Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/30701
標題: 開發以生物晶片系統快速鑑別八種鱗翅目害蟲之方法
Development of a biochip for rapid discrimination of eight lepidopteran pests
作者: Kuo, Hung-Mao
郭竑懋
關鍵字: biochip;鱗翅目害蟲;internal transcribed spacer 2 (ITS2);lepidopteran pests;rapid discrimination;生物晶片;核醣體第二轉錄區間;快速鑑別
出版社: 昆蟲學系所
引用: 林益昇、龔玉惠、方敏男。1999。瓜菜類-絲瓜。蔬菜病蟲害綜合防治專輯,瓜28頁。台灣省政府農林廳。436頁。 洪巧珍、黃振聲、侯豐男。1998。粗腳姬捲葉蛾(Cryptophlebia ombrodelta (Lower))之人工飼料飼育方法及其羽化與交尾行為。植保會刊。40:297-307。 洪巧珍、江碧媛、黃振聲。2002。荔枝細蛾(Conopomorpha sinensis Bradley)之飼育技術及其羽化與交尾行為。植保會刊44:89-99。 洪巧珍、黃振聲、王效岳。2006。荔枝與龍眼鱗翅目害蟲種類與發生情形調查。台灣昆蟲26︰27-44。 張德前、陳慶忠。1989。三種危害豆類蔬菜之鱗翅目害蟲觀察。台中區農改場研究彙報24:21-29。 植物保護手冊。2004。行政院農委會農業藥物毒物試驗所編印。835頁。 黃啟鐘、彭武康、N. S. Talekar。2003。豆莢螟(Maruca vitrata)(鱗翅目:螟蛾科)為害田菁(Sesbania cannabina)之特性。台灣昆蟲23:1-11。 謝進來。1997。番荔枝(釋迦)重要害蟲發生與防治。台東區農業專訊19:8-11。 Apte, S., Smith, P. J., and Wallis, G. P. (2007) Mitochondrial phylogeography of New Zealand freshwater crayfishes, Paranephrops spp. Mol. Ecol. 16: 1897-1908. Armstrong, K. F., Cameron, C. M., and Frampton, E. R. (1997) Fruit fly (Diptera: Tephritidae) species identification: a rapid molecular diagnostic technique for quarantine application. Bull. Entomol. Res. 87: 111-118. Behura, S. K., Sahu, S. C., Rajamani, S., Devi, A., Mago, R., Nair, S., and Mohan, M. 1999. Differentiation of Asian rice gall midge, Orseolia oryzae (Wood-Mason), biotypes by sequence characterized amplified regions (SCARs). Insect Mol. Biol. 8: 391-397. Black,W. C., DuTeau, N. M., Puterka , G. J., Nechols, J. R., and Pettorini, J. M. (1992) Use of the random amplified polymorphic DNA polymerase chain reaction (RAPD-PCR) to detect DNA polymorphisms in aphids. Bull. Entomol. Res. 82: 151-159. Bouchara, J. P., Hsieh, H. Y., Croquefer, S., Barton, R., Marchais, V., Pihet, M., and Chang, T. C. (2009) Development of an oligonucleotide array for direct detection of fungi in sputum samples from patients with cystic fibrosis. J. Clin. Microbiol. 47: 142-152. Brentani, R. R., Carraro, D. M., Almeida, S. V., Reis, E. M., Neves, E.J., de Souza, S. J., Carvalho, A. F., Brentani, H., and Reis, L. F. (2005) Gene expression arrays in cancer research: methods and applications. Crit. Rev. Oncol. Hematol. 54: 95-105. Cenis, J. L., Perez, P., and Farers, A. (1993) Identification of aphid (Homoptera: Aphididae) species and clones by random amplified polymorphic DNA. Ann. Entomol. Soc. Am. 86: 545-550. Chen, Y. H. (2007) Development of a biochip for discrimination of four tephritid fruit flies (Diptera: Tephritidae). Master Thesis, Department of Entomology, National Chung Hsing University. Dankbar, D. M., Dawson, E. D., Mehlmann, M., Moore, C. L., Smagala, J. A., and Shaw, M. W. (2007) Diagnostic microarray for influenza B viruses. Anal. Chem. 79: 2084-2090. Dawson, E. D., Moore, C. L., Dankbar, D. M., Mehlmann, M., Townsend, M. B., Smagala, J. A., Smith, C. B., Cox, N. J., Kuchta, R. D., and Rowlen, K. L. (2007) Identification of A/H5N1 influenza viruses using a single gene diagnostic microarray. Anal. Chem. 79: 378-384. Douglas, L. J., and Haymer, D. S. (2001) Ribosomal ITS1 polymorphisms in Ceratitis capitata and Ceratitis rosa (Diptera: Tephritidae). Ann. Entomol. Soc. Am. 94: 726-731. Ehrenreich, A. (2006) DNA microarray technology for the microbiologist: an overview. Appl. Microbiol. Biotechnol. 