| Grape phylloxera, Daktulosphaira vitifoliae (Homoptera: Phylloxeridae), originated from the east part of Rocky Mountains in North America, is a kind of devastating grape insect. Found on vitis vine, grape phylloxera do harm to vitis grape only. Grape phylloxera, was found in 1935 in China, sopposed to be introduced when importing grape seedlings around 1900 and has been a quarantine pest since 1957 when it was identified to be it in 1954, but vanished (could not to be found) through cut measures according to law or political reasons during 1965-1978, but it has been rediscovered again at suburb vineyards of Shanghai city in 2005. By grafting resistance rootstocks, the pest has been under control for more than 100 years in the word and there is no other equally effective method for substitution or rotating to avoid resisitance. Even worse situation for grape industry in China, most of the grapes are cultivated by cuttage, and resistance rootstocks are rarely used, so the risk of spreading of the pest poses a serious threat to the industry. Routine ground survey is still the main detecting method for grape phylloxera, but this method is time consuming and ineffective. In order to provide a quick, accurate and sensitive method to detect and explore grape phylloxera and population dynamics, a real-time PCR method had been studied, and a new method for control the pest, intercropping management, was preliminarily taken for. The results were as follows:1. A real-time PCR method for detection of grape phylloxera, according to specific primers and TaqMan-MGB probe established by Karen Herbert et al, had been designed based on the internal transcribed space region 2 (ITS2). Primers and probe could be used to detect grape phylloxera in vineyard through the experiment, the recombinant plasmid containing the ITS2 sequence was constructed as a standard control, then an absolute quantitative method of FQ-PCR was developed by optimizing reaction conditions, and sensitivity and reproducibility were carried out. The sensitivity of this method was proved to be 1.625 copies/μl; the coefficient of variation value was less than 5%. The DNA, that is distilling from 0.25 g soil which contain 10 grape phylloxera crawlers, can be detected efficiently, even as few as diluted a thousand times, and rhizosphere soil was detected qualitatively. The TaqMan-MGB probe-based real-time fluorescent PCR assay served as a promising approach for easy, rapid and highly specific and sensitive detection of grape phylloxera.2. Under the selection criteria of having insecticidal activity, economic rationality, shade tolerance, and dwarf plant but more root, Schizone peta tenuifolia (Benth.) Briq,Astragalusmembranaceus (Fisch.) Bunge and Scutellaria baicalensis Georgi were chosen as intercrops to control grape phylloxera. Field trials had been taken for two years. Vine vigour was investigated in maturity stage, and the grape phylloxera population was investigated in 0-15 cm soil layer both in the second peak of root growth stage and overwintering period. The results showed that intercropping of Schizone peta tenuifolia (Benth.) Briq, Astragalusmembranaceus (Fisch.) Bunge and Scutellaria baicalensis Georgi had significant effect on grape phylloxera population control, in which intercropping of Schizone peta tenuifolia (Benth.) Briq had effect on tree vigor recovery. In addition, the maturation root survey found that intercropping of Scutellaria baicalensis Georgi could promoted the growth of grape roots. In conclusion, the intercropping could be an effective method for grape phylloxera controlling, and further research need to be taken to check and explore the mechamism for intercroping. |
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