RFLP markers for cultivar identification in tall fescue (Festuca arundinacea Schreb.)

RFLP molecular markers have been used for the identification of 12 commercial varieties of tall fescue (Festuca arundinacea Schreb.). Of these, 7 could be distinguished on the basis of the presence/absence of specific and unique bands whereas the other 5 could be distinguished on the basis of band combinations. No differences in the RFLP pattern were detected between accessions of the same variety, whether obtained from the same source or from different sources. Bulked DNA probes from 100 plants are recommended as representative sample for allogamic crops. The use of molecular markers for seed certification in tall fescue is discussed.     

Velvet bentgrass (Agrostis canina L.) is the likely ancestral diploid maternal parent of allotetraploid creeping bentgrass (Agrostis stolonifera L.)

Understanding genetic relationships among the three most important Agrostis species will be important in breeding and genomic studies aimed at cultivar improvement. Creeping, colonial, and velvet bentgrasses (Agrostis stolonifera L., A. capillaris L., and A. canina L., respectively) are commercially important turfgrass species often used on golf courses. Velvet bentgrass is a diploid and creeping and colonial bentgrasses are both allotetraploids. A model for the genomic relationships among these species was previously developed from cytological evidence. The genome designations were A₁A₁ for velvet bentgrass, A₁A₁A₂A₂ for colonial bentgrass, and A₂A₂A₃A₃ for creeping bentgrass. Here we used phylogenetic analysis based on DNA sequences of nuclear ITS and protein coding genes and the plastid trnK intron and matK gene to reexamine these relationships. In contrast to the previous model, the DNA sequence analysis suggested that velvet bentgrass was closely related to creeping bentgrass and it is likely the maternal parent of creeping bentgrass. Phylogenetic analysis of some conserved nuclear genes revealed a close relationship of the velvet bentgrass sequences with the A₂ subgenome sequences of creeping bentgrass. We therefore propose that velvet bentgrass be designated as having the A₂ genome, rather than the A₁ genome as in the previous model.     

RAPD and ISSR molecular markers in <Emphasis Type="Italic">Olea europaea</Emphasis> L.: Genetic variability and molecular cultivar identification

Thirty Portuguese and eight foreign olive (Olea europaea L.) cultivars were screened using Random Amplified Polymorphic DNA (RAPD) and Inter-Simple Sequence Repeat (ISSR) markers. Twenty RAPD primers amplified 301 reproducible bands of which 262 were polymorphic; and 17 ISSR primers amplified 204 bands of which 180 were polymorphic. The percentage of polymorphic bands detected by ISSR and RAPD was similar (88 and 87%, respectively). The genetic variability observed was similar in the Portuguese and foreign olive cultivars. Seven ISSR and 12 RAPD primers were able to distinguish individually all 38 olive cultivars. Twenty specific molecular markers are now available to be converted into Sequence Characterised Amplified Region (SCAR) markers. Relationships among Portuguese and foreign cultivars is discussed.     

Selection of efficient vesicular-arbuscular mycorrhizal fungi for wetland rice (Oryza sativa L.)

Summary We tested the response of the wetland rice cultivar Prakash to inoculation with ten vescular-arbucular mycorrhizal (VAM) fungi (three selected from the first screening and seven isolated from local paddy fields) in a pot experiment under flooded conditions in order to select the most efficient mycorrhizal fungi to inoculate the rice nursery. A sandy clay loam soil was used as the substrate, fertilized with the recommended N and K levels (100 kg N ha^–1 as ammonium sulphate and 50 kg K ha^–1 as muriate of potash) and half the recommended level of P (25 kg ha^–1 as super phosphate). The inoculation was made into dry nursery beds and the beds were flooded when the seedlings were about 25 cm high, in 15 days. Twenty-eight-day old seedlings were transferred to pots filled with well puddled soil flooded with 5 cm of standing water. Based on the increase in grain yield and total biomass, Glomus intraradices and Acaulospora sp. were considered efficient and suitable for inoculation into rice nurseries.     

