Windsor M L, Milborrow B V, McFarlane I J
School of Biochemistry, University of New South Wales, P.O. Box 1, Kensington, New South Wales 2033, Australia.
Plant Physiol. 1992 Sep;100(1):54-62. doi: 10.1104/pp.100.1.54.
The uptake of (+)-S- and (-)-R-abscisic acid (ABA) by suspension culture cells of hopbush (Dodonaea viscosa L. Jacqu.) was followed over a range of temperatures, pH values, and time intervals. The natural (+)-S-ABA was taken up about five times faster than the unnatural (-)-R-ABA. Each 10 degrees C rise in temperature from 1 to 31 degrees C increased the rate of uptake (Q(10)) of (+)-S-ABA about 2.2-fold, whereas that of the (-)-R increased with a Q(10) of 1.4. (+)-ABA was taken into the cells by a saturable carrier, but (-)-ABA and both enantiomers of 2-trans-ABA were not; they appeared to enter by passive diffusion. The uptake of (+)-ABA was linear over the first 8 hours but concentrations within the cells decreased after 2 hours to remain constant after 4 hours as rapid metabolism was induced. Electron microscopy of thin sections of the cells, combined with a stereological analysis of their shape, showed that the vacuoles comprised 80% of the cell volume and the cytoplasm plus nucleus comprised 20%. There were no photosynthetically active plastids in the cells. Concentrations of the endogenous ABA in the cytoplasm (pH 7.32) and vacuoles (pH 5.88) were calculated by applying the Henderson-Hasselbalch equation (ABA pK(a) 4.7) so that, provided no active metabolic redistribution occurred, the concentration in the cytoplasm was 7.9 micromolar and that in the vacuole was 0.3 micromolar. In vivo pH was measured by (31)P nuclear magnetic resonance spectroscopy.
在一系列温度、pH值和时间间隔条件下,对粘叶山龙眼(Dodonaea viscosa L. Jacqu.)悬浮培养细胞对(+)-S-脱落酸(ABA)和(-)-R-脱落酸的吸收情况进行了跟踪研究。天然的(+)-S-ABA的吸收速度比非天然的(-)-R-ABA快约五倍。温度从1℃升高到31℃,每升高10℃,(+)-S-ABA的吸收速率(Q10)增加约2.2倍,而(-)-R-ABA的吸收速率以1.4的Q10增加。(+)-ABA通过可饱和载体进入细胞,但(-)-ABA和2-反式-ABA的两种对映体则不然;它们似乎是通过被动扩散进入的。(+)-ABA的吸收在最初8小时内呈线性,但2小时后细胞内浓度下降,4小时后随着快速代谢的诱导而保持恒定。对细胞薄片进行电子显微镜观察,并结合对其形状的体视学分析,结果表明液泡占细胞体积的80%,细胞质加细胞核占20%。细胞中没有光合活性质体。通过应用亨德森-哈塞尔巴尔赫方程(ABA pKa 4.7)计算细胞质(pH 7.32)和液泡(pH 5.88)中内源性ABA的浓度,因此,如果没有活跃的代谢重新分布发生,细胞质中的浓度为7.9微摩尔,液泡中的浓度为0.3微摩尔。通过31P核磁共振光谱法测量体内pH值。