【作者】 刘廷武；

【导师】 郑海雷；

【作者基本信息】 厦门大学 ， 植物学， 2009， 硕士

【摘要】 酸雨作为世界十大环境问题之一,已越来越受到社会及学者的关注;酸雨会引起植物体及生态系统钙流失,从而严重危害植物生长,导致森林衰退。已有研究显示,外源钙的施加可以帮助受酸雨危害过森林的恢复,但不同钙浓度下植物对酸雨抗性如何尚缺乏研究;同时,目前对酸雨危害的研究也多集中在生理水平,少见机理层次的报道。基于以上两方面研究的重要性,本文以酸雨区主要物种马尾松（Pinus massoniana）、柳杉（Cryptomeria fortunei）、杉木（Cunninghamialanceolata）、枫香（Liquidambar formosana）、华山松（Pinus armandi）、锥栗（Fagaceae henryi）种子和幼苗为研究对象,研究其在3种钙水平上对酸雨的响应;并利用荧光差异凝胶电泳（DIGE）技术,分析酸雨处理下马尾松和湿地松（Pinus elliotii）幼苗蛋白质组学的变化。主要结果如下:pH3.0酸雨处理下,6种植物种子的活力、发芽率、发芽指数、胚根长度受到抑制;不同钙浓度对种子的发芽能力影响显著,高钙条件下种子发芽能力得到有效恢复,即钙能够缓解酸雨对植物种子的危害。不同植物对钙的喜好程度不同,恢复程度也存在种间差异。不同钙浓度下,酸雨对6种植物叶片表面伤害、叶绿素含量及净光合速率的影响程度不同,钙离子能够提高幼苗抵抗酸雨的能力;随钙浓度升高,叶片伤斑减小甚至消失,叶绿素含量升高,光合能力增强。经pH3.0酸雨处理马尾松幼苗2月,有35个蛋白的表达变化量在2倍以上;其中,下调蛋白点14个,上调蛋白点21个。对差异蛋白进行归类分析发现,酸雨处理可以下调3个Rubisico大亚基蛋白,致使光合作用效率降低。同时,蔗糖磷酸酶、丙酮酸脱氢酶、磷酸甘油醛脱氢酶、磷酸核酮糖激酶、果糖二磷酸醛缩酶、苯醌氧化还原酶表达量下降,导致糖酵解、磷酸戊糖途径、脂肪酸代谢等生理活动不能正常进行。另一方面,酸雨胁迫可以导致ATP合成酶、颗粒结合型淀粉合成酶、叶绿体苹果酸脱氢酶的表达量升高,上调机体内这些能量相关酶的表达,增加对酸雨胁迫的抗性。此外,酸雨胁迫还可以下调赤霉素氧化酶、异黄酮还原酶、胆色素脱色氨酶、脱氢奎尼酸合成酶等与次生代谢相关酶的表达。最后,酸雨胁迫下,与Ca²⁺有关的单二氢抗坏血酸还原酶、热激蛋白、过氧化物酶的表达量升高,致使机体对酸雨的抗性提高。总之,酸雨胁迫对植物体的影响影响是多途径的,机体则通过降低自身能量损耗及激活其他能量途径来保证能量供应,维持正常生长;钙离子可以诱导植物体过量表达相关蛋白,提高对酸雨胁迫的抗性。更多还原

【Abstract】 As one of the world’s top ten environmental issues,acid rains have attracted more and more attentions of society and scholars.Acid rains could deplete the calcium from the plants or ecosystems,lead to serious negative effects on plant growth,and thus cause the decay of forests.The application of exogenous calcium have previously been reported to aid in the recovery of forest that was damaged by acid rains,however,few studies have been performed to compare the resistance to acid rain under different calcium concentrations.On the other hand,many researchers have devoted their attentions to impacts of acid rains on physiology but not on mechanism levels.Base on the above significances of acid rain effects,this thesis was designed to investigate the effects of acid rains on plant seeds and seedlings of Pinus massoniana,Cryptomeria fortune,Cunninghamia lanceolata,Liquidambar formosana,Pinus armandi and Fagaceae henryi at three calcium concentrations levels.And also,difference gel electrophoresis(DIGE) technology was used to determine the variations of the proteomics in Pinus massoniana and Pinus elliotii under acid rains stress.The main results are as follows:The vigor index,seed germination rate,germination index and radicle length of the six kinds of plant seeds were significantly inhibited by pH 3.0 acid rain treatment.; Calcium play an important role in seed germination with higher germination ability at higher calcium condition,indicating the alleviation of the acid rain induced damage on seed.There was an inter-specific difference of the restoration extent since the plants have different needs for calcium.Different damages of acid rains on the leaf necrosis,chlorophyll content and net photosynthetic rate of the six plants were observed at different calcium concentrations;and the seedling was more resistant to acid rains stress at higher calcium concentration.As an increase of calcium concentration,damage-areas caused by acid rains on leaves reduced or even disappeared and chlorophyll content and photosynthetic capacity increased.After 2 months treated by pH 3.0 acid rains,35 proteins expression with at least a 2-fold changed were observed in seedling of Pinus massoniana,therein 14 proteins being down-regulated and 21 proteins being up-regulated.Due to the acid rains stress, there were 3 down-regulated proteins belonging to Rubisico large subunits;and caused the reduction of the plant photosynthesis.At the same time,several energy metabolism-related proteins were also detected to be down-regulated,such as sucrose-phosphatase,pyruvate dehydrogenase,glyceraldehyde-phosphate dehydrogenase,phosphoribulokinase,fructose-bisphosphate aldolase,Quinone oxidoreductase,which are key enzyme of glycolysis,pentose phosphate and fatty acid metabolism pathway.The above results indicate acid rain can influence the activity of protease and further influence plant’s physiological processes.Acid rain also lead to other energy-related protein expression increased.Such as ATP synthase,granule-bound starch synthase,malate dehydrogenase et al.It means plant rely on more energy supply by other energy pathway in order to increase resistance to acid rain.Acid rain induce Pinus massoniana secondary metabolites enzyme express change,such as caffeate O-methyltransferase,putative gibberellin 20-oxidase,phenylcoumaran benzylic ether reductase,3-dehydroquinate synthase.At last,the resistance-related proteins are up regulated to resist acid rain stress.Such as peroxiredoxin,Hsp70,monodehydroascorbate reductase.In a word,acid rain stress has a great impact on various plants’ metabolic pathways.In order to ensure the normal growth of the seedling,plant can reduce their energy consumption or activate the other pathway of energy.We also find some stress related proteins are over expression by the supply of calcium.It means that calcium can improve the resistance to aicd rain.更多还原