Susceptibility to sharka (Plum pox virus) in Prunus mandshurica×P. armeniaca seedlings

Prunus mandshurica [(Maxim.) Koehne] from Central Asia is a species related to apricot and grown in China and Mongolia. This species has been used in apricot breeding as a source of frost resistance. In addition, P. mandshurica has been suggested as the possible origin of some North American apricot cultivars resistant to sharka (Plum pox virus, PPV). The aim of this work was to transmit the resistance to PPV from P. mandshurica to the Spanish apricot cultivar ‘Currot’ by traditional crossing. The resistance to a Dideron PPV isolate of the descendants of P. mandshuricaבCurrot’ and their progenitors was evaluated under controlled conditions in a greenhouse. The results showed the susceptibility of both, the progenitors and the offspring to PPV, as being much higher than in other apricot seedlings. The results showed that the P. mandshurica accession studied is not a good progenitor in breeding apricot for PPV resistance, but just the opposite. The possible role of P. mandshurica as a source of resistance in apricot resistant cultivars is questioned.     

Resistance of almond cultivars to Plum pox virus (sharka)

Sharka, a disease caused by Plum pox virus (PPV), mainly affects some Prunus species, including apricot, peach and plum, and to a lesser degree, sweet cherry and sour cherry. In almond, different PPV isolates have been transmitted experimentally to the ‘Aï’ cultivar. In this study, the resistance of 10 almond cultivars to a Dideron PPV isolate was evaluated in controlled conditions by grafting the cultivars on to inoculated GF305 peach rootstocks. The results demonstrated a high level of resistance to PPV in all the almond cultivars assayed. They did not show any symptoms and were ELISA and RT‐PCR negative, despite the strong symptoms observed in their GF305 rootstocks. The implications of these results for the dispersion of PPV, and the potential role of almond as a source of resistance to PPV in other Prunus species such as peach, are also discussed.     

Coat protein-mediated protection against plum pox virus in herbaceous model plants and transformation of apricot and plum

Summary The expression of the viral coat protein gene in transgenic plants has been shown to induce tolerance against virus infection (Beachy et al., 1990). Transgenic plants ofNicotiana clevelandii andNicotiana benthamiana- herbaceous host plants for PPV - transformed withAgrobacterium strain LBA 4404 containing the plasmid pBinPPVm, regenerated on selection media containing kanamycin were tested for the expression of the PPV coat protein gene by ELISA and immuno western blot. After rooting and acclimatisation plants were tested for the protection against PPV Following the inoculation plants were investigated for symptom development and virus accumulation. Different lines were identified, according to the different reaction to the mechanical inoculation, ranging from a complete absence to a strong reduction of symptoms. There have not been many reports on transformation of trees in general, and in fruit trees particularly. It is obvious that the major obstacle is the regeneration of transformed plantlets. Attempts to improve crop plants by genetic engineering techniques will always depend very strongly on the availability of reliable protocols for transformation, selection and regeneration (Laimer et al., 1989, 1990). Different systems involving juvenile and adult plant material have been developed allowing the transfer of foreign genes into apricot and plum cultivars. We report the transformation and regeneration ofPrunus armeniaca andPrunus domestica plants withAgrobacterium tumefaciens strain LBA 4404 containing various binary plasmids, pBinGUSint, carrying the marker geneβ-glucuronidase (GUS) and pBinPPVm, carrying the coat protein gene of Plum Pox Virus (PPV), the causal agent of Sharka disease. The marker geneGUS was used for the optical evaluation of the efficiency of different transformation systems involving cotyledons of immature embryos as well as somatic embryos and leaf discs. The coat protein gene of PPV was used to introduce the coat protein mediated resistance against one of the most important pathogens of stone fruit trees in Europe and the whole Mediterranean area.     

RFLP variability in apricot (Prunus armeniaca L.)

