Comparative proteomic analysis of longan (Dimocarpus longan Lour.) seed abortion

Abstract

Two-dimensional gel electrophoresis (2-DE), coupled with mass spectroscopy, was used to study seed abortion in Dimocarpus longan Lour. (cv. Minjiao 64-1) by comparing normal and aborted seeds at three developmental stages. More than 1,000 protein spots were reproducibly detected in 2-DE gels, with 43 protein spots being significantly altered in their intensity between normal and aborted seeds at least at one stage. Thirty-five proteins were identified by matrix-assisted laser desorption ionization-time of flight-tandem mass spectrometry (MALDI-TOF-MS/MS) analysis and protein database searching. Most of the identified proteins were associated with a variety of functions, including energy and metabolism (30%), programed cell death (9%), antioxidative processes (14%), chaperonin (23%), cell division, amino acid metabolism, secondary metabolism, and other functional classes. Furthermore, the expression patterns of HSP70 and cytosolic ascorbate peroxidase (cAPX) were validated by immunoblotting analysis. This study provides a novel, global insight into proteomic differences between normal and aborted seeds in longan. We anticipate that identification of the differentially expressed proteins may lead to a better understanding of the molecular basis for seed abortion in longan.

Regulation of Anthocyanin Biosynthesis in Purple Leaves of Zijuan Tea (Camellia sinensis var. kitamura)

“…Total protein concentration of the tea leaves was extracted in accordance with a previously described phenol extraction method [41]. Protein powder (10 mg) was solubilized in 250 μL of lysis buffer (7 mol·L −1 urea, 2 mol·L −1 thiourea, 40 g·L −1 3-[(3-cholamidopropyl)-dimethylammonio] propanesulfonate (CHAPS), and 40 mmol·L −1 dithiothreitol (DTT)) in a water bath at 37 °C for 2.5 h. The homogenate was centrifuged at 17,000× g for 15 min at room temperature.…”

Proteomic Analysis of Salt-Responsive Proteins in the Leaves of Mangrove Kandelia candel during Short-Term Stress

“…Two-dimensional gel electrophoresis was undertaken according to the method of Liang et al [84] and Liu et al [85] with the Ettan IPGphor system (GE Healthcare, Little Chalfont, UK). The leaf protein (1,500 µg) was loaded onto IPG strips (24 cm, linear gradient pH 4–7; GE Healthcare) in a rehydration tray for 12 h. Isoelectric focusing (IEF) was performed under the following conditions: 200 V for 1 h, 500 V for 1 h, 1 kV for 1 h, gradient to 8 kV for 0.5 h, and, finally, 8 kV for a total of 48,000 Vhs at 20°C.…”

Proteomic analysis of changes in the Kandelia candel chloroplast proteins reveals pathways associated with salt tolerance

“…8, 10 and 13) and phosphoglycerate kinase (spots 17, 23 and 24) were up-regulated after 3-day salt treatment, and up-regulated or unchanged after 6-day salt treatment. It was not surprising that some proteins were represented by more than one spot, which may reflect isoproteins or post-translational modifications of a single protein [45][46][47]. The principal role of RuBisCO activase is to release inhibitory sugar phosphates such as ribulose-1,5-bisphosphate (RuBP) from the active sites of RuBisCO so that CO 2 can activate the enzyme by carbamylation [48].…”