The Prospects of Tissue Culture and Genetic Engineering for Strawberry Improvement

Abstract

Strawberry shoot meristem cultures have been used for producing virus-free strawberry plants. Plantlet regneration from leaf mesophyll protoplasts, and Agrobacterium tumefaciens mediated genetic transformation, mark the beginning of strawberry improvement with biotechnologies. The use of biotechnological tools has a greater potential in producing strawberry plants with traits such as early maturation, frost and disease resistance, mite and nematode resistance, high yield, better quality, suitability for mechanical harvesting, shipping ability, ellgaic acid content, aroma compounds and cost-effective micropropagation. Gene identification and mapping with RFLPs or RAPDs would be desirable. Transgenic strawberries should be tested for their quality before releasing to the consumers.     

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.     

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.     

Comparisons of terrestrial and aquatic bioassays for oil-contaminated soil toxicity

Background  Petroleum products are widely used in various sections of industry and they are one of the most abundant sources of environmental contamination. These products are classified by their physico-chemical properties such as boiling point, density and viscosity. Oil contamination in the environment is primarily evaluated by measuring the chemical concentrations of petroleum products in the solid or water phase. The results of chemical analyses do not correspond directly with the harmful environmental effects of petroleum products on the soil flora and fauna, because the interactions between oil compounds and the production of their methabolites in soil are not measured in chemical assessments. These kinds of effects of complex chemical mixtures in soil can be estimated by bioassays. Therefore, ecotoxicological tests are important for estimating soil quality in the risk assessment of oil-contaminated soil sites. Objectives  The objective of this study was to examine the oil-contaminated soil site of a closed petrol station with both chemical and ecotoxicological methods. The goals of this study were to compare the sensitivity of the terrestrial and aquatic bioassays and to compare the toxicity responses of aquatic bioassays determined from three different extraction procedures. In addition, our aim was to characterise a cost-effective battery of bioassays that could be applied to a comparison of oil-contaminated soils. It was in our interest to investigate oil-contaminated soil with oil concentrations of 2500–12000 mg/kg and to find out the possible differences between terrestrial and aquatic toxicity tests. Methods  Six soil samples from a closed petrol station were examined for toxicity with terrestrial and aquatic tests. Terrestrial tests includedEnchytraeus albidus survival and reproduction assays and seed germination assays using wheat, cress, lettuce, and red clover seeds and growth inhibition assays of onions. The toxicities of the water-extractable fractions of the soil samples obtained from three different extractions were tested with aquatic bioassays based on plants (onion and duckweed growth inhibition tests), microbes (luminescent bacteria test), and enzyme inhibition (reverse electron transport test, RET). Chemical analyses of the solid samples were carried out simultaneously. Results. Oil concentrations ranged from 2500 to 12000 mg/kg, BTEX varied from 300 to 2800 mg/kg, and fuel additives: MTBE and TAME from 0.0 1 to 260 mg/kg. Only the sample contain-ing 12000 mg/kg oil had a significant toxic impact on all test organisms. Soil samples with oil concentrations 2500–6200 mg/ kg had no or only slight adverse effects on the test organisms with one exception, theE. albidus reproduction test. TheE. albidus survival and reproduction tests were the most sensitive bioassays of the terrestrial tests, and the luminescent bacteria test of the aquatic tests.     

Pancreatic Degenerative Atrophy and Chronic Pancreatitis in Dogs a Comparative Study of 60 Cases

During the years 1977–1980 60 cases of non-neoplastic chronic exocrine pancreatic disease in dogs were investigated clinically and pathologically. The disorders were clinically divided into pancreatic degenerative atrophy (PDA) and chronic pancreatitis. Fifty dogs had PDA and 45 of them were German shepherd dogs. The PDA cases formed both clinically and pathologically a homogeneous group except for 1 case. All the dogs had maldigestion and protease activity was absent from the faeces. General inanition and highly atrophic pancreas were the most typical macroscopic findings. Histologically the exocrine pancreas contained atypical acinar tissue and mononuclear cell infiltrations. Five of the dogs died spontaneously, 4 of them had intestinal torsion and 1 had paralytic ileus.There were 10 dogs with chronic pancreatitis. This group was rather heterogeneous both clinically and pathologically. The pancreas was slightly enlarged and the consistency was firm. The histologic picture was one of fibrous tissue proliferation and inflammatory cell infiltrations in the interstitium. The dogs nutritional state as well as faecal protease activity were normal.     