The level of polymorphism of the restriction fragment length polymorphisms (RFLPs) detected by 33 almond genomic and cDNA probes was studied in a set of 52 European and North-American apricot cultivars. Eighteen of these probes were polymorphic and yielded a total of 48 scorable bands, allowing the identification of 45 different phenotypes. Most cultivars (43) had an individually distinguishable RFLP phenotype, and three of the five clusters with the same phenotype contained cultivars that were likely to be synonymous. The group of Spanish cultivars (25) had a lower level of polymorphism than the others, suggesting that bottlenecks may have occurred in the recent history of the apricot that have eroded its genetic variability.     

不同地理区间上家马母系结构的差异分析

为了分析家马在世界各地理区间上的遗传结构差异,对家马遗传资源评价提供有益帮助,并对其母性起源和驯化提供一些分子生物学依据。对4个家马品种（蒙古马、锡尼河马、三河马和纯血马）共36个个体的mtDNA Cytb基因全序列进行了扩增和测序,并结合目前GenBank上登录的世界各地所有家马Cytb全序列进行分析,共检测到86种单倍型80个多态位点,其单倍型多样度为0.946,核苷酸多样度为0.00454,表明家马的遗传多态性比较丰富。首次通过将各品种家马根据地理区域划分为北美、西欧、东欧、中东、中亚、东亚6个类群以Cytb全序列作为标记比较母系遗传结构差异。结果表明：北美类群和亚洲类群之间差异较显著,而其余类群间则差异不显著,一方面说明亚欧大陆各地区马种之间母系基因渗透现象比较严重;另一方面一定程度上支持北美马种来自欧洲的论断。构建的Median Joining网络聚类图显示,所有家马的母系起源于A-G 7个单倍型群,进一步支持家马多重母系起源的观点。并且,家马各单倍型群都是由多个地理区间上分布的家马混杂而成的,即未发现由单独一种地理区间的家马构成的单倍型群,即地理位置、母系构成以及母性起源之间没有明显的相互关系。     

Genetic diversity in apricot cultivars based on AFLP markers

A set of cultivars used as genitors in apricot breeding programs aimed at introducing sharka resistance were examined by AFLP molecular marker analysis. The markers obtained indicated that apricot cultivars resistant to sharka were related to the European cultivars, but they potentially share a common ancestor donor of sharka outside of the European group. Segregation of AFLP and RAPD markers has been tested in two progenies of two crosses, one between a resistant and a susceptible cultivar and one between two susceptible accessions. The heterozygozity of families obtained from crosses between resistant and susceptible cultivars is similar to those obtained from other self-incompatible Prunus species. On the other hand, heterozygosity of a progeny obtained from a cross between European susceptible cultivars was very low. The usefulness of the information provided by AFLPs for apricot breeding programs is discussed. This revised version was published online in August 2006 with corrections to the Cover Date.     

Analysis of genetic profiles of winter wheats from Russia

With the aid of GRIS, the wheat genetic resources database, the retrospective analysis of winter wheat breeding programs in the ex-USSR during its existence, and modern Russia, was conducted. The dynamics of genetic diversity of released cultivars was studied. A significant tendency to reduce the use of local materials was discovered, although a stable set of original ancestors has prevailed over the last 40 years. The modern cultivar genes pool has increased as a result of the utilisation of North American semidwarf varieties. Breeding programs at different breeding centres are distinguished by varying levels of genetic diversity. The need to discover new sources of disease resistance and environmental adaptation are problems that remain. This revised version was published online in August 2006 with corrections to the Cover Date.     