73: 255-273 Engel, E. A., Escobar, P. F., Rojas, L. A., Rivera P. A., Fiore, N., and Valenzuela, P. D. (2010) A diagnostic oligonucleotide microarray for simultaneous detection of grapevine viruses. J. Virol. Meth. 163: 445-451. Fodor, S. P., Read, J. L., Pirrung, M. C., Stryer, L., Lu, A. T., and Solas, D. (1991) Light-directed, spatially addressable parallel chemical synthesis. Science 251: 767-773. Frutos, R. L., Fernández-Espinar, M. T., and Querol, A. (2004) Identification of species of the genus Candida by analysis of the 5.8S rRNA gene and the two ribosomal internal transcribed spacers. Antonie Van Leeuwenhoek 85:175-185 Guschin, D. Y., Mobarry, B. K., Proudnikov, D., Stahl, D. A., Rittmann, B. E., and Mirzabekov, A. D. (1997) Oligonucleotide microchips as genosensors for determinative and environmental studies in microbiology. Appl. Environ. Microbiol. 63: 2397-2402. Han, W., Liu, B., Cao, B., Beutin, L., Krüger, U., and Liu, H. (2007) Plastic polymers for efficient DNA microarray hybridization: Application to microbiological diagnostics. Appl. Environ. Microbiol. 73: 4082-4088. Harrington, C. A., Rosenow, C., and Retief, J. (2000) Monitoring gene expression using DNA microarrays. Curr. Opin. Microbiol. 3: 285-291. He, Z., Wu, L., Li, X., Fields, M. W., and Zhou, J. (2005) Empirical establishment of oligonucleotide probe design criteria. Appl. Environ. Microbiol. 71: 3753-3760. Heinze, B. (1994) RAPD reactions from crude plant DNA. Mol. Biotech. 1: 307-310. Hsu, K., Hua, T., Chang, N. T., and Yeh, W. B. (2005) Molecular identification of important spider mites using multiplex PCR and PCR-RFLP. Formosan Entomol. 25: 243-254. (in Chinese) Jin, L. Q., Li, J. W., Wang, S. Q., Chao, F. H., Wang, X. W., and Yuan, Z. Q. (2005) Detection and identification of intestinal pathogenic bacteria by hybridization to oligonucleotide microarrays. World J. Gastoenterol. 11: 7615-7619. Kane, M. D., Jatkoe, T. A., Stumpf, C. R., Lu, J., Thomas, J. D., and Madore, S. J. (2000) Assessment of the sensitivity and specificity of olgonucleotide (50 mer) microarrays. Nuclecic Acids Res. 28: 4552- 4557. Katsu, M., Kidd, S., Ando A., Moretti-Branchini M. L., Mikami Y., Nishimura K., and Meyer, W. (2004) The internal transcribed spacers and 5.8S rRNA gene show extensive diversity among isolates of the Cryptococcus neoformans species complex. FEMS Yeast Res. 4: 377-388. Kucho, K. I., Yoneda, H., Harada, M., and Ishiura, M. (2004) Determinants of sensitivity and specificity in spotted DNA microarrays with unmodified oligonucleotides. Gens Genet. Syst. 79: 189-197. Liu, H. H., Cao, X., Yang, Y., Liu, M. G., and Wang, Y. F. (2006) Array-based nano-amplification technique was applied in detection of hepatitis E virus. J. Biochem. Mol. Biol. 39:247-252. Loxdale, H. D., and Lushai, G. (1998) Molecular markers in entomology. Bull. Entomol. Res. 88: 577-600. Lu, K. H., Chang, C. M., Chen, C. Y., and Chen, Y. J. (2002) Development of species-specific PCR primers for identification of three key tephritid pests in Taiwan. Plant Prot. Bull. 44: 255-265. (in Chinese) Lu, K. H., Chang, S. C., Hsu, J. Y., and Yang, J. T. (2003) A combination of traditional and modern techniques for insect identification- using the codling moth, Cydia pomonella L. (Lepidoptera: Tortricidae), as an example. Plant Prot. Bull. 44: 359-364. (in Chinese) Lucchini, S., Thompson, A., and Hinton, J. C. D. (2001) Microarrays for microbiologists. Microbiology 147:1403-1441. Majoros, L., Karods, G., Belák, A., Maráz, A., Asztalos, L., Csánky, E., Barta, Z., and Szabó, B. (2003) Restriction enzyme analysis of ribosomal DNA shows that Candida inconspicua clinical isolates can be misidentified as Candida norvegensis with traditional diagnostic procedures. J. Clin. Microbiol. 