植物源抗煙草花葉病毒活性物質(zhì)的篩選、分離及抗病機(jī)理初探.pdf

1、中國(guó)煙草總公司鄭州煙草研究院碩士學(xué)位論文植物源抗煙草花葉病毒活性物姓名：金光輝申請(qǐng)學(xué)位級(jí)別：碩士專業(yè)：食品科學(xué)指導(dǎo)教師：尹啟生;楊軍2010-06-20鄭州煙草研究院碩士學(xué)位論文 英文摘要 Abstract Tobacco mosaic virus (TMV) is a worldwide-occurred tobacco viral dise

2、ase and usually makes big lost in tobacco production. How to inhibit TMV infection and replication has become a hotspot in the field of plant virus disease control. Searching antiviral materials from plants is one of the

3、 methods to prevent and control plant virus disease. Screening and isolating antiviral proteins from plants and its mechanism as well as the physical and chemical properties were studied. The research results were as fol

4、lows: By local lesion assay, crude extracts from 18 plants belonging to 13 families were determined for anti-viral activity against TMV. The results showed that Inhibition activity on TMV infecting Nicotiana glutinosa va

5、ried in different plant extracts. Six extracts exhibited over 50% anti-viral activity. Among them extracts from Spinacia oleracea and Albizzia julibrissin Durazz significantly inhibited TMV infection with 97.44% and 96.9

6、2%, respectively. Crude extracts from Spinacia oleracea and Albizzia julibrissin Durazz were isolated primarily using ammonium sulfate precipitation, and the inhibition activity of the fractions was determined. The resul

7、t showed that anti-viral activities of extract from Spinacia oleracea were found in two fractions. However, only 40%~80% fraction from Albizzia julibrissin Durazz had show severe anti-viral activities. The fractions of 4

8、0%-80% ammonium sulfate precipitation were purified with cation-exchange chromatography (SP-sepharose HP) and gel-filtration chromatography. By local lesion assay, notable inactivation effect of the purified faction was

9、observed. However, such effect did not increase when incubation time was prolonged. The fraction had high antiviral activity of preventing infection against TMV. The fraction was shown to have moderate pH stability. With

10、in the pH range of 5-11, the inhibition rate can stay at high level of 90%. The effect on the inhibition rate of the fraction treated with different temperature varied greatly. However, the antiviral activity can maintai

11、n over 50% within the temperature range of 25℃-60 . ℃ The fractions of 0%-40% ammonium sulfate precipitation was further purified with cation-exchange chromatography (SP-sepharose HP) and activity screening. By local I