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Pest Analysis for the Importation of Potato Seed from France, Netherlands, Russia and Research on the Detection of Potato Virus with RT-PCR

Author: LiuXing
Tutor: NingHong；ZhangMin
School: Sichuan Agricultural University
Course: Of Plant Pathology
Keywords: potato virus pest risk analysis RT-PCR detection
CLC: S435.32
Type: Master's thesis
Year: 2009
Downloads: 169
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Abstract


Potato is the fourth most important crop in the world,which inferior to the rice,wheat and com.China takes the lead in the yield and planting area of potato around theworld.However,due to the serious diseases and no official managements,there is a huge difference of potato quality comparing with the countries developed in potato producing.Importing eximious potato seed from those countries would be a vital way to improve the producing of potato in china.Basing on the rule of FAO and IPPC,it’s necessarily that to do the pest risk analysis before introducing seed to protect the security of agriculture and zoology.So far,more than 30 viruses or viroid were found on potato,and the virus diseases restrict the potato producing.To set up a rapid,accurate and sensitive detection of potato virus will make sure the potato quality and control the virus spread. Through the research of the pest risk analysis for the importation of potato seed from France,Netherlands,and Russia.We forecast the risk of introducing potato seed and establish relative measures to reduce the risk.Establishing the technique by using reverse transcription polymerase chain reaction（RT-PCR） to detect one virus or several viruses.According to guideline for pest risk analysis constituted by FAO,our PRA initiates by spreading pathway of pests,which relate to potato from France,Netherlands and Russia.The number of pests on potato from France is 179,from Netherlands is 152,from Russia is 142.In the light of the geographical distribution,regulatory criteria,economic importance, the possibility for pests spreaded by potato seed,we made a primary assessment and compartmentalize the pests into potential regulated pest（PRP） and non-regulated pest （NRP）.The further assessment made for PRP.Basing on the aspects of introduction potential,establishment potential,spread potential after establishment,potential economic importance,and so on.Research defined 14 kinds of quarantine pest（QP） and 23 kinds of regulated non-quarantine pest（RNQP） form France potato seed,26 kinds of QP and 26 kinds of RNQP form Netherlands,21 kinds of QP and 17 kinds of RNQP form Russia.Basing on the result of PRA,potato virus M,tomato spotted wilt virus and potato spindle tuber viroid were selected to detect by RT-PCR.Two methods which included RNAprep Plant Kit and RNAiso Plus were used to extract potato virus RNA.Both of them could extract pure and intact RNA,and qualified for RT-PCR.Three viruses primers was designed based on the conservative area of complete genome.To detect PVM, TSWV,PSTVd by simplex reverse transcription polymerase chain reaction（RT-PCR） and the amplification fragment were 630bp（PVM）,747bp（TSWV） and 251bp（PSTVd）. The amplified cDNA of PVM、TSWV and PSTVd were extracted from the gel,and sequenced.Compared in NCBI BLAST,and the result showed that the sequences of amplification products were correct.From antisense primer rations of two viruses on RT stage,Mg2+ concentration on PCR stage to research and optimize the simplex,duplex and multiplex RT-PCR system.

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CLC: > Agricultural Sciences > Plant Protection > Pest and Disease Control > Crop pests and diseases and their prevention > Tuber Crops Pests and Diseases > Diseases and insect pests of potatoes ( potatoes)
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