Genetic variation among cultivars of red clover (Trifolium pratense L.) detected by RAPD markers amplified from bulk genomic DNA

Summary The use of random amplified polymorphic DNA (RAPD) markers obtained from bulked samples was investigated for cultivar identification in red clover. Pooled samples were examined in order to minimize variation within cultivars. To determine the appropriate number of individuals to include in the bulked samples representing each cultivar, DNA samples from two, three, four, five, ten and twenty individuals were pooled. Twenty was found to be an appropriate number of red clover individuals per bulk in order to amplify only the DNA sequences shared among most individuals in each cultivar. Fourteen 10-mer primers were used to amplify genomic DNA from combined leaf samples of 15 red clover cultivars from European, Japanese and North American origins. A total of 79 amplified products, of which 55 were polymorphic, was obtained. Cultivar-specific bands were observed with 13 primers. The amplification patterns obtained from two primers could distinguish all 15 red clover cultivars. Rogers' genetic distances for all 105 pairwise comparisons were calculated to evaluate relationships among these cultivars. Cluster analysis based on these genetic distances separated these 15 cultivars into three groups, with two of the groups consisting of a single Japanese cultivar each, while the third group included cultivars from European, North American, and Japanese origins.     

Genetic structure and differentiation in hop (Humulus lupulus L.) as inferred from microsatellites

A set of 67 wild and cultivated hop accessions, representative of hop diversity, was genotyped with 29 SSR markers in order to investigate the population structure and genetic diversity among hop genotypes. A total of 314 alleles was detected, with an average of 10.8 alleles per locus and an average PIC content of 0.607. Model-based clustering placed the accessions into five germplasm groups. A distance-based tree showed good agreement with five germplasm groups, and additionally assigned accessions omitted from model-based analysis into two additional germplasm groups. The 67 hop accessions were thus subdivided in seven germplasm groups, with three corresponding to major breeding groups and four to wild hops. This finding is in accordance with two biogeographically separated hop germplasms (European and North American origin) and with the known history of the accessions. North American hop germplasm was partitioned into native and cultivated germplasm groups. European germplasm was divided into two groups of hop cultivars representing distinguishable European germplasms and three new groups of native hops, which were differentiated for the first time by this analysis. Admixture analysis showed shares of various ancestries in hop cultivars, mostly congruent with pedigree data, and the introgression of various ancestries in some native hops. The above results have so far given the most detailed insight to date into the population structure of hop diversity, which is important for its effective use in hop breeding.     

Genetic relationships in cultivars of hop, Humulus lupulus L., determined by RAPD analysis

Random amplified polymorphic DNA (RAPD) was used to evaluate the genetic variability and relationship of 65 hop cultivars from all the major hop-growing regions in the world. Twenty-eight selected random primers used in the RAPD reaction generated an average of 38.6%) polymorphic fragments, which was sufficient to produce 47 different RAPD profiles among the cultivars examined. The level of genetic variability was much higher than previously reported. Genetic similarity was estimated and UPGMA cluster analysis was performed using the RAPD data. Cluster analysis separated the cultivars into genetically related RAPD groups which were compared with pedigree data and grouping of the hop cultivars by essential oil type. The RAPD groups, strongly supported by pedigree data, gave more precise information on the level and distribution of genetic variability within hop cultivars than characterization by essential oils. Cultivars were divided into American and European groups, supporting the distinction between two geo-graphically distinct hop germplasms. Five genetically distinct groups revealed differences within the European germplasm, reflecting past hop breeding practices which have been adopted in different regions. The use of RAPD markers for hop germplasm characterization and genetic diversity study is discussed.     

