Leaves were used to detect Ca2+ flux using Non‐invasive Micro‐test Technology (NMT) as described previously.[55 (link)
] The Ca2+ content (mg·g−1) was measured using a calcium colorimetric assay kit (Beyotime Biotechnology Co., Ltd. Shanghai, China).
Leaves were incubated in 1 mg ml−1, pH 3.8, DAB‐HCl (Sigma‐Aldrich, USA) in the dark for 8 h. The cotyledons were then cleared by boiling in alcoholic lactophenol (95% ethanol: lactophenol, 2:1 v/v) for 20 min. The reddish color of the cotyledons was used as evidence of H2O2 and visualized using a Nikon D40 camera (Japan). The quantification of H2O2 was conducted using the test kit (G0112W) from Suzhou Grace Biotechnology Co., Ltd (Suzhou, China), following the manufacturer instructions. For SA extraction, plant tissues were ground into fine powder by liquid nitrogen, and 0.1 g was used from each sample following the method of.[26 (link)
] Samples were then analyzed using LC/MS.
] The Ca2+ content (mg·g−1) was measured using a calcium colorimetric assay kit (Beyotime Biotechnology Co., Ltd. Shanghai, China).
Leaves were incubated in 1 mg ml−1, pH 3.8, DAB‐HCl (Sigma‐Aldrich, USA) in the dark for 8 h. The cotyledons were then cleared by boiling in alcoholic lactophenol (95% ethanol: lactophenol, 2:1 v/v) for 20 min. The reddish color of the cotyledons was used as evidence of H2O2 and visualized using a Nikon D40 camera (Japan). The quantification of H2O2 was conducted using the test kit (G0112W) from Suzhou Grace Biotechnology Co., Ltd (Suzhou, China), following the manufacturer instructions. For SA extraction, plant tissues were ground into fine powder by liquid nitrogen, and 0.1 g was used from each sample following the method of.[26 (link)
] Samples were then analyzed using LC/MS.
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