伊氏杀线真菌与苏云金芽孢杆菌对松材线虫的联合毒力研究
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摘要
[目的]探究伊氏杀线真菌与苏云金芽孢杆菌(Bacillus thuringiensis,Bt)对松材线虫的联合毒力。[方法]使用Esteya vermicola孢子悬浮液与Bt发酵液处理松材线虫,通过线虫的形态变化、死亡率及死亡速度3个方面测定E.vermicola与Bt联合对松材线虫的影响。[结果]经过E.vermicola孢子悬浮液处理过的线虫会出现内容物渗漏,体腔收缩、弯曲、断裂的现象;高浓度的E.vermicola孢子悬浮液与Bt发酵液联合处理松材线虫可以明显地提高线虫的死亡率,最高可以达到100%;联合处理的线虫死亡速度较快,随着处理时间的延长,死亡率会一直呈上升的趋势。[结论]应用这两种生防微生物在合适的浓度下混配能够在较短时间内提高松材线虫的死亡率。
[Objective] To explore the toxicities of Esteya vermicola in combination with Bacillus thuringiensis(Bt) against Bursaphelenchus xylophilus. [Method] The joint toxicities of E. vermicola and Bt against pine wood nematodes was measured by the morphological changes, mortality and death rate of nematodes treated with E. vermicola spore suspension and Bt fermentation supernatant. [Result] The results showed that the nematodes treated by E. vermicola spore suspension could cause leakage of body contents and shrinkage, bending and fracture of body cavity. The combination of high concentration of E. vermicola spore suspension and Bt fermentation solution could significantly increase the mortality of nematodes, with a maximum of 100%. The combined treatment of pine wood nematodes died faster, and the mortality rate would continue to rise with the extension of treatment time. [Conclusion] Application of these two bio-control agents at the right concentrations can increase the mortality of pine wood nematodes in a relatively short time.
[Objective] To explore the toxicities of Esteya vermicola in combination with Bacillus thuringiensis(Bt) against Bursaphelenchus xylophilus. [Method] The joint toxicities of E. vermicola and Bt against pine wood nematodes was measured by the morphological changes, mortality and death rate of nematodes treated with E. vermicola spore suspension and Bt fermentation supernatant. [Result] The results showed that the nematodes treated by E. vermicola spore suspension could cause leakage of body contents and shrinkage, bending and fracture of body cavity. The combination of high concentration of E. vermicola spore suspension and Bt fermentation solution could significantly increase the mortality of nematodes, with a maximum of 100%. The combined treatment of pine wood nematodes died faster, and the mortality rate would continue to rise with the extension of treatment time. [Conclusion] Application of these two bio-control agents at the right concentrations can increase the mortality of pine wood nematodes in a relatively short time.
引文
[1] Dwinell L D. The pinewood nematode: regulation and mitigation[J]. Annual Review of Phytopathology, 1997, 35:153-166.
[2] Prasa S S S V, Tilak K V B R,Gollakota K G. Role of Bacillus thuringiensis var. thuringiensis on the larval survivability and egg hatching of Meloidogyne spp., the causative agent of root knot disease [J]. Journal of Invertebrate Pathology, 1972, 20(3): 377-378.
[3] Wei J Z, Hale K, Carta L, et al. Bacillus thuringiensis crystal proteins that target nematodes [J]. Proceedings of the National Academy of Science of the United States of America, 2003, 100(5): 2760-2765.
[4] 罗兰, 谢丙炎, 杨宇红, 等. 具杀线虫活性的苏云金杆菌筛选研究[J]. 植物病理学报, 2007, 37(3): 314-316.
[5] Niu Q H, Huang X W, Zhang L, et al. A Trojan horse mechanism of bacterial pathogenesis against nematodes[J]. Proceedings of the National Academy of Science of the United States of America, 2010, 107 (38): 16631-16636.
[6] 彭双, 闫淑珍, 陈双林. 具杀线虫活性植物内寄生细菌的筛选和活性产物[J]. 微生物学报, 2011, 51(3): 368-376.
[7] Liou J Y, Shih J Y, d Tzena S S. Esteya, a new nematophagous genus from Taiwan, attacking the pinewood nematode (Bursaphelenchus xylophilus)[J]. Mycological Research, 1999, 103(2): 242-248.
[8] 杜婷, 张永安, 王玉珠, 等. 内寄生真菌Esteya vermicola对松材线虫侵染活力的测定[J]. 林业科学研究, 2014, 27(2): 174-178.
[9] Wang Z, Zhang Y A, Wang C Y, et al. Esteya vermicola controls the pinewood nematode, Bursaphelenchus xylophilus, in pine seedlings[J]. Journal of Invertebrate Pathology, 1972, 20(3): 377-378.
[10] 王修清, 王倩, 王亚萍, 等. 杀线虫真菌Sr18发酵液对松材线虫超微结构的影响[J]. 北京林业大学学报, 2017, 39(7) : 69-75.
