Detection And Quarantine Biotechnology For Plant Pest And Development Of Stabilized Diagnostic Reagent Kits

Since 1999 China entered into WTO, there are rapid augment of plant quarantine aliens. This requires effective diagnosis strategy and reagent kits in exit and import aspect to precisely detection of agricultural crop pests such as pathogenic bacteria, fungi, and virus or viroid which usually cause varied sever diseases in agriculture productions. Traditional methods for identification of plant pathogen and disease diagnosis usually based on visual observation of typical symptom of disease caused by certain microbial form inside or outside of plant host; cultivation and isolation of organism in laboratory; Pathogenecity test, serological assay. The disadvantages with routine methods for plant pest detection, are time consume, poor discrimination between microbes with common behavioral features; limited to pathogens with known growth requirements, failure to detect diseases caused by unculturable or slow-growing causal agents, or pathogen exist in lower detectable limitation. Nowadays, new molecular methods based on genetic (DNA or RNA) sequence information to identify plant pathogens with in vitro amplification of polymerase chain reaction (PCR) leads to development of specific and standard PCR methods with rapid, sensitivity and specificity has widely applied to identification and diagnosis of disease both medicine and agriculture aspects.To meet the high requirement of enhancing capability of detection and quarantine technology against plant alien pest in agricultural plant, quarantine target pest which impact security of agricultural production including causal agents of citrus bacterial canker (Xanthomonas axonopodis pv. citri), Huanglongbing (Candidatus Liberibacter asiaticus), wheat dwarf bunt (Tilletia controversa) and potato virus diseases (Potato virus) were selected, designing specific primer pairs and probes based on the unique sequence of target pest, using efficient sample extraction method of filter membrane for removing inhibitors of PCR, establishment of immunological and molecular biology detection approaches of target quarantine pests with rapid, sensitive and accuracy; development of diagnostic reagent kits based on stabilizer of macromolecules, creation of standardized protocols of detection of reagent kits, in order to increment of capability of inspection and quarantine against alien pest of agricultural plant pests invasion and spread, guarantee the safety of agriculture productions...