Analysis of Genetic Diversity in Colonial Bentgrass (<Emphasis Type="Italic">Agrostis capillaris</Emphasis> L.) using Randomly Amplified Polymorphic DNA (RAPD) Markers

Colonial bentgrass (Agrostis capillaris L.) is a cool-season grass, native to temperate Asia and Europe. It has good tolerance to low temperatures and partial shade and is well suited to golf course fairways and tees. Little information is available regarding levels and patterns of genetic variation among populations of colonial bentgrass, which would be useful for breeding programs. To study the genetic relationships among 27 colonial bentgrass accessions obtained from the US National Plant Germplasm System (NPGS), randomly amplified polymorphic DNA (RAPD) markers were scored and analyzed. Out of 80 primers screened, 16 were selected for further analysis, which yielded a total of 120 polymorphic bands used to differentiate the accessions. Dice's similarity coefficients for pair-wise comparisons ranged from 0.23 to 0.84 based on the RAPD data. Since there was no similarity coefficient value close to 1 between any two accessions, there was no apparent duplication among the sampled accessions. A dendrogram constructed on the basis of the Unweighted Pair Group Method with Arithmetic average (UPGMA) clustering algorithm clearly separated 26 of the accessions into three clusters with one accession distinct from the rest. The least similar pair of accessions was PI 204397 from Turkey and PI 628720 from Bulgaria, and the most similar pair was PI 509437 from Romania and PI 491264 from Finland. Clustering patterns based on principal components analysis (PCA) corresponded well with the dendrogram. A high cophenetic correlation (r = 0.82) was found between the RAPD data matrix and cophenetic matrix. The accession PI 628720, from Bulgaria, did not cluster with any other accessions.     

Characterization and identification of Lactuca sativa cultivars and wild relatives with SDS-electrophoresis (Lactuca sect. Lactuca, Compositae)

Summary Patterns of achene proteins of L. sativa cultivars are mutually compared and compared with L. saligna, L. serriola and L. virosa. L. virosa and L. saligna are easy to identify and are characterized by typical banding patterns. L. sativa and L. serriola share the same banding pattern. They differ clearly from L. saligna and L. virosa. L. sativa and L. serriola are closely related (in terms of similarity), they form a common genepool and L. serriola is involved in the domestication process of L. sativa. Cultivar identification was not possible with the SDS-PAGE method.     

Genetic relationships and diversity among Brazilian cultivars and landraces of common beans (Phaseolus vulgaris L.) revealed by AFLP markers

The genetic variation and relationships among 31 accessions of Phaseolus vulgaris L., and two representatives of Vigna unguiculata L., were evaluated by AFLP analysis. A total of 263 DNA fragments across all materials were scored using nine primer combinations, averaging 32 per primer. More than 95% of the amplification products showed polymorphism, indicating high variation at the DNA level among these accessions. Pair-wise genetic similarity (Jaccard's coefficient) ranged from 0.553 to 0.840, with a mean of 0.765. Twenty-three accessions (70%) clustered into three groups. A majority of the commercial cultivars (91%) clustered within a single group, whereas the landraces were distributed along all the variation. An apparent correlation with phaseolin types was detected. Results of this study suggest that Brazilian landraces truly represent the overall genetic variability of Phaseolus vulgaris, confirming the multiple origins of these materials, and their potential as a source of variation for breeding programs.     

RAPD and SSR markers for characterization and identification of ancient cultivars of <Emphasis Type="Italic">Olea europaea</Emphasis> L. in the Emilia region,Northern Italy

The Emilia region (Northern Italy) is characterised by the occurrence of microclimates that permit olive growing. The presence of the species, albeit sporadic, in these territories for several centuries as a fruit crop is well documented, by both archaeological and written testimony, and by a large number of plants well over a century old, located in particular sites, favourable for growth and development of the tree. Olive genetic diversity was studied using RAPD and SSR techniques, on plants growing in the Emilia territory (Reggio Emilia and Parma provinces). For genotype identification comparisons were made with 8 cultivars, some of which from Central Italy. Screening was obtained analysing patterns produced by 20 RAPD primers and 3 SSR primers, developed by other authors; the primers and we were able to discriminate olive cultivars with a sufficient degree of reliability. The dendrograms obtained from the analysis show the genetic relationship among accessions present in the Parma-Reggio Emilia district. Our results demonstrated the reliability of RAPDs and SSRs to identify all studied olive cultivars and to reveal the degree of their relatedness to each other. The analysis also reveals the presence of an interesting amount of genetic diversity among the studied individuals.     

