Identification and relationships of accessions grown in the grapevine (<Emphasis Type="Italic">Vitis vinifera</Emphasis> L.) Germplasm Bank of Castilla y Léon (Spain) and the varieties authorized in the VQPRD areas of the region by SSR-marker analysis

A total of sixty-five accessions from the Spanish region of Castilla y León including those authorized in the VQPRD areas were characterized for six SSR loci. All the samples but one unknown were identified by comparison to other databases. Thirty synonymous samples and three cases of homonymy were found out, confirming in most cases ampelographic expectations. Five unique genotypes belonging to local varieties in risk of extinction were detected. Several parameters were calculated to assess the usefulness of the chosen loci in this work. A dendrogram representing the genetic similarities among the accessions was constructed using the neighbor-joining method to investigate possible parentage relationships in the sample, and to explain them from an historical and cultural point of view.     

Microsatellite analysis of Alpine grape cultivars (<Emphasis Type="Italic">Vitis vinifera</Emphasis> L.): alleged descendants of Pliny the Elder’s <Emphasis Type="Italic">Raetica</Emphasis> are genetically related

According to Pliny the Elder and other Greco-Roman geoponics, Raetica was a famous white grape as well as a white wine produced in Raetia, a Province of the Roman Empire. Does Raetica grape have modern descendants? Etymologically and geographically, the white ‘Rèze’ from Valais (Switzerland) would be the best candidate. Using available microsatellite data, we searched for relatives of ‘Rèze’ in our database containing over 1,700 genotypes of grape cultivars from all over the world. Twelve cultivars showing putative first-degree (parent–offspring or full-siblings) or second-degree (grandparent–grandoffspring, uncle–nephew or half-siblings) relationships with ‘Rèze’ were then analysed at 60 microsatellite markers. Calculation of allele sharing and likelihood ratios between competing relationship categories revealed that four cultivars had parent–offspring relationship with ‘Rèze’: ‘Cascarolo Bianco’ (Piedmont, Italy), ‘Arvine Grande’ (Valais, Switzerland), ‘Groppello di Revò’ and ‘Nosiola’ (Trentino, Italy). Given that some of these are also said to be Raetica descendants, we may well be on the tracks of Pliny the Elder’s Raetica grape. However, there is no evidence about the identity of Raetica. Analysis of ancient DNA of grape pips excavated from archaeological sites of the Roman times might provide key information. Our first attempts were unsuccessful, but analysis of additional samples and optimisation of the method could provide groundbreaking results about the identity of the grapes cultivated in classical antiquity.     

Chloroplast Microsatellites to Investigate the Origin of Grapevine

The origin of the grapevine, Vitis vinifera ssp . sativa L., has been investigated with archaeobotanical–archaeological, cultural and historical data indicating a unique domestication centre located in the Caucasian and Middel-East regions about 6–7000 years ago, but, events leading to the domestication of this species are still an open issue. In this work, eight universal chloroplast microsatellites are used to assess genetic relationships among varieties selected as representatives of four distinct geographical groups from Middle-East to Western European regions. Results show that two out of the eight analysed chloroplast loci are polymorphic within the 142 individuals. Allele variants of the cpSSR loci combine in a total of six different haplotypes. The analysis of haplotypes distribution and haplotype diversity (HD) suggest that only three out of the six haplotypes are represented in the Caucasian and Middle-East samples, with 90% of individuals sharing the same haplotype. Moreover, the presence of all six haplotypes in the European accessions, with a high level of haplotype diversity, suggests varietal influx in these areas. Concerning the Western European varieties, especially in Spanish accessions, half of the individuals share haplotype VI which is completely absent in the Caucasian and Middle-East cultivars. This result opens the discussion about the existence of a unique and common domestication centre, located in the Caucasian and Middle-East area, for all the European cultivars. This work suggests the usefulness of chloroplast genome markers to provide information on haplotype distributions that could help to identify further geographical areas for grapevine varietal evolution.     

