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.     

Genetic diversity and mixed infections of begomoviruses infecting tomato, pepper and cucurbit crops in Nicaragua

Begomoviruses were detected in Nicaraguan fields of tomato ( Lycopersicon esculentum ) and adjacently growing plants of pepper ( Capsicum annuum ), chilli pepper ( C . baccatum ), cushaw ( Cucurbita argyrosperma ) and Mexican fireplant ( Euphorbia heterophylla ) using polymerase chain reaction (PCR) and universal begomovirus primers. All tomato and Mexican fireplant plants showing symptoms were infected with begomoviruses, while only 30–46% of the pepper, chilli pepper and cushaw plants showing symptoms tested virus-positive. No begomoviruses were found in potato. The virus species were provisionally identified by sequencing 533 bp of the viral coat protein gene ( AV1 ). Tomato severe leaf curl virus (ToSLCV), Tomato leaf curl Sinaloa virus (ToLCSinV) and Pepper golden mosaic virus (PepGMV) were found to infect both tomato and pepper. A new provisional species designated Tomato leaf curl Las Playitas virus (ToLCLPV) was detected in a tomato plant. Squash yellow mottle virus (SYMoV) and PepGMV were found in cucurbits, the latter for the first time in this host. Euphorbia mosaic virus (EuMV) was detected in Mexican fireplant. Sequencing of a larger number of PCR-amplified clones from selected plants revealed intraspecific viral sequence variability, and also multiple begomovirus infections which could represent up to three species in a single tomato or cushaw plant. Phylogenetic grouping of virus sequences did not correlate with the host of origin.     

Comparison of Straw Mulch,Insecticides, Mineral Oil,and Birch Extract for Control of Transmission of Potato virus Y in Seed Potato Crops

Potato virus Y (PVY) is a major pathogen of potato and transmitted non-persistently by aphids. Aphis fabae is the main vector of PVY in the High Grade Seed Potato Production Area (HG area) in Finland, where the number of aphids and infection pressure with PVY are rather low, but problems with PVY occur in PVY-susceptible cultivars. The aim of the study was to test straw mulch, mineral oil, birch extract, and insecticides for control of PVY in small-scale field experiments and, additionally, at farm level in growers’ fields in the HG area of Finland. The insecticide esfenvalerate reduced the incidence of PVY in the progeny tubers by 29% in one of the 3 years, whereas other chemical treatments or birch extract had no significant effect on PVY incidence. Spraying foliage with mineral oil (Sunoco 11 E/3) reduced the incidence of PVY in 2 years by 43 to 58%, respectively. Straw mulch spread to the field at the time of plant emergence reduced PVY incidence in all 3 years by 50–70%. At farm level, straw mulch reduced the incidence of PVY in the progeny tubers by 25–47%, respectively, in both years tested; however, combining application of straw mulch and mineral oil did not further reduce incidence of PVY. Successful control of PVY in the HG area of Finland using straw mulch may be explained by transmission of PVY early in the growing season at the time of plant emergence and the relatively low number of vector aphids.     

Forest management inspired by natural disturbance dynamics (DISTDYN) – a long-term research and development project in Finland

A large-scale and long-term research and development project Forest management inspired by natural disturbance dynamics (DISTDYN) was initiated in Finland in 2009. The aim is to examine how variation in spatial and temporal scales of felling affects forest structure and biota. Harvesting methods applied in this project include selection, gap, partial and clear felling with variable retention. Rehabilitation is also applied as dead wood is intentionally increased while harvesting. The motivation of the project lies in the growing interest in using natural disturbances as an inspiration of forest-ecosystem management and in the recognition that mid- to small-scale disturbances are a characteristic feature of unmanaged Fennoscandian forests. The effects of felling methods are studied at the stand scale. However, a unique aspect of the project is the focus on landscape-scale impacts of different harvesting alternatives by comparing ca. 150-ha landscape units with varying harvesting regimes. Overall, the project aims at creating a long-lasting multidisciplinary research infrastructure. It also addresses core issues of forest-ecosystem management, including ecological, timber-production, economic and socio-cultural performances of alternative harvesting practices.     

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.     

Is the soil microbial community related to the basal area of trees in a Scots pine stand?

