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.     

Low Level of Linkage Disequilibrium at the COMT (Caffeic Acid O-methyl Transferase) Locus in European Maize (<Emphasis Type="Italic">Zea mays</Emphasis> L.)

During recent decades, the whole plant yield of European maize (Zea mays L.) has increased substantially. However, during the same period of time cell wall digestibility, and consequently fodder quality, has decreased. Alleles for digestible cell walls have thus been lost, either during breeding for stalk standability or by genetic drift during breeding for grain yield. The brown-midrib 3 mutant (bm3) in maize (Z. mays L.), controlled by the caffeic acid O-methyl transferase (COMT) locus, has a positive influence on maize fodder quality. In this study, 42 European maize lines were used to evaluate the nucleotide diversity and linkage disequilibrium (LD) pattern across 2.3 kb of this locus. In agreement with what has been found for other maize loci, we found high diversity values (π = 0.00834), rapid LD decay even in an elite line sample, and indication of selection as well as of recombination events for seven site combinations. The diversity values differed slightly between inbreds of the Flint and Dent pools and most haplotypes were specific for one of the two heterotic groups. The polymorphisms identified at this single locus enable the construction of single nucleotide polymorphisms (SNP) and indel markers for haplotype identification and a rough discrimination of inbreds into Flint and Dent heterotic groups. The results reported here document the degree of allelic diversity in breeding materials and allow for targeted search of novel alleles in genetic resources.     

Searching for New Sources of Pink Stem Borer Resistance in Maize (<Emphasis Type="Italic">Zea mays</Emphasis> L.)

The pink stem borer (Sesamia nonagrioides Lef.) is the main corn (Zea mays L.) pest in the Mediterranean area. Although, screening for resistance to this pest has been successful, the level of resistance shown by the most resistant varieties is not high. The objectives of the present work were: (i) the evaluation for pink stem borer resistance of the nontested inbred lines, field and popcorn materials, from the collection maintained at the Misión Biológica de Galicia and (ii) the study of the performance, under pink stem borer infestation, of inbreds selected for resistance to pink stem borer in hybrid combination. Forty four inbred lines nontested yet for resistant to pink stem borer along with five inbred lines previously reported as resistant to stem and ear attack, were evaluated in 2002 and 2003. In adjacent experiments, hybrids EP79× W552, EP77× B93, and A661× EP42 were tested along with a resistant hybrid, MEB531-Bt, and a susceptible hybrid, INRA 260. New sources of resistance to pink stem borer have been detected among inbred lines improving the level of resistance presented by previously tested inbreds. Some of these lines were successfully developed by pedigree selection for resistance to pink stem borer.     

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.     

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.     

Analysis of Genetic Variation Among Accessions of <Emphasis Type="Italic">Pueraria montana</Emphasis> (Lour.) Merr. var. <Emphasis Type="Italic">lobata</Emphasis> and <Emphasis Type="Italic">Pueraria phaseoloides</Emphasis> (Roxb.) Benth. based on RAPD Markers

Pueraria montana var. lobata and P. phaseoloides originating from tropical Asia and parts of Oceania are ecologically and economically important legumes that are used as green manure, cover crop or forage plants. Conservation and use of plant genetic resources require an understanding of the extent and distribution of genetic diversity in any given region. In this study, genetic variation of five P. montana var. lobata and 16 P. phaseoloides accessions was analysed developing a random amplified polymorphic DNA (RAPD) marker methodology for Pueraria species and thereby creating basic data for follow-up research and the development of conservation strategies. Seeds were collected from native populations in Bac Kan Province, a mountainous region in Northeast Vietnam. P. montana var. lobata presented a high level of variation with 54.3% of the detected markers being polymorphic, whereas P. phaseoloides exhibited an intermediate to high level of variation (45.5%). The P. montana var. lobata accessions clustered in congruence with their eco-geographical origin. For P. phaseoloides no correspondence between sampling sites and genetic differentiation was found. Inter-population differentiation was measured as Jaccard's similarity coefficient (JSC). Mean JSC amounted to 0.35 in P. montana var. lobata and 0.52 in P. phaseoloides. Results are compared to other genetic studies of herbaceous legumes and conservation strategies are suggested.     

