美洲黑杨对冬季水淹的性别特异性响应

发布时间：2018-01-12 04:12

本文关键词：美洲黑杨对冬季水淹的性别特异性响应　出处：《中国科学院研究生院(武汉植物园)》2015年硕士论文　论文类型：学位论文

【摘要】：三峡水库受“冬季蓄水,夏季泄洪”人工水位调节的影响,形成了垂直落差达30 m的水库消落带,造成植物多样性急剧下降,从而引起一系列的生态问题。消落带防护林对维护库岸稳定、减少水土流失、保持生态景观以及水体净化等方面起着重要作用,因此在三峡水库上部消落带构建生态防护林具有重要的意义。三峡水库消落带生态防护林建设的适宜树种必须具备耐冬季水淹、适应夏季干旱、改良土壤的能力。杨树具有适应性强,生长速度快和丰产等特性,具有重要的生态及经济价值,常作为极端环境生态修复的先锋树种。前人研究表明,美洲黑杨(Populus deltoides)具备较强的耐水淹能力。另外,杨树为雌雄异株植物,前人研究表明其雌雄植株对非生物胁迫存在性别差异响应。本研究以美洲黑杨为研究材料,模拟三峡水库消落带冬季水淹环境条件,研究雌雄植株对冬季水淹的性别差异性响应。相关研究成果为三峡库区消落带生态防护林建设提供理论依据,主要结果如下：1通用提取缓冲液对胁迫相关生理生化指标提取效率的研究为节约试验材料和人工成本,研制一种适用于多种生理指标的通用提取液十分有必要。由于大部分生理指标提取液为磷酸缓冲液或Tris-HCl缓冲液且差异不大,因此比较通用磷酸缓冲液、通用Tris-HCl缓冲液及专一性提取液对9种生理指标的提取效率,研究发现100 mmol/L的通用磷酸缓冲液适用于GR, GPx, CAT, SOD, O2-,可溶性蛋白和游离脯氨酸等大部分生理指标的提取测定。2冬季水淹下美洲黑杨雌雄植株生理生化的响应差异经过130天冬季水淹胁迫后,所有的雌雄植株能完全存活,表明美洲黑杨具备较强的耐水淹能力。但是,雌雄植株在生长发育、生理生化方面的响应存在性别差异。正常水分状况下,雌株在还原性糖,O2-, .OH, APx, CAT, A及WUEi本底含量优于雄株；而在水淹状况下,雌株在Ci, H2O2, MDA,株高和基径等方面受到的负面影响大于雄株；相反,雄株在GSH, SOD和POD对水淹胁迫正反馈优于雌株；另外,雌雄植株在叶绿素荧光方面对水淹胁迫前后的差异不显著。3冬季水淹下美洲黑杨雌雄植株的蛋白质组学分析利用同位素标记相对和绝对定量(isobaric tags for relative and absolute quantitation, iTRAQ)技术,对冬季水淹条件下美洲黑杨雌雄植株的叶片全蛋白质组学进行了分析,通过两两比较,共鉴定了757个至少在一个组合中有显著性差异的蛋白。其中25个是雌株特异性蛋白,在水淹条件下,雄株中表达量比雌株高,而且在雌株中受到水淹诱导后上调。另有80个蛋白在水淹以后,雄株中表达量比雌株低,而且在雌株中受水淹诱导下调。这105个蛋白有可能直接参与了雌雄植株对水淹胁迫的应答。相关鉴定的蛋白主要参与了Transcription, Replication, Recombination and Repair, Post Modification, Protein Turnover, Chaperones, Coenzyme Transport及Metabolism等途径。以上结果表明,100 mmol/L的通用磷酸缓冲液适用于同时提取GR, GPx, CAT, SOD, O2-,可溶性蛋白和游离脯氨酸等大部分生理指标。美洲黑杨在130天水淹处理以后,所有的雌雄植株能完全存活,表明美洲黑杨具备较强的耐水淹能力。但是,雌雄植株在生长发育、生理生化及蛋白等方面的响应存在性别差异,雄株比雌株有着更强的耐水淹能力,该研究成果能为三峡水库消落带杨树生态防护林建设提供科学指导。
[Abstract]:The Three Gorges Reservoir by water in winter, the summer flood "artificial water level regulation, forming a vertical drop of 30 m reservoir riparian zone, resulting in plant diversity decreased rapidly, which caused a series of ecological problems. With Shelterbelts on the maintenance of stability of reservoir drawdown, reduce soil erosion, maintain plays an important role the ecological landscape and water purification and so on, so the upper riparian zone in the Three Gorges reservoir ecological protection forest construction has important significance. A suitable tree species of the Three Gorges reservoir, the construction of ecological protection forest must have resistance to winter flooding, summer drought, soil improvement ability. Poplar has strong adaptability, fast growth and high yield characteristics that has important ecological and economic value, often as a pioneer species in the extreme environment of ecological restoration. Previous studies showed that America Black Poplar (Populus deltoides) have stronger resistance to flooding. On the other Outside, poplar is dioecious, previous studies showed that there are gender differences in the responses of male and female plants to abiotic stress. In this study, black poplar of America as research materials, simulation of the Three Gorges reservoirtotal winter flooding conditions of winter flooding on gender differences in male and female plants response. Related research results with the construction of ecological protection forest to provide a theoretical basis for the Three Gorges Reservoir area, the main results are as follows: 1 Research on the general extraction buffer on the extraction efficiency of stress related physiological and biochemical indexes in order to save