Study On RT-PCR Detection And Elimination Technique Of Citrus Tatter Leaf Disease

Citrus tatter leaf virus disease (CTLV) is an important citrus disease. In the field, the orange or trifoliate orange citrus rootstocks obviously infected by tatter leaf virus, while they showed no or slightly symptoms in the rootstocks of other plants. Subsequently, tatter leaf disease was easy to produce and spread virus. Detection of Citrus tatter leaf disease present methods are indicator plants, polyacrylamide gel electrophoresis and molecular detection, including RT-PCR is a rapid detection method of citrus tatter leaf disease, China has been applied, but not yet launched in Guangxi research in this area reported. Guangxi is China's main citrus producing areas, and some citrus varieties, such as days of the grass by the serious damage of citrus tatter leaf virus affecting citrus production in our region. In order to quickly understand the field of citrus plants, the introduction of citrus varieties and seedlings infected with broken leaf disease situation, urgent need to establish a rapid and accurate detection of citrus tatter leaf disease. Heat treatment at home and abroad with tip grafting has been shown to removal of citrus tatter leaf disease advanced and effective method. However, the traditional method of heat treatment equipment, potted plants and space are higher, more complicated steps. In order to find a simple and effective method of detoxification treatment carried out this study using growth regulators to conduct high temperature and humidity cuttings for detoxification of culture. The main result of this paper is as follows:1,The RNA OF lots of high quality samples were extracted by means of modified technology of Bugos. In this research, the RT-PCR was optimized in the sides of Primer, Primer concentration, dNTP concentration, Annealing temperature, etc. After that, the optimized RT-PCR can detect the positive control CTLV steadily and have the advantages of high sensitivity and specificity strongly. It was Suitable for the Fast detection of the large number of Field samples.2,Part of The cultivated varieties in Guangxi, Vietnamese varieties, varieties of Sino-vietnamese border, the seedling of no Huanglong and the scion of Removal illness huanglong disease were Fast detected by RT-PCR technology. The results showed that, there were 2 varieties having disease in 6 Guangxi Vietnamese varieties,14 varieties havingno disease in Vietnamese varieties,1 varieties having disease in 6 varieties of Sino-vietnamese border. In the GongChuan, gongben, shiwen,shangxiahong, renanyihao, shanchuan and xinjin, there are 2 xinjin having CTLV。The 33 varieties of citrus which already removal huanglong disease were grafted on the Poncirus trifoliate seeding from a unit of guangxi. And found the nanfeng mandarin orange and xinjin had CTLV.3,During The cultivation of tube plants, striped the skin of seeding can improve the sterile germination of germination Poncirus trifoliate significantly. The treatment of divesting the inside and outside skin of seeding was best. The Germination rate was 96.7%, the Poncirus trifoliate seeds which treated by 150 mg/L IBA having higher Germination rate, it was 99.4%.4. the cuttings were treated by soaked of different growth regulator and high temperature in lighting incubator for 40 days, the germination rate of Cuttings were 53% and 90%. The highest was the soaked of IBA growth regulator, it was 90%, and the Rooting rate was 57%. The heat treatment process of virus-free for Citrus cuttings can increase the rate of sprout, and to achieve the purpose of heat treatment. In this test, the method of growth regulator and heat treatment was a heat treatment method simply, effectively and practicably.