Ha Phuc T, He Ruifeng, Killiny Nabil, Brown Judith K, Omsland Anders, Gang David R, Beyenal Haluk
Gene and Linda Voiland School of Chemical Engineering and Bioengineering, Washington State University, Pullman, WA, 99163, USA.
Institute of Biological Chemistry, Washington State University, Pullman, WA, 99163, USA.
Biofilm. 2019 Sep 11;1:100005. doi: 10.1016/j.bioflm.2019.100005. eCollection 2019 Dec.
Inability to culture the phloem-restricted alpha-proteobacterium " Liberibacter asiaticus" (" L. asiaticus") or the closely related species (" Liberibacter americanus" and " Liberibacter africanus") that are associated with Huanglongbing (HLB) hampers the development of effective long-term control strategies for this devastating disease. Here we report successful establishment and long-term maintenance of host-free " L. asiaticus" cultures, with the bacterium growing within cultured biofilms derived from infected citrus tissue. The biofilms were grown in a newly designed growth medium under specific conditions. The initial biofilm-based culture has been successfully maintained for over two years and has undergone over a dozen subcultures. Multiple independent cultures have been established and maintained in a biofilm reactor system, opening the door to the development of pure culture of L. asiaticus and the use of genetics-based methods to understand and mitigate the spread of HLB
无法培养与黄龙病（HLB）相关的韧皮部限制性α-变形菌“亚洲韧皮杆菌”（“L. asiaticus”）或其密切相关的物种（“美洲韧皮杆菌”和“非洲韧皮杆菌”），这阻碍了针对这种毁灭性疾病制定有效的长期控制策略。在此，我们报告了无宿主“L. asiaticus”培养物的成功建立和长期维持，该细菌在源自受感染柑橘组织的培养生物膜内生长。这些生物膜在特定条件下于新设计的生长培养基中生长。基于生物膜的初始培养物已成功维持了两年多，并进行了十多次传代培养。已在生物膜反应器系统中建立并维持了多个独立培养物，为亚洲韧皮杆菌纯培养物的开发以及使用基于遗传学的方法来理解和减轻HLB的传播打开了大门。
