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1.
Globodera pallida is a parasitic root cyst nematode of potato, which causes reduction of crop yield and quality in infested fields. Field populations of G. pallida containing mixtures of pathotypes Pa2 and Pa3 (Pa2/3) are currently most relevant for potato cultivation in middle Europe. Genes for resistance to G. pallida have been introgressed into the cultivated potato gene pool from the wild, tuber bearing Solanum species S. spegazzinii and S. vernei. Selection of resistant genotypes in breeding programs is hampered by the fact that the phenotypic evaluation of resistance to G. pallida is time consuming, costly and often ambiguous. DNA-based markers diagnostic for resistance to G. pallida would facilitate the development of resistant varieties. A tetraploid F1 hybrid family SR-Gpa segregating for quantitative resistance to G.␣pallida was developed and evaluated for resistance to G. pallida population ‘Chavornay’. Two subpopulations of 30 highly resistant and 30 susceptible individuals were selected and genotyped for 96 single nucleotide polymorphism (SNP) markers tagging 12 genomic regions on 10 potato chromosomes. Seven SNPs were found significantly linked to the nematode resistance, which were all located within a resistance ‘hotspot’ on potato chromosome V. A haplotype model for these seven SNPs was deduced from the SNP patterns observed in the SR-Gpa family. A PCR assay ‘HC’ was developed, which specifically detected the SNP haplotype c that was linked with high levels of nematode resistance. The HC marker was only found in accessions of S.␣vernei. Screening with the HC marker 34 potato varieties resistant to G. pallida pathotypes Pa2 and/or Pa3 and 22 susceptible varieties demonstrated that the HC marker was highly diagnostic for presence of high levels of resistance to G. pallida pathotype Pa2/Pa3.Amirali Sattarzadeh and Ute Achenbach contributed equally to the work  相似文献   

2.
Fourteen somatic hybrids generated by electrofusion of mesophyll protoplasts from a non-flowering dihaploid S. tuberosum clone, DHAK-11, and a male-sterile dihaploid clone S. tuberosum, DHAK-33, were grown in the greenhouse and subjected to morphological assessments and tests for fertility and resistance to the white potato cyst nematode Globodera pallida pathotypes Pa2 and Pa3. The ploidy level of the hybrids ranged from 38 to 63 chromosomes. All hybrids developed flowers with violet petals except for one, hy-56, that possessed red petals. The colour of the tuber skin was purple in all hybrids except in hy-56 where the tuber skin was red. All of the hybrids were female fertile and generated viable seeds. Near-tetraploid hybrids produced the highest number of seeds per fruit and these seeds had a normal size. Hybrids with 58 or more chromosomes produced smaller seeds and less seeds per fruit. The germination frequency of the seeds was not influenced by the chromosome number of the hybrids. Pollen viability was determined and the male fertility of three hybrids was tested. Pollination with these three hybrids gave rise to fruit development, but only one produced viable seeds. The hybrids were tested for resistance to G. pallida pathotypes Pa2 and Pa3. A high level of resistance to Pa3, inherited from one parental clone, DHAK-11, and a high level of resistance to Pa2, inherited from the other parental clone, DHAK-33, was combined in four hybrids. These results demonstrate, that protoplast fusion is an efficient method for restoring the fertility of somatic hybrids generated from sterile parent clones, and is a powerful procedure for the complementation of multigenetic disease resistance traits in potato breeding lines.  相似文献   

3.
Using electrofusion of protoplasts from different dihaploid potato breeding clones, 308 somatic hybrids, i.e. 16.7% of the totally regenerated plants, were obtained. The results indicate that factors such as ‘combining ability’ strongly influence hybrid formation in these intraspecific fusion experiments. Hybrid identification was predominantly carried out by isoenzyme analysis of the peroxidases and/or esterases in polyacrylamide-gradient gel electrophoresis. Further confirmation of the hybrid character was obtained by scoring phenotypic markers (petal colour, stem anthocyanin production, tuber characteristics, etc.) and, in some combinations, by evaluating tolerance against the herbicide metribuzin and nematode resistance against Globodera rostochiensis (Ro 1). The successful inheritance of metribuzin tolerance and nematode resistance confirm that monogenic dominant inherited traits are combined in the hybrids.  相似文献   

