Plant Physiol. 2007 November; 145(3): 853–862.
Suberin LocalizationTumor cross sections as used for ABA immunolocalization were stained with a saturated solution of Sudan-III in 92% ethanol (w/v) for 10 min at 70°C, then washed with glycerol:water (1:1, v/v). Sections were examined using a Leica DMR microscope with bright-field illumination or fluorescence excitation of 488 nm using a two-dimensional blue filter (Leica).
Plant Physiol. 2007 November; 145(3): 853?62.
Determination of ABA ContentFree ABA and ABA released from its conjugates was analyzed by ELISA as described earlier (Jiang et al., 2004). Recoveries of ABA during the purification procedures were checked routinely using radioactive ABA and found to be more than 95%. The immunochemicals were generously supplied by Professor Weiler, Ruhr Universit鋞 Bochum (Germany).
Plant Physiol. 2007 November; 145(3): 853–862.
Agrobacterium-Mediated Transient TransformationFor transient transformation of Arabidopsis, Agrobacterium GV3101 (pMP90; Koncz and Schell, 1986) was used. This strain harbored the binary plasmid pMDC164 for expression of GUS under control of the 2× cauliflower mosaic virus 35S promoter and was mixed with strain 19 K (Latz et al., 2007) before infiltration to prevent gene silencing. Growth of agrobacteria and infiltration (agroinfiltration) into leaves of 2- to 3-month-old Arabidopsis leaves or injection into the base of inflorescence stalks of 3- to 4-month-old plants was carried out as described in Zipfel et al. (2006) with minor modifications. Qualitative and quantitative measurements of GUS activity 4 to 7 d after infiltration with agrobacteria were performed according to Jefferson et al. (1987). For more detailed protocol see Supplemental Materials and Methods S1.
Plant Physiol. 2006 January; 140(1): 302–310
Germination AssayGermination rates were compared between seed lots that were produced, harvested, and stored under identical conditions. Before planting, seeds were surface sterilized with 70% ethanol for 1 min, then with 2% hypochlorite for 5 min, and rinsed five times with sterile deionized water. Fifty to 100 seeds from wild type (Col-0), rgs1-2, gpa1-3, and 35S-RGS1 seeds were stratified at 4°C for 48 h and planted in petri dishes on half-strength Murashige and Skoog (MS) 0.8% phytoagar medium lacking Suc and Gamborg vitamins at 22°C under continuous white light in triplicate. Seeds were considered germinated when the radicles completely penetrated the seed coat.
Plant Physiol. 2006 January; 140(1): 302–310
Hormonal and Sugar Treatment The effects of ABA on seed germination were studied by determining the germination rates of 70 to 100 seeds pretreated with deionized water or 100 μm fluridone and planted in triplicate on medium containing ABA (mixed isomers; Sigma). The effects of Glc, Suc, mannitol, and sorbitol were studied in a similar manner. Sterilized seeds were stratified at 4°C for 48 h and sown on plates containing different concentrations or types of sugars (d-Glc, Suc, d-mannitol, and d-sorbitol) in triplicate. For direct comparisons of germination rates, each plate was subdivided and all seed lines were planted on the same plate.