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.     

Genetic diversity among Northwestern Argentinian cultivars of common bean (Phaseolus vulgaris L.) as revealed by RAPD markers

The genetic diversity of 10 commercial cultivars of common beans, developed in Northern Argentina was analyzed based on RAPD markers. Sixteen primers were assayed and among them only 4 showed polymorphisms. A similarity matrix was generated by applying three different association coefficients, Simple Matching, Jaccard and Dice. By the UPGMA method dendrograms were generated and also the principal coordinate analysis was performed. The similarity values found were higher than 40% suggesting that genetic diversity is low. Both cluster analysis and principal coordinates analysis associated commercial cultivars either to the Andean or the Mesoamerican gene pool.     

Oligotrophic pseudomonads in the rhizosphere: Suppressiveness to Pythium damping-off of cucumber seedlings (Cucumis sativus L.)

Summary Pseudomonads recovered from the cucumber rhizosphere were evaluated for their ability to suppress Pythium ultimum damping-off. Full-strength and 250-fold diluted selective media formulations with King's B medium (KB and KB/250, respectively) as the basal medium were used as the recovery media. Eight per cent of the isolates recovered (11/138) could induce suppression to Pythium damping-off and 45% of these biocontrol agents produced fluorescent pigments. No differences in the ability to induce suppression among the isolates recovered on full-strength and diluted media were detected. The growth rates of the selected isolates indicated that growth in KB broth was faster or equal to that in KB/10. Therefore, no obligate oligotrophs were recovered. Additionally, growth was observed in 15 000-fold diluted KB for 10 of 11 test isolates, which indicated that these isolates were facultative oligotrophs. In general, the faster growing isolates were more likely to induce suppression when used as seed treatments. Finally, the in vitro antifungal properties of test isolates against P. ultimum indicated that 4 of the 11 isolates inhibited P. ultimum.     

不同化感潜力水稻品种对低钾的生理与分子响应

选择国际公认的化感水稻品种“P1312777”和非化感水稻品种“Lemont”为材料,在K元素为5mg&#183;L^-1（低K）和40mg&#183;L^-1（正常K）2个水平的营养液中培养,通过水稻形态学指标（根长、株高、根冠比和生物量）、生理生化指标（SOD、POD、CAT、根系活力及植株中N、P、K含量）,评价不同化感潜力水稻品种对低K胁迫的生理响应,并采用实时荧光定量PCR（Realtime Fluorescent Quantitative PCR,FQ-PCR）,分析了低K胁迫下水稻根和叶中与N、P、K吸收利用相关的12个关键酶的基因表达差异。结果表明,低K促进了化感水稻“P1312777”根的生长,根冠比加大,生物量增加,但对株高的影响不显著;而该条件下非化感水稻“Lemont”的上述指标均受到不同程度的抑制。低K对两种水稻的保护酶系（SOD、POD、CAT）和根系活力均有一定程度的抑制作用,植株中N、P、K含量降低,但非化感水稻“Lemont”受抑制的程度远大于化感水稻“P1312777”。FQ-PCR检测结果表明,低K胁迫下两种水稻根、叶中的12个关键酶的基因均呈现上调表达,而化感水稻“P1312777”的基因表达上调倍数均明显大于非化感水稻“Lemont”。低K胁迫下两种水稻品种的形态学差异、生理与分子响应均表明,化感水稻“P1312777”比非化感水稻“Lemont”具有更强的适应K匮乏的能力。     

Time-course of nitrogen fixation in two bambara groundnut (Vigna subterranea L. Verdc.) cultivars

