The effect of gamma irradiation on potato microtuber production in vitro

The effects of low doses of gamma irradiation and potato (Solanum tuberosum L.) cultivar on the production of microtubers in vitro were investigated. Nodal segments from virus free explants of three potato cultivars (cv.) were placed on tuberization inducing medium and irradiated with 4 doses of gamma radiation (2.5, 5, 10, 15 Gy). Cv. Diamant produced the highest number of microtubers followed by Draga and Spunta. Irradiation of the explants with 2.5 Gy of gamma radiation led to a significant increase in the number of microtubers (38% increase over the control). Average weight of microtubers was not significantly influenced by low doses of gamma irradiation. Draga microtubers were the largest followed by Diamant and Spunta. Microtubers resembled mature tubers in shape (Spunta was oval and Draga and Diamant were spherical). Size of microtubers was crucial for sprouting in vivo. It is suggested that only microtubers larger than 5 mm in diameter (250 mg) be used to produce minitubers in vivo. Since 2.5 Gy is a low irradiation dose, it can be used to enhance tuberization in vitro without fear of genetic changes in the used cultivars. This revised version was published online in June 2006 with corrections to the Cover Date.     

Genetic differentiation of wild and cultivated Lens based on molecular markers

Genetic diversity among 83 lentil genotypes including 23 wild types, 19 indigenous varieties, 5 exotic lines and 36 advanced breeding lines was studied using molecular markers. A total of 112 amplicons were produced using 15 RAPD and 8 SSR markers. Dendrogram based on Jaccard similarity coefficient and UPGMA analysis revealed two major clusters and one minor cluster. Cluster I comprised 21 wild accessions of L. orientalis and 1 L. ervoides subspecies. Nineteen Indian varieties grouped together in subcluster IIA indicating their narrow genetic base. Subcluster IIB consisted of 41 genotypes including 5 exotic and 36 advanced breeding lines mainly derived from exotic genotypes. The narrow genetic base of released cultivars and germplasm lines emphasized the need for broadening of genetic base of breeding material using exotic collections and wild species to ascertain genetic improvement upon existing cultivars.     

The R1 gene for late blight resistance in early and late maturing potato cultivars

The method of polymerase chain reaction was used to amplify a fragment of the LZ-NBS-LRR receptor kinase gene R1; the gene was transferred into potato (Solanum tuberosum) from its wild-growing relative S. demissum and confers the race-specific recognition of the pathogen Phytophthora infestans. To verify this method as a test for the presence of the late blight resistance gene R1, the amplified genome fragment was cloned from the potato hybrid comprising the germplasm of S. demissum. The primary structure of this fragment, which corresponded to the receptor domain of kinase, did not practically differ from the matching sequence in S. demissum. In addition, the method was verified by scoring the set of plant differentials, wherein the presence of R1 was established with race-specific Phytophthora isolates. By screening 70 potato cultivars, we established a significant relationship between the presence of the gene R1 fragment and the phenotypic characters of late blight resistance and late maturity. This evidence supports the idea that R1 was introgressed from short-day S. demissum into potato plants together with some gene(s) conferring late transition to flowering.     

Assessing genetic potential in germplasm collections of crop plants by marker-trait association: a case study for potatoes with quantitative variation of resistance to late blight and maturity type

Genetic diversity of crop plants resulting from breeding and selection is preserved in gene banks. Utilization of such materials for further crop improvement depends on knowledge of agronomic performance and useful traits, which is usually obtained by phenotypic evaluation. Associations between DNA markers and agronomic characters in collections of crop plants would (i) allow assessment of the genetic potential of specific genotypes prior to phenotypic evaluation, (ii) identify superior trait alleles in germplasm collections, (iii) facilitate high resolution QTL mapping and (iv) validate candidate genes responsible for quantitative agronomic characters. The feasibility of association mapping was tested in a gene bank collection of 600 potato cultivars bred between 1850 and 1990 in different countries. The cultivars were genotyped with five DNA markers linked to previously mapped QTL for resistance to late blight and plant maturity. Specific DNA fragments were tested for association with these quantitative characters based on passport evaluation data. Highly significant association with QTL for resistance to late blight and plant maturity was detected with PCR markers specific for R1, a major gene for resistance to late blight, and anonymous PCR markers flanking the R1 locus at 0.2 Centimorgan genetic distance. The marker alleles associated with increased resistance and later plant maturity were traced to an introgression from the wild species S. demissum. These DNA markers are the first marker that are diagnostic for quantitative agronomic characters in a large collection of cultivars.     

