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为了明确自然种群白背飞虱Sogatella furcifera中Wolbachia和Cardinium的感染情况以及Wolbachia与其特有的WO噬菌体之间的关系,以采自中国7个省区9个地点的白背飞虱为研究材料,运用PCR检测的方法调查了Wolbachia,Cardinium以及WO噬菌体在各飞虱种群中的感染率和组织分布特点。结果表明:白背飞虱广泛双重感染Wolbachia和Cardinium,并且都表现出很高的感染率。白背飞虱各种群Cardinium的感染率几乎均为100%;Wolbachia的感染率也较高,但雌雄虫感染率差异较大,雌虫的感染率几乎均为100%,而雄虫的感染率从22.2%～95.0%不等。另外,通过不同DNA聚合酶、不同提取方法的对比,揭示了DNA粗提样品在基于PCR技术的胞内共生菌检测中的不足之处。对白背飞虱头部、胸部、腹部、足和翅5个不同部位组织的检测结果表明,不仅在含有生殖组织的腹部有这两类共生菌的感染,在其他非生殖组织中同样也感染了这两类共生菌;虽然Wolbachia和Cardinium在寄主的各个组织中均有分布,但是两者在白背飞虱成虫(尤其是雄虫)阶段的动态变化有明显的差异。进一步对Wolbachia宿主特异性WO噬菌体的检测结果表明,自然种群雄虫中Wolbachia的感染率与不感染个体中WO噬菌体的比率呈明显的负相关。因此推测,雄虫中Wolbachia感染率相对较低的原因可能是由于Wolbachia基因组中溶原性的WO噬菌体受到某种因素的诱导已转化为裂解性噬菌体。研究结果为进一步揭示Wolbachia和Cardinium双重感染条件下对寄主的生殖调控作用及其机制、垂直传播规律、两者之间的相互关系以及进一步的应用研究等方面提供了重要的理论基础。
In order to clarify the infection status of Wolbachia and Cardinium in Sogatella furcifera and the relationship between Wolbachia and its own WO phage, the whitebacked planthoppers collected from 9 sites in 7 provinces and autonomous regions in China were used as the research materials PCR detection method to investigate the Wolbachia, Cardinium and WO phage in each plant population infection rate and tissue distribution characteristics. The results showed that Woolbachia and Cardinium were widely infected by WBPH and all showed high infection rates. The prevalence of Cardinium in each population of whitebacked planthoppers was almost 100%. The infection rates of Wolbachia were also high, but the infection rates of male and female were quite different. The infection rates of females were almost 100% Rates ranged from 22.2% to 95.0%. In addition, comparison of different extraction methods by different DNA polymerases reveals the shortcomings of crude DNA samples in the detection of intracellular viable bacteria based on PCR technology. Test results on the tissues of the head, chest, abdomen, foot and wing in five different parts of whitebacked planthopper showed that not only the infection of these two types of commensal bacteria in the abdomen containing reproductive tissues but also other non-reproductive tissues These two types of commensal bacteria; although Wolbachia and Cardinium are distributed in all tissues of the host, there are obvious differences between the two species in the dynamics of whitebacked planthopper (especially male) stage. Further testing of Wolbachia host-specific WO phages showed a significant negative correlation between the Wolbachia infection rates in natural populations of males and the proportion of WO phages in uninfected individuals. Therefore, it is speculated that the relatively low prevalence of Wolbachia in male may be due to the fact that the lysogenic WO phage in the Wolbachia genome have been transformed into lytic phages by induction of a certain factor. The results provide an important theoretical basis for further revealing the reproductive regulatory role of the host and its mechanism, the vertical transmission law, the relationship between the two and further application research under the dual infection of Wolbachia and Cardinium.