjiang rice wheat, one accession (i.e. Daomai 2) carried subunits 2.1 + 10.1 encoded by Glu-D1, which is very rare in common wheat.     

Analysis of genetic diversity in a sweet potato (Ipomoea batatas L.) germplasm collection from Tanzania as revealed by AFLP

Sweet potato (Ipomoea batatas L.) is the fifth most important crop in the developing countries after rice, wheat, maize and cassava. The amplified fragment length polymorphism (AFLP) method was used to study the genetic diversity and relationships of sweet potato accessions in the germplasm collection of Sokoine University of Agriculture, Morogoro and Sugarcane Research Institute, Kibaha, Tanzania. AFLP analysis of 97 sweet potato accessions using ten primer combinations gave a total of 202 clear polymorphic bands. Each one of the 97 sweet potato accessions could be distinguished based on these primer combinations. Estimates of genetic similarities were obtained by the Dice coefficient, and a final dendrogram was constructed with the un-weight pair-group method using arithmetic average. AFLP-based genetic similarity varied from 0.388 to 0.941, with a mean of 0.709. Cluster analysis using genetic similarity divided the accessions into two main groups suggesting that there are genetic relationships among the accessions. Principal Coordinate analysis confirmed the pattern of the cluster analysis. Analysis of molecular variance revealed greater variation within regions (96.19%) than among regions (3.81%). The results from the AFLP analysis revealed a relatively low genetic diversity among the germplasm accessions and the genetic distances between regions were low. A maximally diverse subset of 13 accessions capturing 97% of the molecular markers diversity was identified. We were able to detect duplicates accessions in the germplasm collection using the highly polymorphic markers obtained by AFLP, which were found to be an efficient tool to characterize the genetic diversity and relationships of sweet potato accessions in the germplasm collection in Tanzania.     

Temporal variation of diversity in Italian durum wheat germplasm

The aim of this work is to analyse the temporal change of genetic diversity in Italian durum wheat germplasm. The germplasm deployed in this study (158 accessions), belonging to 5 different historical classes, was characterised for its microsatellite and gliadin markers. The level of genetic diversity (He), based on gliadin and SSR markers results – on average – greater in indigenous landraces present in Italy before 1915, with the exception of pure line material which had been selected from landraces (showing highest level of heterozigosity for gliadin markers). Genotypes obtained from crosses or mutagenesis (referring to the 1950–1960 period) along with those resulting from crosses between CIMMYT lines and old materials (1970s and beyond) were also genetically more diverse. Forty-nine percent of indigenous landraces were genetically heterogeneous. Nine out of 53 landrace accessions were able to capture 4 different SSR private alleles. It is speculated that the reduction of allele richness is an indicator of the genetic erosion of the pre-breeding germplasm and it is pointed out that the implementation of appropriate methods of genetic conservation of this germplasm is a priority for breeding and food safety.     

Collection of Lupinus angustifolius L. Germplasm and Characterisation of Morphological and Molecular Diversity

Lupinus angustifolius L. is a Mediterranean species, domesticated in the 20th century, representing an important grain legume crop in Australia and other countries. This work is focused on the collection of wild germplasm and on the characterisation of morphological and molecular diversity of germplasm accessions. It reports the collection of 81 wild L. angustifolius accessions from the South and Centre of Portugal, available at the ‘Instituto Superior de Agronomia Gene Bank’, with subsequent morphological and molecular characterisation of a selection of these and other accessions. A multivariate analysis of morphological traits on 88 L. angustifolius accessions (including 59 wild Portuguese accessions, 15 cultivars and 14 breeding lines) showed a cline of variation on wild germplasm, with plants from Southern Portugal characterised by earlier flowering, higher vegetative development and larger seeds. AFLP and ISSR molecular markers grouped modern cultivars as sub-clusters within the wider diversity of wild germplasm, revealing the narrow pool of genetic diversity on which domesticated accessions are based. The importance of preserving, characterising and using wild genetic resources for L. angustifolius crop improvement is outlined by the results obtained.     