41:5250-5253 Martin, C., Roberts, D., Van Der Weide, M., Rossau, R., Jannes, G., Smith, T., and Maher, M. (2000) Development of a PCR-based line probe assay for identification of fungal pathogens. J. Clin. Microbiol. 38: 3735-3742. Merseburger, A. S., Anastasiadis, A. G., Hennenlotter, J., Schilling, D., Simon, P., Machtens, S. A., Serth, J., Stenzl, A., and Kuczyk, M. A. (2006) Tissue microarrays: applications in urological cancer research. World J. Urol. 24: 579-584. Paran, I., and Michelmore, R. W. (1993) Development of reliable PCR- based markers linked to downy mildew resistance genes in lettuce. Theor. Appl. Genet. 9: 85-93. Perez, T., Albornoz, J., and Dominguez, A. (1998) An evaluation of RAPD fragment reproducibility and nature. Mol. Ecol. 7: 1347-1357. Rudi, K., Skulberg, O. M., Skulber, R., and Jaskobsen, K. S. (2000) Application of sequence-specific labeled 16S rRNA gene oligonucleotide probes for genetic profiling of cyanobacterial abundance and diversity by array hybridization. Appl. Environ. Microbiol. 66: 4004-4011. Rudolf, M., Shiyang, K., Vaidya, G., and Ng, P. K. L. (2006) DNA barcoding and taxonomy in Diptera: A tale of high intraspecific variability and low identification success. Syst. Biol. 55: 715-728. Shi, J., Wu, Y., Cai, M., and Shang, S. (2009) Rapid diagnosis of herpetic encephalitis in children by PCR-microarray technology for simultaneous detection of seven human herpes viruses. Eur. J. Pediatr. 169: 421-425. Shih, H. T., Ng, P. K. L., Schubart, C. D., and Chang, H. W. (2007) Phylogeny and phylogeography of the genus Geothelphusa (Crustacea: Decapoda, Brcahyura, Potamidae) in southwestern Taiwan based on two mitochondrial genes. Zool. Sci. 24: 57-66. Sip, M., Bystricka, D., Kmoch, S., Noskova, L., Hartmannova, H., and Dedic, P. (2010) Detection of viral infections by an oligonucleotide microarray. J. Virol. Meth. 165: 64-70. Small, J., Call, D. R., Brockman, F. J., Straub, T. M., and Chandler, D. P. (2001) Direct detection of 16S rRNA in soil extracts by using oligonucleotide microarrays. Appl. Environ. Microbiol. 67: 4708-4716. Sonja, J. S., Lewis, M. L., and Joshi, R. C. (2006) DNA barcoding applied to invasive leafminers (Diptera : Agromyzidae) in the Philippines. Ann. Entomol. Soc. Am. 99: 204-210. Sugita, T., Nishikawa, A., and Shinoda, T. (1998) Identification of Trichosporon asahii by PCR based on sequences of the internal transcribed spacer regions. J. Clin. Microbiol. 36: 2742-2744. Tan, Z. J., Hu, X. G., Cao, G. U. S., and Tang, Y. (2003) Analysis of gene expression profile of pancreatic carcinoma using DNA micarray. World J. Gastroenterol. 9: 818-823. Tiberini, A., Tomassoli, L., Barba, M., and Hadidi, A. (2010) Oligonucleotide microarray-based detection and identification of 10 major tomato viruses. J. Virol. Meth. 168: 133-140. Tinker, A. V., Boussioutas, A., and Bowtell, D. D. L. (2006) The challenges of gene expression microarrays for the study of human cancer. Cancer Cell 9: 333-339. Tobler, N. E., Pfunder, M., Herzog, K., Frey, J. E., and Altwegg, M. (2005) Rapid detection and species identification of Mycobacterium spp. using real-time PCR and DNA-microarray. J. Microbiol. Meth. 9: 1-9. Tóth, M., Talekar, N.S., Szöcs, G. (1996) Optimization of blends of synthetic pheromone components for trapping male limabean pod borers (Etiella zinckenella Tr.) (Lepidoptera: Phycitidae): preliminary evidence on geographical differences. Bioorg