Mori Kentaro, Goto-Yamamoto Nami, Kitayama Masahiko, Hashizume Katsumi
National Research Institute of Brewing, Higashi-Hiroshima, Hiroshima 739-0046, Japan.
J Exp Bot. 2007;58(8):1935-45. doi: 10.1093/jxb/erm055. Epub 2007 Apr 23.
To determine the mechanism of inhibition of anthocyanin accumulation in the skin of grape berries due to high temperature, the effects of high temperature on anthocyanin composition and the responses in terms of gene transcript levels were examined using Vitis vinifera L. cv. Cabernet Sauvignon. High temperature (maximum 35 degrees C) reduced the total anthocyanin content to less than half of that in the control berries (maximum 25 degrees C). HPLC analysis showed that the concentrations of anthocyanins, with the exception of malvidin derivatives (3-glucoside, 3-acetylglucoside, and 3-p-coumaroylglucoside), decreased considerably in the berries grown under high temperature as compared with the control. However, Affymetrix Vitis GeneChip microarray analysis indicated that the anthocyanin biosynthetic genes were not strongly down-regulated at high temperature. A quantitative real time PCR analysis confirmed this finding. To demonstrate the possibility that high temperature increases anthocyanin degradation in grape skin, stable isotope-labelled tracer experiments were carried out. Softened green berries of Cabernet Sauvignon were cut and aseptically incubated on filter paper with 1 mM aqueous L-[1-(13)C]phenylalanine solution for 1 week. Thereafter, the changes in (13)C-labelled anthocyanins were examined under different temperatures (15, 25, and 35 degrees C). In the berries cultured at 35 degrees C, the content of total (13)C-labelled anthocyanins that were produced before exposure to high temperature was markedly reduced as compared with those cultured at 15 degrees C and 25 degrees C. These data suggest that the decrease in anthocyanin accumulation under high temperature results from factors such as anthocyanin degradation as well as the inhibition of mRNA transcription of the anthocyanin biosynthetic genes.
为了确定高温导致葡萄浆果表皮花青素积累受抑制的机制,以赤霞珠葡萄(Vitis vinifera L. cv. Cabernet Sauvignon)为材料,研究了高温对花青素组成的影响以及基因转录水平的响应。高温(最高35℃)使总花青素含量降至对照浆果(最高25℃)的一半以下。高效液相色谱分析表明,与对照相比,高温下生长的浆果中除了锦葵色素衍生物(3-葡萄糖苷、3-乙酰葡萄糖苷和3-对香豆酰葡萄糖苷)外,其他花青素的浓度均显著降低。然而,Affymetrix葡萄基因芯片微阵列分析表明,花青素生物合成基因在高温下并未强烈下调。定量实时PCR分析证实了这一发现。为了证明高温会增加葡萄皮中花青素降解的可能性,进行了稳定同位素标记示踪实验。将赤霞珠软化的绿色浆果切开,无菌培养在含有1 mM L-[1-(13)C]苯丙氨酸水溶液的滤纸上1周。此后,在不同温度(15、25和35℃)下检测(13)C标记的花青素的变化。与在15℃和25℃培养的浆果相比,在35℃培养的浆果中,高温处理前产生的总(13)C标记花青素含量显著降低。这些数据表明,高温下花青素积累的减少是由花青素降解以及花青素生物合成基因mRNA转录受抑制等因素导致的。