RFLP diversity within and between major groups of barley in Europe

Restriction fragment length polymorphism (RFLP) diversity has been determined and analyzed as expressed by 33 single‐ or low‐copy clone/ enzyme combinations at 32 loci distributed over all chromosomes of the barley genome within a sample of 223 European barley accessions comprised of pure line (single‐head progenies) genotypes. The accessions have been selected to include landraces and widely grown cultivars derived from crossbreeding during the 20th century in North‐, West‐ and Central European countries. Genetic diversity obtained from 83 alleles across all accessions is characterized by the diversity index H = 0.385. The diversity indices determined for landraces and cultivars were almost equal, with the difference between spring (H = 0.260) and winter (H = 0.415) barley approaching statistical significance, while comparisons of other groupings only revealed statistically insignificant trends. A more detailed analysis based on differences in allele frequency distributions at each locus (clone/enzyme combinations resp.) revealed very clear differences related to the existence, continuity and dynamics of changes in group‐specific RFLP profiles. With the majority (69%) of RFLP alleles at 23 out of 32 loci on all barley chromosomes involved, contributions from chromosomes 1H, 3H, 4H and 5H seem to be of special importance. Differences in the overall average of abundance indicate higher levels of genetic diversity within both groups of winter barley compared with both groups of spring barley, from which the most frequent alleles at 15 (2‐rowed spring barley) and 17 (6‐rowed spring barley) RFLP loci approach fixation. The results of this study are discussed in relation to the history of barley cultivation and barley breeding in Europe, and possible explanations for group‐specific differences in the RFLP profiles of landraces and cultivars as well as for the high levels of (nearly) fixed alleles of both subsets of spring barley, and with respect to progress in barley breeding that it has been possible to obtain within the rather narrow RFLP profiles.     

亚洲不同生态区水稻群体遗传多样性与特异性分析

研究不同生态区水稻群体的遗传多样性及其遗传结构分化、群体特异性及其相互关系,为水稻起源及亲本遴选提供遗传基础。本研究利用119对SSR标记对太湖、黑龙江省和越南的440种水稻品种进行分析。结果在三个不同生态区中的太湖流域水稻中检测到783个等位基因,平均多态信息含量（PIC）为0.49;在黑龙江水稻中检测到296个等位基因,平均多态信息含量（PIC）为0.21;在越南水稻中共检测到374个等位基因,平均多态信息含量（PIC）为0.28,这表明越南水稻和黑龙江水稻的多样性低于太湖水稻。特缺等位变异数最多的是黑龙江亚群（132）,最少的是太湖亚群（4）;特有等位变异数最多的是太湖亚群（284）,最少的是黑龙江亚群（25）。从亚群间补充等位变异数来看：其补充等位变异数最多的是太湖亚群对黑龙江亚群的补充（447）,最少的是黑龙江亚群对太湖亚群的补充（29）。利用Structure2.2软件将三个地区的440个品种分为7类血缘,发现每个血缘都不是独立的而是相互渗透的。     

Genotyping apricot cultivars for self-(in)compatibility by means of RNases associated with S alleles

In previous work the existence of proteins with RNase activity associated with S alleles in apricot was demonstrated. These proteins were inherited as described previously for the inheritance of self‐compatibility in this species. In this study, new cultivars have been genotyped for self‐compatibility using this method and it has been demonstrated that in all self‐compatible cultivars examined, the self‐compatibility allele is the same and is associated with an RNase with high activity. Homozygous self‐compatible individuals have been detected among established cultivars as well as among seedlings following breeding activity. This germplasm is of great value within the breeding programme because only self‐compatible seedlings will be produced. The number of S alleles in apricot appears to be low and only eight different alleles have been found in the large number of different cultivars screened. Furthermore, there are alleles present in the Spanish population that are also found in the genetic pool of North American cultivars. The screening of a progeny from the cross between the American cultivar ‘Goldrich’ and the Spanish cultivar ‘Pepito’ demonstrated the existence of the common allele S₂ (detected previously by examining RNases), which was confirmed by the segregation of self‐compatibility in the progeny.     