TheA-value method, involving the application of a higher¹⁵N rate to a reference non-N₂-fixing plant, was used to assess the magnitude of N₂ fixation in two bambara groundnut cultivars at four growth stages [vegetative, 0–47 days after planting (DAP); early pod-filling, 47–99 DAP; mid-pod-filling, 99–120 DAP; physiological maturity, 120–148 DAP). The cultivars were Ex-Ada, a bunchy type, and CS-88-11, a slightly spreading type. They were grown on a loamy sand. Uninoculated Ex-Ada and CS-88-11 were used as reference plants to measure the N₂ fixed in the inoculated bambara groundnuts. In this greenhouse study, soil was the major source of N in bambara groundnuts during vegetative growth, and during this period it accounted for over 80% of the N accumulaed in the plants. However, N₂ fixation became the major source of plant N during reproductive growth. There were significant differences between the two cultivars in the ability to fix N₂, and at physiological maturity, almost 75% of the N in CS-88-11 was derived from the atmosphere compared to 55% in Ex-Ada. Also, the total N fixed in CS-88-11 at physiological maturity was almost double that in Ex-Ada. Our data indicate that the higher N₂ fixation in CS-88-11 was due to two factors, a higher intensity of N₂ fixation and a longer active period of N₂ fixation. The results also suggest that bambara groundnut genotypes could be selected for higher N₂ fixation in farining systems.     

Evaluation of genetic diversity and relationships within an on-farm collection of Perilla frutescens (L.) Britt. using microsatellite markers

The present study demonstrates utilization of 11 microsatellite markers to explore genetic diversity held in Perilla frutescens (L.) Britt. landrace accessions growing on farms in different parts of Korea and Japan and to assess their genetic relationships. All microsatellite loci were polymorphic and produced a total of 96 alleles ranging from 4 to 20, with an average of 8.7 alleles per locus. Of the 96 alleles found, a total of 15 unique landrace-specific alleles were observed at 9 different loci. The locus GBPFM203 provided the highest number of alleles (20), of which five were unique and each specific to a particular landrace accession. The occurrence of unique, accession-specific alleles presented molecular evidence for the generation of new alleles within on-farm collection of Perilla. The mean values of observed (H _O) and expected heterozygosity (H _E) were 0.39 and 0.68, respectively, indicating a considerable amount of polymorphism within this collection. A genetic distance-based phylogeny grouped the two Perilla varieties, var. frutescens and var. crispa (Thunb.) Decne into two distinct groups. Accessions belonging to var. frutescens could also be divided into two subgroups at a close genetic distance (GD = 0.432). The overall clustering pattern did not strictly follow the grouping of accessions according to their geographic origins. These observations are indicative of extensive germplasm exchange among farms from different geographical regions. The genetic similarity observed among the Perilla landraces may be useful for future Perilla crop variety identification, conservation, and improvement programs.     

Studying the extent of genetic diversity among Gossypium arboreum L. genotypes/cultivars using DNA fingerprinting

Genetic diversity is an area of concern for sustaining crop yield. Information on genetic relatedness/diversity among Gossypium arboreum L. cultivars/genotypes is scanty. We have used random amplified polymorphic DNA (RAPD) analysis to assess the genetic divergence/relationship among 30 genotypes/cultivars of G. arboreum. Of 45 primers surveyed, 63% were polymorphic. Out of the total number of loci amplified, 36% were polymorphic. The calculated genetic similarity between the cultivars/genotypes was in the range of 47.05–98.73%. Two genotypes, HK-244 and Entry-17, were the most distantly related. The average genetic relatedness among all the genotypes was 80.46%. However, most of the cultivated varieties showed a close genetic relationship, indicating a narrow genetic base in comparison to the non-cultivated germplasm. The calculated coefficients were used to construct a dendrogram using the unweighted pair group of arithmetic means (UPGMA) algorithm, which grouped the genotypes/cultivars into two major and three smaller clusters. The study is the first comprehensive analysis of the genetic diversity of G. arboreum germplasm and identifies cultivars that will be useful in extending the genetic diversity of cultivated varieties and future genome mapping projects.     

