低温驯化对海滨斯氏线虫生化物质含量及其生存活力的影响
|
摘要
【目的】本文为明确海滨斯氏线虫Steinernema litorale开发应用的潜能和价值,研究了由本实验室从哈尔滨市香坊区原生态榆树根际土壤中分离到的海滨斯氏线虫的适低温特性。【方法】采用25℃(6 d)、10℃(6 d)、4℃(6 d)和25℃(2 d)-10℃(2 d)-4℃(2 d)处理海滨斯氏线虫,分析该线虫冷冻存活率、不同温度处理后的贮存稳定性以及体内生化物质—可溶性糖、糖原、海藻糖、脂肪与蛋白质含量的影响。【结果】海滨斯氏线虫经过25℃(6 d)、10℃(6 d)、4℃(6 d)和25℃(2 d)-10℃(2 d)-4℃(2 d)低温驯化,于﹣20℃冷冻36 h后的存活率分别为0.9%、23.6%、20.0%和49.2%,经25℃(2 d)-10℃(2 d)-4℃(2 d)处理的线虫冷冻存活率明显高于其它3组,阶段性降温显著提高了线虫耐寒力;将上述4组低温驯化后的海滨斯氏线虫置于4℃贮存7个月,其存活率差异不显著(P<0.05),均为65%以上;25℃(6 d)驯化的线虫杀虫活性显著低于其他3个处理组线虫的杀虫活性。海滨斯氏线虫经25℃(2 d)-10℃(2 d)-4℃(2 d)阶段降温驯化后体内可溶性糖、脂肪、海藻糖含量最高且显著高于25℃(6 d)驯化的线虫体内可溶性糖、脂肪、海藻糖含量;25℃(6 d)驯化的线虫体内蛋白质、糖原含量与其他3个处理组差异不显著。【结论】阶段降温的低温驯化更有利于提高海滨斯氏线虫耐寒能力,并能用于后期长期贮存。
[Objectives] To clarify the potential value of S. litorale as a biological control agent, we studied the appropriate low temperature characteristics of this species in rhizosphere soil from under elms in the Xiangfang district, Harbin city. [Methods] S. litorale were exposed to several temperature treatments, including 25 ℃(6 d), 10 ℃(6 d), 4 ℃(6 d) and 25 ℃(2 d)-10 ℃(2 d)-4 ℃(2 d), and their freezing survival rate, low temperature dormancy, and biochemical(soluble sugar, glycogen, trehalose, lipid and protein) content, measured and compared. [Results] The freezing survival rates of S. litorale were 0.9%, 23.6%, 20.0% and 49.2% after cold acclimation at 25 ℃(6 d), 10 ℃(6 d), 4 ℃(6 d) and 25 ℃(2 d)-10 ℃(2 d)-4 ℃(2 d), respectively, followed by a 36 h-20 ℃ freezing treatment. The freezing survival rates of S. litorale after 25 ℃(2 d)-10 ℃(2 d)-4 ℃(2 d) was significantly higher than those of the other three temperature treatments. Staged cooling significantly improved cold tolerance. The survival rates of nematodes in the four treatment groups were all above 65% and were not significantly different(P < 0.05) after they had been kept at 4 ℃ for 7 months. Mortality in the 25 ℃(6 d) treatment group was significantly lower than that in the other three treatment groups. The soluble sugar, lipid and trehalose contents of the 25 ℃(2 d)-10 ℃(2 d)-4 ℃(2 d) treatment groups were the highest; significantly higher than those of the 25 ℃(6 d) treatment group. The protein and glycogen content of the 25 ℃(6 d) treatment group were not significantly different from those of the other three treatment groups. [Conclusion] Cold acclimation by staged temperature reduction improves the cold tolerance of S. litorale and can be used to maintain these animals in a dormant state for prolonged periods.
