【作者】 周自云；

【导师】 胡景江；

【作者基本信息】 西北农林科技大学 ， 植物学， 2004， 硕士

【摘要】 利用生物的或者物理、化学的因子处理植株，改变植物对病害的反应，产生局部或系统抗性的现象称为诱导抗病性。能够诱导植物产生抗性的物质叫激发子。诱导抗病性是生物中广泛存在的免疫现象。低聚糖是一类生物活性分子，具有调节植物生长发育、形态发生、果实成熟及抗逆性等作用。自上世纪七十年代中期美国学者Albersheim发现低聚糖诱导植物抗病性的功能以来，糖生物学开始兴起，关于低聚糖诱导植物抗性的研究已经成为当前生物学领域内一大热点。本研究以杨树溃疡病菌菌丝体提取物和甲壳几丁质为原料，制备成三种激发子诱导杨树愈伤组织。通过检测植物材料中的PAL、几丁质酶、β-N-乙酰氨基葡萄糖苷酶、SOD、POD、CAT、PPO活性的变化和HRGP、绿原酸含量的变化，筛选激发子，探讨植物诱导抗病的可能机制。研究表明，用溃疡病菌提取物制备的激发子I，最佳浓度其含糖量为21μg/ml；酸解的溃疡病菌提取物激发子II最佳制备方法为加入激发子10%（v/v）的1mol/L HCl酸解30min；用甲壳几丁质制备激发子III制备方法为将甲壳几丁质以1:30（w/v）的比例加入到2%醋酸溶液中，搅拌使其溶解，然后用H2O2进行氧化降解，降解条件为H2O2浓度3%，用量30%，反应温度75℃，时间3h。对诱导处理愈伤组织保护酶活性的测定发现：经三种激发子处理后，毛白杨愈伤组织中的SOD、POD、CAT、PPO活性均能被有效诱导，其中CAT在诱导处理的2h达到最大活性；SOD在诱导处理的2.5h达到峰值；PPO在诱导处理的3.5h达到最大活性； POD活性在6～12h达到最大。毛白杨愈伤组织经三种激发子处理后，几丁质酶、β-N-乙酰氨基葡萄糖苷酶、HRGP、PAL和绿原酸也都能被有效诱导，其中几丁质酶、β-N-乙酰氨基葡萄糖苷酶活性变化呈现相同规律，在4h达到最大活性随后降低； PAL活性、绿原酸含量变化呈现相同规律，在48h达到最大随后降低；HRGP含量持续增加。更多还原

【Abstract】 The phenomena that a plant shows partial or systemic resistance to some organic, physical or chemical factors calls induction resistance. The factor causing the induction resistance in plants calls elicitor. Induction resistance is an immunization in organism. Oligosaccharide is a kind of biological active molecular that can modulate the growth and development of plants. Albersheim, an American researcher, discovered that oligosaccharides could induce resistance in plants at middle of 1970’s. Since then, it becomes a hot point in biological research field. The Populus tomentosa callus was reduced by three elicitors, which were produced by Dothiorella gregaria and chitin. In order to obtain the best elictor, the changes of PAL, chitinase, β-N-acetyle-D-glucosaminidase, SOD, POD, CAT, PPO, HRGP and chlorogenic acid were tested. The reason that causes the induction resistance in plant was also discussed.The optimal concentration of the elicitor I, which was produced by Dothiorella gregaria, corresponding to 21μg/ml saccharide. The most suitable method for producing elicitor II was hydrolyzing the elicitorⅠin 10%（v/v）1mol/L HCl for 30min. The elicitorⅢ was made from chito-chitin by dissolving the chitin in 1:30（w/v）2% acetic acid and hydrolyzing the solution in 1:0.3(v/v) 3% H2O2 75℃ for 3h.SOD, POD, CAT, and PPO could be induced by the three elicitors. Induced by the three elicitors, the maximum of CAT appeared at 2 hour later, SOD at the 2.5 hour later, PPO at the 3.5 hour later, and POD at the 6～12 hour later.Chitinase, β-N-acetyle-D-glucosaminidase, HRGP, PAL and chlorogenic acid could be induced by the three elicitors. Chitinase and β-N-acetyle-D-glucosaminidase reached the maximum at 4 hour later, PAL and chlorogenic acid reached the maximum at 48 hour later, and the content of HRGP was increasing after inducing.更多还原