4.
A Cytosolic Phospholipase A2 from Potato Tissues Appears to Be Patatin   总被引:3,自引:0,他引:3  
Phospholipase (PL) A2 is involved in signal transduction inthe resistance reaction that is induced in potato by inoculationof an incompatible race of Phytophthora infestans, the lateblight fungus, or by treatment with fungal elicitor hyphal wallcomponents (Kawakita et al. 1993). In this study, PLA2 in thesoluble fraction from potato tuber was purified. The followingresults suggested that the enzyme was, in fact, patatin: (1)the molecular mass of the purified enzyme was 40 kDa, the sameas that of patatin; (2) the pI of the purified enzyme was approximately4.75, which corresponds to that of patatin; and (3) the amino-terminalamino acid sequence of the purified enzyme showed a high degreeof homology to that of patatin. Patatin is known as a storageprotein of the potato tuber and it has been shown to have esteraseactivity. However, other enzymatic activities and the function(s)of patatin are unknown. We investigated the PLA activities ofthe purified patatin. The PLA2 activity of the patatin was muchhigher than the PLA1 activity, even though the protein exhibitedboth activities. The PLA2 activity of the enzyme was particularlyapparent when phosphatidylcholine with linoleic acid at thesn-2 position was used as substrate. Lower activity was observedwith phosphatidylcholine with palmitic acid, oleic acid andarachidonic acid at the sn-2 position. (Received October 5, 1995; Accepted February 9, 1996)  相似文献   

5.
Summary Following both chemical and electrical fusion of protoplasts of a dihaploid line of potato (Solanum tuberosum), (PDH40), with those of the wild species, Solanum brevidens, 11 and 40 somatic hybrid plants, respectively were obtained. Fifteen of these somatic hybrid genotypes and the two parents were studied further in a small field trial to assess field performance and phenotypic variability. In the UK, somatic hybrid plants are classified as genetically engineered organisms, and the UK Advisory Committee on Genetic Manipulation have imposed various restrictions on field experiments. Examination of the somatic hybrids in the field showed extensive phenotypic variability, and no two genotypes were identical. Some of the variation reflected changes in chromosome numbers, but other factors were also involved. Half the somatic hybrid genotypes produced tubers in the field, although the tubers were smaller and differed morphologically from those of PDH40. The results of the study suggest that the extent of somaclonal variation manifested in somatic hybrids is greater than that found in protoplast-derived plants of potato. The implications of this observation and the current regulations concerning field experiments of somatic hybrid plants in the UK are discussed.  相似文献   

6.
Seven trials conducted over four years on sites naturally infested with the white potato cyst nematode established that potato clones bred for resistance to Globodera pallida allowed significantly less nematode multiplication than conventional cultivars under field conditions. Nematode multiplication was inversely related to initial infestation level. The nematicide, aldicarb, significantly reduced nematode multiplication. However, nematode multiplication on nematicide treated susceptible cultivars was greater than on untreated partially resistant clones, indicating that resistance may offer more effective control of G. pallida than chemical treatment. Integration of host plant resistance and nematicide treatment is discussed.  相似文献   

7.
The effects of oxamyl applied to the seedbed and growing partially resistant potatoes in controlling potato pale cyst-nematode, Globodera pallida, were assessed in eleven field experiments on sandy, peaty and silty loam soils in England from 1986 to 1988. Standardised procedures allowed valid comparisons to be made between data from the three experiments in 1987 and from the seven experiments in 1988. In soil moderately or heavily infested with G. pallida, oxamyl frequently increased tuber yields of susceptible cv. Désirée and of partially resistant potatoes and lessened crude nematode increase (Pf/Pi) significantly at some sites but not at others. Although 2.8 kg oxamyl ha“1 often increased tuber yields significantly, it was significantly less effective than 5.6 kg in controlling G. pallida at one site and at another site G. pallida increased more than in untreated plots. Potatoes partially resistant to G. pallida were more effective that oxamyl in controlling nematode increase at some sites but not at others. Combined use of oxamyl and partially resistant potatoes was generally more effective in controlling G. pallida than either measure alone. As measured by a ‘control coefficient’ (weight of tubers over 40mm diameter (t ha“1) 4-crude nematode increase (Pf/Pi)), the most effective integrated control of G. pallida was obtained by growing cvs Glenna, Morag or Santé in soil treated with 5.6 kg oxamyl ha-1.  相似文献   