[Objectives] To clarify the potential value of S. litorale as a biological control agent, we studied the appropriate low temperature characteristics of this species in rhizosphere soil from under elms in the Xiangfang district, Harbin city. [Methods] S. litorale were exposed to several temperature treatments, including 25 ℃(6 d), 10 ℃(6 d), 4 ℃(6 d) and 25 ℃(2 d)-10 ℃(2 d)-4 ℃(2 d), and their freezing survival rate, low temperature dormancy, and biochemical(soluble sugar, glycogen, trehalose, lipid and protein) content, measured and compared. [Results] The freezing survival rates of S. litorale were 0.9%, 23.6%, 20.0% and 49.2% after cold acclimation at 25 ℃(6 d), 10 ℃(6 d), 4 ℃(6 d) and 25 ℃(2 d)-10 ℃(2 d)-4 ℃(2 d), respectively, followed by a 36 h-20 ℃ freezing treatment. The freezing survival rates of S. litorale after 25 ℃(2 d)-10 ℃(2 d)-4 ℃(2 d) was significantly higher than those of the other three temperature treatments. Staged cooling significantly improved cold tolerance. The survival rates of nematodes in the four treatment groups were all above 65% and were not significantly different(P < 0.05) after they had been kept at 4 ℃ for 7 months. Mortality in the 25 ℃(6 d) treatment group was significantly lower than that in the other three treatment groups. The soluble sugar, lipid and trehalose contents of the 25 ℃(2 d)-10 ℃(2 d)-4 ℃(2 d) treatment groups were the highest; significantly higher than those of the 25 ℃(6 d) treatment group. The protein and glycogen content of the 25 ℃(6 d) treatment group were not significantly different from those of the other three treatment groups. [Conclusion] Cold acclimation by staged temperature reduction improves the cold tolerance of S. litorale and can be used to maintain these animals in a dormant state for prolonged periods.
引文
Ali F,Wharton DA,2009.Cold tolerance of entomopathogenic nematodes.New Zealand Plant Protection,62(1):396.
Ash C,Atkinson HJ,1986.Nematodirus battus:development of cold hardiness in dormant eggs.Experimental Parasitology,62(1):24-28.
Brown IM,Gaugler R,1996.Cold tolerance of Steinernematid and Heterorhabditid nematodes.Journal of Thermal Biology,21(2):115-121.
Dutky SR,Thompson JV,Cantwell GE,1964.A technique for the mass propagation of the DD-136 nematode.Journal of Insect Physiology,6:417-422.
Forge TA,Macguidwin AE,1990.Cold hardening of Meloidogyne hapla second-stage juveniles.Journal of Nematology,22(1):101-105.
Grewal PS,Jagdale GB,2002.Enhanced trehalose accumulation and desiccation survival of entomopathogenic nematodes through cold preacclimation.Biocontrol Science&Technology,12(5):533-545.
Grewal PS,2015.Enhanced ambient storage stability of an entomopathogenic nematode through anhydrobiosis.Pest Management Science,56(5):401-406.
Jagdale GB,Grewal PS,2003.Acclimation of entomopathogenic nematodes to novel temperatures:trehalose accumulation and the acquisition of thermotolerance.International Journal for Parasitology,33(2):145-152.
Jagdale GB,Grewal PS,Salminen SO,2005.Both heat-shock and cold-shock influence trehalose metabolism in an entomopathogenic nematode.Journal of Parasitology,91(5):988-994.
Li CJ,Tan GZ,Wang Y,Xu YL,2014.Optimizing freezing storage conditions for the entomopathogenic nematode isolated from the cold region in China.Plant Protection,40(5):49-54.[李春杰,谭国忠,王义,许艳丽,2014.寒区昆虫病原线虫冷冻储存条件优化.植物保护,40(5):49-54.]
Li CJ,Wang LF,Pan FJ,Xu YL,2015.Effect of pretreatment method on freezing survival of entomopathogenic nematodes.Journal of Northwest A&F University(Natural Science Edition),43(2):198-202.[李春杰,王丽芳,潘凤娟,许艳丽,2015.预处理方式对昆虫病原线虫冷冻存活的影响.西北农林科技大学学报:自然科学版,43(2):198-202.]
Liu R,Liu QZ,2016.Study on the biological characteristics of Steinernema litorale.Chinese Journal of Biological Control,32(1):101-105.[刘瑞,刘奇志,2016.海滨斯氏线虫生物学特性研究.中国生物防治学报,32(1):101-105.]
