孔扩散-CAS法检测铁载体及其在捕食线虫真菌中的作用

Detection of Siderophore by Agar Well Diffusion CAS(WD-CAS)Method and Its Application in Nematode Fungi

基于传统CAS检测法,改良原有操作,规避CAS检测液对真菌生长的抑制作用,以实现真菌产铁载体的有效定性分析。以捕食线虫真菌少孢节丛孢菌(Arthrobtrys oligospora)为研究对象,通过比较三种不同处理方式下,即覆盖CAS(O-CAS)法、CAS区域法以及孔扩散CAS(WD-CAS)法,少孢节丛孢菌生长及铁载体检出情况,确定最佳铁载体检测方法。对少孢节丛孢菌不同生活方式条件下产铁载体情况进行分析,验证该方法在解决实际生物学问题中的应用效果,并初步探究少孢节丛孢菌产铁载体特征。结果表明,三种处理方式均能实现对少孢节丛孢菌中铁载体的有效检测,相比O-CAS法,CAS区域法以及WD-CAS法能够更好地规避CAS检测液对真菌生长的抑制作用,其中的WD-CAS法整体表现效果最佳,能很好的在少孢节丛孢菌从腐生生活方式转变为寄生生活方式后实现对铁载体合成情况的有效监测。少孢节丛孢菌具有多种铁载体合成能力,且合成的铁载体种类因培养时间和培养基组成的不同而有所差异。WD-CAS法不仅能够避免因CAS检测液对微生物生长抑制造成的检测困难,还可以实现检测液的实时添加,保证检测结果的准确。研究结果为微生物中铁载体的高效检测及进一步研究铁载体在捕食线虫真菌捕杀线虫中的功能提供参考。

Based on the traditional CAS detection method,the original operation was improved to avoid the inhibition of CAS detection solution on the growth of fungi,so as to achieve effective qualitative analysis of fungal siderophore.The nematode-preying fungi Arthrospora oligospora were selected as a research object.The growth of A.oligospora and the detection of siderophore conditions were determined by comparing three modes of different treatments,namely covering CAS(O-CAS)method,CAS region method and agar well diffusion CAS(WD-CAS)method.The siderophore of A.oligospora under different lifestyle conditions were analyzed,the application effect of this method in solving practical biological problems was verified,and the characteristics of A.oligospora were initially explored.All the three treatments could effectively detect the features of siderophore in A.oligospora.Compared with O-CAS method,CAS region method and WD-CAS method could better avoid the inhibition effect of CAS detection solution on the fungal growth,and among them WD-CAS method had the best overall performance,and could effectively detect the posture of siderophore synthesis in A.oligospora during its lifeway changeover from saprophytic one to parasitic one.And A.oligospora possesses multiple synthesis abilities of siderophore,and the types of synthesized siderophore category varied to some extent with different culture time and medium composition.The results showed that WD-CAS method can not only avoid the detection difficulties caused by the inhibition of microbial growth caused by CAS detection solution,but also could realize the real-time addition of detection solution to ensure the accuracy of detection results.The correlated results provided a reference for the efficient detection of siderophore in microorganisms,and provide a basis for further research on the function of siderophore in the preying of nematodes by nematode fungi.