Genetic diversity of 38 spinach (<Emphasis Type="Italic">Spinacia oleracea</Emphasis> L.) germplasm accessions and 10 commercial hybrids assessed by TRAP markers

Target region amplification polymorphism (TRAP) markers were used to assess genetic variability among 38 germplasm accessions and 10 commercial hybrids of spinach (Spinacia oleracea L.), an economically important leafy vegetable crop in many countries. Germplasm accessions with different geographic origins and 10 commercial hybrids were examined. For assessing genetic diversity within accessions, DNA was extracted from 12 individual seedlings from six germplasm accessions and two hybrids. A relatively high level of polymorphism was found within accessions based on 59 polymorphic TRAP markers generated from one fixed primer derived from the Arabidopsis-like telomere repeat sequence and two arbitrary primers. For evaluating interaccession variability, DNA was extracted from a bulk of six to 13 seedlings of each accession. Of the 492 fragments amplified by 12 primer combinations, 96 (19.5%) were polymorphic and discriminated the 48 accessions from each other. The average pair-wise genetic similarity coefficient (Dice) was 57.5% with a range from 23.2 to 85.3%. A dendrogram indicated that the genetic relationships among the accessions were not highly associated with the geographic locations in which the germplasms were collected. The seven commercial hybrids were grouped in three separate clusters, suggesting that the phenotype-based breeding activities tended to reduce the genetic variability. This preliminary study demonstrated that TRAP markers are effective for fingerprinting and evaluating genetic variability among spinach germplasms. Mention of trade names or commercial products in this article is solely for the purpose of providing specific information and does not imply recommendation or endorsement by the US Department of Agriculture. The U.S. Government's right to retain a non-exclusive, royalty-free license in and to any copyright is acknowledged.     

Genetic diversity trend of common wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis> L.) in China revealed with AFLP markers

A total of 242 accessions of common wheat (Triticum aestivum L.) released in China since the 1940s were evaluated with AFLP (amplified fragment length polymorphism) technique. Genetic diversity was analyzed using five pairs of polymorphic primer combinations with 245 polymorphic bands. The highest genetic diversity was found in the accessions of the 1950s, and in the next place was that in the 1940s. The genetic diversity began to descend in the 1960s, and fell to the lowest in the 1970s. After that, the genetic diversity came back to some extent in the 1980s, however, it became much lower in the 1990s compared with that in the 1940–1950s. Landraces and introduced accessions from foreign countries showed greater genetic diversity in comparison to improved varieties. In addition, greater genetic diversity was observed in winter wheat. It was emphasized that great attention should be paid on further exploration of genetic diversity in wheat breeding program.     

Variation in salt tolerance within a Georgian wheat germplasm collection

Bread wheat Triticum aestivum L. possesses a genetic variation for the ability to survive and reproduce under salt stress conditions. Durum wheat (T. durum Desf.) is in general more sensitive in comparison to bread wheat, however, exceptions can be found showing the same extent of salt tolerance. Endemic wheats in general are characterised by a high adaptability to their environment. The level and variability of salt tolerance were assessed in a germplasm collection of 144 winter and spring wheat accessions from Georgia comprising Triticum aestivum L., T. durum Desf., T. dicoccon Schrank, T. polonicum L. and Georgian endemics: T. carthlicum Nevski, T. karamyschevii Nevski, T. macha Dekapr. et Menabde, T. timopheevii (Zhuk.) Zhuk. and T. zhukovskyi Menabde et Ericzjan. The accessions were tested for salt tolerance at the germination stage. Large variability in salt tolerance within the Georgian germplasm was found among the different wheat species. The endemic hexaploid winter wheat T. macha and the endemic tetraploid wheat T. timopheevii were among the most tolerant materials, thus presenting promising donors for salt tolerant traits in future breeding efforts for salinity tolerance in wheat.     

Characterisation of germplasm accessions of Napier grass (pennisetum purpureum and P. purpureum×P. glaucum Hybrids) and comparison with farm clones using RAPD