Dispersal of durum wheat [<Emphasis Type="Italic">Triticum turgidum</Emphasis> L. ssp. <Emphasis Type="Italic">turgidum</Emphasis> convar. <Emphasis Type="Italic">durum</Emphasis> (Desf.) MacKey] landraces across the Mediterranean basin assessed by AFLPs and microsatellites

A comprehensive characterization of crop germplasm is critical to the optimal improvement of the quality and productivity of crops. Genetic relationships and variability were evaluated among 63 durum wheat landraces from the Mediterranean basin using amplified fragment length polymorphisms (AFLPs) and microsatellites markers. The genetic diversity indices found were comparable to those of other crop species, with average polymorphism information content (PIC) values of 0.24 and 0.70 for AFLP and microsatellites, respectively. The mean number of alleles observed for the microsatellites loci was 9.15. Non-metric multi-dimensional scaling clustered the accessions according to their geographical origin with the landraces from the South shore of the Mediterranean Sea closely related. The results support two dispersal patterns of durum wheat in the Mediterranean basin, one through its north side and a second one through its south side.     

Morphological variability and resource allocation in the annual teosinte (<Emphasis Type="Italic">Zea mays</Emphasis> ssp.<Emphasis Type="Italic"> mexicana</Emphasis> (Schrader) Iltis) of the Toluca Valley,Mexico

This study explores the morphological variability and resource allocation of weedy, annual teosinte (Zea mays ssp. mexicana (Schrader) Iltis, Chalco race) from the Valley of Toluca, central Mexico, and compares it with data reported for maize. One hundred mature and fertile teosinte plants were selected for variation in size and measured. The extremes found for fertile plants were 0.12 and 3.5 m in height and 0.09 vs. 694 g in weight. Fruit were polymorphic, with significant correlation between color and weight. Small plants invested slightly more biomass in the leaves and tall plants invested more in the stem. The minimum mass necessary to produce fertile plants was much lower in teosinte than in maize. Very small teosinte plants formed only female inflorescences, whereas maize produces a tassel. The harvest index was similar to landrace maize, and did not decrease in taller plants, as teosinte can increase the reproductive sink. Toluca teosinte retains several important characteristics of weeds: size variability, adaptation of fruit quantity to environmental conditions, and fruit polymorphism.     

Conservation in a changing world: <Emphasis Type="Italic">in situ</Emphasis> conservation of the giant maize of Jala

Using survey data from Jala, Mexico, this case-study evaluates in situ maize conservation of the variety ‘Jala’ (Zea mays L.). Though historically ‘Jala’ was the dominant variety grown in the valley of Jala, today less than 20% of farmers grow it on only 5% of the maize area. Younger growers of the ‘Jala’ variety specialize in it, growing relatively large amounts for niche markets. Older, diversified farmers grow small areas for household use and to compete in a local contest. Conservation of the ‘Jala’ variety has been heavily influenced by shifting ideal concepts of maize, as determined by market and consumption demands and by a contest designed to promote in situ conservation. The current move away from nationalized purchasing may favor ‘Jala’s’ continued conservation.     

The Inheritance of Morphological and Biochemical Traits Introgressed into Common Wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis> L.) from <Emphasis Type="Italic">Aegilops speltoides</Emphasis> Tausch

The genetic control of morphological characters and gliadin composition was studied in two bread wheat lines with introgressed segments from Aegilops speltoides Tausch. It was found that the transferred trait of leaf hairiness is controlled by one dominant gene, non-allelic to the known gene, Hl1. It was localized in 7B chromosome. Whole plant non-glaucousness is under the control of an inhibitor gene, allelic to the gene W1^I of wheat located on chromosome 2B. This gene was found to be epistatic to the gene controlling spike waxlessness. The introgressed gene for spike glume color was found to be allelic to the Rg1 gene located on 1BS of common wheat, but it was linked with another allele of the gliadin locus Gli-B1.     

Diversity in White Lupin (<Emphasis Type="Italic">Lupinus albus</Emphasis> L.) Landraces from Northwest Iberianplateau