2007—2008年度冀鲁豫川四省小麦区试品种（系） 遗传多样性和抗条锈性分析

为了解2007—2008年度河北、河南、山东和四川四省小麦区试品种（系）的遗传多样性和抗条锈性,采用亲缘系数系统分析了参试的75个小麦品种（系）的遗传多样性,并接种条锈菌混合菌系（CYR33、CYR32、CYR31、CYR30、CYR29、CYR17、Su-1和V26）进行抗条锈性鉴定与评价。结果表明,在供试品种（系）的2775个组合中,284个组合存在遗传相似性,所有供试品种（系）的COP值在0.0000～0.5000,亲缘系数总和为216.4531。通过对COP值聚类分析,将供试材料分为10大类;河北、河南、山东3省参试品种（系）间遗传相似性较高,对小麦条锈病抗性相对较差,四川参试品种（系）遗传相似性较低,对小麦条锈病的抗性较好。从参试品种（系）系谱看,利用前一个时期选育的品种作亲本,特别是主要推广品种‘豫麦2号’作为亲本的利用率较高是河南省参试品种（系）遗传相似性较高的重要原因;四川省参试品种（系）遗传多样性相对较高,抗条锈性相对较好,主要由于该省广泛利用了当地和其他地区的种质资源。因此,不断发掘小麦新的种质资源,并提高其利用水平,是当前高产稳产新品种选育的当务之急。     

High genetic diversity of Spanish common wheats as judged from gliadin alleles

Gliadin alleles were identified in 100 common wheat cultivars registered and/or grown in Spain during the last 40 years. A very high level of genetic polymorphism was found: in total, 103 allelic variants including one null‐allele were found at the six major Gli loci in the Spanish wheats studied. An average genetic diversity for these six loci was found to be higher (H=0.844) than in any group of wheat cultivars studied previously. Spanish wheats bred in Spain demonstrated even higher genetic diversity (H=0.868), probably because of the occurrence in this group of some landraces (local varieties) assumed to be strongly differentiated to fit local environments. The high level of genetic diversity of wheats grown in Spain was maintained by the introduction of distantly related wheat germplasm from different sources, especially from Italy and CIMMYT. A slight decrease of genetic diversity in recently registered cultivars might be caused by the excessive introduction of French wheats. Thirteen new alleles found in Spanish wheats were catalogued, including Gli‐D2w which encodes the first Gli‐D2‐controlledγ‐gliadin to be found.     

瓜尔豆品种遗传关系分析

瓜尔豆[Cyamopsis tetragonoloba (L.) Taub.]为一年生抗旱耐热豆类植物,是Cyamopsis属中经济性最为重要的一个种。有关商业品种的遗传关系的信息对于瓜尔豆育种至关重要。本文依据6个瓜尔豆,1个C. senegalensis 种和1个 C. serrata种的引种系的RAPD和植物学性状数据采用UPGMA方法 进行了聚类分析。供试的三个种共扩增出29条带,其中26条具有多态性。每个引物扩增出的多态性片段为2至6条,平均3.3条。基于这26条多态性RAPD片段的Jaccard相似系数,聚类分析将这3个种分为2组：第一组包括C. serrata 和 C. senegalensis 2个野生种,第二组包括Cyamopsis tetragonoloba种的6个品种。在第二组中,‘Kinman’与其他5个品种的关系较远。用这8个材料形态性状非相似系数进行的聚类分析获得了类似的结果,但用形态性状比之RAPD检测到组内更多的变异。     

Genetic variation in the olive tree (Olea europaea L.) cultivated in Morocco

Summary Genetic variation was studied using enzyme polymorphism at 8 loci in ancient olive trees. These were being cultivated in 10 sites along a transect from North (Pre-Rif) to South (Anti-Atlas), involving the main ecological areas where the species is cultivated in Morocco. For the 328 trees studied, 16 alleles and 68 multilocus genotypes, of which one was present in more than half the individuals analysed, were detected. Eighty seven per cent of the total genetic diversity was attributable to within site variation and showed a high proportion of local genotypes. The highest values for both genotype number and heterozygosity were observed in the South of Morocco. Such high variation may be due to the partial domestication of olive material which may be derived from crosses between cultivars or between cultivars and feral or wild olive (oleaster) trees growing frequently in the Southern region. Genetic diversity in Moroccan olive, constitutes an important genetic resource which must be conserved for further breeding.     

