Genetic structure and history of Swiss maize (<Emphasis Type="Italic">Zea mays</Emphasis> L. ssp. <Emphasis Type="Italic">mays</Emphasis>) landraces

Between 1930 and 2003 with emphasis on the 1940s maize landraces (Zea mays L. ssp. mays) from all over Switzerland were collected for maintenance and further use in a new Swiss breeding program. The genetic relationship and diversity among these accessions stored in the Swiss gene bank is largely unknown. Our hypothesis was that due to the unique geographic, climatic, and cultural diversity in Switzerland a diverse population of maize landraces had developed over the past three centuries. The aims were to characterize the genetic diversity of the Swiss landraces and their genetic relationship with accessions from neighbouring regions as well as reviewing their history, collection, and maintenance. The characterization and grouping was based on analyses with ten microsatellite markers. Geographic, cultural, and climatic conditions explained a division in two distinct groups of accessions. One group consisted of landraces collected in the southern parts of Switzerland. This group was related to the Italian Orange Flints. The other group contained accessions from northern Switzerland which were related to Northern European Flints in particular German Flints. Historic evidence was found for a frequent exchange of landraces within the country resulting in a lack of region-specific or landrace-specific genetic groups. The relatively large separation between the accessions, indicated by high F _ST (0.42), might be explained partly by a bottleneck during the collection and maintenance phase as well as by geographical and cultural separation of north and south of the country. Due to the high genetic diversity, the accessions here are a potential resource for broadening the European flint pool.     

Genetic diversity of Swiss maize (<Emphasis Type="Italic">Zea mays</Emphasis> L. ssp. <Emphasis Type="Italic">mays</Emphasis>) assessed with individuals and bulks on agarose gels

About 65 years ago, more than 150 Swiss maize landraces (Zea mays L. ssp. mays) of the flint type were collected and conserved ex situ. Due to the climatically and culturally diverse environment of the Alps, a considerable genetic diversity of this material was assumed. To prove this, an efficient method was required to carry out genetic profiling of all the accessions in the Swiss Gene Bank. Simple sequence repeat marker (SSR) profiling in combination with the visualization of the polymerase chain reaction (PCR) products on agarose gels was chosen. Here a set of 19 different landrace accessions was analyzed to: (i) investigate their genetic diversity, (ii) investigate and display the population structure and (iii) determine whether DNA bulks rather than single plants can be used for such analyses. Four repeated samples of one accession were found to be much closer to one another than to the rest of accessions. Furthermore, specific alleles were identified for several accessions. The PCR products of the bulked DNA samples represented only a small part of the variation revealed by the analysis of individuals. Loci with four base repeat motifs performed better in the analysis of bulks than loci with other repeat motifs. The correlation between genetic distance matrices, based on the analysis of individuals and bulks, respectively, was significant. Thus, the single plant approach allowed for sufficient differentiation of accessions, and DNA bulks visualized on agarose gels led to correlated genetic distances although a limited number of alleles were detected. Although the limited resolution of agarose gels likely causes some bias, profiling of larger sets with the individual plant approach appears feasible and more informative compared to the bulk analysis we conducted.     

Maintenance of Squash (<Emphasis Type="Italic">Cucurbita</Emphasis> spp.) Landrace Diversity by Farmers' Activities in Mexico

Squash (Cucurbita spp.) is a common component in traditional cropping systems in Mexico, mainly in the agroecosystem known as the “milpa”, in which squash is cultivated in association with maize (Zea mays), the main crop. Using a questionnaire, 80 farmers were interviewed about crop production and selection practices in order to understand how these factors affect genetic diversity of local squash populations. We found that the most of the farmers who cultivate squash were elderly 59.8 ± 14.5 (mean ± SD; n = 78) years old. Squash varieties in the area were exclusively locally adapted landraces, and had not been replaced by modern squash cultivars. Two cultivated squash species, C. argyrosperma ssp. argyrosperma and C. moschata, had been grown intercropped with maize by 97.5% of the interviewed farmers, but only 50.0% were still producing squash at the time of the study. Farmers recognize typical characteristics of particular varieties within each of the local cultivated squash species, and selection is directed to maintain their identity. Nearly two thirds of the farmers (62.0%) had exchanged seeds of squash for planting, a practice that serves to increase genetic variability in the populations. All of the interviewed farmers were conscious of the possible hybridization between the wild gourd (C. argyrosperma ssp. sororia) and their cultivated squash. Despite various natural and human managed factors identified as contributing to enhancement of genetic diversity in these populations, results of the study show that genetic erosion of Cucurbita is likely in the region in the near future.     