Selection at the Ms locus in open pollinated onion (Allium cepa L.) populations possessing S-cytoplasm or mixtures of N- and S-cytoplasms

The bulb onion (Allium cepa L.) was historically seed propagated by open pollination (OP). Cytoplasmic-genic male sterility (CMS) and protandry encourage outcrossing among individual onion plants. The most common source of CMS in onion is conditioned by the interaction of sterile (S) cytoplasm with a single nuclear male-fertility restoration (Ms) locus. We previously reported that the majority of OP onion populations possess normal (N) male-fertile cytoplasm and varying frequencies of the dominant Ms allele. It was unclear why N-cytoplasmic onion populations often possess relatively high frequencies of the Ms allele, which has no obvious function. We used computer simulations to estimate changes in allelic frequencies at Ms for onion populations possessing S-cytoplasm or a mixture of N- and S-cytoplasms, and to determine if frequencies of the Ms allele stay constant or change due to failure of male gamete production from male-sterile (S msms) plants. The models revealed selection against the recessive ms allele over generations in onion populations possessing S-cytoplasm and varying amounts of self pollination and inbreeding depression. These models were consistent with field and molecular analyses documenting that N-cytoplasm and the dominant Ms allele predominate in OP onion populations.     

Differential responses of chickpea (Cicer arietinum L.) — Rhizobium combinations to saline soil conditions

Summary Chickpea cultivars (Cicer arietinum L.) and their symbiosis with specific strains of Rhizobium spp. were examined under salt stress. The growth of rhizobia declined with NaCl concentrations increasing from 0.01 to 2% (w : v). Two Rhizobium spp. strains (F-75 and KG 31) tolerated 1.5% NaCl. Of the 10 chickpea cultivars examined, only three (Pusa 312, Pusa 212, and Pusa 240) germinated at 1.5% NaCl. The chickpea — Rhizobium spp. symbiosis was examined in the field, with soil varying in salinity from electrical conductivity (EC) 4.5 to EC 5.2 dSm^-1, to identify combinations giving satisfactory yields. Significant interactions between strains and cultivars caused differential yields of nodules, dry matter, and grain. Four chickpea — Rhizobium spp. combinations, Pusa 240 and F-75 (660 kg ha^-1), Pusa 240 and IC 76 (440 kg ha^-1), Pusa 240 and KG 31 (390 kg ha^-1), and Pusa 312 and KG 31 (380 kg ha^-1), produced significantly higher grain yields in saline soil.     

Identification of sweet cherry cultivars (Prunus avium L.) and analysis of their genetic relationships by chloroplast sequence-characterised amplified regions (cpSCAR)

Ten cpSCAR markers that show polymorphism in Prunus avium were used to fingerprint sweet cherry cultivars. The purpose of the study was also to contribute to identification and to help determine their genetic interrelationships. Samples of ‘0900 Ziraat’, a superior Turkish variety, which were collected in several locations all over Turkey, had identical cpSCAR patterns, and they resembled a common European haplotype, A. ‘Sweetheart’, ‘Summit’ and ‘Canada Giant’ and their haplotype are intermediate between the previously described haplotypes A and B, which were originally found in Central and Eastern European sweet and wild cherries, and those from Northern Turkey, respectively. The data therefore suggests a local maternal descent (within Europe and Asia Minor) of the cultivars analysed. Our results show that chloroplast DNA analysis is a straightforward way to classify cherry cultivars. We compare our results to others previously reported for sweet cherry cultivars, and conclude that cpSCAR diversity data could be considered for phylogenetic studies in this group.     

Collection and characterization of naturalized populations of white clover (Trifolium repens L.) in Argentina

A collection formed from 53 accessions of white clover stolonswas carried out in naturalized populations from Argentina betweenparallels 26° and 37° South latitudes and 58° to 62°West longitude. Almost half of them (43%) wereobtained from grazed sites. Collected samples were grown in the fieldat Pergamino Experimental Station and compared with a local cultivar.Attributes scored or measured were petiole length, width and lengthof central leaflet, relation width/length, height, growing habit,leaf mark, stolon thickness and density, days to flowering,cyanogenesis, seed yield and winter herbage yield. There weresignificant correlations among seven pairs of characters. Allpopulations were cyanogenic within a range from 40 to 100%.Populations from grazed sites had prostrate habit and lower canopyheight than those from non grazed sites. The collection was clusteredin seven groups. Differences among groups were attributed toagroecological differences in sites of collection. Results obtainedencouraged in Argentina further collection missions for naturalizedpopulations of white clover.     

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

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