Identification of somatic hybrids of dihaploid Solanum tuberosum lines and S. brevidens by species specific RAPD patterns and assessment of disease resistance of the hybrids

Summary Symmetric somatic hybrids were produced by electrofusion of protoplasts of two dihaploid tuber-bearing potato (Solanum tuberosum L.) lines and Solanum brevidens Phil., a diploid non-tuber-bearing wild potato species. A total of 985 plants was obtained. Verification of nuclear hybridity of putative hybrids was based on additive RAPD patterns, general morphological characteristics and chromosome counts. 53 (90%) calli regenerated into plants which were identified as somatic hybrids. Most of the hybrids were aneuploids at the tetraploid (4×) or hexaploid (6×) level. The 20 hybrids tested expressed a high level of resistance to potato virus Y (PVY ^N ) characteristic of the S. brevidens parent. Resistance to late blight (Phytophthora infestans (Mont.) de Bary) varied between hybrids, but was on average better than that of the fusion parents. Resistance of hybrids to bacterial stem rot (Erwinia carotovora subsp. atroseptica (van Hall) Dye) was not superior to that of commercial potato cultivars.     

Phytotoxin produced by the netted scab pathogen, <Emphasis Type="Italic">Streptomyces turgidiscabies</Emphasis> strain 65, isolated in Sweden

Streptomyces spp. are a highly diverse group of bacteria most of which are soil-inhabiting saprophytes. A few are plant pathogens that produce a family of phytotoxins called thaxtomins and cause significant economic losses, e.g., by reducing the marketability of potato tubers (Solanum tuberosum). In northern Europe, S. scabies, S. turgidiscabies and S. europaeiscabiei are the most common plant pathogenic species. In this study, a Streptomyces strain isolated from a netted scab lesion on a tuber of potato cv. Bintje in northern Sweden was identified as S. turgidiscabies but was found to differ in the genomic region carrying genes required for thaxtomin biosynthesis. Our results showed that the strain did not produce thaxtomin but rather phytotoxin fridamycin E, which is an anthraquinone novel to plant pathogenic Streptomyces spp. Fridamycin E was shown to reduce or inhibit sprouting of potato microtubers in vitro. While fridamycin E is known to have antibiotic activity against Gram-positive bacteria, the inhibitory activity of fridamycin E on plant growth is a novel finding.     

Enhanced production of dihaploid lines via anther culture of tetraploid potato (Solanum tuberosum L. ssp.tuberosum) clones

A total of 1000 anther-derived plants was regenerated from tetraploid potato (Solanum tuberosum L.) genotypes. Capacity to undergo androgenesis was analysed in 41 potato cultivars and 7 clones grown either in the greenhouse or in the field. Of the 48 genotypes, 33 produced embryos and 23 regenerated shoots from embryos. Anther-derived plantlet production was determined in genotypes 86110, Agria, Calgary, Escort, Helios, Idole, JO 0982 JO 1432, Kainuun Musta, Kardal, KE48, Matilda, Nicola, Petra, Pito, Rustica, Stirling, Torridon, Ute, Van Gogh, Vebeca, Vento and White Lady. The highest number of shoots (24 shoots/100 anthers) was obtained from cv. Calgary, when anthers were isolated from field-grown donor plants. Incubating anthers at 28 C, rather than at 20 C or 24 C, enhanced embryo production in four genotypes tested. However, shoot production was improved only in cv. Pito cultured at 28 C. When anthers of cv. Petra were cultured at 28 C for four weeks, followed by reduction of culture temperature to 24 C, a high rate of shoot production was recorded (14 shoots/100 anthers). The ratio between dihaploids and tetraploids varied among the anther-derived plants of the different genotypes. The number of dihaploids was highest in potato clone JO1432 (100%) and in cv. Calgary (93%) and lowest in cvs. Pito (21%) and Torridon (6%).     

Production and characterization of “second generation” somatic hybrids derived from protoplast fusion between interspecific somatohaploid and dihaploidSolanum tuberosum L.