Genetic diversity and phylogenetic relationships among accessions of hop, Humulus lupulus, as determined by amplified fragment length polymorphism fingerprinting compared with pedigree data

DNA fingerprinting using amplified fragment length polymorphisms (AFLPs) was successfully employed to detect genetic relationships and variability among 90 hop cultivars and breeding lines comprising a collection of the world's hop germplasm. Seven AFLP primer combinations produced a total of 347 fragments of which 151 (43.5%)) were polymorphic. One‐hundred and thirty informative, highly reproducible DNA polymorphisms were used to estimate the genetic similarity (GS) which varied between 1.0 (e.g. ‘Saazer’ vs. ‘Tettnanger’) and 1.17 (‘Columbus’ vs. ‘Tettnanger’, ‘Spalter’ and ‘Saazer’). UPGMA (unweighted pair‐group method with arithmetic averages) clustering revealed two main clusters, reflecting the two main sources of origin and the two main breeding objectives: one cluster of mainly European origin representing the aroma pool and a second cluster associating accessions with European germplasm infiltrated by wild American genes with less aroma quality, but a higher bittering potential. Each main branch was composed of four or three subclusters with subgroups, respectively. Assignment of almost all genotypes in the dendrogram was consistent with the pedigree data as far as they are known. Consequently, AFLPs are shown to be suitable for assessing the genetic variability in hop germplasm and are useful for describing the genetic relationships among cultivars and accessions, which allows phylogenetic questions to be addressed.     

Exotic barley germplasms in breeding for resistance to soil-borne viruses

Summary Soil-borne mosaic inducing viruses, i.e., barley mild mosaic virus (BaMMV), barley yellow mosaic virus (BaYMV), and BaYMV-2, cause one of the most important diseases of winter barley in Western Europe. Since resistance of all commercial European barley cultivars is due to a single recessive gene (ym4) which is not effective against BaYMV-2, exotic barley germplasms (Hordeum vulgare L., H. spontaneum Koch) were screened for resistance to the different viruses and analyzed for genetic diversity concerning BaMMV resistance. In these studies it turned out that resistance to BaMMV is entirely inherited recessively and that a high degree of genetic diversity concerning resistance is present within the barley gene pool at least to BaMMV. Therefore, exotic barley germplasms are a very useful source for the incorporation of different resistance genes into barley breeding lines, thereby enabling the breeder to create cultivars adapted to cultivation in the growing area of fields infested by soil-borne viruses. Furthermore, in order to obtain more information on these germplasms they were evaluated for agronomic traits and isozyme, RFLP and RAPD analyses were carried out on these varieties to detect markers linked to the respective resistance genes and to obtain information on the genetic similarity between yellow mosaic resistant barley accessions derived from different parts of the world. Actual results of these studies are briefly reviewed.     

Molecular characterization of genetic diversity in European germplasm of perennial ryegrass

Summary Perennial ryegrass (Lolium perenne L.) is the most important grass species for temperate grassland agriculture. The level and distribution of genetic variation in gene bank ecotype collections is still largely unknown but of great interest for the planning of breeding programs. The objectives of this study were to (i) assess the molecular diversity of Polish ecotypes of perennial ryegrass, and (ii) compare the relationship between this group and German ecotypes and European cultivars investigated previously. A total number of 166 polymorphic marker bands were detected among the 171 individual plants of the 9 Polish ecotypes. In a joint analysis with 9 Polish and 22 German ecotypes, and 22 European cultivars 172 polymorphic RAPD markers could be found. Genetic distance among the Polish ecotypes ranged from 0.31 to 0.51, while for all 53 populations a broader range was detected (0.25–0.67). An analysis of molecular variance (AMOVA) revealed a much larger variation within populations (71%) than among them (29%). The Polish ecotypes contained the highest within population variation (74%). The largest among group difference (15%) was found between the Polish ecotypes versus all other accessions. We conclude that the Polish ecotypes represent a valuable genetic resource for enlarging the genetic variation in the West European germplasm pool of perennial ryegrass.