Med Chem.4: 495-497. Trosts, A., Graf, B., Eucker, J., Sezer, O., Possinger, K., Göbel, U. B., and Adam, T. (2004) Identification of clinically relevant yeasts by PCR/RFLP. J. Microbiol. Meth. 56: 201-211. Varga, J. (1993) Restriction fragment length polymorphisms in the mitochondrial DNAs of the Aspergillus niger Aggregate. Mycol. Res. 97: 1207-1212. Vidigal, T. H., Spatz, L., Kissinger, J. C., Redondo, R. A., Pires, E. C., Simpson, A. J., and Carvalho, O. S. (2005) Analysis of the first and second internal transcribed spacer sequence of the ribosomal DNA in Biomphalaria tenagophila complex (Mollusca: Planorbidae). Mem. Inst. Osealdo. Cruz. 99: 153-158. Wang, L. C., Pan, C. H., Severinghaus, L. L., Liu, L. Y., Chen, C. T., Pu, C. E., Huang, D., Lir, J. T., Chin, S. C., Cheng, M. C., Lee, S. H., and Wang, C. H. (2008) Simultaneous detection and differentiation of Newcastle disease and avian influenza viruses using oligonucleotide microarrays. Vet. Microbiol. 127: 217-226. Wang, R. F., Kim, S. J., Robertson, L. H., and Cernigila, C. E. (2002) Development of a membrane-array method for the detection of human intestinal bacteria in fecal samples. Mol. Cell. Probes 16: 341-350. Wheeler, D. B., Bailey, S. N., Guertin, D. A., Carpenter, A. E., Higgins, C. O., and Sabatini, D. M. (2004) RNAi living-cell microarrays for loss-of-function screens in Drosophila melanogaster cell. Nat. Methods 1: 1-6. Williams, J. G. K., Kubelik, A. R., Rafalski, J. A., and Tingey, S. V. (1990) DNA polymorphisms amplified by arbitrary primers are useful genetic markers. Nucleic Acid Res. 18: 6531-6535. Yang, M. M. (1996) New techniques in pest identification: the application of biochemical, computer, and acoustic analysis. Proceeding of A Symposium on New Technique of Plant Protection. Taiwan Agric. Res. Inst., Spec. Publ. 57: 11-22. (in Chinese) Yeh, H. T., Ko, C. C., Hsu, T. C., and Shih, C. J. (2005) PCR-RFLP technique for molecular identification of Rhopalosiphum (Aphididae). Formosan Entomol. 25: 33-46. (in Chinese) Yeh, W. B., Ho, C. L., Hui, C. F., and Ho, C. C. (2000) Genetic diversity of Amblyseius longispinosus and A. wamersleyi (Acari: Phytoseiidae) using RAPD analysis. Chinese J. Entomol. 20: 335-345. (in Chinese). Zabeau, M. and Vos, P. (1993) Selective restriction fragment amplification: a general method for DNA fingerprinting. European Patent Application EP 534858. Zhang, Y., Yin, J., Li, G., Li, M., Huang, X., Chen, H., Zhao, W., and Zhu, S. (2010) Oligonucleotide microarray with a minimal number of probes for the detection and identification of thirteen genera of plant viruses. J. Virol. Meth. 167: 53-60. Zhao, Z., Peytavi, R., Diaz-Quijada, G. A., Picard F. J., Huletsky, A., Leblanc, E., Frenette, J., Boivin, G., Veres, T., Dumoulin, M. M., and Bergeron, M. G. (2008) Plastic polymers for efficient DNA microarray hybridization: Application to microbiological diagnostics. J. Clin. Microbiol. 46: 3752-3758. Zheng, Z. B., Wu, Y. D., Yu, X. L., and Shang, S. Q. (2008) DNA microarray technology for simultaneous detection and species identification of seven human herpes viruses. J. Med. Virol. 80: 1042-1050. Zhong, D., Menge, D., Temu, E., Chen, H., and Yan, G. (2006) Amplified fragment length polymorphism mapping of quantitative trait loci for malaria parasite susceptibility in the yellow fever mosquito Aedes aegypti. Genetics 173: 1337-1345. Zhou, J. (2003) Microarrays for bacterial detection and microbial community analysis. Curr. Opin. Microbiol. 6: 288-294.