Application of DNA markers linked to the potato <Emphasis Type="Italic">H1</Emphasis> gene conferring resistance to pathotype Ro1 of <Emphasis Type="Italic">Globodera rostochiensis</Emphasis>

Ninety-one potato genotypes (cultivars and breeding lines) selected as resistant or susceptible to pathotype Ro1 of Globodera rostochiensis were screened for the presence of two PCR markers, 0.14 and 0.76 kb in length. Both PCR markers were linked with the H1 gene, located at the distal end of the long arm of chromosome V, and were present in 88 to 100% of the resistant cultivars and breeding lines. The 0.76 kb PCR marker was detected in all resistant genotypes and in approximately 86% of susceptible breeding lines as well as in all susceptible cultivars. The 0.14 kb marker was detected in 88% of resistant breeding lines and in 94% of resistant cultivars. Most of the susceptible genotypes tested (91% of cultivars, but only 50% of breeding lines) did not show the presence of the 0.14 kb marker. We conclude that the 0.14 kb H1 marker is likely to be useful for the proper selection of potato genotypes resistant to the Ro1 pathotype of G. rostochiensis.     

Role of certain potato tubers constituents in their resistance to bacterial soft rot caused by Erwinia carotovora pv. carotovora

Erwinia soft rot causes destructive and serious damages to many vegetable crops including potato, in the field, transit and storage periods. The role of certain potato tuber constituents in the physiology of disease resistance has been investigated. Pectin substances and calcium contents of potato tuber had a pronounced role in the physiology of disease resistance. Alpha and Santa (less susceptible cultivars) contained the higher amount of pectin and calcium compared by Mirkal, Diamant, Askort, Geganite and Nicola cultivars (more susceptible cultivars). Tubers extracts of all healthy tested potato tubers cultivars contained fructose except Santa cultivar and glucose except Alpha and Mirkal cultivars. Tuber extracts of the more susceptible cultivars (Nicola and Askort) contained a higher concentration of glucose and fructose than those of less susceptible cultivars (Geganite, Mirkal, Santa, Alpha and Diamant).     

Resistance to cucumber mosaic virus in potato

Reactions to two subgroup I isolates (Fny-CMV and Pf-CMV) and two subgroup II isolates (A9-CMV and LS-CMV) of cucumber mosaic virus (CMV) were studied in three non tuber-bearing wild potato species (Solanum spp.) of the series Etuberosa, and in two tuber-bearing interspecific potato hybrids and four potato cultivars using graft-inoculation. Three classes of phenotypic reactions (susceptible, hypersensitive, extreme resistance) were observed in the tuber-bearing genotypes. Susceptible genotypes developed mosaic or severe mosaic with leaf malformation and had high CMV titres. Hypersensitive genotypes developed either top necrosis or vein necrosis and/or necrotic spots on apical leaves, and had low CMV titres. Extremely resistant genotypes had no symptoms and no CMV was detected. The hybrid 87HW13.7 (S. tuberosum×S. multidissectum) developed top necrosis specific to infection with Fny-CMV. The hybrid ‘A6’ (S. demissum×S. tuberosum cv. Aquila) was hypersensitive to all CMV isolates tested. Extreme resistance was not functional against all CMV isolates. Neither hypersensitivity nor extreme resistance were related to the CMV subgroup.     

