Sequences Essential for Transmission of Spiroplasma citri by Its Leafhopper Vector, Circulifer haematoceps, Revealed by Plasmid Curing and Replacement Based on Incompatibility
Sequences Essential for Transmission of Spiroplasma citri by Its Leafhopper Vector, Circulifer haematoceps, Revealed by Plasmid Curing and Replacement Based on Incompatibility
复制标题
DOI：
10.1128/aem.00181-10
复制
发表时间：
2010-05-01
影响因子：
4.4
通讯作者：
Renaudin, Joel
中科院分区：
文献类型：
--
作者：
Breton, Marc;Duret, Sybille;Renaudin, Joel
柑橘螺原体GII 3含有高度相关的低拷贝数质粒pSci 1至pSci-6。尽管它们的复制区域之间有很强的相似性，但这些质粒在螺原体细胞中共存，表明它们是相互相容的。pSci 1 - 6质粒编码称为S.柑橘粘附相关蛋白（ScARPs）（pSci 1 - 5）和亲水性蛋白P32（pSci 6），它们暂时与昆虫传播有关，因为它们在非昆虫传播菌株中未检测到。为了进一步研究质粒编码决定簇在昆虫传播中的作用，我们构建了S。通过开发基于具有相同复制区的质粒的不相容性的质粒消除/替换策略，将不同的柑橘突变株的质粒含量区分开来。实验传播这些S.通过注射到叶蝉载体Circulifer haematoceps中的citri质粒突变体揭示了pSci 6，更准确地说，编码未知功能的蛋白质的pSci6_06编码序列对于传播是必需的。与此相反，ScARPs和P32在叶蝉媒介的螺原体的获得和传播中均被忽略，即使S.获得了缺乏pSci 1至pSci-5（编码ScARP）的柑橘突变体，并以比野生型菌株GII 3更低的效率传递。
Spiroplasma citri GII3 contains highly related low-copy-number plasmids pSci1 to -6. Despite the strong similarities between their replication regions, these plasmids coexist in the spiroplasma cells, indicating that they are mutually compatible. The pSci1 to -6 plasmids encode the membrane proteins known as S. citri adhesion-related proteins (ScARPs) (pSci1 to -5) and the hydrophilic protein P32 (pSci6), which had been tentatively associated with insect transmission, as they were not detected in non-insect-transmissible strains. With the aim of further investigating the role of plasmid-encoded determinants in insect transmission, we have constructed S. citri mutant strains that differ in their plasmid contents by developing a plasmid curing/replacement strategy based on the incompatibility of plasmids having identical replication regions. Experimental transmission of these S. citri plasmid mutants through injection into the leafhopper vector Circulifer haematoceps revealed that pSci6, more precisely, the pSci6_06 coding sequence, encoding a protein of unknown function, was essential for transmission. In contrast, ScARPs and P32 were dispensable for both acquisition and transmission of the spiroplasmas by the leafhopper vector, even though S. citri mutants lacking pSci1 to -5 (encoding ScARPs) were acquired and transmitted at lower efficiencies than the wild-type strain GII3.