摘要: 
Different stages of insect pests, i.e., eggs, larvae, pupae, and adults, frequently appear in the field. Traditionally, discrimination of pest species is based on the morphological characters of adults; yet, it is much difficult to be identified in other stages. In this study, a biochip technology was applied to develop the method for rapid species identification of eight lepidopteran pests, including Conopomorpha sinensis, Adoxophyes sp., Cryptophlebia ombrodelta, Cydia notanthes, Diaphania indica, Maruca vitrata, Anonaepestis bengalella Ragonot., and Etiella sp. The internal transcribed spacer 2 (ITS2) genes of eight lepidopteran pests were selected for this study. First, a pair of universal primers was obtained for stable PCR amplification of ITS2 fragments from the genomic DNA of each species. And then, after DNA sequencing and alignment, several potential species-specific oligonuleotide probes were designed (with 20~30 nts in length of the most variable regions of each species) and screened. After various tests, a optimal standard operating procedure (SOP) has been set up, which is 1 J for UV cross linking, 52oC for hybridization temperature, and 30 cycles for PCR amplification, and 100~10 ng of template DNA is used for the identification. Finally, this biochip possesses high species specificity and sensitivity for rapid discrimination of at least these eight lepidopteran pests.

鱗翅目害蟲常會以卵、幼蟲、蛹或是成蟲等不同時期出現於田間;傳統上害蟲種類鑑定多是以成蟲之外部形態為主要依據,因此要以其他各個時期之外部形態鑑定出正確的種類是較為困難的。本研究利用生物晶片技術,選用了八種鱗翅目害蟲,包括荔枝細蛾(Conopomorpha sinensis)、茶姬捲葉蛾(Adoxophyes sp.)、粗腳姬捲葉蛾(Cryptophlebia ombrodelta)、花姬捲葉蛾(Cydia notanthes)、瓜螟(Diaphania indica)、豆莢螟(Maruca vitrata)、番荔枝斑螟蛾(Anonaepestis bengalella)及豆莢斑螟蛾(Etiella sp.)之核醣體第二轉錄區間(internal transcribed spacer 2, ITS2)基因為材料,開發一套可快速鑑定鱗翅目害蟲之系統。首先設計通用性引子(universal primer),並以PCR增幅各蟲種ITS2之DNA片段,經定序及比對後,針對每一種蟲序列之特異區段,設計數組長度約20~30個鹼基之探針,並經過篩選得到數組專一性之探針。經過數種晶片條件之測試與調整,最後得到此晶片最適之標準使用範圍為:以1 J紫外光能量固定探針於晶片上,PCR增幅循環次數為30次,雜合溫度為52℃,而樣本DNA量範圍為100~10 ng。此生物晶片具有專一性及高靈敏度,可快速鑑別此八種不同之鱗翅目害蟲。
URI: http://hdl.handle.net/11455/30701
其他識別: U0005-2208201116211400
Appears in Collections:昆蟲學系

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