A Comparison of AFLP and RAPD Markers for Genetic Diversity and Cultivar Identification in Cotton

AFLP and RAPDmarkers were employed in sixteen diploid cotton (Gossypium sp) cultivars for genetic diversity estimation and cultivar identification. Polymorphism information content (PIC) and percent polymorphism were found to be more for AFLP markers as compared to RAPD markers. Average Jaccard’s genetic similarity index was found to be almost similar using either AFLP or RAPD markers. All the cultivars could be distinguished from one another using AFLP markers and also by the combined RAPD profiles. Cultivar identification indicators like resolving power, marker index and probability of chance identity of two cultivars suggested the usefulness of AFLP markers over the RAPD markers. AFLP and RAPD analyses revealed limited genetic diversity in the studied cultivars. Cluster analysis of both RAPD and AFLP data produced two clusters, one containing cultivars of G. herbaceum and another containing cultivars of G. arboreum species. Highly positive correlation between cophenetic matrices using RAPD and AFLP markers was observed. AFLP markers were found to be more efficient for genetic diversity estimation, polymorphism detection and cultivar identification.     

RAPD polymorphisms in spring wheat cultivars and lines with different level of Fusarium resistance

Random amplified polymorphic DNA (RAPD) markers have been used to characterize the genetic diversity among 35 spring wheat cultivars and lines with different levels of Fusarium resistance. The objectives of this study were to determine RAPD-based genetic similarity between accessions and to derive associations between Fusarium head blight (FHB) and RAPD markers. Two bulked DNA from either highly resistant lines or susceptible lines were used to screen polymorphic primers. Out of 160 screened primers, 17 primers generated reproducible and polymorphic fragments. Genetic similarity calculated from the RAPD data ranged from 0.64 to 0.98. A dendrogram was prepared on the basis of a similarity matrix using the UPGMA algorithm, which corresponded well with the results of principal component analysis and separated the 35 genotypes into two groups. Association analysis between RAPD markers and the FHB index detected three RAPD markers, H19(1000), F2(500) and B1(2400), significantly associated with FHB-resistant genotypes. These results suggest that a collection of unrelated genotypes can be used to identify markers linked to an agronomically important quantitative trait like FHB. These markers will be useful for marker-assistant breeding and can be used as candidate markers for further gene mapping and cloning.     

Effects of Initial Nematode Density on Population Dynamics of Globodera rostochiensis on Resistant and Susceptible Potatoes

The influence of resistant and susceptible potato cultivars on Globodera rostochiensis population density changes was studied at different nematode inoculum levels (Pi) in the greenhouse and field. Soil in which one susceptible and two resistant cultivars were grown and fallow soil in pots was infested with cysts to result in densities of 0.04-75 eggs/cm³ soil. A resistant cultivar was grown in an infested field with Pi of 0.7-16.7 eggs/cm³ soil. Pi was positively correlated with decline of soil population densities due to hatch where resistant potatoes were grown in the greenhouse and in the field but not in fallow soil. However, Pi was not correlated with in vitro hatch of G. rostochiensis cysts in water or potato root diffusate. Under continuous culture o f a resistant cultivar, viable eggs per cyst declined 60-90% per plant growth cycle (4 weeks) and the number of cysts containing viable eggs had decreased by 77% after five cycles. The rate of G. rostochiensis reproduction on both resistant and susceptible cultivars was negatively correlated with Pi. These data were used to predict the effect of resistant and susceptible potato cultivars on G. rostochiensis soil population dynamics.     

