Apex cryopreservation of several strawberry genotypes by two encapsulation-dehydration methods.

This paper presents results from a study to develop cryopreservation procedures for apices of several strawberry genotypes. Five Fragaria x ananassa Duch. cultivars and two wild species (F. chiloensis and F. virginiana) have been screened using the encapsulation-dehydration method and/or a protocol which compromises vitrification and encapsulation-dehydration. Apices were encapsulated in an alginate gel, precultured on media containing high levels of sucrose (0.8 M, conventional protocol), or a combination of 0.4 M sucrose and 2 M glycerol. Recovery rates varied among genotypes (23-63%). The latter method reduced considerably the time needed for the cryogenic procedure by eliminating the pre-treatment with 0.8 M sucrose for 19 h prior to dehydration, as required by the conventional procedure.

Chemical Alternatives to Methyl Bromide in Spanish Strawberry Nurseries.

Strawberry runners are a high-value cash crop in Spain that requires vigorous transplants free of pathogens. Preplant soil fumigation with methyl bromide, or with mixtures of methyl bromide and chloropicrin, is a standard practice for controlling soilborne diseases. Soil fumigants chloropicrin, 1,3-dichloropropene, dazomet, metam-sodium, metam potassium, and dimethyl disulfide were evaluated in combination with different plastic films as alternatives for methyl bromide soil fumigation of strawberry nurseries. The studies were conducted over a 4-year period, with fumigant applications prior to planting. Verticillium wilt (caused by Verticillium spp.) and crown rot (caused by Phytophthora cactorum) were the main diseases. Chloropicrin, 1,3-dichloropropene, and dazomet compared well with methyl bromide fumigation for control of strawberry nursery diseases. Furthermore, 1,3-dichloropropene and methyl bromide, applied at 50% rate under virtually impermeable film, provided effective disease control in strawberry nurseries. Fumigant effects on fungal soil populations are discussed.

Shoot regeneration and Agrobacterium-mediated transformation of Fragaria vesca L.

An efficient and reliable method for shoot regeneration from leaf disks of Fragaria vesca L. has been developed. This protocol has been successfully employed to obtain transformed plants using Agrobacterium tumefaciens as gene vector. Murashige and Skoog basal medium supplemented with benzyladenine (4 mg/l) and indole-3-butyric acid (0.25 mg/l) induced the maximum percentage of shoot regeneration (98%) and the highest number of shoot colonies per explant (4.6) after 8 weeks of culture. Isolated shoots would elongate and proliferate when the benzyladenine concentration was lowered to 0.5 mg/l. The established protocol for shoot regeneration was employed to transform leaf disks using Agrobacterium tumefaciens carrying the plasmid pBI121. A 7.7% of the inoculated explants showed kanamycin resistance after 10 weeks of selection in a medium containing 25 mg/l of this antibiotic. The transgenic shoots obtained were rooted in the presence of 25 mg/ kanamycin and successfully acclimatized. The final percentage of transformation obtained based on beta-glucuronidase expression was 6.9%.