Characterization of Portuguese populations of <Emphasis Type="Italic">Vitis vinifera</Emphasis> L. ssp. <Emphasis Type="Italic">sylvestris</Emphasis> (Gmelin) Hegi

Wild populations of Vitis vinifera L.␣have been located in Portugal. Morphological characterization was carried out in three populations located in Alcácer do Sal, Castelo Branco, and Montemor-o-Novo, and then compared using multivariate discriminant analysis. These populations were from three different hydrological basins, therefore cross-pollination was not possible. It was verified that in each population all plants were different. The data suggest that the frequency of female and male plants is rather variable in wild populations. The morphology of the adult leaf, from the Alcácer do Sal population, had particular features when compared with Castelo Branco and Montemor-o-Novo populations, which were more homogeneous. The length of teeth compared with width at the end of the base, and the density of prostrate hairs between and on main veins (lower side) were the variables which allowed the best discrimination among populations.     

Arbuscular mycorrhizal fungi-mediated nitrogen transfer from vineyard cover crops to grapevines

Cover crops are often planted in between vineyard rows to reduce soil erosion, increase soil fertility, and improve soil structure. Roots of both grapevines and cover crops form mutualistic symbioses with arbuscular mycorrhizal (AM) fungi, and may be interconnected by AM hyphae. To study nutrient transfer from cover crops to grapevines through AM fungal links, we grew grapevines and cover crops in specially designed containers in the greenhouse that restricted their root systems to separate compartments, but allowed AM fungi to colonize both root systems. Leaves of two cover crops, a grass (Bromus hordeaceus) and a legume (Medicago polymorpha), were labeled with 99 atom% ¹⁵N solution for 24 h. Grapevine leaves were analyzed for ¹⁵N content 2, 5, and 10 days after labeling. Our results showed evidence of AM fungi-mediated ¹⁵N transfer from cover crops to grapevines 5 and 10 days after labeling. N transfer was significantly greater from the grass to the grapevine than from the legume to the grapevine. Possible reasons for the differences between the two cover crops include lower ¹⁵N enrichment in legume roots, higher biomass of grass roots, and/or differences in AM fungal community composition. Further studies are needed to investigate N transfer from grapevines to cover crops and to determine net N transfer between the two crops throughout their growing seasons, in order to understand the significance of AM fungi-mediated interplant nutrient transfers in the field.     

多胺对共生条件下丛枝菌根真菌及宿主植物生长发育的影响

为探讨多胺对共生条件下丛枝菌根真菌及其宿主植物生长发育的影响,本研究以丛枝菌根真菌(Gigaspora margarita)为试验材料,通过施用不同浓度的多胺(Polyamine,PA)及其生物合成抑制剂[Methylglyoxal bis(guanylhydrazone),MGBG]处理接种丛枝菌根真菌的葡萄微繁苗,研究共生培养条件下外源多胺及多胺合成抑制剂对丛枝菌根真菌孢子萌发、芽管菌丝及其宿主植物生长发育的影响.试验结果表明,共生培养条件下,一定浓度的外源PA对丛枝菌根真菌及其宿主植物的生长发育具显著促进作用,丛枝菌根真菌孢子数、菌丝长度、侵染率、丛枝丰富度及菌根化葡萄幼苗生长势均显著提高.MGBG则表现较强的抑制作用.且该抑制作用可被外源PA部分解除,证明外源多胺对菌根化葡萄微繁苗生长发育的促进作用是通过活化根系土壤中丛枝菌根真菌,促进微繁苗丛枝菌根共生体的良好发育,最大程度地发挥菌根化效应得以表现的.     