Molecular Analysis of Rhynchosporium secalis Population Collected from Barley Cultivars having Different Resistance Specificities

Molecular analysis was performed to detect genetic diversity in 106 Rhynchosporium secalis isolates collected from different regions of Canada using random amplified polymorphic DNA (RAPD) markers. The isolates collected from barley cultivars having different resistance specificity to R. secalis and grown in geographically distinct regions, exhibited reproducible variation for 2–3 polymorphic PCR products per decamer primer. Analysis of 1960 RAPD markers data obtained with five primers formed 5 groups with different genetic similarity. High genetic variation was observed in R. secalis isolates obtained from resistant and susceptible cultivars of barley. Isolates collected from susceptible cultivars showed a tendency to group together, whereas isolates from resistant cultivars were divergent. R. secalis isolates infecting different barley cultivars released as resistant to the barley scald formed a specific group with UPGMA, even though all these isolates were collected from the same epidemiological region. Analysis of 15 isolates collected from one resistant cultivar Duke formed three clusters with low bootstrap values indicating high genetic diversity among the isolates present on a single host cultivar.     

Homologs of APETALA1/FRUITFULL in Solanum plants

The MADS-box APETALA1 genes control plant transition to flowering and the floral morphogenesis proper. The experimental evidence of APETALA1 overexpression presumes that this class of genes can also directly affect time to flowering. We therefore cloned and compared homologs of APETALA1 class genes from potato (Solanum tuberosum cultivars adapted to long day conditions) and its wild relative Solanum demissum, a short-day subtropical species. The homologs isolated from these plants belong to the subclass FRUITFULL. The inconsiderable variations in the primary structure of these homologs cannot explain the diverse photoperiodic reactions of particular Solanum genotypes.     

Components of Resistance to Early Blight in Four Potato Cultivars: Effect of Leaf Position

Components of early blight resistance were quantified in leaves of different ages in four potato cultivars. The components of resistance: incubation period (IP), lesion number (LN), early blight severity, lesion expansion rate (LER), latent period (LP) and spore production by lesion area (SPLA), were evaluated separately in the lower, middle and upper leaves of four potato cultivars. Plants of cultivar Aracy (resistant), Delta (moderately resistant), Desirée (susceptible) and Bintje (susceptible) were inoculated with an Alternaria solani isolate at the beginning of the flowering stage. Disease severity varied in different plant parts. In all cultivars, regardless of resistance, the smallest values of LN, and severity were recorded on the upper leaves, suggesting that young tissues are less susceptible. In cultivar Aracy, the IP was long, with small values of LN and LER and consequently, low values of early blight severity in all leaf positions were recorded. Although IP was long in cultivar Aracy, no differences between the moderately resistant cultivar Delta and the susceptible cultivars Bintje and Desirée could be detected for this component. The IP was only influenced by leaf position in cultivar Aracy. Clear differences in resistance levels among cultivars could be detected regarding LN, severity and LER. However, neither LP nor SPLA were associated with resistance level of cultivars or with leaf position. Analyses according to plant part suggest that evaluations on leaves of the middle third part are most suitable for screening for early blight resistance in potato.     

马铃薯抗晚疫病基因Rpi-blb2的LRR区域多态性及分子进化分析

马铃薯抗晚疫病基因Rpi-blb2是来源于马铃薯野生种Solanum bulbocastanum中的一个具有广谱抗性的NBS-LRR类抗病基因。文章用PCR的方法从20个高抗晚疫病的马铃薯栽培种和20个高感晚疫病的马铃薯栽培种以及7个马铃薯野生种中克隆了Rpi-blb2基因的LRR区段。采用生物信息学方法对这些序列的相似性、多态性位点、核酸多样性指数等参数进行了分析,发现Rpi-blb2的LRR区域在核酸水平上变异程度很高,而且存在多处热点突变位点;通过对该区域的Ka/Ks值进行估算,发现Rpi-blb2的LRR区域总体上受到纯化选择,功能保守,但是LRR区域的不同部位所受到的选择压力却不尽相同。同时,从核酸水平来看,Rpi-blb2基因的LRR区域在马铃薯栽培种和马铃薯野生种之间没有发现明显的分化。     

Nucleotide polymorphism and molecular evolution of the LRR region in potato late blight resistance gene Rpi-blb2