Collecting maize (<Emphasis Type="Italic">Zea mays</Emphasis> L. convar. <Emphasis Type="Italic">mays</Emphasis>) with potential technological ability for bread making in Portugal

The Portuguese maize bread (“broa”) manufactured from traditional maize landraces still plays an important economic and social role on Central and Northern rural communities of the country. However the traditional maize landraces agricultural systems are changing. Local maize landraces are in risk of disappearing because of the progressive adoption of hybrid varieties not suitable for bread production. These changes are contributing to a major loss of genetic diversity. An expedition took place in the Central region of Portugal (Beira Interior and Beira Litoral) with the purpose of collecting enduring maize landraces with technological ability for bread production and to access the possibility of establishing a participatory plant breeding and conservation program. A total of 51 different maize landraces and 175 other varieties of associated crops were collected. Maize landraces showed to maintain high diversity and potential for improvement. The production relayed on small farms with polycrop, quality oriented, sustainable systems. A participatory plant breeding and conservation program would be possible on this region with the proviso that local authorities would be involved. This program would allow a higher valuation of these maize populations, contributing to halt the current loss of these unique Portuguese maize landraces.     

Population clonal diversity and fine-scale genetic structure in <Emphasis Type="Italic">Oryza officinalis</Emphasis> (Poaceae) from China,implications for <Emphasis Type="Italic">in situ</Emphasis> conservation

Population clonal diversity and fine-scale genetic structure of Oryza officinalis Wall. ex Watt, an endangered species in China recently experiencing habitat degradation, was estimated using inter simple sequence repeat markers. We analyzed the genetic variations of 440 samples exhaustively collected from nine O. officinalis populations. Relatively rich clonal diversity and poor genetic variation were found in the extant populations. We found that the number of genets, the percentage of polymorphic loci, and gene diversity decreased with population decline, suggesting that habitat degradation will lead to further genetic depletion of O. officinalis populations. A pronounced spatial genetic structure occurs at both the ramet and genet levels in several larger populations, which is the result of clonal growth and concomitant inbreeding. The in situ conserved population PS holds much more genotypes than other populations with the similar population size, which might have more seedling recruitments from the soil seed bank due to habitat disturbance, suggesting a moderate disturbance combined with habitat degradation-avoiding measures are effective for in situ conservation of this species.     

Social and environmental influences on tartary buckwheat (<Emphasis Type="Italic">Fagopyrum tataricum</Emphasis> Gaertn.) varietal diversity in Yunnan,China

Effective conservation strategies aimed to protect crop genetic resources require multiple sources of information. We used a combination of AFLP genotyping and farmer surveys to understand the extent, distribution and management of tartary buckwheat (Fagopyrum tataricum Gaertn.) diversity in its center of origin in Yunnan Province, China. We found genetic evidence of gene flow in tartary buckwheat throughout the study area, with small but statistically significant regional and village-level components. We also found genetic differentiation by seed color. Although most farmers reported exchanging seed in localized kinship networks, our results imply homogenizing gene flow is occurring. Yi ethnic farmers tend to plant more buckwheat than non-Yi farmers, and we found that in some communities, Yi farmers serve as seed sources for farmers of other ethnicities. Different tartary buckwheat varieties did not have different end uses; rather farmers maintained varietal diversity in order to protect crop yield and quality. Individual farmers’ seed exchange practices reflect their ideas about components of seed quality, as well as priorities in protecting buckwheat yield. From the standpoint of genetic resources conservation, the presence of a culturally rich farmer exchange network and hierarchical structuring of tartary buckwheat genetic diversity demonstrates the importance of maintaining an interlinked community of tartary buckwheat farmers in Yunnan.     