Protoplasts were fused to produce somatic hybrids between a triploid (2n=3x=32-34) interspecific somatohaploid betweenSolanum brevidens Phil. andS. tuberosum L., and a dihaploid (2n=2x=24) anther-derived line ofS. tuberosum cv. Van Gogh. A total of 265 plants were regenerated from protoplast fusion derived calli and their hybridity was verified using fusion partner specific RAPD markers. These “second generation” somatic hybrids were aneuploid pentaploids (2n=5x=51-65) with a 2C DNA content ranging from 3.36 to 4.43 pg, which corresponded to the sum of the 2C values of each of the fusion partners (somatohaploid: 2.22 pg; and the dihaploid line of cv. Van Gogh: 1.87 pg). Most of the “second generation” somatic hybrids were vigorous, but variable in morphology. They were extremely resistant to PLRV and they had tolerance to PVY infection derived from the somatohaploid fusion partner. Even though most of the “second generation” hybrids tuberized, the tuber morphology was variable and most were poorly shaped. InErwinia soft rot resistance tests, the tubers showed higher level of resistance than the tetraploidS. tuberosum cultivars, the dihaploidS. tuberosum fusion partners and the hexaploid somatic hybrids betweenS. brevidens andS. tuberosum. The “second generation” somatic hybrids were all male sterile and failed to produce berries or seeds.     

Aerosol particles in the developing world; a comparison between New Delhi in India and Beijing in China

In developing countries, aerosol particles damage the health of hundreds of millions of people. Migration from the country side to megacities increases emissions and exposure to particles. Some countries have started to limit emissions based on particulate mass, but this may increase particle number concentrations. In this study we discuss some earlier measurements carried out in the developing world and compare results from one-week measurement campaigns concerning the particle number size distribution and PM₁₀ mass concentrations in New Delhi, India and Beijing, China. Our results show that submicron particle concentrations are high in both places. The average PM₁₀ concentration was 360 μg/m³ in New Delhi and 120 μg/m³ in Beijing. The corresponding total particle number concentrations in the size range 3–800 nm were 63 000 cm^?3 and 35 000 cm^?3. Number and mass concentrations and their characteristics showed significantly different behaviour between these two locations, which stresses the importance of long-term simultaneous measurements of both quantities in different types of megacities.     

Influence of incorporated wild Solanum genomes on potato properties in terms of starch nanostructure and glycoalkaloid content

Interspecific somatic hybrids produced by protoplast fusion between two wild Solanum species (S. acaule, acl; S. brevidens, brd) and cultivated potato Solanum tuberosum (tbr) were analyzed in terms of the starch nanometer-range structure and glycoalkaloid (GA) contents. The crystallinity of starch granules, the average size of starch crystallites, and the lamellar distances were obtained from tuber samples using wide-angle and small-angle X-ray scattering methods. These measurements showed that incorporation of wild genomes from either nontuberous (brd) or tuberous (acl) Solanum species caused no significant modifications of the nanostructure of potato starch. In contrast, the GA profiles of the hybrids, which were analyzed by LC-ESI-MS in both tuber and foliage samples, differed considerably from those of cultivated potato. Regardless of the low total tuber GA concentrations (approximately 9 mg/100 g of fresh weight), the somatic hybrids contained GAs not detected in the parental species. A high proportion of spirotype GAs consisting of 5,6-dihydrogenated aglycons, for example, alpha-tomatine and tomatidine bound with solatriose, and chacotriose were found in the hybrids. In conclusion, the foliage of interspecific hybrids contained a higher variation in the structures of GAs than did the tubers.     

Examination of the leaf-drop symptom of virus-infected potato using anther culture-derived haploids

ABSTRACT Necrotic lesions and vein necrosis characteristic of the hypersensitive response (HR) controlled by the dominant resistance gene Ny develop in potato cv. Pito after infection with potato virus Y ordinary strain (PVY masculine) at a low temperature (16/18 degrees C night/day). In contrast, at high temperatures (19/24 degrees C night/day), large coalesced lesions develop in the lower infected leaves, which wither and remain hanging from stems forming the leaf-drop symptom; mosaic symptoms with no necrosis also develop in the top leaves. The genetic basis of the leaf-drop symptom and its dependence on temperature were examined using a novel approach involving 58 haploids (2n = 24) derived from 'Pito' (2n = 48) through anther culture. These haploids and 'Pito' were graft-inoculated with PVY(O) at 19/24 to 25 degrees C (night/day). Necrotic symptoms were expressed in 28 haploids, of which 18 haploids (phenotype class N) developed top necrosis, vein necrosis, or both and necrotic lesions that are characteristic of HR. Ten haploids showed leaf drop similar to 'Pito' (phenotype class LD). Thirty haploids were susceptible and showed only mosaic symptoms (phenotype class S). These data indicated that necrosis was induced by a single dominant gene, Ny, in the simplex condition. However, the three distinct phenotypic classes (N, LD, and S) among the haploids grown under the same environmental conditions showed that another locus (gene) was involved in modifying the HR triggered by Ny. Data suggested that this locus contains a dominant temperature-dependent modifier (Tdm) gene that alters the expression of PVY-induced HR at higher temperatures, resulting in leaf drop.     

