Virus Elimination On Chrysanthemum Morifolium (Ramat.)Tzvel.’Chuju’ And Evaluation Of Bioactive Components Of Virus-free Plants

Chuzhou chrysanthemum(Chrysanthemum morifolium(Ramat.)Tzvel.’Chuju’)is a genus Asteraceae perennial herb.Because of its unique and good medicinal properties,so well-known ranked the first of four Chinese medicinal Chrysanthemum,the first in the famous herbal medicine in Anhui Province.Since ’Chuju’ cultivars are typically propagated vegetatively,the risk of disseminating viral pathogens is high.A number of viruses are known to infect ’Chuju’;some are benign,affecting flower or leaf color,but others weaken the plant or reduce the value of the product by distorting the flowers or by discoloring the foliage.’Chuju’ quality degradation,yield and quality to drop,resulting serious economic losses to crop production worldwide.We from virus detection,determining the type of virus,comparison of three different virus-free way to eliminate virus to identification of virus-free seedlings of genetic stability,established an efficient virus-free system and study on the quality of ’Chuju’,provided a theoretical references for extensive application of virus-free’Chuju’.The main results are as follows:1.In the field exhibiting symptoms of the virus ’Chuju’ for the material,we detected eight virus by electron microscopy and nested RT-PCR method,and ultimately determine’Chuju’ mainly containing Chrysanthemum virus B(CVB)and Chrysanthemum stunt viroid(CSVd);here,a highly specific and sensitive nested PCR-based assay is described for detecting Chrysanthemum virus B.The coat protein gene of the virus as specific primers for reverse transcription to obtain cDNA.At the same time nested amplification PCR is employed to detect it.The sensitivity is up to the 10"9,greatly improved the sensitivity.The assay is ideal for performing an early stage diagnosis of the disease.2.Comparison of three different virus-free way to build ’Chuju’ rapid propagation of virus-free system and the quality of research,Combined thermotherapy and tissue culture;The results showed that:a shoot-tip meristem culture of the highest survival rate for 78.78%;when ribavirin concentration was 10 mg/mL,the survival rate of shoot is higher;combined chemotherapy and tissue culture representing a significant difference from tissue culture.3.On DNA molecular level,12 pairs of primers of SRAP that were selected which was used to determine the genetic stability of virus-free test-tube plantlets.There were no difference bands among fragments detected.it proved that three ’different methods of virus-free seedlings didn’t change genetic stability of virus-free ’Chuju’.At the same time we compared the content of total flavonoids,chlorogenic acid,luteolin,chlorogenic acid A of three different methods of virus-free seedlings and control,the results show that the total flavonoids,chlorogenic acid,luteolin,chlorogenic acid of virus-free seedlings of’Chuju’had a significantly higher than the control.Heat treatment combined with tissue culture of virus-free seedlings respectively,an increase of 1.93,4.46,2.02,14.58 times higher than field seedlings.