Functional Analysis of a Putative Type III Secretion System in Stress Adaption by CCNWXJ12-2.

CCNWXJ12-2, isolated from root nodules of the desert plant , contains two type III secretion systems (T3SSs). T3SSs are specialized machinery with wide distribution in bacteria that inject effector proteins into target cells. Our previous study showed that the expression of T3SS1 is upregulated in high-salt conditions. Here, phylogenetic analysis of T3SS1 using the core protein RhcU suggested that T3SS1 belongs to the α-Rhc II subgroup of the Rhc T3SS family. To elaborate the function of CCNWXJ12-2 T3SS1 in stress adaption, two T3SS1 mutants ( and ) were constructed and analyzed. β-galactosidase transcriptional fusion assays showed that activity of the promoter of T3SS1 was induced by salts. Mutant was more sensitive to NaCl and LiCl than the wild-type, but was not. Both mutants were more sensitive to KCl than the wild-type. The intracellular Na concentration in in high-NaCl conditions (0.4 M) increased by 37% compared to that of the wild-type strain, while the Na concentration in increased by 13%. The K concentration in both mutants increased by 16% compared to the wild-type in high-KCl conditions (0.3 M). Strain showed decreased survival compared to the wild-type after treatment with HO, while the survival rate of was almost the same as that of the wild-type. Antioxidant enzyme activities in were lower than those in the wild-type strain, but this was not the case for . Our data elucidate the beneficial effects of T3SS1 in the adaption of CCNWXJ12-2 to stress.