Ectopic expression of a grape aspartic protease gene, AP13, in Arabidopsis thaliana improves resistance to powdery mildew but increases susceptibility to Botrytis cinerea

Expression of the Grape VqSTS21 Gene in Arabidopsis Confers Resistance to Osmotic Stress and Biotrophic Pathogens but Not Botrytis cinerea

“…Botrytis cinerea was obtained from tomato and cultured on potato dextrose agar medium at 25°C in the dark. Conidial spore suspensions (2 × 10 7 conidia/ml) were prepared with 14-day old cultures using sterile, distilled water as described previously ( Guo et al, 2016 ). Fifty leaves from 24 four-week old plants, including the three transgenic lines and untransformed controls, respectively, were rinsed with distilled water and 10 μl conidial suspension was applied, after which time the leaves were placed on 1% agarose overlaid with wet filter paper and the glass Petri dishes were then sealed.…”

“…Disks were rinsed with sterile water three times, homogenized in 100 μl 10 mM MgCl 2 , and the solution was gradually diluted. One hundred microliters of the resulting diluted solution were then plated onto King’s B agar plates supplemented with 50 μg/ml rifampicin and 50 μg/ml kanamycin for 48 h at 28°C ( Guo et al, 2016 ).…”

“…In the case of NBT staining, inoculated leaves were incubated in HEPES buffer (pH 7.5) containing 6 mM NBT for 2∼3 h ( Tu et al, 2016 ). Trypan blue staining was carried out as described previously ( Guo et al, 2016 ). Briefly, infected leaves were soaked in boiled trypan blue solution (a 1:1:1:1:1 ratio of trypan blue, phenol, glycerol, lactic acid, and water) for 2–3 min, and were subsequently depigmented in 2.5 g/ml chloral hydrate for 1–2 days.…”

“…To determine the inhibitory concentrations of NaCl and mannitol for cotyledon greening rates, both transgenic and Col-0 seeds were sown on MS medium containing different concentrations of NaCl and mannitol, respectively. Subsequently, 120 sterilized seeds from each line were sown on MS medium supplemented with 130 mM NaCl or 250 mM mannitol to stimulate salt and drought stress, respectively ( Guo et al, 2016 ; Tu et al, 2016 ), and were grown at 21∼23°C with a 16 h/8 h photoperiod (100 μmol m -2 s -1 photon flux density). Cotyledon greening rates were recorded daily.…”

Ectopic Expression of Grapevine Gene VaRGA1 in Arabidopsis Improves Resistance to Downy Mildew and Pseudomonas syringae pv. tomato DC3000 But Increases Susceptibility to Botrytis cinerea

“…Solutions of 100 mM SA (Tokyo, Japan) and 50 mM methyl jasmonate (MeJA, Haarlem, The Netherlands) were sprayed on grape leaves with the same growth conditions, and aseptic ultra-pure water was sprayed as the mock [70]. The expression level of VaRGA1 was determined at 0, 1, 3, 6, 12, 24, and 48 h post-treatment.…”