Toxic Effects Of Resistance Associated Proteins(Cs-cystatin,Cs-Rip) Of Tea On Empoasca Vitis

Empoasca vitis (Hemiptera, Cicadellidae) is a major insect pest threatening tea production throughout China. E.vitis cause a decreased of15%in tea yield, even up to50%. Thus, study how to effectively control the pest is the research hotspot currently. Nowadays, there are many studies about this species but the classification of this species has been controversial. In addition, it is difficult to give consideration to ecological safety and efficacy at the same time. Accordingly, the species were redescribed by morphological and molecular identification technology in this paper. The toxic effects from two kinds of tea plant proteins (Cs-Cystatin, Cs-RIP) on E.vitis were investigated by molecular biology and tissue pathology technique. In order to provide technological sustentation for developing a new control technology and provide sample standards for related research work.Result showed:(1) Morphological description:The whole body of E. vitis is green or yellow-green. There are no white spots in the head and prothorax backboard. The forewing is yellowish green, the wing side is transparent. The whole latter wing is transparent, the Wing veins is faint yellow. Foot and body are the same color. The metathoracic tibia end is grey or brown. In The end of the female abdomen with stylomere, Male with a triangular bone plate (Base flap). Molecular identification:We get the28S rRNA sequence of E.vitis, phylogenetic analysis showed that species has the closest relationship with Typhlocyba serrate. In addition, we cloned ITS2sequence, and confirmed this is the first time to get this sequence in Cicadellidae insects by sequence alignment.(2) Cs-Cystatin and Cs-RIP gene was cloned from tea plant. Sequence of Cs-Cystatin consisted of306bp, encoding a protein of101aa. The Cs-RIP gene consisted of1713bp, encoding a protein of570aa. We successfully obtained Cs-Cystatin and Cs-RIP proteins by construction of vitro expression system. The results from feeding the E. vitis with these two resistant proteines on the culture mixed, showed that Cs-RIP significantly reduced the survival rate, but the Cs-Cystatin has no effects. Observation by paraffin section showed that esophagus contained the E.vitis had feeding with Cs-RIP on the culture mixed had obvious collapsed and the lumen totally occluded. The fore-and mid-gut obviously shrinked, the villus broke down and fell into the lumen; the flight muscle in the thorax also dissolved with the increase of the feeding time. But no gastrointestinal lesions were found in the tissue sections feeding Cs-Cystatin. The results suggested that Cs-RIP has significant toxic effects on E.vitis, with potential application in pest control.