The main energy sources of soil microorganisms are litter fall, root litter and exudation. The amount on these carbon inputs vary according to basal area of the forest stand. We hypothesized that soil microbes utilizing these soil carbon sources relate to the basal area of trees. We measured the amount of soil microbial biomass, soil respiration and microbial community structure as determined by phospholipid fatty acid (PLFA) profiles in the humus layer (FH) of an even-aged stand of Scots pine (Pinus sylvestris L.) with four different basal area levels ranging from 19.9 m² ha⁻¹ in the study plot Kasper 1 to 35.7 m² ha⁻¹ in Kasper 4. Increasing trend in basal respiration, total PLFAs and fungal-to-bacterial ratio was observed from Kasper 1 to Kasper 3 (basal area 29.2 m² ha⁻¹). The soil microbial community structure in Kasper 3 differed from that of the other study plots.     

Diversity of Nordic Landrace Potatoes (<Emphasis Type="Italic">Solanum tuberosum</Emphasis> L.) Revealed by AFLPs and Morphological Characters

The management of genebank collections of cultivated potato is costly due to the need for in vitro maintenance and virus eradication. Therefore, it is important to set up conservation strategies, which prevent duplicates entering the collections. In this study, 32 Nordic potato landraces were studied for 57 morphological traits and analysed for amplified fragment length polymorphism (AFLP). Most landraces could be distinguished based on the morphological characters, except five accessions. Using five primer combinations to generate 114 reproducible AFLPs, of which 63 (55%) were polymorphic, the five morphologically indistinguishable accessions were placed into two groups with identical AFLP patterns, suggesting that some of the accessions were redundant for long-term preservation. The AFLP data showed that the Nordic collection of potato landraces is composed of genetically different clones, and morphological analysis revealed a wide range of variability. This variability seems to be distributed randomly over the Nordic region since the cluster analysis based on AFLPs and morphological traits revealed no grouping based on the country of origin. Principal component analysis suggests that fewer morphological traits than used in this study will be sufficient to discriminate between different genotypes of cultivated potato (Solanum tuberosum L.). Future possibilities for rationalising potato collections are discussed.     

Phoma–Didymella complex on hybrid arctic bramble with wilting symptoms

Pycnidia containing conidia characteristic of Phoma spp. and pseudothecia containing ascospores characteristic of Didymella applanata were isolated from edges of expanding stem lesions and dead stems of wilted cultivated hybrid arctic bramble plants ( Rubus arcticus nothossp. stellarcticus ) in Sweden in 1998 and 1999. The fungi were morphologically similar when grown on culture media, but some differences in growth rate were observed. They were also similar to the reference isolates of D. applanata (anamorph Phoma argillacea ). However, they were different from an isolate of Phoma sp. (HPP 38) isolated from cultivated arctic bramble ( R. arcticus ssp. arcticus ) in Finland in 1980, and from reference isolates of Phoma glomerata isolated from other hosts. Multivariate analysis of growth rate data and conidial dimensions measured in vitro indicated that the fungi isolated from hybrid arctic bramble in Sweden were not distinguishable from D. applanata , but were clearly distinct from P. glomerata and P. exigua. Furthermore, they had identical ITS1 and ITS2 sequences, and were placed in a phylogenetic clade very closely related to the clade that contained isolates of D. applanata isolated from raspberry ( Rubus idaeus ). In contrast, isolate HPP 38 from Finland was placed in a clade with P. exigua. These data indicate that the Swedish isolates infecting arctic bramble belong to a strain of D. applanata that differs from the isolate infecting raspberry only by two common nucleotide substitutions in ITS2. Fungi of the Phoma–Didymella complex on wild and cultivated arctic bramble (a total of 291 plants showing symptoms sampled from 37 sites) were detected by a PCR-based assay and found to be common in northern Sweden, but rare, albeit widely distributed, in Finland.     

Resistance of potato to cucumber mosaic virus appears related to localization in inoculated leaves

Although CMV can cause extensive damage to solanaceous crops like tomato, tobacco and pepper, it has little if any impact on the related crop, potato. To better understand this phenomenon, we have tested 32 potato genotypes for susceptibility or resistance to local and systemic Fny-CMV infection. Using ELISA, all genotypes appeared to show virus replication in leaves that were mechanically inoculated. In contrast, only ten genotypes exhibited systemic spread as expressed by significant CMV titers in new growth. These data suggest that, although CMV can replicate in potato cells and appears to spread short distances cell-to-cell, long distance movement appears to be blocked in most potato genotypes. Therefore, the general resistance of potato crops to CMV infection may be related to virus localization at points of entry.     

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.