the material and labor costs, it is necessary to develop a suitable for various physiological indicators of general extract. Due to extract most of the physiological indexes of phosphate buffer or Tris-HCl buffer the liquid and the difference is not so common, phosphate buffer, Tris-HCl buffer and the specificity of the extracted effect on 9 kinds of physiological index rate The study found that general, 100 mmol/L phosphate buffer for GR, GPx, CAT, SOD, O2-, extraction of most physiological indexes of soluble protein and free proline determination response of Populus deltoides male and female plants physiological and biochemical.2 winter flooding difference after 130 days of winter flooding stress, male and female plants can all survived completely. That America black poplar have strong waterlogging tolerance. However, male and female plants in growth and development, there are gender differences in response to physiological and biochemical aspects. The normal water condition, the female plant in the reducing sugar, O2-,.OH, APx, CAT, A and WUEi is better than that of the background content of male plants; while in the flooded condition. The female plant in Ci, H2O2, MDA, negative effects on plant height and basal diameter as much more than the male plants; on the contrary, the male plant in GSH, SOD and POD on the flooding stress positive feedback is better than the female line; in addition, male and female plants in the chlorophyll fluorescence of water stress No significant difference before and after the forced.3 winter flooding under Populus deltoides male and female plants proteomic analysis using isobaric tags for relative and absolute quantitation (isobaric tags for relative and absolute quantitation iTRAQ) technology, the winter flooding conditions of Populus deltoides male plant leaf proteome was analyzed by comparison of 22, 757 proteins were identified at least significant difference in a group. 25 of them are female specific protein, in flooded conditions, the expression in the male plants than female plants, and induced by flooding after increase in female plants. Another 80 protein in flooded after expression in the male plants than female plants, and in female plants by flooding induced down-regulation. These 105 proteins may be directly involved in the male and female plants to flooding stress response. The identified proteins mainly involved in Transcription, Repl Ication Recombination, and Repair, Post Modification, Protein Turnover, Chaperones Coenzyme, Transport and Metabolism. The results above indicated that the general 100 mmol/L phosphate buffer was suitable for the simultaneous extraction of GR, GPx, CAT, SOD, O2-, most physiological indexes of soluble protein and free proline. Deltoides in 130 days flooded after all, male and female plants can survive, that America black poplar have strong waterlogging tolerance. However, male and female plants in growth and development, physiological and biochemical aspects of protein response due to gender differences, male plants than female plants have a stronger flooding-tolerant ability, the research results can provide scientific guidance for the Three Gorges reservoirtotal poplar ecological shelterbelt construction.

【学位授予单位】：中国科学院研究生院(武汉植物园)
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S792.11

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