Fifty six germplasm accessions of the important East African fodder crop Napier grass, Pennisetum purpureum, and its hybrids with P. glaucum, were characterised using 67 random amplified polymorphic DNA (RAPD) fragments. No or very low intra-accession variation was found for 49 of the accessions examined, confirming field observations that this species is predominantly clonally propagated. Comparison of intra and inter-accession variation identified several groups of identical/similar accessions that could be targeted if the collection is to be rationalised, and also highlighted two misplantings of germplasm material during transfer to a field trial site. A neighbour joining dendrogram of Jaccard's similarity estimates, clearly separated 50 accessions of P. purpureum from three P. glaucum individuals, and placed six hybrid accessions in an intermediate position. These groupings were well supported by a nested AMOVA (P<0.001; 29.5% of total variance due to taxonomic delineation). The main group of P. purpureum individuals could be further differentiated into five sub-groups (designated East Africa, Southern Africa, USA1, USA2 and Miscellaneous, to reflect the majority membership of sub-groups) and examination of the within P. purpureum component of the nested AMOVA, found them to be significantly different (P<0.001; 18.8% of variance). Genetic diversity across all accessions was found to be fairly high (Shannon's diversity index 0.306) and thus the collection probably represents a wide genetic base for this species. In addition to germplasm accessions, 25 Kenyan farm clones were also analysed. A principal coordinate analysis found that all but one of the clones clustered with the main P. purpureum group of accessions, indicating that the majority are probably not interspecific hybrids. The origin and pedigree of clones is discussed based on genetic similarity amongst clones and to germplasm accessions.     

(英文)

对来源于美、中、俄及埃塞阿比亚等22个国家的142份硬粒小麦材料的种子贮藏蛋白位点及遗传变异进行了研究。供试的硬粒小麦(Triticum durum Desf )材料共检测出37条醇溶蛋白条带，无1条带纹为所有材料共有，多态性达到100％，说明硬粒小麦具有丰富的醇溶蛋白等位变异。聚类分析将142份供试材料分为3个大类，材料间遗传差异大小在不同的国家有所不同，表明醇溶蛋白带型与地理来源有一定关系。高分子量谷蛋白电泳共分离出14种亚基和15种亚基组合，但是优质亚基所占比例不高，这可能是因为硬粒小麦加工用途的特殊性，使得多年的育种并未太多改变硬粒小麦高分子量谷蛋白亚基等位变异的频率，促成优质亚基的累计。     

Evaluating landraces of bread wheat <Emphasis Type="Italic">Triticum aestivum</Emphasis> L. for tolerance to aluminium under low pH conditions

Bread wheat (Triticum aestivum L.) landraces held within ex situ collections offer a valuable and largely unexplored genetic resource for wheat improvement programs. To maximise full utilisation of such collections the evaluation of landrace accessions for traits of interest is required. In this study, 250 accessions from 21 countries were screened sequentially for tolerance to aluminium (Al) using haematoxylin staining of root tips and by root regrowth measurement. The staining test indicated tolerance in 35 accessions, with an intermediate response to Al exhibited in a further 21 accessions. Of the 35 accessions classified as tolerant, 33 also exhibited increased root length following exposure to Al. The tolerant genotypes originated from Bulgaria, Croatia, India, Italy, Nepal, Spain, Tunisia, and Turkey. AFLP analysis of the 35 tolerant accessions indicated that these represent diverse genetic backgrounds. These accessions form a valuable set of germplasm for the study of Al tolerance and may be of benefit to breeding programs for expanding the diversity of the gene pool from which tolerant cultivars are developed.     

Extent and Pattern of Genetic Diversity for Morpho-agronomic Traits in Ethiopian Highland Pulse Landraces: I. Field Pea (<Emphasis Type="Italic">Pisum sativum</Emphasis> L.)

An experiment was conducted in 2001 at Holetta and Kulumsa, Ethiopia, to study the extent and pattern of genetic diversity in Ethiopian field pea (Pisum sativum L.) landraces. One hundred forty-eight germplasm accessions were grown in an alpha lattice design with 2 replications. Data on 12 traits were collected and analyzed. Differences among the accessions were significant for most of the traits (except number of seeds/ pod) at each location even though differences pooled over location were mostly non-significant. The accessions were grouped into five clusters of different sizes. Accessions from the southern part of the country (Arsi) distributed overall clusters while those from the northern half (North and South Wello, North Gonder and North Shewa) fell into clusters C₁ to C₃. Cumulative effects of a number of characters dictated differentiation of the accessions into clusters. There was no definite relationship between geographic diversity and genetic diversity as overlapping was encountered in clustering pattern among accessions from different parts of the country. Accessions from different regions might have similar genetic background and those from the same origin might also have different genetic background. Therefore, geographic diversity should not necessarily be used as an index of genetic diversity and parental selection should be based on a systematic study of genetic diversity in a specific population. Genetic distances among most of the clusters were significant that crosses between parents selected out of them are expected to generate desirable genetic recombination. Selection should also consider the special advantages of each cluster and each accession within a cluster. Future germplasm collection, conservation and breeding efforts should focus not only on inter-regional diversity but also on intra-regional diversity.     

