生物氧化是生物化学代谢部分的重要内容,因其复杂性和抽象性不易被学生理解。采用非变性聚丙烯酰胺凝胶电泳(Native polyacrylamide gel electrophoresis, Native-PAGE)构建了适用于本科实验教学的“线粒体复合体酶I活性方法”。从锥虫...生物氧化是生物化学代谢部分的重要内容,因其复杂性和抽象性不易被学生理解。采用非变性聚丙烯酰胺凝胶电泳(Native polyacrylamide gel electrophoresis, Native-PAGE)构建了适用于本科实验教学的“线粒体复合体酶I活性方法”。从锥虫中提取线粒体粗体物,以10μg/孔样品蛋白上样量,进行非变性聚丙烯酰胺凝胶电泳, NADH脱氢酶酶促反应时间为30分钟,可得到分辨率高、条带清晰的特异性酶活性条带。将Native-PAGE方法应用到本科生化实验中,可有效拓展教学内容,有助于教学紧密联系科研,对本科生化实验教学的开展有借鉴意义。展开更多
Tree endophytic fungi play an important role in reducing insect herbivory,either by repelling them or kill-ing them directly.Identifying which fungi show such activ-ity could lead to new environmentally friendly pesti...Tree endophytic fungi play an important role in reducing insect herbivory,either by repelling them or kill-ing them directly.Identifying which fungi show such activ-ity could lead to new environmentally friendly pesticides.In this study,the Mediterranean basin climate conditions are projected to harshen in the next decades,will increase vulnerability of tree species to pest invasions.Endophytic fungi were isolated from wood and leaves of Quercus pyr-enaica,Q.ilex and Q.suber and tested for virulence against adults of the mealworm beetle,Tenebrio molitor L.using a direct contact method.Only 3 of 111 sporulating isolates had entomopathogenic activity,all identified as Lecanicillium lecanii.The pathogenicity of L.lecanii on T.molitor resulted in a median lethal time(TL50)of 14-16 d.Compared with commercial products,L.lecanii caused faster insect death than the nematode Steinernema carpocapsae and nuclear polyhedrosis virus(no effect on T.molitor survival),and slower than Beauveria bassiana(TL50=5),Beauveria pseu-dobassiana(TL50=8d)and Bacillus thuriengensis(80%mortality first day after inoculation).Mortality was also accelerated under water stress,reducing TL50 by an addi-tional 33%.Remarkably,water stress alone had a comparable effect on mortality to that of L.lecanii isolates.This study confirms T.molitor as a good model insect for pathogenicity testing and agrees with management policies proposed in the EU Green Deal.展开更多
基金supported by LIFE project MYCORESTORE“Innovative use of mycological resources for resilient and productive Mediterranean forests threatened by climate change,LIFE18 CCA/ES/001110”projects VA178P23 and VA208P20 funded by JCYL(Spain),both co-financed by FEDER(UE)budget.
文摘Tree endophytic fungi play an important role in reducing insect herbivory,either by repelling them or kill-ing them directly.Identifying which fungi show such activ-ity could lead to new environmentally friendly pesticides.In this study,the Mediterranean basin climate conditions are projected to harshen in the next decades,will increase vulnerability of tree species to pest invasions.Endophytic fungi were isolated from wood and leaves of Quercus pyr-enaica,Q.ilex and Q.suber and tested for virulence against adults of the mealworm beetle,Tenebrio molitor L.using a direct contact method.Only 3 of 111 sporulating isolates had entomopathogenic activity,all identified as Lecanicillium lecanii.The pathogenicity of L.lecanii on T.molitor resulted in a median lethal time(TL50)of 14-16 d.Compared with commercial products,L.lecanii caused faster insect death than the nematode Steinernema carpocapsae and nuclear polyhedrosis virus(no effect on T.molitor survival),and slower than Beauveria bassiana(TL50=5),Beauveria pseu-dobassiana(TL50=8d)and Bacillus thuriengensis(80%mortality first day after inoculation).Mortality was also accelerated under water stress,reducing TL50 by an addi-tional 33%.Remarkably,water stress alone had a comparable effect on mortality to that of L.lecanii isolates.This study confirms T.molitor as a good model insect for pathogenicity testing and agrees with management policies proposed in the EU Green Deal.