Geographical distribution and diversity of maize (<Emphasis Type="Italic">Zea mays</Emphasis> L. subsp. <Emphasis Type="Italic">mays</Emphasis>) races in Mexico

The center of origin of maize is in Mexico and maize diversity is very high in many parts of the country. Here we describe and analyze the geographic distribution and diversity of maize using a database of 18,176 georeferenced observations of maize races. The number of observations by race follows the common “hollow curve” distribution, with six races comprising 54 % of the records and 37 races only 10 %. Few races are widespread, and some have a highly localized distribution. The average maximum distance (D _max) between two records of the same race was 1169 km, but seven races had a D _max lower than 200 km. The average circular area range size (CA₂₅) was 51, but 14 races had a CA₂₅ lower than 10. Some races have very few observations but were found over a large area. These races may not be valid and their classification should be reviewed. Areas of high race richness and diversity are in mountainous northwest, south central, and southern Mexico in the states of Oaxaca, Jalisco, Michoacán and Chiapas. Six contiguous maize regions were defined using hierarchical clustering. Despite the high sample size, further sampling could improve estimates of race diversity in several regions.     

Maize diversity in southern Brazil: indication of a microcenter of <Emphasis Type="Italic">Zea mays</Emphasis> L.

‘Microcenters of diversity’ are restricted geographical areas within which a significant diversity of genetic resources is accumulated. Maize genetic variability, one of the largest among cultivated species, particularly in far western Santa Catarina, southern Brazil, has shown a rich store of landraces, conserved on farm by small-scale farmers. Thus, this study aimed to characterize the diversity of Zea mays L. landraces in two municipalities of this micro-region in Santa Catarina, as well as its geographic distribution, developing the census of diversity as a methodological proposal. Diagnosis was made according to the farmers’ knowledge. The field survey was conducted in 70 rural communities involving 2049 farms. The methodology allowed identifying 136 populations of wild relatives, of which some belong to the species Zea luxurians (Durieu and Ascherson) Bird, and 1513 populations of maize landraces, comprising 1078 of popcorn, 337 of common maize, 61 of sweet maize and 37 of flour maize. The identification of 59 morphological groups, the high use, adaptive and agronomic values, and the Shannon index (H′), estimated according to the grain shape (0.79), endosperm type (0.73), size (0.87), color (1.40), and morphological group (3.16), showed at set a considerable diversity distribution conserved on farm in this micro-region. Landrace richness and the presence of wild relative species, associated with the local human activity and sociocultural aspects, allowed to characterize the far western Santa Catarina as a ‘microcenter of diversity’ of Zea mays L.     

Biodiversity of <Emphasis Type="Italic">Secale strictum</Emphasis> in Iran measured using microsatellites

The mountain rye Secale strictum is native to the Middle East and is the progenitor of the cultivated rye. Regarding lack of information about the genetic diversity of this species in Iran, this study was aimed to evaluate its genetic variation and to examine the patterns of diversity related to the varieties and geography. Fifteen wheat and rye derived microsatellite markers were used to achieve this aim. High levels of diversity, with an average number of 6.1 alleles per locus (ranging up to 11) and high level polymorphism with polymorphism rate averaging 0.624 (between populations) and 0.357 (within populations) were observed among 125 individuals from 19 populations collected from various regions of Iran. The Northwestern populations showed the highest and the Northern populations showed the lowest polymorphism and diversity. One population of the Northwest of the country was notably closer in its allele range to the S. cereale accessions used as outgroup. No taxon or geographic specific marker was detected, suggesting high gene flow between the populations, however some groupings which can be related to the geographic regions and varieties, were evident. The analysis of molecular variance attributed same portions of genetic diversity to the within and between populations with no significant variation among different geographic regions. The results of this study indicated that the Iranian genepool of Secale strictum is valuable to search for new useful alleles for crop improvement.     