8.
Recent studies have shown that living and heat-killed cells of the rhizobacterium Rhizobium etli strain G12 induce in potato roots systemic resistance to infection by the potato cyst nematode Globodera pallida. To better understand the mechanisms of induced resistance, we focused on identifying the inducing agent. Since heat-stable bacterial surface carbohydrates such as exopolysaccharides (EPS) and lipopolysaccharides (LPS) are essential for recognition in the symbiotic interaction between Rhizobium and legumes, their role in the R. etli-potato interaction was studied. EPS and LPS were extracted from bacterial cultures, applied to potato roots, and tested for activity as an inducer of plant resistance to the plant-parasitic nematode. Whereas EPS did not affect G. pallida infection, LPS reduced nematode infection significantly in concentrations as low as 1 and 0.1 mg ml−1. Split-root experiments, guaranteeing a spatial separation of inducing agent and challenging pathogen, showed that soil treatments of one half of the root system with LPS resulted in a highly significant (up to 37%) systemic induced reduction of G. pallida infection of potato roots in the other half. The results clearly showed that LPS of R. etli G12 act as the inducing agent of systemic resistance in potato roots.  相似文献   

9.
Interspecific somatic hybrids between the dihaploid Solanum tuberosum and the wild species S. pinnatisectum Dun. were produced via protoplast fusion. Protoplast isolation, electrofusion, culture of post-fusion products and regeneration of calli/shoots were undertaken following optimized protocols. Regenerants were characterized for hybridity, ploidy and resistance to Phytophthora infestans (Mont.) de Bery, causal fungal pathogen of late blight disease. From a total of 126 regenerated macrocalli, 12 somatic hybrids were confirmed by possessing species-specific diagnostic bands of their corresponding parents as revealed by RAPD, SSRs and cytoplasmic-DNA analyses. Tetraploid status of the 12 hybrids was determined using flow cytometry analysis. Intermediate phenotypes for leaf, flower, and tuber characteristics and high male fertility were observed in field-grown hybrid plants. Hybrids were highly resistant to foliage late blight based on field assessment for two seasons. In contrast, moderate level of resistance to foliage blight was observed in hybrids based on the detached leaf assay under laboratory conditions. Overall, somatic hybrids with moderate levels of resistance to foliage blight were identified, and these will be useful for in situ hybridization in potato breeding efforts.  相似文献   

10.
Laboratory, pot and field experiments investigated the effects of the fungus Zygorrhynchus moelleri on the growth of potato and on the reproduction of the potato cyst nematodes (PCN), Globodera pallida and G rostochiensis. Preliminary laboratory tests showed that Z. moelleri growth was favoured by temperatures and pH ranges commonly present in field soils. The fungus colonised potato roots in vitro and in compost or field soil. It also stimulated in vitro root growth of three potato cultivars. In pot experiments Z. moelleri stimulated potato growth, particularly in the presence of PCN attack. In field plots infested with a mixture of G pallida and G. rostochiensis, tuber yields were not increased after application of the fungus but, in G pallida‐infested plots, yields were significantly increased after drills were inoculated with Z. moelleri. The application of Z. moelleri had no apparent effects on nematode reproduction. Factors influencing the interactions between Z. moelleri, potato and potato cyst nematodes are discussed and the potential role of the fungus as a plant growth promoter in organic potato production considered.  相似文献   

