【作者】 张楠；

【导师】 孙治强；

【作者基本信息】 河南农业大学 ， 设施栽培， 2008， 硕士

【摘要】 植物寄生线虫已成为一类重要病原物,线虫病害已成为农、林生产上的重要问题,国内外均非常重视寻找合适、高效、无污染的控制方法,越来越多的植物源物质被发现有杀线活性。本文以丰富的大豆子实脱壳后的副产品为材料,提取、纯化大豆荚壳中具杀线活性的成分,并进行杀线活性测定和杀线机理研究,旨在开发新型的能有效防治根结线虫的杀线乳剂。试验结果如下:1.用不同溶剂对类黄酮进行提取,以75 %乙醇对类黄酮提取率最高,可达5.60 %,且室内抑杀根结线虫活性较强;以乙醇为溶剂进一步对影响提取率4因素做正交试验,得到最佳提取工艺条件为:温度70℃,10倍量80 %乙醇回流提取24 h。此条件下类黄酮的提取率为8.58 %,显著高于次佳工艺。2.通过液-液分配萃取法对大豆荚壳中的类黄酮活性成分进行初步分离,发现其主要分布于氯仿、乙酸乙酯、正丁醇和水层萃取物中。其中,在正丁醇层中含量最高,处理24、48、72 h后,线虫的校正死亡率均可达50 %以上;通过柱层析和薄层层析法对正丁醇萃取层做进一步分离、纯化,合并Rf值得到8种组分,其组分2和7的杀线活性较高,处理24、48、72 h后,线虫的校正死亡率均可达50 %以上。3.对进一步分离所得的杀线效果较好的组分2和7进行杀线机理的研究,结果表明:处理24、48h后,其对线虫体内的蛋白质含量和糖含量均有一定的抑制作用,其含量显著低于对照,线虫体内谷胱甘肽转硫酶的比活力也显著低于对照。处理24 h后,线虫体内过氧化氢酶的比活力与对照差异不显著,但处理48 h后,显著低于对照。这就说明:组分2和7中的杀线活性成分可能阻碍了虫体的新陈代谢,导致线虫体内氧化和抗氧化作用失衡,致使线虫死亡。且活性成分可能影响线虫虫体角质层的形层,导致虫体内含物外渗,也是导致线虫死亡一个原因。4.确定了大豆荚壳乳油的最佳配方为大豆荚壳浸膏(15%)+95%乙醇(65%)+二甲苯(10%)+ OP-10(10%),进行杀线活性测定的结果表明:200倍液室内杀线效果最好;而100倍液的盆栽、大田杀线效果最明显,且乳油质量检测结果表明该乳油符合农药商品的要求。更多还原

【Abstract】 Plant-parasitic nematodes had become an important kind of pathogenyt and it is a problem for agriculture and forestry now.All over the world want to find some efficient and environment friendly means to control it. More and more plants were found to have nematicidal activity.This experiment extracted and purificated the nematicidal fractions form the soybean hull and studied the nematicidal activity of soybean hull and the mechanism by choosing the abundant byproduct of soybean as material.The results are as follows:1. Extracting flavonoids by different solvents , the extractive ratio was highest by 75% ethanol,it was 5.60%. and its nematicidal activity was strongest. Choosed the ethanol as solvent and did orthogonal plan for 4 facters which could effect the extractive ratio.The optimium technics was 70℃, 10:1 of 80% ethanol to soybean hull, heating and refluxing for 24 hours. The extractive ratio of optimium technics could be 8.58% and it was higher than Hypo-optimal technics significantly.2. Preliminary extracted the fractions of nematicidal activity by liquid-liquid. There were most flavonoids form soybean hull in the four fractions such as chloroform, ethyl acetate,n-butanol and water.N-butanol had most,the revised mortality were above 50% after 24 hours ,48 hours and72 hours. Ulteriorly extracted the fractions of nematicidal activity by column chromatography and TLC and amalgamated the Rf of the sample of n-butanol into 8 fractions.Fraction 2 and fraction7 had high nematicidal effect, the revised mortality were above 50% after 24 hours,48 hours and72 hours.3. Study the mechanism of fraction 2 and fraction7, the results indicated that: it could inhibit the protein and sugar and them were lower than CK evidently.The Specific activity of GST were lower than CK evidently after 24 hours and 48 hours. The specific activity of CAT were lower than CK not evidently after 24 hours but lower than CK evidently after 48 hours. It showed:the nematicidal activity of fraction 2 and fraction7 could baffle it’s metabolism and unbalance their oxidation and antioxygenation and make them to die.And the nematicidal activity may effect the cuticle of nematodes, make the substance of nematodes extravasating,it could make them to die too. 4. Choosed the optimium condition was soybean hull condensate(15%),95% of ethanol(65%),xylene(10%) and of OP-10(10%). The nematicidal result saied:the 200 times was best in room and the 100 times was best in land. Did some detection for the EC and the result saied:the EC accords with the pesticide.更多还原