Lv Y,Han LL,Zhao KJ,Song J,Zhu MH,2014.Isolation and identification of entomopathogenic nematodes and their symbiotic bacteria from Harbin(Northeast China)and infectivity of entomopathogenic nematodes in low temperature.Chinese Journal of Biological Control,30(4):558-563.[吕洋,韩岚岚,赵奎军,宋捷,朱明贺,2014.哈尔滨两种斯氏线虫及其共生菌的分离鉴定和低温侵染力分析.中国生物防治学报,30(4):558-563.]
Patricia M,Scott AIH,Gregor G,Andrew YG,Andrew R,2007.An explicit test of the phospholipid saturation hypothesis of acquired cold tolerance in Caenorhabditis elegans.Proceedings of the National Academy of Sciences of the United States of America,104(13):5489-5494.
Qian XJ,2014.Entomological nematodes resources of Gansu province and its adaptation to abiotic stress.Doctoral dissertation.Gansu:Gansu Agricultural University.[钱秀娟,2014.甘肃省昆虫病原线虫资源及其对非生物胁迫的适应性研究.博士学位论文.甘肃:甘肃农业大学.]
Smith HJ,1987.Factors affecting preconditioning of Trichinella spiralis nativa larvae in musculature to low temperatures.Canadian Journal of Veterinary Research,51(2):169-173.
Wang LF,Xu YL,Li CJ,2011.The research of entomopathogenic nematodes cold tolerance.System Sciences and Comprehensive Studies in Agriculture,27(4):490-494.[王丽芳,许艳丽,李春杰,2011.昆虫病原线虫耐寒性研究进展.农业系统科学与综合研究,27(4):490-494.]
Watanabe M,2006.Anhydrobiosis in invertebrates.Applied Entomology&Zoology,41(1):15-31.
Wharton DA,Judge KF,Worland MR,2000.Cold acclimation and cryoprotectants in a freeze-tolerant Antarctic nematode,Panagrolaimus davidi.Journal of Comparative Physiology B,170(4):321-327.
Wharton DA,Goodall G,Marshall CJ,2003.Freezing survival and cryoprotective dehydration as cold tolerance mechanisms in the Antarctic nematode Panagrolaimus davidi.Journal of Experimental Biology,206(2):215-221.
White GF,1927.A Method for obtaining infective nematode larvae from cultures.Science,66(1709):302-303.
Yoshida M,2004.Steinernema litorale n.sp.(Rhabditida:Steinernematidae),a new entomopathogenic nematode from Japan.Nematology,6(6):819-838.
Zhang AB,Nan Gong ZY,Song P,Wang QY,2015.Eficacy of entomopathogenic nematodes against Agrotis segetum larvae.Journal of Environmental Entomology,37(3):591-597.[张安邦,南宫自艳,宋萍,王勤英,2015.昆虫病原线虫对黄地老虎致病力的研究.环境昆虫学报,37(3):591-597.]
Ash C,Atkinson HJ,1986.Nematodirus battus:development of cold hardiness in dormant eggs.Experimental Parasitology,62(1):24-28.
Brown IM,Gaugler R,1996.Cold tolerance of Steinernematid and Heterorhabditid nematodes.Journal of Thermal Biology,21(2):115-121.
Dutky SR,Thompson JV,Cantwell GE,1964.A technique for the mass propagation of the DD-136 nematode.Journal of Insect Physiology,6:417-422.
Forge TA,Macguidwin AE,1990.Cold hardening of Meloidogyne hapla second-stage juveniles.Journal of Nematology,22(1):101-105.
Grewal PS,Jagdale GB,2002.Enhanced trehalose accumulation and desiccation survival of entomopathogenic nematodes through cold preacclimation.Biocontrol Science&Technology,12(5):533-545.
Grewal PS,2015.Enhanced ambient storage stability of an entomopathogenic nematode through anhydrobiosis.Pest Management Science,56(5):401-406.
Jagdale GB,Grewal PS,2003.Acclimation of entomopathogenic nematodes to novel temperatures:trehalose accumulation and the acquisition of thermotolerance.International Journal for Parasitology,33(2):145-152.