Combined use of gliadins and SSRs to analyse the genetic variability of the Spanish collection of cultivated diploid wheat (Triticum monococcum L. ssp. monococcum)

This work studied the combined use of gliadins and SSRs to analyse inter- and intra-accession variability of the Spanish collection of cultivated einkorn (Triticum monococcum L. ssp. monococcum) maintained at the CRF-INIA. In general, gliadin loci presented higher discrimination power than SSRs, reflecting the high variability of the gliadins. The loci on chromosome 6A were the most polymorphic with similar PIC values for both marker systems, showing that these markers are very useful for genetic variability studies in wheat. The gliadin results indicated that the Spanish einkorn collection possessed high genetic diversity, being the differentiation large between varieties and small within them. Some associations between gliadin alleles and geographical and agro-morphological data were found. Agro-morphological relations were also observed in the clusters of the SSRs dendrogram. A high concordance was found between gliadins and SSRs for genotype identification. In addition, both systems provide complementary information to resolve the different cases of intra-accession variability not detected at the agro-morphological level, and to identify separately all the genotypes analysed. The combined use of both genetic markers is an excellent tool for genetic resource evaluation in addition to agro-morphological evaluation.     

Extent and Pattern of Genetic Diversity for Morpho-agronomic Traits in Ethiopian Highland Pulse Landraces II. Faba Bean (<Emphasis Type="Italic">Vicia faba</Emphasis> L.)

A field experiment was conducted in 2001 at Holetta and Kulumsa, Ethiopia, to study genetic diversity in Ethiopian faba bean (Vicia faba L.) landraces. One hundred sixty random germplasm accessions were grown in an alpha lattice design with two replications. Data on 12 traits were collected and analyzed. Significant differences were observed among the accessions for most of the traits (except number of pods/podding nodes) at each location even though differences pooled over location were mostly non-significant. Cluster analysis distinguished seven diversity classes of different sizes. Accessions from the northern half of the country (North and South Wello, North Gonder and North Shewa) were closely related while those from the southern part of the country (Arsi) were highly diverse. Cumulative effects of a number of characters dictated differentiation of the accessions into clusters. Some overlapping were encountered between accessions from the northern and those from the southern parts of the country. The study revealed that accessions from different regions might have similar genetic background and those from the same origin might also have different genetic background. Therefore, geographic diversity should not necessarily be used as an index of genetic diversity and parental selection should be based on a systematic study of genetic diversity in a specific population. Genetic distances between most of the clusters were significant that crosses between parents selected out of them are expected to generate desirable progenies. Future germplasm collection, conservation and utilization strategies should put more focus not only on inter-regional diversity in the country as a whole but also on intra-regional diversity in Arsi.     

Genetic diversity of wild soybean (<Emphasis Type="Italic">Glycine soja</Emphasis> Sieb. et Zucc.) and Japanese cultivated soybeans [<Emphasis Type="Italic">G. max</Emphasis> (L.) Merr.] based on microsatellite (SSR) analysis and the selection of a core collection

Wild soybeans, Glycine soja, are a source of genetic variation to improve soybeans. To improve the efficiency evaluation of conserved germplasm a core or mini-core collection approach that maximizes allelic diversity in a proportion of the whole collection has frequently been advocated. The genetic diversity of a wild soybean collection (1,305 accessions) plus Japanese cultivated soybeans (53 accessions) were analyzed at 20 SSR marker loci. Higher levels of allelic diversity were found in wild soybeans (28 alleles per locus) than Japanese cultivated soybean (five alleles per locus). The genetic distance between wild soybeans from different regions reflected their proximity. Accessions from Russia consisted of a diverse array of alleles resulting in accessions being spread further apart in a PCA plot than accessions from other regions. Accessions of wild soybean from Korea included many rare alleles and thus had a high representation in the core collection. The two core collections developed here, traditional and mini, consisted of 192 accessions with 97% of the allelic diversity (14% of the whole collection) and 53 accessions with 62.4% of the allelic diversity (5% of the whole collection), respectively.     

