Small RNA Deep Sequencing Identifying Viruses And Rapid Propagation Technology Optimizing Of Strawberry

Strawberry is an important herb fruit trees. Because of its fruit have beautiful appearance, rich fragrance and abundant nutrition, tasting sweet, sour, soft and succulent, stawberries are welcomed by consumers at home and abroad. However, vegetative propagation that spreading strawberry virus which causes serious economic damage, reducing productivity as high as 80% per year. Therefore, rapid and effective strawberry virus detection and detoxification technology is of great significance to ensure the high quality and efficient detoxication of detoxicated strawberry seedlings, and also to ensure the quantity of the strawberry.Samples with viral disease-like symptoms were collected from strawberry plants in the fields in Changsha. Optimized the total RNA extraction method of the strawberry. This study combined RT-PCR with siRNA deep sequencing detecting virus in Changsha. Rapidly and accuratly detecting strawberry virus in Changsha.Finally, the test also simply optimization rapid propagation technology of strawberry. The main contents results are as follows:1?Compared the effect of Trizol UP method, Trizol Plant method, CTAB-LiCl method method and RNAprep Pure Plant Kit method of extracting strawberry total RNA and use the electrophoresis to detect the integrity of the RNA. The method of RNAprep Pure Plant Kit can extract RNA effectly with 18S,28S and 5S stripes, the brightness of the 28S stripe is 1.5-2 times to18S and did not appear degradation dimmers. This method can effectively remove polysaccharide, extract high quality, good integrity, higher yield RNA of strawberry with, high yield, can meet the requirements of small RNA deep sequencing; The CTAB-LiCl method could extract the RNA with 28S and 18S stripe, but had degradation dimmers, could meet the requirement of RT-PCR. But in terms of its cost, and method of CTAB-LiCl cost lower than the RNAprep Pure Plant Kit. Trizol UP method and Trizol Plant methods failed to run out of RNA bands.2?Using the small RNA deep sequencing technology to sequencing and bioinformatics analysis of mixed strawberry samples with viral disease-like symptoms in Changsha, CMV, SVBV, LySV and PRV are found.We only found SVBV and CMV by RT-PCR verification, and did not found PRV and LySV.3?Using RT-PCR method detecting virus of 53 strawberry samples in Changsha. The result shows,42 strawberry samples are separate infect by CMV, are with positive rate of viruses is 79.25%. This is the first time that people found cucumber Mosaic virus on strawberry plants in Changsha. Also, is the second time found Cucumber Mosaic virus in China on the strawberry.10 strawberry samples are infect by SVBV, while the positive rate is 18.87%, all of them are mixed infection with CMV. The 53 samples are failed to detect PRV and LySV.4?To optimized strawberry proliferation medium, The test shows that the best proliferation concentration for strawberries is MS+6-BA 1mg/L+NAA 0.10 mg/L. The growth coefficient is 10.91.