Nitrogen capture by grapevine roots and arbuscular mycorrhizal fungi from legume cover-crop residues under low rates of mineral fertilization

The influence of mineral fertilization on root uptake and arbuscular mycorrhizal fungi-mediated ¹⁵N capture from labeled legume (Medicago polymorpha) residue was examined in winegrapes (Vitis vinifera) in the greenhouse, to evaluate compatibility of fertilization with incorporation of cover-crop residue in winegrape production. Plants grown in marginal vineyard soil were either fertilized with 0.25× Hoagland’s solution or not. This low fertilization rate represents the deficit management approach typical of winegrape production. Access to residue in a separate compartment was controlled to allow mycorrhizal roots (roots + hyphae), hyphae (hyphae-intact), or neither (hyphae-rotated) to proliferate in the residue by means of mesh core treatments. Leaves were weekly analyzed for ¹⁵N. On day 42, plants were analyzed for ¹⁵N and biomass; roots were examined for intraradical colonization; and soils were analyzed for ¹⁵N, inorganic N, Olsen-P, X-K, and extraradical colonization. As expected, extraradical colonization of soil outside the cores was unaffected by mesh core treatment, while that inside the cores varied significantly. ¹⁵N atom% excess was highest in leaves of roots + hyphae. In comparison, leaf ¹⁵N atom% excess in hyphae-intact was consistently intermediate between roots + hyphae and hyphae-rotated, the latter of which remained unchanged over time. Fertilization stimulated host and fungal growth, based on higher biomass and intraradical colonization of fertilized plants. Fertilization did not affect hyphal or root proliferation in residue but did lower %N derived from residue in leaves and stems by 50%. Our results suggest that even low fertilization rates decrease grapevine N uptake from legume crop residue by both extraradical hyphae and roots.     

Microsatellite markers and genetic diversity assessment in Lolium temulentum

Lolium and Festuca are two important genera of cool-season forage and turf grasses worldwide. Lolium temulentum L. (darnel ryegrass) has been proposed as a model species for genomics studies of cool-season forage and turf grasses. A study with 41 darnel ryegrass, three tall fescue (Festuca arundinacea Schreb.), two tetraploid fescue (F. glaucescens), and two meadow fescue (F. pratensis) genotypes was initiated to (i) identify a set of microsatellite (simple sequence repeats) markers useful for L. temulentum L., and (ii) to utilize such markers for assessing the genetic variability of L. temulentum accessions collected from different geographical regions of the world. A total of 40 tall fescue (TF) EST-SSRs and 60 Festuca–Lolium (F × L) genomic SSRs were screened on a subset of eight genotypes. The selected 30 tall fescue EST-SSRs and 32 F × L genomic SSRs were used for further analysis of genotypes. The TF-EST- and the F × L genomic-SSRs identified 10.3 and 9.3 alleles per marker, respectively with an average polymorphic information content (PIC) value of 0.66. The phenogram based on 319 EST-SSR and 296 genomic SSR fragments, grouped L. temulentum accessions into three major clusters except for accession ABY-BA 8892.78. Lolium temulentum accession ABY-BA 8892.78 did not cluster with any other accession. The Festuca clusters were distantly related with darnel ryegrass clusters with a similarity coefficient of 0.26. The selected set of tall fescue EST- and F × L genomic SSRs were useful in assessing L. temulentum genetic diversity and could benefit the genetic improvement of members of the Festuca–Lolium complex.     

Analysis of Microsatellite Diversity in Ethiopian Tetraploid Wheat Landraces

The extent and patterns of microsatellite diversity in 141 Ethiopian tetraploid wheat landraces consisting of three species Triticum durum Desf., T. dicoccon Schrank and T. turgidum L. were analyzed using 29 microsatellite markers. A high level of polymorphism and a large number of alleles unique for each species were detected. Compared to emmer (T. dicoccon) and poulard (T. turgidum) wheats, a higher genetic diversity was observed in T. durum. The A-genome was more polymorphic than the B-genome in all the three species. Microsatellites with (GA) _n -repeats had a higher number of alleles than (GT) _n -repeats. A species pairwise comparison was made to determine the percentage of shared alleles and a large number of common alleles among species were observed. Average gene diversity, across the 29 microsatellite loci, was 0.684 for T. durum, 0.616 for T. dicoccon and 0.688 for T. turgidum. Genetic distances were lower between T. durum and T. turgidum (0.26) than between T. durum and T. dicoccon (0.34) or between T. turgidum and T. dicoccon (0.38). A significant correlation (p < 0.01) was found between the number of alleles per locus and the gene diversity in all the three species. Allelic frequency variation was highest between T. turgidum and T. dicoccon (10.62%) and lowest between T. durum and T. turgidum (4.86%). A genetic similarity coefficient of 0.34, 0.46 and 0.37 was found in T. durum, T. dicoccon, and T. turgidum, respectively. The dendogram, which was constructed on the basis of a similarity matrix using the UPGMA algorithm, distinguished all accessions represented in the study.     