Jagdale GB,Grewal PS,Salminen SO,2005.Both heat-shock and cold-shock influence trehalose metabolism in an entomopathogenic nematode.Journal of Parasitology,91(5):988-994.
Li CJ,Tan GZ,Wang Y,Xu YL,2014.Optimizing freezing storage conditions for the entomopathogenic nematode isolated from the cold region in China.Plant Protection,40(5):49-54.[李春杰,谭国忠,王义,许艳丽,2014.寒区昆虫病原线虫冷冻储存条件优化.植物保护,40(5):49-54.]
Li CJ,Wang LF,Pan FJ,Xu YL,2015.Effect of pretreatment method on freezing survival of entomopathogenic nematodes.Journal of Northwest A&F University(Natural Science Edition),43(2):198-202.[李春杰,王丽芳,潘凤娟,许艳丽,2015.预处理方式对昆虫病原线虫冷冻存活的影响.西北农林科技大学学报:自然科学版,43(2):198-202.]
Liu R,Liu QZ,2016.Study on the biological characteristics of Steinernema litorale.Chinese Journal of Biological Control,32(1):101-105.[刘瑞,刘奇志,2016.海滨斯氏线虫生物学特性研究.中国生物防治学报,32(1):101-105.]
Lv Y,Han LL,Zhao KJ,Song J,Zhu MH,2014.Isolation and identification of entomopathogenic nematodes and their symbiotic bacteria from Harbin(Northeast China)and infectivity of entomopathogenic nematodes in low temperature.Chinese Journal of Biological Control,30(4):558-563.[吕洋,韩岚岚,赵奎军,宋捷,朱明贺,2014.哈尔滨两种斯氏线虫及其共生菌的分离鉴定和低温侵染力分析.中国生物防治学报,30(4):558-563.]
Patricia M,Scott AIH,Gregor G,Andrew YG,Andrew R,2007.An explicit test of the phospholipid saturation hypothesis of acquired cold tolerance in Caenorhabditis elegans.Proceedings of the National Academy of Sciences of the United States of America,104(13):5489-5494.
Qian XJ,2014.Entomological nematodes resources of Gansu province and its adaptation to abiotic stress.Doctoral dissertation.Gansu:Gansu Agricultural University.[钱秀娟,2014.甘肃省昆虫病原线虫资源及其对非生物胁迫的适应性研究.博士学位论文.甘肃:甘肃农业大学.]
Smith HJ,1987.Factors affecting preconditioning of Trichinella spiralis nativa larvae in musculature to low temperatures.Canadian Journal of Veterinary Research,51(2):169-173.
Wang LF,Xu YL,Li CJ,2011.The research of entomopathogenic nematodes cold tolerance.System Sciences and Comprehensive Studies in Agriculture,27(4):490-494.[王丽芳,许艳丽,李春杰,2011.昆虫病原线虫耐寒性研究进展.农业系统科学与综合研究,27(4):490-494.]
Watanabe M,2006.Anhydrobiosis in invertebrates.Applied Entomology&Zoology,41(1):15-31.
Wharton DA,Judge KF,Worland MR,2000.Cold acclimation and cryoprotectants in a freeze-tolerant Antarctic nematode,Panagrolaimus davidi.Journal of Comparative Physiology B,170(4):321-327.
Wharton DA,Goodall G,Marshall CJ,2003.Freezing survival and cryoprotective dehydration as cold tolerance mechanisms in the Antarctic nematode Panagrolaimus davidi.Journal of Experimental Biology,206(2):215-221.
White GF,1927.A Method for obtaining infective nematode larvae from cultures.Science,66(1709):302-303.
Yoshida M,2004.Steinernema litorale n.sp.(Rhabditida:Steinernematidae),a new entomopathogenic nematode from Japan.Nematology,6(6):819-838.
Zhang AB,Nan Gong ZY,Song P,Wang QY,2015.Eficacy of entomopathogenic nematodes against Agrotis segetum larvae.Journal of Environmental Entomology,37(3):591-597.[张安邦,南宫自艳,宋萍,王勤英,2015.昆虫病原线虫对黄地老虎致病力的研究.环境昆虫学报,37(3):591-597.]