All the accessions (35) of white lupin (Lupinus albus L.) landraces collected from northwest Iberian plateau and maintained at the Spanish germplasm collection (CRF of INIA), were studied with the general objective of assessing the biodiversity of these landraces and to ascertain their value as genetic resources for the development of germplasm adapted to the areas where they were collected with long and chilly winters. The characterization study comprised 51 qualitative and 50 quantitative characters. Quantitative parameters were analyzed by Principal Components Analysis (PCA). The 2-dimensional plot (49.3% of cumulative variability) formed a main group of accessions and 4 outliers (#1, #9, #27 and #28) separated in the first PC. The characters with a higher contribution to the first PC were inflorescence length, leaf petiole length and leaf central foliole area. The presence of alkaloids, the percentage of plants killed by soil borne fungal diseases, the phenology, the yield per plant and yield components were also assessed. Twenty six accessions (the main group in the PCA plot) showed high homogeneity, with the following highlights: bitterness, indeterminate growth habit, early ripeness and spring sowing with the exception of #18, in which seeding date is unknown and it had a later phenology. By contrast accessions #9 and #27 were bitter of autumn sowing with determinate growing habit and they had the longest values of primary inflorescence length and a later phenology when they were sown in spring. Average values for yield and yield components showed a broad range of variation between accessions. The main yield component was the number of pods per plant. Accession #22 had the highest mean seed mass, although this had not significant influence in yield. Accession #17 showed the highest yield and #1 the lowest. This last also had the lowest values of inflorescence length, leaf petiole length and leaf central foliole area. No significant differences of resistance to soil-borne fungi were found between accessions. The year had a significant effect in the phenology, yield and yield components except for number of seeds per pod. The studied material might be of interest for the development of spring sowing germplasm adapted to north Iberian peninsula.     

Variability Among Maize (Zea mays L.) Inbred Lines for Seed Longevity

Genotypes conserved in active collections may suffer genetic erosion and modifications. The objective of this work was to investigate changes in germination and vigour in maize (Zea mays L.) inbred lines during cold storage in an active collection. Seeds of 16 maize inbreds produced along 16 years were evaluated for emergence and seedling vigour in a growth chamber. Linear and quadratic regressions of vigour and viability-related traits over seed age were calculated and tested for homogeneity. The seed of five production years of five inbreds that behaved differently in the regression analysis was multiplied in 1998, and original and renewed seed were evaluated in a growth chamber in 1999. Viability and vigour decreased linearly with age for most inbreds, particularly for B84 and EP10, varied at random for a few inbreds, and remained high for EP56 and A295. Aging caused reduction of vigour and loss of viability in most inbreds. There was variability for seed longevity among inbred lines; longevity was highest for inbred EP56. During storage, some seeds of each inbred died, while enduring seeds, when multiplied, produced new seed with enhanced viability and vigour compared to the average seed of the inbred, suggesting natural selection for viability and vigour during storage within inbred lines.     

Diversity among landraces of Indian snapmelon (<Emphasis Type="Italic">Cucumis melo</Emphasis> var. <Emphasis Type="Italic">momordica</Emphasis>)

Diversity among 36 snapmelon landraces, collected from 2 agro-ecological regions of India (9 agro-climatic sub-regions), was assayed using RAPD primers, morphological traits of plant habit and fruit, 2 yield-associated traits, pest and disease resistance and biochemical composition (TSS, ascorbic acid, titrable acidity). Typical differences among accessions were observed in plant and fruit characteristics and snapmelon germplasm with high titrable acidity and possessing resistance to downy mildew, Cucumber mosaic virus, Zucchini yellow mosaic virus, Papaya ringspot virus, Aphis gossypii and Meloidogyne incognita was noticed in the collection. RAPD based grouping analysis revealed that Indian snapmelon was rich in genetic variation and region and sub-region approach should be followed across India for acquisition of additional melon landraces. Accessions of var. agrestis and var. momordica clustered together and there was a separate cluster of the accessions of var. reticulatus. Comparative analysis of the genetic variability among Indian snapmelons and an array of previously characterized reference accessions of melon from Spain, Israel, Korea, Japan, Maldives, Iraq, Pakistan and India using SSRs showed that Indian snapmelon germplasm contained a high degree of unique genetic variability which was needed to be preserved to broaden the genetic base of melon germplasm available with the scientific community. N. P. S. Dhillon and Ranjana contributed equally to this work and are considered the first authors.     