Validity of Phyllometric Parameters Used to Differentiate Locan Vitis Vinifera L. Cultivars

Phyllometry (measurement of parameters in leaves) was studied in 13 local vinifera grapevine cultivars from theampelographic collection in Vipava (Slovenia). The main purpose of the study was to find out which parameterswere stable in vines of the same cultivars. Altogether 71 parameters were measured and/ or calculated. Amongthem 25 parameters with the least difference among the same cultivars were identified and processed by theCluster Analysis. The dendrogram obtained presents vines of the same cultivars together. A correct groupingwithin cultivars confirms an accurate grouping between cultivars. Most of selected parameters are the sizes ofangles and rations between the basic parameters, as they are not influenced by the leaf size but the leaf shape. Suchmethod makes procedures easier and results more effective.     

Detection of genetic diversity in Libyan wheat genotypesusing wheat microsatellite markers

Twenty-four wheat microsatellites (WMS) wereused to estimate the extent of genetic diversity among 15 Libyanwheat genotypes. The WMS used determined 26 loci located on 20different chromosomes, and were capable of detecting 116 alleles withan average of 4.5 alleles per locus. Only two markers located on 2DSand 4DL, were monomorphic. The results indicated that the B genome(5.9 alleles per locus) was more variable than the A and Dgenomes (4.1 and 2.7 alleles per locus, respectively).Furthermore, the results obtained suggest that a relatively smallnumber of primers can be used to distinguish all genotypes used andto estimate their genetic diversity. Genetic dissimilarity valuesbetween genotypes, calculated by the WMS derived data, were used toproduce a dendrogram. The diversity within the analysed germplasm isdiscussed.     

Genetic and Phenotypic Diversity in Downy-mildew-resistant Sorghum (Sorghum bicolor (L.) Moench) Germplasm

Genetic and phenotypic diversity among randomly selected 36 downy-mildew-resistant sorghum accessions were assessed, the former using 10 simple sequence repeat (SSR) marker loci and the latter using 20 phenotypic traits. The number of alleles (a _j ) at individual loci varied from five to 14 with an average of 8.8 alleles per locus. Nei's gene diversity (H _j ) varied from 0.59 to 0.92 with an average of 0.81 per locus. High gene diversity and allelic richness were observed in races durra caudatum (H _j = 0.76, a _j = 4.3) and guinea caudatum (H _j = 0.76, a _j = 3.8) and in east Africa (H _j = 0.78, a _j = 7.2). The regions were genetically more differentiated than the races as indicated by Wright's F _st. The pattern of SSR-based clustering of accessions was more in accordance with their geographic proximity than with their racial likeness. This clustering pattern matched little with that obtained from phenotypic traits. The inter-accession genetic distance varied from 0.30 to 1.00 with an average of 0.78. Inter-accession phenotypic distance varied from 0.01 to 0.55 with an average of 0.33. Eleven accession-pairs had phenotypic distance of more than 0.50 and genetic distance of more than 0.70. These could be used as potential parents in a sorghum downy mildew resistance-breeding program.     

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.     

Spore production by the vesicular-arbuscular mycorrhizal fungus Glomus monosporum as related to host species,root colonization and plant growth enhancement

Summary The vesicular-arbuscular mycorrhizal fungus Glomus monosporum was inoculated on grapevine (Vitis vinifera), red clover (Trifolium pratense), meadow grass (Poa pratensis) and onion (Allium cepa) as hosts in two experiments carried out in different environments. Grapevine and clover showed the largest growth response and spore production. Mycorrhizal infection was lowest in meadow grass. Very poor correlations were observed, on an overall basis, between spore production and per cent root infection or infected root length. Spore production per unit infected root length for each host species was a comparatively stable parameter; it was largest for grapevine and smallest for meadow grass in both experiments. Sporulation was positively correlated with growth enhancement by mycorrhizal plants, and growth increments per unit infected root followed the same trend as spore numbers per unit infected root, i.e. largest for grapevine and lowest for meadow-grass. It is concluded that the ability of G. monosporum to produce spores and to enhance plant growth per unit infected root length depends on the host plant species.     

