【作者】 白海群；

【导师】 刘华山；

【作者基本信息】 河南农业大学 ， 作物栽培学与耕作学， 2009， 硕士

【摘要】 本实验通过盆栽、室内和大田试验,从生理生化指标、超微结构变化、体外钝化和烟叶生长等几方面,研究了不同植物提取液对烟草普通花叶病的疗效和对烟叶品质的影响。结果如下:1.植物提取液对感染TMV烟株叶片叶绿素、保护酶活性和丙二醛(MDA)含量及的影响感染TMV烟株叶片经植物提取液处理后,均不同程度的提高了叶片抗氧化酶活性,降低了膜脂过氧化程度。经过植物提取液处理的烟叶片中叶绿素和类黄酮含量与发病对照相比明显增加,SOD、CAT和POD活性与发病对照相比明显提高,MDA含量与发病对照相比有所降低。说明植物提取液可以通过增加叶片叶绿素含量、提高SOD、CAT和POD活性、降低MDA含量来增强植物的抗病性,与健康株相比,提取液处理后的烟株叶片生理生化指标均达到或接近健康株。各种提取液的效果为:配伍制剂>莲>白花蛇舌草。2.植物提取液对感染烟草叶片总酚、类黄酮和核酸含量的影响喷施植物提取液后能够提高感病烟株体内总酚和类黄酮含量,在15d时喷施配伍的制剂分别提高了16%和76%,莲提高了14%和53%,白花蛇舌草提高了9%和52%。另外,喷施植物提取液能够提高感病株的DNA含量,降低RNA含量,在第20d时,喷施配伍制剂、莲和白花蛇舌草叶片内DNA比阳性对照分别提高了15%、11%和11%,RNA降低了32%、29%和18%。3.植物提取液对TMV体外活性的影响植物提取液对TMV有强烈的钝化作用,且以配伍制剂最好,在3h时抑制率可达到92.13%,而白花蛇舌草只有50.77%;并且,作用时间越长,抑制效果越显著,在1h时,配伍制剂水提取的抑制率是50%,而在3h时,达到92.13%;水提取法对TMV的抑制作用略好于乙醇提取法,在3h时,白花蛇舌草水提取为60.61%,而白花蛇舌草乙醇提取为50.77%。但经透析处理后,致病性有所提高。电镜观察发现,在体外钝化过程中植物提取液能使病毒粒体断裂和凝集,配伍制剂的作用效果好于莲荷白花蛇舌草;作用时间越长,病毒粒子破坏越严重。植物提取液对TMV的增殖也有抑制作用,48h后制剂水提取液的抑制率为38.4%。4.植物提取液对感染烟草叶片超微结构的影响利用透射电镜研究了植物提取液处理的烟株在细胞和亚细胞水平上的变化,处理烟株的叶片细胞中病毒粒体和病毒结晶体显著减少,由病毒侵染而发生的细胞破坏程度显著减轻,薄壁细胞中叶绿体等各种细胞器基本完好,仅少数细胞有轻度病变。5.植物提取液对感染烟草叶片病程相关蛋白(PR)的影响电泳实验结果表明,经提取液处理的烟株叶片,产生新的病程相关蛋白量有差异。除了一些共有的蛋白相继产生和消失外,还产生一些各自特有的新蛋白,其中在处理后的第5d,制剂多出Rf值分别为:0.219、0.31、0.407、0.422和0.434的几条带,而少了Rf值分别为:0.659、0.743和0.951。在处理后的第20d,病程相关蛋白数量明显减少,接近健康株。6.植物提取液对烟叶品质的影响烟叶化学成分检测结果表明,配伍制剂处理能协调烟叶的化学成分,改善烟叶品质。钾、总糖和还原糖含量分别提高了29.71%、9.39%和4.86%,而烟碱、总氮和氯分别降低了18.23%、9.04%和37.02%,其中钾/氯、总糖/烟碱、总氮/烟碱的比值分别为5.12、13.36和1.05,均处于比较适宜的范围。7.植物提取液对烟苗生长的影响在苗床喷施配伍制剂能使烟苗鲜重,干重增加,尤其是根的鲜重和干重明显增加,并且发叶速度快,在育苗后70天调查,处理的叶数、根的鲜重和干重分别是7.25片/株、0.143克/株和0.008克/株,而对照为6.5片/株、0.113克/株和0.005克/株。更多还原

