南中国海扁枝滨珊瑚白化综合症病原的生物标志鉴定及快速检测方法建立

Coral reef is the most important basis of marine territory, fishery sources and tour sources in the South China Sea. Porites andrewsi White Syndrome (PAWS) was one of the severe disease for coral reef there. Two Vibrio alginolyticus strains, XSBZ03 and XSBZ14, were proved to be the pathogenic agent for PAWS in that sea area. However, their epidemic pattern, pathogenic mechanism and hazard rating to the corals are unknown now. Thus, it is the inevitable course of coral disease prevention and successful coral transplantation to character their nucleic acid molecular biomarker and then develop the rapid diagnosis method. In this study, the genomes of strains XSBZ03 and XSBZ14 will be sequenced by Roche 454 sequencing technology and then their specific biomarkers will be identified by pan-genomic analysis and southern blot. With the molecular biomarker, PCR and real-time PCR will be developed for primary rapid detection and final detection of the two strains. The results of the study will be very useful for the quick diagnosis of the disease. Of course, the results will be helpful to survey their epidemiological pattern, explore the propagation mechanism of the disease, evolve the safety and effective control measure, evaluate the hazard rating of the two strains and develop the specific pathogen-free coral transplantation.

珊瑚礁是南中国海的海上疆土、渔业资源和旅游资源的基石，扁枝滨珊瑚白化综合症是当前南中国海珊瑚礁白化的最主要疾病之一。项目申请人已确定特异遗传型溶藻弧菌菌株XSBZ03和XSBZ14是该海域扁枝滨珊瑚白化综合症的病原，但还不清楚这两株病原的流行规律、致病机理和危害程度。因此，开展这两株病原的分子生物标志鉴定并建立相应的快速检测方法，是当前开展南中国海珊瑚疾病快速诊断和确保珊瑚成功移植的必由之路。本项目拟采用454技术测定病原XSBZ03和XSBZ14的全基因组，利用泛基因组分析技术和Southern杂交技术确定其生物标志，采用PCR和荧光定量PCR技术分别建立其初诊与确诊的快速检测方法。项目成果可直接为该疾病快速诊断提供可靠依据，还可为开展该疾病流行病学调查，探寻该疾病传播机制，制订安全高效控制措施，评估上述病原对南中国海珊瑚礁危害程度和建立无特定病原珊瑚移植方法奠定基础，意义重大。

珊瑚礁是南中国海的海上疆土、渔业资源和旅游资源的基石，扁枝滨珊瑚白化综合症是当前南中国海珊瑚礁白化的最主要疾病之一。本项目采用454高通量测序技术，测定了南中国海扁枝滨珊瑚病原溶藻弧菌菌株XSBZ03和XSBZ14的基因组；利用Newbler序列拼接软件、 泛基因组比较技术和perl脚本单拷贝基因家族序列分析技术，确定菌株XSBZ03和XSBZ14的基因组草图和特异基因生物标志；以XSBZ03的IGS序列为靶序列，构建了XSBZ03的普通PCR检测方法；分别以XSBZ03的单拷贝基因S1、XSBZ14的单拷贝基因S2为靶序列，构建了珊瑚病原菌株XSBZ03和XSBZ14的双重PCR检测方法；以XSBZ03的单拷贝基因S3为靶序列，构建珊瑚病原菌株XSBZ03的荧光定量PCR检测方法；以XSBZ14单拷贝基因S2为靶序列构建珊瑚病原菌株XSBZ14的荧光定量PCR检测方法。本项目成果可直接为该疾病快速诊断提供可靠依据，还可为开展该疾病流行病学调查，探寻该疾病传播机制，制订安全高效控制措施，评估上述病原对南中国海珊瑚礁危害程度和建立无特定病原珊瑚移植方法奠定基础，意义重大。