11.
Six potato cultivars with different levels of resistance to the white potato cyst nematode (PCN) Globodera pallida Pa2 were grown for three seasons in field plots to which G. pallida Pa2 cysts had been introduced earlier. There were two planting times, corresponding to early and maincrop commercial planting times, and two initial PCN population densities, high and low. The effect of cultivar on PCN population density was far greater than the effect of planting time or initial nematode population. The final PCN populations for the cultivars Ilam Hardy, Wha, 4696A(2), Sovereign, D40/6 and V390 were 151, 74, 27, 1.4, 0.2 and 0.06 eggs per g of soil respectively. It is concluded that resistant potato cultivars can be very effective in controlling G. pallida Pa2 in the field.  相似文献   

12.
Differential expression of potato tuber protein genes   总被引:2,自引:1,他引:1       下载免费PDF全文
  相似文献   

13.
Non-resistant but tolerant cv. Cara and non-resistant but relatively intolerant cv. Pentland Dell were grown in split plots encompassing a range of population densities of potato cyst nematode, Globodera pallida. Light interception and its efficiency of conversion were estimated by regular ground cover measurements and plant harvests. It was concluded that increasing levels of infestation with G. pallida only slightly decreased the efficiency of utilisation of intercepted radiation. Heavy infestation of G. pallida initially decreased the top growth and light interception of both cultivars by similar proportions, but in later harvests, this adverse effect markedly decreased for Cara whereas it slightly increased for Pentland Dell. This difference was due to the heavily infested Cara eventually achieving and maintaining 100% ground cover whereas the equivalent Pentland Dell never exceeded 75% ground cover. Consequently, final tuber yields were decreased much more for Pentland Dell than for Cara though the decreases in tuber yield were less than those for top growth. The importance of nematode effects on top growth, and hence on light interception, with regard to both yield losses and tolerance differences, were clearly demonstrated. Both linear and logarithmic models were used to describe the relationship between the initial population density of G. pallida and yield, and the implications of differences in tolerance on the parameters in the logarithmic model are discussed.  相似文献   

14.
Three field experiments were made to determine the effectiveness of small-plot trials in detecting differences between potato cultivars/clones in their tolerance of damage by potato cyst-nematodes. A nematicide (aldicarb) was applied at three rates to decrease nematode damage. The largest rate of aldicarb increased tuber yields most but the relationship between yield response and nematicide rate was not linear. The yield increases of the cultivars and clones differed, indicating that they have different degrees of tolerance of potato cyst nematodes. The results were analysed in several ways and the untreated yield as a proportion of the treated provided the best means of expressing and comparing tolerance; but whichever method was used the tolerance rankings of the cultivars and clones were similar. At two sites infested with Globodera rostochiensis, the rankings of the 10 cultivars and clones were similar but at a third site, heavily infested with G. pallida, they were different. Aldicarb decreased the nematode population density after harvest at the G. pallida site but was less effective at the G. rostochiensis sites, which were less heavily infested. Growing resistant or partially resistant potatoes usually prevented nematode increase, and the more resistant cultivars and clones decreased population densities markedly.  相似文献   

