几丁质对南方根结线虫的生物防治研究

青岛农业大学

毕业论文（设计）

题目：刀孢蜡蚧菌Lecanicillium psalloitae CFCC84958产生的几丁质酶对南方根结线

虫卵的破坏作用

姓名： XXXXX

学院：农学与植物保护学院

专业：植物保护

班级： XXXX 学号： XXXXX

2011 年 5 月 20 日

目录

中文摘要 (1)

Abstrac t (2)

引言 (3)

1材料与方法 (6)

1.1供试菌株 (6)

1.2供试试剂 (6)

1.3产几丁质酶培养条件 (6)

1.4几丁质酶活性测定 (6)

1.4.1几丁质降解酶系活性测定 (6)

1.4.2 N-乙酰葡萄糖胺（NAG）标准曲线 (7)

1.4.3几丁质外切酶活性测定 (7)

1.4.4对硝基苯酚（pNP）标准曲线 (7)

1.5刀孢蜡蚧菌产生的几丁质酶对根结线虫卵的破坏 (8)

1.5.1根结线虫卵悬液的制备 (8)

1.5.2卵孵化抑制率及破坏率的测定 (8)

2结果与分析 (9)

2.1几丁质降解酶系活性测定的结果 (9)

2.2几丁质外切酶活性测定的结果 (9)

2.3刀孢蜡蚧菌培养滤液对卵孵化的影响 (10)

2.4南方根结线虫卵壳的形态变化 (11)

2.5结论 (12)

3讨论与展望 (13)

3.1讨论 (13)

3.2展望 (13)

3.2.1安全性评价 (13)

3.2.2根结线虫生物防治展望 (13)

致谢 (15)

参考文献 (16)

刀孢蜡蚧菌Lecanicillium psalloitae CFCC84958产生的几丁质酶对南方根结线虫卵的破坏作用

植物保护专业 XXXX

指导教师 XXX

摘要：根结线虫卵壳主要是由几丁质和蛋白质构成。根结线虫卵寄生或产毒真菌产生几丁质酶活性是评价食线虫菌物生物防治潜力的重要生化指标之一。本研究分别采用根结线虫卵寄生真菌刀孢蜡蚧菌，测定其产生的几丁质降解酶活性，该菌的培养滤液从第2d具有几丁质降解酶系活性，第4-6d就达到酶活性高峰值3.98 μmol /h mL；从第2d开始就产生几丁质外切酶，第4d酶活性为0.26umol/h mL。表明刀孢蜡蚧菌CFCC84958菌株具有较高产生几丁质酶的活性。该菌4d的培养滤液对根结线虫卵孵化和破坏作用随浓度的增加而增强，7d后孵化抑制率和破坏率分别为94.52%和84.32%。显微观察线虫卵壳变形和破坏情况的结果表明，刀孢蜡蚧菌CFCC84958产生的几丁质酶可以引致根结线虫卵壳的裂解，抑制根结线虫卵的孵化。关键词：根结线虫卵；刀孢蜡蚧菌；几丁质酶；卵破坏；卵孵化

The Destructive Effect of Chitinase Activities produced by Lecanicillium psalloitae CFCC84958 on Meloidogyne incognita

eggs

Student majoring in Plant Protection XXXX

Tutor XXX

Abstract: The eggshells of Meloidogyne spp. are mainly composed of chitin and protein. The chitinase activity of fungi which parasitize eggs or produce toxins for Meloidogyne spp. is one of the most important biochemical characters. It is used for evaluating the biocontrol potential of nematophagous fungi. The endochitinase and exodchitinase of Lecanicillium psalloitae CFCC84958were tested with NAG andpNP.The maximum activity of endochitinase was 3.98 μmol/h ml during4d-6d and the activity of exodchitinase was 0.26μmol/h ml at the 4 d. The results showed that L.psalloitae CFCC84958 had high chitinase activity. After 7 days of incubation with the chitinase produced by L.psalloitae CFCC84958, rates of hatching inhibitory and destruction of eg gs achieved 94.52% and 84.32%, respectively. Microscopic observations demonstrated that the eggshell of Meloidogyne spp. was deformed and destroyed. The reseach indicated that chitinase produced by L. psalloitae CFCC84958 caused the lysis of Meloidogyne spp. eggshells and resulted in the inhibition of egg hatching i n vitro.

Key words : Meloidogyne spp. ; Lecanicillium psalloitae; Chitinases；egg destruction; egg hatching