为了探讨热激蛋白在低温锻炼诱导的耐热性形成过程中的保护功能,进一步分析葡萄幼苗对温度逆境产生交叉适应性的细胞学机制,试验以葡萄(Vitis vinifera L.cv.Jingxiu)幼苗为试材,采用胶体金免疫电镜定位技术,定位观察了小分子热激蛋白17.6(sHSP17.6)在葡萄叶片中的亚细胞分布情况,结果表明,低温锻炼预处理可增强其在高温胁迫条件下的表达水平,细胞核和叶绿体中代表sHSP17.6的免疫金颗粒密度比相应的对照细胞明显增多,尤其是胁迫处理3 h时.这一结果为sHSP17.6参与低温锻炼诱导的耐热性提供了更直观的细胞学证据.
In order to invest igate the protect ing role of small heat shock protein 17. 6( sHSP17. 6) in the heat tolerance induced by cold acclimat ion( CA),and then to explore the mechanism of cross adaptation to different temperature stresses in grape plants. Leaves from two-year-old young grape ( Vitis vinif era L. cv. Jingxiu) plants were used as exper-imental materials.The subcellular localizations of sHSP17. 6 within mesophyll cells were observed using immuno-gold labeling electron microscopic technique. The results showed that CA pretreatment could enhance the expression level of sHSP17. 6 within heat stress. Compared to non-acclimated plants, the density of sHSP17. 6 gold particles was obviously increased in the nucleus and chloroplast of the CA leaves, especially 3 h after heat stress. Which offered the cytological evidence of sHSP17. 6 was contributed to the heat tolerance induced by CA pretreatment in grape plants.
[1] 曾韶西,王以柔,李美如.不同胁迫预处理提高水稻幼苗抗寒性期间膜保护系统的变化比较[J].植物学报,1997,39(4):308-314.
[2] Gong M,Chen B,Li Z G,et al.Heat shock induced cross adaptation to heat,chilling,drought and salt stress in maize seedlings and involvement of HO2[J].Journal of Plant Physiology,2001,158:1125-1130.
[3] Wang W X,Vinocur B,Altman A.Plant responses to drought,salinity and extreme temperatures:towards genetic engineering for stress tolerance[J].Plants,2003,218:1-14.
[4] Roach J M,Bingham S E,Sommerfeld M R.Protein expression during heat stress in thermo-intolerance and thermo-tolerance diatoms[J].Journal of Experimental Marine Biology and Ecology,2004,306:231-243.
[5] Jinn T L,Chiu C C,Song W W,et al.Azetidine-induced accumulation of class Ⅰ small heat shock proteins in the souble fraction provides thermotoleranee in soybean seedlings[J].Plant and Cell Physiology,2004,45:1759-1767.
[6] Keeler S J,Boettger C M,Haynes J G,et al.Acquired thermotolerance and expression of the HSP100/CI pB genes of Lima bean[J].Plant Physiology,2000,123:1121-1132.
[7] Zhang J H,Huang W D,Liu Y P,et al.Effect of temperature acclimation pretreatment on the ultrastrueture of mesophyll cell in young grape plants under cross temperature-stresses[J].Journal of Integrative Plant Biology,2005,47:959-970.
[8] Zhang J H,Liu Y P,Pan Q H,et al.Changes in membraneassociated H+-ATPase activities and amounts in young grape plants during the cross adaptation to temperature stresses[J].Plant Science,2006,170:768-777.
[9] 张俊环,张国强,刘悦萍,等.温度逆境交叉适应过程中葡萄幼苗质膜Ca2+-ATPase的细胞化学定位与活性变化[J].中国农业科学,2006,39(8):1617-1625.
[10] 张俊环,黄卫东.葡萄幼苗在温度逆境交叉适应过程中活性氧及抗氧化酶的变化[J].园艺学报,2007,34(5):1073-1080.
[11] Zhang J H,Wang L J,Pan Q H,et al.Accumulation and subcellular localization of heat shock proteins in young grape leaves during cross-adaptation to temperature stresses[J].Scientia Horticulturae,2008,117(3):231-240.
[12] Zhang D P,Lu Y M,Wang Y Z,et al.Acid invertase is predominantly localized to cell walls of both the practically symplasmically isolated sieve element/companion cell complex and parenchyma cells in developing apple fruits[J].Plant Cell and Environment,2001,24:691-702.
[13] 吴扬,张伟,徐代根,等.热应激蛋白的诱导、定位与再分布研究[J].同济医科大学学报,1995,24(6):419-421.
[14] Scgui-Simarro J M,Testillano P S,Risueno M C.Hsp70 and Hsp90 change their expression and subcellular localization after microspore embryoganesis induction in Brassica napus L[J].Journal of Structural Biology,2003,142:379-391.