The current status of knowledge on the cellular biology of potato

Summary Conventional potato breeding does not make full use of the existing biodiversity within theSolanum genus. Moreover breeding at the 4x-4x level is slow. The potential for breeding at the diploid and dihaploid levels has therefore been explored. This requires use of novel techniques to overcome deviations from the desired Endosperm Balance Number. Somatic hybridization approaches include symmetric and asymmetric hybridizations and cybridization. Interesting traits have been successfully transferred through these techniques. Introgression and chromosome elimination have profited from the recent and rapid development of analytical techniques such as RFLP and RAPD. Cellular approaches in potato breeding may be combined with conventional breeding by a stepwise reduction of the ploidy level followed by resynthesis of a new heterozygous tetraploid clone. Such schemes have been used to include virus or nematode resistance. Haploids may be derived from different sources or obtained through different techniques. Dihaploid-dihaploid breeding programmes may be especially interesting. Because of this potential, cellular biology of potato deserves the continued interest of the scientific community.     

Using multi-source data to map and model the predisposition of forests to wind disturbance

Natural disturbances such as wind are known to cause threats to ecosystem services as well as sustainable forest ecosystem management. The objective of this research was to better understand and quantify drivers of predisposition to wind disturbance, and to model and map the probability of wind-induced forest disturbances (P_DIS) in order to support forest management planning. To accomplish this, we used open-access airborne light detection and ranging (LiDAR) data as well as multi-source National Forest Inventory (NFI) data to model P_DIS in southern Finland. A strong winter storm occurred in the study area in December 2011. High spatial resolution aerial images, acquired after the disturbance event, were used as reference data. Potential drivers associated with P_DIS were examined using a multivariate logistic regression model. The model based on LiDAR provided good agreement with detected areas susceptible to wind disturbance (73%); however, when LiDAR was combined with multi-source NFI data, the results were more promising: prediction accuracy increased to 81%. The strongest predictors in the model were mean canopy height, mean elevation, and stem volume of the main tree species (Norway spruce and Scots pine). Our results indicate that open-access LiDAR data can be used to model and map the probability of predisposition to wind disturbance, providing spatially detailed, valuable information for planning and mitigation purposes.     

Seasonal acclimation of photosystem II in Pinus sylvestris. I. Estimating the rate constants of sustained thermal energy dissipation and photochemistry

Acclimation of the partitioning of absorbed light energy in Photosystem II (PSII) between photochemical and non-photochemical processes includes short-term adjustments that are rapidly reversed in the dark and seasonal acclimation processes that are unaffected by dark acclimation. Thus, by using dark-acclimated leaves to study the seasonal acclimation of PSII, the confounding effect of short-term adjustments is eliminated. The maximum quantum yield of photochemistry, estimated by chlorophyll fluorescence analysis as F(v)/F(m), where F(v) = (F(m) - F(o)), and F(m) and F(o) are maximum and minimum chlorophyll fluorescence, respectively, has been widely used to follow the seasonal acclimation of PSII, because it is measured in dark-acclimated leaves. Seasonal changes in F(v)/F(m) can be caused by adjustments in either the photochemical capacity in PSII, or the capacity of thermal dissipation in PSII, or both. However, there is a lack of chlorophyll fluorescence parameters that can distinguish between these processes. In this study, we introduce two new parameters: the rate constants of sustained thermal energy dissipation (k(NPQ)) and of photochemistry (k(P)). We estimated k(NPQ) and k(P) from dark-acclimated F(o) and F(m) measured during spring recovery of photosynthesis in Scots pine (Pinus sylvestris L.) trees. We suggest that k(NPQ) and k(P) be used to study the mechanisms underlying the observed seasonal acclimation in PSII, because these parameters provide quantitative data that complement and extend F(v)/F(m) measurements.