Biological nitrogen fixation in maize (<Emphasis Type="Italic">Zea mays</Emphasis> L.) by <Superscript>15</Superscript>N isotope-dilution and identification of associated culturable diazotrophs

The nitrogen-fixing capacity of a range of commercial cultivars of maize (Zea mays L.) was evaluated by the ¹⁵N isotope-dilution method. Biological nitrogen fixation (BNF) expressed as percent nitrogen derived from air (Ndfa) ranged from 12 to 33 regardless of nitrogen fertilization. BNF was not affected by mineral nitrogen fertilization except on cultivar Topacio and PAU-871 cultivars. Subsequently, culturable bacterial diazotrophs were isolated from endophytic tissue of maize: seed, root, stem, and leaf. All isolates were able to grow on N-free semisolid medium. Eleven bacteria isolates showed nitrogen-fixing capacity by the reduction of acetylene to ethylene and confirmed by PCR the presence of nifH gene in their genome. Identification of the 11 isolates was performed by bacteriological methods, 16S rRNA gene sequences, and phylogenetic analysis, which indicated that the bacteria isolated were closely related to Pantoea, Pseudomonas, Rhanella, Herbaspirillum, Azospirillum, Rhizobium (Agrobacterium), and Brevundimonas. This study demonstrated that maize cultivars obtain significant nitrogen from BNF, varying by maize cultivar and nitrogen fertilization level. The endophytic diazotrophic bacteria isolated from root, stem, and leaf tissues of maize cultivars may contribute to BNF in these plants.     

RAPD and ISSR fingerprinting in cultivated chickpea (<Emphasis Type="Italic">Cicer arietinum</Emphasis> L.) and its wild progenitor <Emphasis Type="Italic">Cicer reticulatum</Emphasis> Ladizinsky

Detection of genetic relationships between 19 chickpea cultivars and five accessions of its wild progenitor Cicer reticulatum Ladizinsky were investigated by using RAPD and ISSR markers. On an average, six bands per primer were observed in RAPD analysis and 11 bands per primer in ISSR analysis. In RAPD, the wild accessions shared 77.8% polymorphic bands with chickpea cultivars, whereas they shared 79.6% polymorphic bands in ISSR analysis. In RAPD analysis 51.7% and 50.5% polymorphic bands were observed among wild accessions and chickpea cultivars, respectively. Similarly, 65.63% and 56.25% polymorphic bands were found in ISSR analysis. The dendrogram developed by pooling the data of RAPD and ISSR analysis revealed that the wild accessions and the ICCV lines showed similar pattern with the dendrogram of RAPD analysis. The ISSR analysis clearly indicated that even with six polymorphic primers, reliable estimation of genetic diversity could be obtained, while nearly 30 primers are required for RAPD. Moreover, RAPD can cause genotyping errors due to competition in the amplification of all RAPD fragments. The markers generated by ISSR and RAPD assays can provide practical information for the management of genetic resources. For the selection of good parental material in breeding programs the genetic data produced through ISSR can be used to correlate with the relationship measures based on pedigree data and morphological traits to minimize the individual inaccuracies in chickpea.     

Geographic variation for isozymes in cherimoya (<Emphasis Type="Italic">Annona cherimola</Emphasis> Mill.)

Cherimoya (Anonna cherimola Mill.) is a fruit tree which originated in Peru and Ecuador and is now cultivated in several subtropical areas of the world. The characterization of cherimoya cultivars at allozyme level has been previously reported, but the geographic distribution and organization of this variation have not been fully characterized. In this study, we assessed the relationships among 206 cherimoya and four atemoya (A. cherimola ×A. squamosa) cultivars based on allozyme polymorphism. We have confirmed the genetic differences between atemoya and cherimoya cultivars, and showed that cherimoya accessions from Madeira, Bolivia and Spain form homogeneous groups of cultivars. Accessions from Chile and California form heterogeneous groups, probably due to their mixed origins. Cultivars from Peru and Ecuador showed a wide range of allelic variation, as is expected for accessions from the center of origin of this species.     

Genetic diversity of Syrian pistachio (<Emphasis Type="Italic">Pistacia vera</Emphasis> L.) varieties evaluated by AFLP markers

Pistachio (Pistacia vera L.) is a strategic nut tree species in the Middle East which holds comparative advantage over other fruit trees in view of its hardiness, income generation opportunities and benefits for the ecosystem. Yet pistachio cultivation depends on a very narrow genetic base, in spite of the existence of many varieties still marginally exploited. Syria is an important center of diversity for pistachio. A country wide ecogeographic survey in this country was carried out to determine the extent of pistachio genetic diversity and its use. As a whole, 114 accessions were collected from 37 farms to assess diversity at morphological and molecular level. Molecular evaluation was carried out using Amplified Fragment Length Polymorphism (AFLP) technique and performed using seven primer pair combinations. Results from the studies allowed the identification of 25 pistachio female varieties in Syria, some of which unique and described for the first time. Three groups of pistachio diversity were identified by cluster analysis which provides useful information about the distribution of genetic diversity in Syria for enhanced use and sustainable conservation.