Diversity and differentiation of <Emphasis Type="Italic">Oryza sativa</Emphasis> and <Emphasis Type="Italic">O. rufipogon</Emphasis> in Indonesia

Analysis of the genetic structure of Indonesian Oryza sativa and O. rufipogon using neighbour-joining trees based on single nucleotide polymorphism (SNP) and simple sequence repeat (SSR) markers revealed that O. sativa in Indonesia is separated from O. rufipogon. Accessions of O. sativa in this study were differentiated into two major groups, indica and tropical japonica, excluding some varieties. SSR and SNP markers revealed the high value of differentiation (F _ST) and genetic distance (D) between indica and tropical japonica and we discovered four loci by SNP markers and one locus by SSR markers that play a role in differentiation between indica and tropical japonica. Interestingly, genetic diversity (H) in O. rufipogon was lower than that in O. sativa, however H in O. rufipogon was the highest and H in tropical japonica was the lowest when O. sativa was divided into two groups. Inbreeding coefficient (Fst) showed evidences that gene flow (Nm) between species and within species might be one of the mechanisms related to the diversification and differentiation of Indonesian rice germplasm by asymmetric pattern between species and within O. sativa as revealed by SSR and SNP markers. In addition, we found evidences on stabilizing selection in Indonesian rice germplasm and they might be the reasons why Indonesian rice germplasm did not differentiate due to source location of landrace. However, we found a weak relation between SSR and SNP markers probably due to highly polymorphic in SSR and the different properties of both markers.     

Salt tolerance in <Emphasis Type="Italic">Zea mays</Emphasis> (L). following inoculation with <Emphasis Type="Italic">Rhizobium</Emphasis> and <Emphasis Type="Italic">Pseudomonas</Emphasis>

This study aimed to investigate the effect of inoculation with plant growth-promoting Rhizobium and Pseudomonas species on NaCl-affected maize. Two cultivars of maize (cv. Agaiti 2002 and cv. Av 4001) selected on the basis of their yield potential were grown in pots outdoors under natural conditions during July. Microorganisms were applied at seedling stage and salt stress was induced 21 days after sowing and maintained up to 50% flowering after 120 days of stress. The salt treatment caused a detrimental effect on growth and development of plants. Co-inoculation resulted in some positive adaptative responses of maize plants under salinity. The salt tolerance from inoculation was generally mediated by decreases in electrolyte leakage and in osmotic potential, an increase in osmoregulant (proline) production, maintenance of relative water content of leaves, and selective uptake of K ions. Generally, the microbial strain acted synergistically. However, under unstressed conditions, Rhizobium was more effective than Pseudomonas but under salt stress the favorable effect was observed even if some exceptions were also observed. The maize cv. Agaiti 2002 appeared to be more responsive to inoculation and was relatively less tolerant to salt compared to that of cv. Av 4001.     

Genotypic and phenotypic changes in wild barley (<Emphasis Type="Italic">Hordeum vulgare</Emphasis> subsp. <Emphasis Type="Italic">spontaneum</Emphasis>) during a period of climate change in Jordan

Climate change and other anthropogenic disturbances can lead to the loss of genetic variation and thereby affect evolutionary potential and survival of plant populations in the wild. We examined these predictions in the primary wild relative of barley, Hordeum vulgare L. subsp. spontaneum (K. Koch) Thell., within its center of diversity, in Jordan. Changes in genotypic and phenotypic diversity were assessed using seed samples collected in 1981 and 2012 from the same 18 sites across Jordan. The overall population structure was conserved, but we observed an increase of within population genetic diversity and a reduction in population differentiation. Phenotypic variation differed among years and sites but the magnitude and direction of change variated among sites. While the sampled region became significantly hotter and drier during this period, simple correlation models did not support association between measures of climate change and the observed genetic and phenotypic changes. Agricultural activities that promote disturbance and demographic fluctuations may affect crop wild relatives that grow in agricultural landscapes, in unexpected ways. The observed increase in genetic diversity within populations might be explained by increased migration or by an advantage of increased genetic variation in the face of variable environmental conditions. This study provides a new perspective on the range of possible responses of crop wild relatives to environmental pressures.     

