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Journal of applied entomology : Zeitschrift für angewandte Entomologie v.141 no.1/2, 2017년, pp.61 - 66   SCI SCIE
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Effective molecular detection of Diaphorina citri Kuwayama (Hemiptera: Liviidae) in bulk insect samples from sticky traps

Fujiwara, K. (National Agriculture and Food Research Organization, Tsukuba, Ibaraki, Japan ); Uechi, N. ( National Agriculture and Food Research Organization, Tsukuba, Ibaraki, Japan ); Shimizu, Y. ( Okinawa Prefectural Plant Protection Center, Naha, Okinawa, Japan ); Toda, S. ( National Agriculture and Food Research Organization, Tsukuba, Ibaraki, Japan ); Inoue, H. ( National Agriculture and Food Research Organization, Tsukuba, Ibaraki, Japan ); Yamaguchi, T. ( Oshima Branch Kagoshima Prefectural institute for Agricultural Development, Uragami‐ ); Iwanami, T. (chou Amami, Amami, Kagoshima, Japan ); Fujikawa, T. ( National Agriculture and Food Research Organization, Tsukuba, Ibaraki, Japan );
  • 초록  

    Abstract Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Liviidae), is the principal vector of citrus greening (huanglongbing) disease. Invasion of new areas by the vector increases the risk of further spread of the disease and has economic impacts on the global citrus industry. Effective implementation of vector surveys is essential to contain disease outbreaks. This is especially true in countries such as Japan, where most of the major citrus‐producing areas are free from citrus greening. Recently, vector surveys have been routinely conducted to maintain ‘disease‐free’ and ‘disease‐ and vector‐free’ areas in Japan, and improvement of methods that can detect D.?citri in native insect populations is imperative. Here, we developed a method of using conventional and real‐time PCR to detect D.?citri among bulk insects captured in sticky traps without the need for preliminary differentiation steps based on morphology. DNA fragments of D.?citri were specifically detected by both conventional and real‐time PCR in a mixture of a 10 −3 dilution (ca. 0.008–0.009?ng/ μ l) of D.?citri DNA and 100?ng/ μ l of bulk insect DNA, indicating that small body parts such as pieces of leg or parts of wings of D.?citri were detectable in the bulk insect samples. No misleading amplification of fragments from the other psyllid species and citrus pests we used occurred under our PCR conditions. Our results suggest that the technique is applicable to extensive surveys of D.?citri in early warning programmes.


  • 주제어

    bulk insect populations .   citrus greening disease .   Diaphorina citri Kuwayama.  

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