Rpi-blb2, which is originally derived from Solanum bulbocastanum, is a broad-spectrum potato late blight resistance gene and belongs to the NBS-LRR family. Here, the LRR homologues of Rpi-blb2 were cloned with PCR method from 40 potato cultivars (including 20 resistant potato cultivars and 20 susceptible ones) and 7 wild potato populations. Then, the similarities of the sequences, polymorphic (segregating) sites, and nucleotide diversities were estimated by bioinformatic methods. The results showed that high nucleotide polymorphism and some hot-spot mutations existed in the LRR region of Rpi-blb2. The test of Ka/Ks ratio showed that the function of LRR was conserved because of the purifying selection, although different positions of the Rpi-blb2 LRR region were under different selection pressures. Moreover, the LRR region of Rpi-blb2 had no clear differentiation between the cultivated and wild potatoes.     

葡萄感霜霉病基因的分子标记(英文)

 在葡萄抗病育种中 ,幼苗期排除感霜霉病的后代具有特别重要的意义 .用 BSA,RAPD和SCAR方法研究了葡萄感霜霉病基因的分子标记 .分析了两个种间杂交组合 [毛葡萄 (抗病 )×欧洲葡萄 (感病 ) ]88- 1 1 0和 88- 84与 88- 1 1 0的 F1代自交或互交所得的 3个 F2 代 ,以及欧洲葡萄品种和中国野生葡萄种 .共筛选了 2 80个随机引物 .引物 OPO1 0产生了一个 RAPD标记 OPO1 0 - 80 0与葡萄感霜霉病主效基因紧密联锁 .将该 DNA片段克隆并测序 .OPO1 0 - 80 0的实际长度为 835bp,所以 OPO1 0 - 80 0应为 OPO1 0 - 835.据其两端序列 ,设计了一对长度为 2 6bp和 2 8bp的特异引物分别扩增上述试材 ,获得了与该 RAPD标记相同大小的一条带 ,将 RAPD标记转化为 SCAR标记SCO1 0 - 835.并证实了此 SCAR标记的通用性 ,该 SCAR标记可用于葡萄抗病育种中杂种后代对霜霉病的抗病与感病性鉴定 .     

Detecting pathogenic bacterial wilt disease of potato using biochemical markers and evaluate resistant in some cultivars

Bacterial wilt caused by Ralstonia solanacearum (Smith), is one of the chief severe diseases of potato in warm temperate regions, tropics and subtropics of the world. The study was conducted to isolate and identify bacterial pathogens and select the most resistant cultivars and avoid the decrease in the total value of Egyptian potato exports to the European Union (EU) due to the quarantine restrictions imposed by the EU on potato tubers exported from Egypt affected by bacterial wilt. The results of traditional identification through morphological and serological studies showed that the five isolates were isolated and identified as Ralstonia solanacearum. Furthermore, the results illustrated that RS5 isolate showed the lowest percentage of disease incidence reduction on the three tested potatoes cultivar Bellini, Spunta and Mondial recorded 9.64%, 15.41% and 34.12%, respectively. While, RS8 isolate exhibited the highest effective one the percentage of disease reduction on all tested potato cultivars. This isolate reduced disease incidence 60.60%, 63.21% and 71.66%, compering to the healthy control treatment. The result of molecular identification represent that the probe used in Taq-man (PCR) was of the type (B2) capable to detect only biovar 2 of R. solanacearum bacterial wilt. Furthermore, primer and probe are specific for detection of the race 3 biovar 2 strain. Positive results were obtained in all assays used including IFAS, protein content and SDS-PAGE with all five isolates. So the isolate (RS5) was the most virulence one, followed by RS1, RS3, RS2 and RS8, registered that the tested isolates were R. solanacearum race 3, biovar 2. Also, studies focused on the form of genetic distances and similarities based on pathogenic and plant growth parameters. The results illustrate that the highest genetic similarity (0.998) was found between Bellini and Spunta cultivars as the closest but the lowest value (0.946) was found between Mondial and Bellini as most distant. These results were similarity with genetic distances and SDS-PAGE profile of the three tested potato cultivars.