Diversity in cowpea (<Emphasis Type="Italic">Vigna unguiculata</Emphasis> (L.) Walp.) local populations from Greece

Cowpea cultivation in many countries around the Mediterranean Basin depends on a number of locally adapted populations conserved on-farm at a small scale, rather than on the use of modern varieties. Documentation, characterization and exploitation of traditional local populations could contribute to their conservation and utilization as sources of desirable characteristics. Therefore, a study was conducted to (a) characterize, (b) assess diversity and (c) classify 23 on-farm conserved local cowpea populations based on 32 agro-morphological traits. Investigations on diversity of characteristics related to seed yield, mineral and seed crude protein content as well as on correlations among them were carried out. A relatively high phenotypic diversity was observed. In particular, a high level of within population diversity was found (\( \bar{H}s \) = 0.34) exceeding that among populations’ diversity (Gst = 0.27). Principal component analysis classified the majority of local populations into two groups (mainly according to populations’ seed coat color and eye color), further divided into six subgroups regardless of the populations’ geographical origin. Significant differences were also observed among the populations studied for potassium and calcium, as well as for their seed crude protein content which ranged from 22.14 to 28.37 %. The results show appreciable levels of intra- and inter-phenotypic diversity in on-farm conserved cowpea populations, which indicates the existence of a valuable gene pool for future exploitation in breeding programs.     

Origin and evolution of Chinese waxy maize: evidence from the <Emphasis Type="Italic">Globulin-1</Emphasis> gene

Waxy maize is a special type of cultivated maize and has grown in China for long history. However, the evolution and origin of waxy maize still remain unknown. We analyzed the origin and evolution of waxy maize by sampling DNA sequences from four taxa with eight populations: waxy maize and other maize cultivars from Southwest China or America (Zea mays L. ssp. mays), parviglumis (Z. mays L. ssp. parviglumis Iltis et Doebley), three more distant species within this genus (Z. luxurians (Durieu et Ascherson) Bird, Zea perennis (Hitchcock) Reeves et Mangelsdorf, and Zea diploperennis Iltis, Doebley et Guzman), and a representative of sister genus (Tripsacum dactyloides L.). We sequenced 20 sequences and downloaded 26 sequences from NCBI for the glb1 locus, which encodes a nonessential seed storage protein. Within the Zea genus samples, the waxy maize has the minimum sequence diversity, which contains 31.1% of the level of diversity of parviglumis and 58.5% of the level of diversity of normal maize from Southwest China. Sequence variation within glb1 locus is consistent with neutral evolution in all four taxa according to Tajima’D test. From the NJ tree for glb1 sequences waxy maize formed two main groups which are intermixed with normal maize sequences. These results suggest that the Chinese waxy maize originate from a single gene mutation from normal maize. Combined with the history of maize dispersal in China we can even think that Chinese waxy maize was divergenced from Chinese flint maize.     

Exploitation of <Emphasis Type="Italic">Malus</Emphasis> EST-SSRs and the utility in evaluation of genetic diversity in <Emphasis Type="Italic">Malus</Emphasis> and <Emphasis Type="Italic">Pyrus</Emphasis>

A total of 8117 suitable SSR-contaning ESTs were acquired by screening from a Malus EST database, among which dinudeotide SSRs were the most abundant repeat motif, within which, CT/TC followed by AG/GA were predominant. Based on the suitable sequences, we developed 147 SSR primer pairs, of which 94 pairs gave amplifications within the expected size range while 65 pairs were found to be polymorphic after a preliminary test. Eighteen primer pairs selected randomly were further used to assess genetic relationship among 20 Malus species or cultivars. As a result, these primers displayed high level of polymorphism with a mean of 6.94 alleles per locus and UPGMA cluster analysis grouped twenty Malus accessions into five groups at the similarity level of 0.6800 that were largely congruent to the traditional taxonomy. Subsequently, all of the 94 primer pairs were tested on four accessions of Pyrus to evaluate the transferability of the markers, and 40 of 72 functional SSRs produced polymorphic amplicons from which 8 SSR loci selected randomly were employed to analyze genetic diversity and relationship among a collection of Pyrus. The 8 primer pairs produced expected bands with the similar size in apples with an average of 7.375 alleles per locus. The observed heterozygosity of different loci ranged from 0.29 (MES₉₆) to 0.83 (MES₁₃₈), with a mean of 0.55 which is lower than 0.63 reported in genome-derived SSR marker analysis in Pyrus. The UPGMA dendrogram was similar to the previous results obtained by using RAPD and AFLP markers. Our results showed that these EST-SSR markers displayed reliable amplification and considerable polymorphism in both Malus and Pyrus, and will contribute to the knowledge of genetic study of Malus and genetically closed genera.     