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.     

Genetic diversity and intra-specific phylogeny of Triticum turgidum L. subsp. dicoccon (Schrank) Thell. revealed by RFLPs and SSRs

Today, emmer wheat, T. turgidum subsp. dicoccon, widely grown in the past is a candidate crop for sustainable agriculture in Italy. As part of a research project aimed at the enhanced use of the hulled wheat germplasm, molecular characterization was carried out to understand the genetic structure of the crop and to identify accessions of interest. A collection of 194 accessions was analyzed with 15 microsatellite loci (SSRs), while only a sample of 38 accessions was tested with 19 RFLP probes. The marker loci were selected on the basis of their independent genomic distribution. Genetic distances and allelic frequencies were calculated for each marker class. The genetic relationships were visualized with dendrograms. RFLP loci were, on average, less polymorphic than SSRs. An average Dice's genetic distance of 0.22 for RFLPs vs 0.38 for SSRs was detected, while an expected average heterozygosity per locus of 0.23 for RFLPs vs 0.26 for SSRs was also estimated. With a least number of 10 loci per marker class it was possible to identify each genotype. The most diverse accessions had different geographic origins. Germplasms from Italy and Ethiopia appear to belong to a more primitive genepool, given that a group of accessions from these countries were genetically differentiated from a Russian-Iranian group.     

Genetic diversity, conservation, and utilization of Theobroma cacao L.: genetic resources in the Dominican Republic

Cacao (Theobroma cacao L.) is a significant agricultural commodity in the Dominican Republic, which ranks 11th in the world for cacao exports. To estimate genetic diversity, determine genetic identity, and identify any labeling errors, 14 SSR markers were employed to fingerprint 955 trees among cacao germplasm accessions and local farmer selections (LFS). Comparisons of homonymous plants across plots revealed a significant misidentification rate estimated to be 40.9 % for germplasm accessions and 17.4 % for LFS. The 14 SSRs amplified a total of 117 alleles with a mean allelic richness of 8.36 alleles per locus and average polymorphism information content (PIC) value of 0.67 for the germplasm collection. Similar levels of variation were detected among the LFS where a total of 113 alleles were amplified with a mean of 8.07 alleles per locus and PIC of 0.57. The observed heterozygosity (H_obs) was 0.67 for the germplasm collection and 0.60 for LFS. Based on population structure analysis 43.9 % of the germplasm accessions and 72.1 % of the LFS are predominantly of the Amelonado ancestry. Among these Amelonado, 51.7 % for the germplasm collection and 50.6 % for LFS corresponded to Trinitario hybrid lineage. Criollo ancestry was found in 7.6 and 9.5 % of the germplasm accessions and LFS, respectively. The Contamana, Nacional, and Iquitos backgrounds were also observed in both populations, but the Curaray background was only detected in the germplasm accessions. No Purús or Guiana ancestry was found in either of the populations. Overall, significant genetic diversity, which could be exploited in the Dominican Republic breeding and selection programs, was identified among the germplasm accessions and LFS.     

(英文)

用SDS-PAGE方法测定了我国10个小麦主产省份171份小麦品种和高代品系的高分子量麦谷蛋白亚基（HMW-GS）组成。鉴定出18种HMW-GS，40种HMW-GS组成形式，其中20种亚基其组成形式只在一个品种（系）中出现。Glu-A1位点亚基1和Null出现最多，Glu-B1位点7＋8和7＋9亚基对占绝对优势，Glu-D1位点2＋12亚基对出现频率最高。Null、7＋9、2＋12、Null，7＋8，2＋12，1、7＋8，2＋12，1、7＋9、2＋12等亚基组成形成出现频率最高，占分析品种的49.71％。与前人研究相比，新育成品种HMW-GS亚基组成发生了明显变化，面包优质亚基（对）1、5＋10出现的频率显著升高，亚基多态性增加，组成形式明显改善，这些对于品质改良和品种选育是非常有利的，新育成品种Glu-1品质评分已超过7。尽管个别品种亚基组成好，品质优良，但总体上看，我国小麦品种与其它国家相比品质还存在一定差距，提高5＋10、17＋18等优质亚基的频率是改善我国小麦面包品质的重要措施。