The Establishment Of PCR Detection System Of Watermelon Quarantine Diseases

Watermelon in China inspection and quarantine objects including subspecies of melon Bacterial fruit blotch caused by acid bacteria watermelon(Acidovorax avenae subsp.citrulli, Aac) and cucumber green mottle mosaic virus (Cucumber green mottle mosaic virus, CGMMV) disease. These two diseases are seed-borne diseases bring great harm to the watermelon and no treatment can completely remove pathogens after the outbreak, often cause significant economic losses.The living standards of residents improved with the development of economy, as economic crop, the consumption of watermelon increased year by year, the seed quantity and cultivated area of watermelon also increased. Enhanced detection of pathogenic bacteria in the watermelon seed prevent the outflow of carrying pathogens seed ensure high quality of watermelon seeds on the market is the basis of keeping agricultural market economic stability. In view of the above two kinds of inspection and quarantine diseases, a test method is stable, simple, fast, economy and directly use the watermelon seed for testing materials is necessary and have a great effect on the agricultural production.(1) A pair of primers were designed according to the conserved region of coat protein gene sequence.Using suspected of watermelon leaf and seed infection CGMMV as material, Suspected susceptible materials were amplified target band while the negative control did not amplify the target strip. The target bands cloned and sequenced, prove the target band is a fragment of the coat protein gene the results show that using the watermelon seeds as material through RT-PCR method can be successful detection of CGMMV.(2)Based on RT-PCR detection method, a rapid detection system of CGMMV was established for large quantities of watermelon seeds, research results show that the maximum seed number is50the CGMMV can be detected when use the watermelon seed for the detection materials.(3)Using suspected of watermelon leaf and seed infection BFB as material, a rapid detection system of BFB was established for large quantities of watermelon seeds, results show that the maximum seed number is1200that the BFB can be detected when use the watermelon seed for the detection materials and the shortest time of preparing templates is100minutes.(4)Using the standard detection system to detect the sample, the testing results coincided with the local epidemic, it illustrated the detection system is stable and reliable.