Genetic diversity in Ethiopian hexaploid and tetraploid wheat germplasm assessed by microsatellite markers

The genetic diversity of a subset of the Ethiopian genebank collection maintained at the IPK Gatersleben was investigated applying 22 wheat microsatellites (WMS). The material consisted of 135 accessions belonging to the species T. aestivum L. (69 accessions), T. aethiopicum Jacubz. (54 accessions) and T. durum Desf. (12 accessions), obtained from different collection missions. In total 286 alleles were detected, ranging from 4 to 26 per WMS. For the three species T. aestivum, T. aethiopicum and T. durum on average 9.9, 7.9 and 7.9 alleles per locus, respectively, were observed. The average PIC values per locus were highly comparable for the three species analysed. Considering the genomes it was shown that the largest numbers of alleles per locus occurred in the B genome (18.4 alleles per locus) compared to A (10.1 alleles per locus) and D (8.2 alleles per locus) genomes. Genetic dissimilarity values between accessions were used to produce a dendrogram. All accessions could be distinguished, clustering in two large groups. Whereas T. aestivum formed a separate cluster, no clear discrimination between the two tetraploid species T. durum and T. aethiopicum was observed.     

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.     

Coconut (<Emphasis Type="Italic">Cocos nucifera</Emphasis> L.) DNA studies support the hypothesis of an ancient Austronesian migration from Southeast Asia to America

The centre of origin of coconut extends from Southwest Asia to Melanesia. Nevertheless, its pre-Columbian existence on the Pacific coast of America is attested. This raises questions about how, when and from where coconut reached America. Our molecular marker study relates the pre-Columbian coconuts to coconuts from the Philippines rather than to those of any other Pacific region, especially Polynesia. Such an origin rules out the possibility of natural dissemination by the sea currents. Our findings corroborate the interpretation of a complex of artefacts found in the Bahía de Caraquez (Ecuador) as related to South-East Asian cultures. Coconut thus appears to have been brought by Austronesian seafarers from the Philippines to Ecuador about 2,250 years BP. We discuss the implications of molecular evidence for assessing the possible contribution of early trans-pacific travels to and from America to the dissemination of domesticated plants and animals.     

磁化处理促进施氮条件下葡萄氮素的代谢和分布

以‘夏黑’葡萄扦插苗为试验材料,采用盆栽试验方法,分析了磁化水灌溉后葡萄叶片、茎和根系中不同形态氮素含量、氮素代谢关键酶活性以及不同氮源的贡献率,探讨磁化作用对‘夏黑’葡萄扦插苗生长以及氮素吸收、分配和利用的影响。以~(15)N为外源氮肥,分3次施入土壤中。试验设置4个处理,包括:磁化水灌溉处理、非磁化水灌溉处理、磁化水灌溉+施氮处理、非磁化水灌溉+施氮处理。磁化处理组中利用磁化装置处理灌溉水。结果表明:1)施氮条件下,与非磁化处理相比,磁化处理后葡萄叶片、根系和全株的全氮量提高,但是肥料中~(15)N对不同器官中氮素的贡献率无显著差异;叶片和根系的氮素利用率显著提高;全氮在叶片中分配率显著提高,在茎中的分配率则显著降低。2)与非磁化处理相比,磁化处理后葡萄叶片中谷氨酰胺合成酶和谷氨酸合酶活性显著提高,根系中显著降低。3)与单独施氮相比,磁化水灌溉+施氮提高了土壤氮含量;氮肥中~(15)N利用率提高,损失率降低。由以上研究结果可以看出,磁化水灌溉不仅可提高氮素代谢关键酶活性,而且可提高不同器官中氮素营养的吸收和利用,从而改变了氮素在不同器官中的分布。