Abstract
In this study, cell-suspension culture of strawberry (Fragaria × ananassa), cultivars Camarosa, and Sweet Charlie has been established. Embryogenic callus was induced by incubating the in vitro juvenile leaf explants on medium, containing 2-mg/l picloram at dark. Suspension culture was initiated from 4-week-old embryogenic calli in the liquid MS medium with 1-mg/l 2,4-D and 2-mg/l picloram. Suspension culture was maintained by sub-culturing each 3 weeks into a fresh medium. At week 9 after third sub-cultures, torpedo and cotyledonary embryo stages were observed. Embryos were then developed into shoots on medium 1 mg/l of each BA and IBA. Obtained shoots were successfully rooted on 1-mg/ml GA3, 0.5-mg/ml BA, and 1-mg/ml IBA. To enhance the resistance availability in strawberry plants, elicitation was applied by adding the JA and SA elicitors to the suspension culture with two doses (0.5 and 1 mM) individually and in combination, in addition to the fungal homogenate of Macrophomina phasiolena at concentration of 106 spor/ml. The fawrky-1-Camarosa gene, which has defense-related function, was detected in the different elicited strawberry tissues and isolated via RT-PCR. The isolated gene was submitted to GenBank with accession number (KX096885).
Highlights
The cultivated strawberry (Fragaria 9 ananassa Duch.), a member of the Rosaceae, is the most economically important soft fruit worldwide (Parikka 2004; Debnath et al 2007)
Embryogenic callus induction: the 4-week-old in vitro juvenile leaf explants started to form callus 10 days postculturing on the three tested media, within 3 weeks, the embryogenic calli were produced
Highest callus formation was observed on medium MSCII either at light or dark, followed by MSCI
Summary
The cultivated strawberry (Fragaria 9 ananassa Duch.), a member of the Rosaceae, is the most economically important soft fruit worldwide (Parikka 2004; Debnath et al 2007). Rhizoctonia spp., Pythium spp, Fusarium spp., and Macrophomina phaseolina are the fungal species that associate with black root rot diseases which leading to limit fruit production worldwide (Browne et al 2002; Millner 2006). Plants have evolved numerous complex defense mechanisms to survive of the fungal and microbial pathogen attacks. HR is associated with the accumulation of salicylic acid and several classes of pathogenesis-related (PR) proteins, many of which exhibiting antimicrobial activity which is enhancing the plant defensive capacity against a broad spectrum of pathogens. This resistance is mostly expressed locally and in distal, uninfected tissues, which is
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