15.
The potato cyst nematode, Globodera pallida, is one of the most important pests of potato worldwide. Owing to regulatory considerations and potential environmental impact, control options for this nematode are becoming increasingly limited. Solanum sisymbriifolium and biological control agents offer viable alternative options for controlling G. pallida. Therefore, experiments were conducted to determine the effect of the nematode trap crop S. sisymbriifolium, alone or in combination with the biocontrol agents Trichoderma harzianum or Plectosphaerella cucumerina, on population decline of G. pallida. Experiments were conducted for three different ‘cropping systems’: potato (Solanum tuberosum), S. sisymbriifolium, or soil only (fallow), each followed by a potato crop. Soil was amended with P. cucumerina, T. harzianum or left unamended, and then infested with nematodes at a rate of five eggs g?1 of soil. After 16 weeks in the greenhouse, plants were removed and the soil containing cysts was refrigerated at 4°C for 8 weeks, and then planted to potato. Cysts of G. pallida were counted after an additional 16‐week period. The Pf/Pi of G. pallida was significantly reduced by 99% in potato following S. sisymbriifolium compared to both the potato‐following‐fallow and the potato‐following‐potato treatments. Amendment of soil with T. harzianum significantly reduced Pf/Pi of G. pallida by 42–47% in the potato‐following‐potato but not in either the potato‐after‐fallow nor in the potato‐after‐S. sisymbriifolium cycles which supports evidence that the plant species may play a role in the biocontrol activity of this fungus. Addition of the fungus P. cucumerina resulted in a 64% decrease in Pf/Pi in the potato‐following‐fallow in one experiment, and an 88% decrease in Pf/Pi in potato‐following‐potato but the decrease in Pf/Pi was not consistent over all experiments. However, both biocontrol fungi resulted in lower numbers of progeny cysts after an initial 16‐week incubation with potato. To look at the effect of varied population density of the nematode on efficacy of S. sisymbriifolium to reduce G. pallida populations, potato, S. sisymbriifolium, or barley were planted into soil infested with G. pallida at rates of 5, 20 or 40 eggs g?1 soil applied as cysts (20, 80 or 160 cysts pot?1). After 16 weeks, numbers of cysts produced in each treatment were determined for each infestation rate. No new cysts were recovered from either S. sisymbriifolium or barley treatments, confirming that neither plant is a host for G. pallida. High numbers of cysts were recovered with potato. Soil from each treatment (containing original cysts and newly‐formed cysts when present) were then planted with potato. After an additional 16 weeks, few cysts were found in the potato‐after‐ S. sisymbriifolium treatments regardless of initial infestation rate. When potato followed barley, numbers of cysts were similar to those found after a single cycle of potato, indicating that the barley crop had no effect on the survival of initial inoculum. Overall, these results suggest that S. sisymbriifolium has potential to significantly reduce G. pallida populations, and also that the cropping system (i.e. the sequence of non‐host and host plants) may play a significant role in the efficacy of fungal biological control agents.  相似文献   

16.
The damage caused by the parasitic root cyst nematode Globodera pallida is a major yield-limiting factor in potato cultivation . Breeding for resistance is facilitated by the PCR-based marker ‘HC’, which is diagnostic for an allele conferring high resistance against G. pallida pathotype Pa2/3 that has been introgressed from the wild potato species Solanum vernei into the Solanum tuberosum tetraploid breeding pool. The major quantitative trait locus (QTL) controlling this nematode resistance maps on potato chromosome V in a hot spot for resistance to various pathogens including nematodes and the oomycete Phytophthora infestans. An unstructured sample of 79 tetraploid, highly heterozygous varieties and breeding clones was selected based on presence (41 genotypes) or absence (38 genotypes) of the HC marker. Testing the clones for resistance to G. pallida confirmed the diagnostic power of the HC marker. The 79 individuals were genotyped for 100 single nucleotide polymorphisms (SNPs) at 10 loci distributed over 38 cM on chromosome V. Forty-five SNPs at six loci spanning 2 cM in the interval between markers GP21-GP179 were associated with resistance to G. pallida. Based on linkage disequilibrium (LD) between SNP markers, six LD groups comprising between 2 and 18 SNPs were identified. The LD groups indicated the existence of multiple alleles at a single resistance locus or at several, physically linked resistance loci. LD group C comprising 18 SNPs corresponded to the ‘HC’ marker. LD group E included 16 SNPs and showed an association peak, which positioned one nematode resistance locus physically close to the R1 gene family. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.  相似文献   

17.
Globodera rostochiensis and G. pallida responded similarly to hatch stimulation by potato root leachate, but proportionally more second-stage juveniles (J2s) of G. rostochiensis hatched than of G. pallida in response to picrolonic acid, sodium thiocyanate, alpha-solanine, and alpha-chaconine. Fractionation of the potato root leachate identified hatching factors with species-selective (active toward both species but stimulating greater hatch of one species than the other), -specific (active toward only one species), and -neutral (equally active toward both species) activities. In a comparison of two populations of each of the two potato cyst nematode (PCN) species, however, greater similarity in response to the individual hatching factors was observed among populations of different species produced under the same conditions than among different populations of the same PCN species. Smaller numbers of species-specific and species-selective hatching factor stimulants and hatching inhibitors than of hatching factors were resolved. In a study to determine whether the different hatching responses of the two species to the same root leachate were associated with different ratios of species-selective and species-specific hatching factors, G. rostochiensis pathotype Ro1 exhibited greater hatch than did G. pallida pathotype Pa2/3 in response to leachate from older plants (more than 38 days old), while G. pallida exhibited greater hatch in response to leachate from younger plants (less than 38 days old); the response of G. pallida pathotype Pal with respect to plant age was intermediate between the other two populations. Combined molecular exclusion-ion exchange chromatography of the root leachates from plants of different ages revealed an increase in the proportion of G. rostochiensis-specific and -selective hatching factors as the plants aged.  相似文献   