【Abstract】 On indicators of physiological and biochemical and vitro passivation and quality of tobacco seedlings,pots and indoor and field experiments were conducted to study the effects of extracts from different plant and inhibitor on tobacco mosaic virus . The f result are as followeds :1. The effects of plant extracts on Chlorophyll and MDA contents and the activity of antioxidant enzymesAfter being treated by plant extracts,the protecting enzymes activities in leaves infected with TMV were all increased in some content, lipid peroxidation levels were decreased. he content of Chlorophyll, MDA, Flavonoid, SODand POD in Nicotiana tabacum was investigated. The results indicated that the compound could remarkedly increase the content of chlorophyll and flavonoid compared with check, enhances the activity of SODand POD compared with check, and the content of MDA was remarkably reduced than check. Explained that the compound. A may through increase the content of chlorophyll and flavonoid, enhances the activity of SOD, PPOand POD, and reduces the content of MDA of lamina to strengthen the disease resistance of Nicotiana tabacum. compared with the healthy plant, the physiologic and biochemical indexes in leaves induced by extracts and inhibitor were nearly or reached to it. the integrated effect all of the agents were that: Compound preparation>Nelum bonucifera Garetu>Oldenlan diadiffusa.2. The effects of plant extracts on total polyphenol、flavonoid and nucleic acid contents of flue-cured with TMVAfter being treated by Compound preparation,contents of total polyphenol and flavonoid in tobacc leaves increase 16% and 76%,and 14% and 53% by Nelum bonucifera Garetu, and 9% and 52% by Oldenlan diadiffusa on 15d.Other ,contents of DNA increase,but contents of RNA decrease by plant extracts, contents of DNA increase 15%、11% and 11%, contents of RNA decrease 32%、29% and 18% by Compound preparation、Nelum bonucifera Garetu and Oldenlan diadiffusa on 20d.3. The effects of plant extracts on inactivating TMV in vitro.extracts from plants had strong effects to inactivating TMV in vitro. pathogenecity of purified TMV particles was greatly decreased when mixed with extracts after three hour.As revealed by electron microscopy,virus particles were broken to small pieces and cohesion.other, extracts from plants had effects to inhibiting on multiplication of TMV , the inhibiting rate of Compound preparation by water was 38.4% after fourty eight hour .4. The effects of plant extracts on ultrastructure in tobacco leave cell with TMVThe cytological and ultrastructural changes in extracts treated plants infected by TMV.The virus particle and virus crystal were reduced significantly in mesophyll cell.The pathological change caused by virus infection in cells,was weakened greatly.Chloroplasts and other organelles in most parenchyma cells remained normal,only a few of them were slightly distorted. 5. The effects of plant extracts on PRs in tobacco leave cell with TMVThe result based on routine PAGE showed that, There are new Pathogenesis-related proteins(PR) in the leaves treated with extracts and inhibitor. On the 5th day after treatment, there were a lot of PRs , Its Rf are 0.219、0.31、0.407、0.422and0.434,but lossing of PRs that Rf are 0.659、0.743 and 0.951。On 20th day after treatment, the number of PR was decrease Obviously, closeing to the health of tobacco.6. The effects of plant extracts on chemistry ingredient of flue-cured tobaccoThe chemistry ingredient in tobacco leaf showed that, the quality of leaf disposed by inhibitor was more harmonized and content was in reason, the usability of leaf is increased obviously. the content of K2O、total sugar and deoxidize sugar were increased 29.71%、9.39%and4.86% to contrast, and nicotine、total nitrogen and Cl were diseased 18.23%、9.04% and 37.02%to contrast;and K2O/ Cl、total sugar/nicotine and total nitrogen/ nicotine are 5.12、13.36 and 1.05.7. The effects of Compound preparation on t tobacco seedling grothBeing treated tobacco seedling with extracts,fresh weight and dry weight were increased, fresh weight and dry weight of root were increased especially. Investigating after growing seedlings on 70d, leaves、fresh weight and dry weight of root are 7.25 tablet /strain、0.143 g/strain and 0.008 g/strain,and contrast are 6.5 tablet /strain、0.113 g/strain and 0.005 g/strain.更多还原