Screening Of Primers And Analysis Of Genetic Diversity Of Huanglong Pathogen In Hunan Province

Citurs Huanglongbing(HLB)is the most important disease of Citrus disease,and it is also an important quarantine pest in Citrus disease.The pathogen is phloem(Candidatus Liberibacter,spp),Citrus industry has huge economic losses every year due to the influence of Huanglong disease.China first discovered the Citrus Huanglongbing is in the region of Guangdong Province,a few years ago in Hunan province also belongs to the occasional area of Citrus Huanglongbing occurred,with the more convenient logistics information and regional transport of toxic seedlings at the same time,with the increase of global temperature to expand the scope of activities of the citrus psyllid,our province has Citrus Huanglongbing into the area Regional nature exists,and even spread to the whole province.In order to determine the current Hunan province Citrus Huanglongbing specific distribution,detection method,the most suitable and different bacteria Huanglongbing pathogen in different areas of the host of citrus.In order to determine the current distribution of Citrus Yellow Dragon disease in Hunan Province,the most suitable detection method,and the difference between the pathogen of Citrus Yellow Dragon disease in different hosts.A pair of primers with high specificity and sensitivity were screened.The samples of the suspected Citrus Yellow Dragon disease in Hunan province were detected by different methods such as conventional PCR,nested PCR and LAMP,and the sensitivity and application were analyzed.Finally,the pathogen 16 S r RNA,gyr B and OMP of Citrus Yellow Dragon disease in different regions and different hosts were analyzed.The difference in gene sequence.The results are as follows:1、 by conventional PCR F3/B3,F1/B1,primer screening of gyr1/gyr2,OI1/OI2 and 16s3/16s4 were 5 pairs of primers,get the best sensitivity and accuracy of application of primer F1/B1;by nested primers F3/B3,F1/B1 and PCR were OI1/OI2 and 16s3/16s4,get the best accuracy and sensitivity using primers F3/B3 and F1/B1;finally,compared to the conventional PCR nested PCR and LAMP different methods for detection of suspected Citrus Huanglongbing samples,LAMP >nested PCR > PCR conventional.2、 analysis of different regions and different hosts of Citrus Huanglongbing pathogen 16 S r RNA,gyr B and omp gene sequences obtained from different hosts,clustering of Citrus Huanglongbing pathogen 16 S r RNA gene sequence of the more significant source of clustering in different parts of Citrus Huanglongbing pathogen gyr B gene sequence is more significant,and the clustering of Citrus Huanglongbing pathogen omp gene sequence by region and host at the same time,can better reflect the province of Citrus Huanglongbing pathogen evolution level.3、1111 samples of suspected Citrus Yellow Dragon disease were tested by PCR in 37 cities and counties of Hunan province and 2 provincial agricultural units.Analysis of systematic investigation,determine the distribution of Citrus Huanglongbing in the whole territory of Yongzhou city and Chenzhou City,Shaoyang city and Huaihua city in southern province,Citrus Huanglongbing disease line: West Huaihua Huitong County-Xupu County-Loudi County of Xinhua city-Shaoyang City,Shaodong County,Hengyang city Qidong County Chang Ning City-Leiyang city-Zhuzhou city-Youxian east to Zhuzhou County of Yanling city.