[11] Viglierchio D R, Schmitt R V. On the methodology of nematode extraction from field samples: Baermann funnel modifications[J]. Journal of Nematology, 1983, 15(3): 438-444.
[12] 杨宝君, 潘宏阳, 汤坚,等. 松材线虫病 [M]. 北京: 中国林业出版社, 2003: 141-148.
[13] Guo Q Q, Du G C, Qi H T, et al. A nematicidal tannin from Punica granatum L.rind and its physiological effect on pine wood nematode (Bursaphelenchus xylophilus)[J]. Pesticide Biochemistry Physiology, 2017, 135: 64-68.
[14] Puntener W. Manual For field Trials in Plant Protection[M].Second ed. Ciba-Geigy Limited, Basle, 1981.
[15] Wang Y B, Wang C Y, Wang Z, et al. Laboratory studies on the development of a conidial formulation of Esteya vermicola[J]. Biocontrol Science and Technology, 2012, 22(11): 1362-1372.
[16] Wang C Y, Fang Z M, Wang Z, et al. Biological control of the pinewood nematode Bursaphelenchus xylophilus by application of the endoparasitic fungi Esteya vermicola[J]. Biological Control, 2011, 56: 91-100.
[17] Wang Z, Wang C Y, Yang Z H, et al. Viability and pathogenicity of Esteya vermicola in pine trees[J]. Biocontrol Science and Technology, 2011, 21(4):387-393.
[2] Prasa S S S V, Tilak K V B R,Gollakota K G. Role of Bacillus thuringiensis var. thuringiensis on the larval survivability and egg hatching of Meloidogyne spp., the causative agent of root knot disease [J]. Journal of Invertebrate Pathology, 1972, 20(3): 377-378.
[3] Wei J Z, Hale K, Carta L, et al. Bacillus thuringiensis crystal proteins that target nematodes [J]. Proceedings of the National Academy of Science of the United States of America, 2003, 100(5): 2760-2765.
[4] 罗兰, 谢丙炎, 杨宇红, 等. 具杀线虫活性的苏云金杆菌筛选研究[J]. 植物病理学报, 2007, 37(3): 314-316.
[5] Niu Q H, Huang X W, Zhang L, et al. A Trojan horse mechanism of bacterial pathogenesis against nematodes[J]. Proceedings of the National Academy of Science of the United States of America, 2010, 107 (38): 16631-16636.
[6] 彭双, 闫淑珍, 陈双林. 具杀线虫活性植物内寄生细菌的筛选和活性产物[J]. 微生物学报, 2011, 51(3): 368-376.
[7] Liou J Y, Shih J Y, d Tzena S S. Esteya, a new nematophagous genus from Taiwan, attacking the pinewood nematode (Bursaphelenchus xylophilus)[J]. Mycological Research, 1999, 103(2): 242-248.
[8] 杜婷, 张永安, 王玉珠, 等. 内寄生真菌Esteya vermicola对松材线虫侵染活力的测定[J]. 林业科学研究, 2014, 27(2): 174-178.
[9] Wang Z, Zhang Y A, Wang C Y, et al. Esteya vermicola controls the pinewood nematode, Bursaphelenchus xylophilus, in pine seedlings[J]. Journal of Invertebrate Pathology, 1972, 20(3): 377-378.
[10] 王修清, 王倩, 王亚萍, 等. 杀线虫真菌Sr18发酵液对松材线虫超微结构的影响[J]. 北京林业大学学报, 2017, 39(7) : 69-75.
[11] Viglierchio D R, Schmitt R V. On the methodology of nematode extraction from field samples: Baermann funnel modifications[J]. Journal of Nematology, 1983, 15(3): 438-444.
[12] 杨宝君, 潘宏阳, 汤坚,等. 松材线虫病 [M]. 北京: 中国林业出版社, 2003: 141-148.
[13] Guo Q Q, Du G C, Qi H T, et al. A nematicidal tannin from Punica granatum L.rind and its physiological effect on pine wood nematode (Bursaphelenchus xylophilus)[J]. Pesticide Biochemistry Physiology, 2017, 135: 64-68.
[14] Puntener W. Manual For field Trials in Plant Protection[M].Second ed. Ciba-Geigy Limited, Basle, 1981.
[15] Wang Y B, Wang C Y, Wang Z, et al. Laboratory studies on the development of a conidial formulation of Esteya vermicola[J]. Biocontrol Science and Technology, 2012, 22(11): 1362-1372.
[16] Wang C Y, Fang Z M, Wang Z, et al. Biological control of the pinewood nematode Bursaphelenchus xylophilus by application of the endoparasitic fungi Esteya vermicola[J]. Biological Control, 2011, 56: 91-100.
[17] Wang Z, Wang C Y, Yang Z H, et al. Viability and pathogenicity of Esteya vermicola in pine trees[J]. Biocontrol Science and Technology, 2011, 21(4):387-393.