The Distribution of <Emphasis Type="Italic">Perilla</Emphasis> Species

Perilla (Lamiaceae) contains one tetraploid species, P. frutescens (L.) Britt. and three diploid species, P. citriodora (Makino) Nakai, P. hirtella Nakai and P. setoyensis G. Honda. Tetraploid species have been traditionally cultivated in Asia for their seed oil and for their fragrant leaves that are used as medicine or as a garnish for fish. The center of diversity is still obscure. To conserve the genetic resources, it is important to know the diversity of the tetraploid species. The three diploid species, which are possible parents of the tetraploid species, are all believed to be indigenous to Japan. Their distribution in China and Korea was clarified on the basis of herbarium and field surveys. The tetraploid species is assumed to have originated somewhere around the mid-to downstream area of the Changjiang River. Though Perilla is not cultivated as often in these areas as in northern China, Korea, the Himalayan region, or Myanmar, these areas should also be important for the conservation of genetic resources of tetraploid Perilla crops because of the expected high genetic diversity.     

Inferring geographic origin of barley (<Emphasis Type="Italic">Hordeum vulgare</Emphasis> L. subsp. <Emphasis Type="Italic">vulgare</Emphasis>) accessions using molecular markers

The USDA Agricultural Research Service (ARS) National Small Grains Collection (NSGC) has 207 landrace barleys obtained from a nursery grown in the Ukraine in 1930 by N.I. Vavilov, many of which have multiple resistance (MR) to disease similar to accessions from Ethiopia. The original collection locations of the accessions from the Vavilov nursery are unknown. The purpose of this study was to determine, using molecular markers, if the MR accessions of unknown origin from the Vavilov nursery are genetically related to MR accessions from Ethiopia. The genetic relatedness among susceptible accessions of unknown origin and among a selection of landrace accessions from the NSGC barley core subset was also assessed. Simple sequence repeat (SSR) and Diversity Array Technology PL (DArT) marker data were used to generate similarity matrices with Dice’s similarity coefficients. Cluster analysis from these results showed that the unknown-origin MR accessions grouped with accessions from Ethiopia. Susceptible accessions of unknown origin were genetically similar to accessions from western Asia, especially near the Caucasus, and from both southern and northern Europe. Based on similarity of marker profiles, some accessions from the core collection are likely duplicates. Future work will seek to identify the probable origin of the remainder of the unknown origin accessions and to more thoroughly characterize the genetic diversity within NSGC barley core subset.     

Genetic diversity in oil and vegetable mustard (<Emphasis Type="Italic">Brassica</Emphasis><Emphasis Type="Italic">juncea</Emphasis>) landraces revealed by SRAP markers

Mustard (Brassica juncea) is an important crop in both ancient and modern world. It has a broad resource of genetic diversity that is used primarily as oilseed but as vegetables, condiment and medicines also. Its superior tolerance to adverse environments, e.g., drought, high temperature and low fertility suggests its better adaptability in future possible harsh environments. Chinese vegetable mustard displays a wide spectrum of morphotypes. A collection of 95 accessions of B. juncea representing oil and vegetable mustards from China, France, India, Pakistan, and Japan were assessed to determine diversity at the molecular level using sequence-related amplified polymorphism (SRAP). Eight SRAP primer combinations identified a total of 326 scorable fragments of which 161 were polymorphic (49.39%). The percentage of polymorphism for each primer combination varied from 21.88 to 66.67%. Both Shannon-Weaver and Simpson genetic diversity index indicated that the level of genetic diversity within vegetable mustard is much higher than within oil mustard, and also winter oil mustards are genetically more diverse than spring oil mustards. Based on the Cluster and Principal Coordinates analysis, which were conducted on the similarity matrix of SRAP marker data, vegetable, spring oil and winter oil mustard were clearly divided into three distinct groups and among these three groups, spring and winter oil mustard are geneticlly closer than vegetable mustard. This suggests that bilateral gene exchange between oil and vegetable gene pools in the breeding program will effectively elevate the genetic potential in developing higher yields, more disease resistance, better quality and better adapted lines.