Infection of plant material derived from Solanum acaule with Clavibacter michiganensis ssp. sepedonicus: temperature as a determining factor in immunity of S. acaule to bacterial ring rot

The development of potato ( Solanum tuberosum ) cultivars immune (completely free of infective agents) to bacterial ring rot disease, caused by Clavibacter michiganensis ssp. sepedonicus , would be a significant step toward eradication of the disease. Earlier results suggested that the wild potato species Solanum acaule , acc. PI472655 (genotype 7-8), was immune to C. michiganensis ssp. sepedonicus . The goal of the present study was to investigate the possibility of transferring this desirable trait to cultivated potato through interspecific somatic hybridization. Eight different somatic hybrids between S. acaule and S. tuberosum , with three different genome ratios, were tested for susceptibility to infection in the glasshouse using two C. michiganensis ssp. sepedonicus strains. All the somatic hybrids expressed symptoms of ring rot and were susceptible to infection as determined by IFAS (indirect immunofluorescent antibody staining) tests. The genome compositions of the hybrids influenced bacterial titres. Most of the hybrids with a higher proportion of the S. acaule genome contained lower numbers of C. michiganensis ssp. sepedonicus cells than hybrids with lower proportions of the S. acaule genome. In growth chamber tests, temperature was found to be a determining factor in the expression of immunity in the tetraploid S . acaule 7-8 line. At 21°C, S . acaule 7-8 was immune to infection by C. michiganensis ssp. sepedonicus , but at 15°C, plants supported large populations of the pathogen. However, none of the S. acaule plants expressed disease symptoms. Thus, S. acaule exhibits temperature-dependent immunity to infection by C. michiganensis ssp. sepedonicus and should be considered tolerant rather than immune.     

Potato Shoot Tip Cryopreservation. A Review

Potato is one of the most important crops worldwide. Genetic resources of potato (Solanum tuberosum L. ssp. tuberosum) and related cultivated species are conserved through storage of tubers, in vitro plants and in cryopreservation. Cryopreservation, storage in or above liquid nitrogen, is the best option to maintain vegetatively propagated plants in the long term. The present review gives comprehensive information about various cryopreservation techniques for potato published from 1977 until the present. It discusses factors that affect the process and success of cryopreservation, such as donor culture conditions, preculture, cooling, warming and post-culture treatments. Studies are presented that analyse the histological and ultrastructural changes after different cryopreservation steps and the morphological pathways during regeneration of plants after rewarming. The maintenance of genetic stability in potato after cryopreservation has also been demonstrated by various phenotypic and molecular methods. The first thermal analyses on potato shoot tips are presented using differential scanning calorimetry to analyse the state of water during cooling and warming. Biochemical analyses of different compounds, such as soluble sugars and proteins, have been performed to understand and improve existing cryogenic methods. Potato is an example where successful virus elimination has been obtained via cryopreservation of shoot tips (cryotherapy). There are already cryopreserved collections of potato shoot tips in Germany, Peru, Czech Republic, South Korea and USA, but additional experiments on fundamental aspects of potato cryopreservation will help to improve understanding of the different cryopreservation methods, start new collections in other countries and also build up existing cryocollections of potato.     

Competitiveness of mechanized tree planting in Finland

Economic pressures and labor shortage are forcing forest owners to minimize silvicultural costs and manage their forests more intensively to enhance wood production and profitability. The need to improve the cost-efficiency of tree planting is spurring its mechanization. The cost-competitiveness and time consumption of mechanized tree planting in Finland were compared to manual planting (MP) in spot mounds formed with a mounding blade and with a continuously advancing spot mounder. The results suggest that mechanized planting must increase its current productivity by 25% and 100% in order to compete with spot mounding or continuously advancing mounder followed by MP, respectively. However, in the hands of skilled operators in optimal conditions, machines can be cost-effective. On average, mechanized planting required 20% less time than MP and excavator-based spot mounding, whereas MP and continuously advancing spot mounder required 30% less time. Effective use of modern machines requires a careful evaluation of the worksite and skilled operators applying optimal operational models.