18.
Using standard hybridoma technology and hierarchical screening, monoclonal antibodies (MAbs) were obtained with specific reactivity against two developmental stages of Globodera pallida. The procedure was based on enzyme-linked immunosorbent assay (ELISA) with homogenates prepared from second-stage juveniles, young adult females, and potato roots. Hybridomas were formed by fusing myelomas with splenocytes derived from mice immunized with either infective juveniles or females of G. pallida. About 600 hybridoma lines were screened from the fusion involving the mouse immunized with juveniles. Two MAbs (LJMAbl &2) were identified with high reactivity toward second-stage juveniles but no reactivity with either potato roots or females of G. pallida. A total of 630 cell lines was screened from the corresponding fusion involving the spleen of a mouse receiving immunogens from adult female nematodes. One MAb (LFMAbl) was obtained with the required specificity against only adult female G. pallida. This work extends the application of monoclonal antibodies in nematology from valuable probes for research and species identification to recognition of developmental stages. These specific MAbs have potential value in plant breeding programs for screening for resistant lines unable to support nematode development.  相似文献   

19.
A new technique is described for establishing different numbers of the potato cyst-nematode Globodera rostochiensis in field soil, which leaves the soil homogeneous in nutrient status. Field plots established in this way were used to compare yield losses in four potato cultivars (Maris Piper, Pentland Crown, Pentland Dell and Désirée) associated with different numbers of G. rostochiensis. Over the range of 7.4 to 148.4 eggs g-1 soil at planting, yield losses were 18.7% (Maris Piper), 53.2% (Désirée), 55.7% (Pentland Crown) and 63.5% (Pentland Dell). Similar results were obtained in another experiment on the same field in a different year using only lightly and heavily infested plots. Treating the seedbed soil with oxamyl before planting prevented significant injury to potatoes by G. rostochiensis but increased the yield of Pentland Dell and perhaps Désirée (but not Maris Piper or Pentland Crown) more than expected from nematode control alone. Treating heavily infested soil with such a nematicide cannot therefore be recommended as part of a valid procedure for establishing lightly and heavily infested plots for comparing tolerances of attack by potato cyst-nematodes in a range of potato genotypes. In peaty loam soils moderately or heavily infested with G. pallida, oxamyl at 5.6 kg a.i. ha-1 incorporated into the seedbed before potatoes were planted generally increased tuber yields, though the effects varied considerably with the cultivar grown. Increase of G. pallida in these soils was controlled better by growing potatoes bred for resistance to it (ZB 35 – 29, Caxton, Santé, Morag, 11233 ab 22, Fingal, A27/23, Cromwell). Increase of G. pallida on susceptible cultivars varied greatly and Romano increased G. pallida no more than the resistant Morag. G. pallida is probably controlled best in peaty loam by growing a resistant cultivar in soil treated with a granular (non-fumigant) nematicide.  相似文献   

20.
Changes in relative abundance of the two potato cyst nematode species Globodera rostochiensis and Globodera pallida were studied during the 1983/84 season at two different population levels in small pots in the glasshouse and at a single population density on plants grown outdoors in 2 litre terylene cloth bags. In both environments G. rostochiensis was the more successful species. Although the ratio of the two species changed and G. pallida was at a lower level at the end of the experiment it was never eliminated. However, when the number of G. pallida in the mixture was small it did better than expected and demonstrated a frequency dependent response.  相似文献   

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