发光和非发光同基因弧菌中毒力基因的表达及对无菌卤虫(Artemia franciscana)的毒力。
Expression of virulence genes in luminescent and nonluminescent isogenic vibrios and virulence towards gnotobiotic brine shrimp (Artemia franciscana).
机构信息
Laboratory of Aquaculture and Artemia Reference Center, Ghent University, Gent, Belgium.
出版信息
J Appl Microbiol. 2011 Feb;110(2):399-406. doi: 10.1111/j.1365-2672.2010.04892.x. Epub 2010 Nov 23.
AIMS
This study aimed to evaluate the expression levels of virulence gene regulators (luxR and toxR) and virulence factors (serine protease, metalloprotease and haemolysin) in luminescent and nonluminescent isogenic Vibrio harveyi and Vibrio campbellii.
METHOD AND RESULTS
Nonluminescent variants have been reported before to become dominant in cultures of luminescent vibrios when grown under static conditions in the dark. Wild-type V. harveyi BB120, V. campbellii LMG 21363, quorum sensing mutants of V. harveyi BB120 and their previously reported nonluminescent isogenic counterparts were used in this study. The expression level of the virulence genes srp serine protease, vhp metalloprotease and vhh haemolysin, the quorum sensing master regulator gene luxR and the virulence regulator gene toxR in isogenic luminescent and nonluminescent strains were quantified using reverse transcriptase real-time PCR. These experiments revealed that the nonluminescent strains produced lower levels of the quorum sensing master regulator gene luxR and the vhp metalloprotease gene (which is known to be regulated by quorum sensing). Finally, challenge tests with gnotobiotic brine shrimp (Artemia franciscana) larvae revealed that the nonluminescent strains are less virulent than their luminescent isogenic counterparts.
CONCLUSION
Nonluminescent variants of V. harveyi and V. campbellii strains produce lower levels of the quorum sensing master regulator gene luxR and the vhp metalloprotease gene and are less virulent to brine shrimp than their isogenic luminescent counterparts.
SIGNIFICANCE AND IMPACT OF THE STUDY
These results indicate that adaptation of luminescent vibrios to specific growth conditions that result in a dominant nonluminescent phenotype is accompanied by a decreased adaptation to a host environment because of altered virulence gene regulation.
目的
本研究旨在评估发光和非发光同源哈维氏弧菌和坎贝尔氏弧菌中毒力基因调控因子(luxR 和 toxR)和毒力因子(丝氨酸蛋白酶、金属蛋白酶和溶血素)的表达水平。
方法和结果
先前已有报道称,在黑暗中静态培养时,非发光变体在发光弧菌培养物中变得占优势。本研究使用野生型 V. harveyi BB120、V. campbellii LMG 21363、V. harveyi BB120 的群体感应突变体及其先前报道的非发光同源物。使用逆转录实时 PCR 定量检测同源发光和非发光菌株中毒力基因 srp 丝氨酸蛋白酶、vhp 金属蛋白酶和 vhh 溶血素、群体感应主调控基因 luxR 和毒力调控基因 toxR 的表达水平。这些实验表明,非发光菌株产生的群体感应主调控基因 luxR 和 vhp 金属蛋白酶基因(已知受群体感应调控)的水平较低。最后,用无菌卤虫(Artemia franciscana)幼虫进行攻毒试验表明,非发光菌株的毒力比其发光同源物低。
结论
哈维氏弧菌和坎贝尔氏弧菌菌株的非发光变体产生较低水平的群体感应主调控基因 luxR 和 vhp 金属蛋白酶基因,并且对卤虫的毒力比其同源发光变体低。
研究的意义和影响
这些结果表明,发光弧菌适应导致主导非发光表型的特定生长条件的过程伴随着毒力基因调控改变,从而降低了对宿主环境的适应能力。