Rapid Species-specific PCR Identification of <i>Bactrocera tau</i> (Diptera: Tephritidae)

HUANG Zhen; HOU Youming; GUO Qiongxia; CHEN Shaoping

doi:10.19303/j.issn.1008-0384.2021.02.012

Volume 36 Issue 2

Feb. 2021

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Article Navigation > Fujian Journal of Agricultural Sciences > 2021 > 36(2): 215-220

HUANG Z, Hou Y M, GUO Q X, et al. Rapid Species-specific PCR Identification of Bactrocera tau (Diptera: Tephritidae) [J]. Fujian Journal of Agricultural Sciences，2021，36（2）：215−220 doi: 10.19303/j.issn.1008-0384.2021.02.012

Citation:

HUANG Z, Hou Y M, GUO Q X, et al. Rapid Species-specific PCR Identification of Bactrocera tau (Diptera: Tephritidae) [J]. Fujian Journal of Agricultural Sciences，2021，36（2）：215−220 doi: 10.19303/j.issn.1008-0384.2021.02.012

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Rapid Species-specific PCR Identification of Bactrocera tau (Diptera: Tephritidae)

doi: 10.19303/j.issn.1008-0384.2021.02.012

HUANG Zhen^{1, 2
,},
HOU Youming^{1
,
,},
GUO Qiongxia^{3
,
,},
CHEN Shaoping^{1, 3}

1.
College of Plant Protection, Fujian Agriculture and Forestry University, Fuzhou, Fujian 　350002, China
2.
Fuzhou Changle Airport Customs, Fuzhou, Fujian 　350209, China
3.
Fuzhou Customs Technology Center, Fuzhou, Fujian 　350001, China

Received Date: 2019-07-01
Rev Recd Date: 2020-11-20

Available Online: 2021-02-08

Publish Date: 2021-02-28

Abstract

Abstract

Objective A rapid method to detect Bactrocera tau (Walker) was established for inspection on produce to prevent potential introduction and spread of the invasive pest. Method Based on the species-specific PCR (SS-PCR) technology, B. tau as control and 20 common fruit flies of similar morphology were used to extract the genomic DNA templates, and the homology of the specimens checked with the COI mt DNA sequence. A rapid and accurate identification method applying the B. tau-specific primers NF404 and NR610 was designed and verified by PCR amplification and electrophoresis analysis. Result The newly developed SS-PCR detection method clearly amplified a single 207 bp band only on the target B. tau, not on any other fruit flies. Comparisons on the test results on the insects or body parts of all fruit flies with homogenous band sequence to B. tau validated the new methodology. Conclusion The newly established SS-PCR method exhibited high repeatability, specificity, and accuracy in detecting B. tau. It was considered applicable for the entry/exit and quarantine inspection on the invasive insect, larvae or dead body parts found on vegetables and fruits.
- Bactrocera tau (Walker),
- species-specific primers,
- species-specific PCR,
- mt DNA COI,
- rapid identification

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References(20)

References

[1]	中华人民共和国国家质量监督检验检疫总局. 南瓜实蝇检疫鉴定方法: SN/T 4176—2015[S]. 北京: 中国标准出版社, 2015.
[2]	安坤鹏, 吴宝峰, 申科, 等. 南亚寡鬃实蝇生物学特性观察及特性研究 [J]. 长江蔬菜, 2011(20):7−13. AN K P, WU B F, SHEN K, et al. Observation and study on biological characteristics of Bactrocera tau [J]. Journal of Changjiang Vegetables, 2011(20): 7−13.(in Chinese)
[3]	SHEN K, HU J, WU B, et al. Competitive interactions between immature stages of Bactrocera cucurbitae (Coquillett) And Bactrocera tau (walker) (Diptera: Tephritidae) under laboratory conditions [J]. Neotropical Entomology, 2014, 43(4): 335−343. doi: 10.1007/s13744-014-0224-y
[4]	SINGH S K, KUMAR D, RAMAMURTHY V V. Biology of Bactrocera (Zeugodacus) Tau (walker) (Diptera: Tephritidae) [J]. Entomological Research, 2010, 40(5): 259−263. doi: 10.1111/j.1748-5967.2010.00296.x
[5]	蒋小龙. 云南边境检疫性实蝇风险分析研究 [J]. 西南农业大学学报, 2002, 24(5):402−405, 421. doi: 10.3969/j.issn.1673-9868.2002.05.006 JIANG X L. Risk analysis of quarantine fruit flies along Yunnan borber [J]. Journal of Southwest Agricultural University, 2002, 24(5): 402−405, 421.(in Chinese) doi: 10.3969/j.issn.1673-9868.2002.05.006
[6]	WU B F, SHEN K, AN K P, et al. Effect of larval density and host species on preimaginal development of Bactrocera tau (Diptera: Tephritidae) [J]. Journal of Economic Entomology, 2011, 104(6): 1840−1850. doi: 10.1603/EC11154
[7]	黄振. 南瓜实蝇的形态特征、危害、寄主与其传入的风险 [J]. 武夷科学, 2017, 33(1):42−46. HUANG Z. The Morphological characteristics, harm, host and imported risk of Bactrocera (Zeugodacus) tau (Walker) [J]. Wuyi Science Journal, 2017, 33(1): 42−46.(in Chinese)
[8]	黄可辉, 郭琼霞, 虞赟, 等. 分子标记法在昆虫学研究中的应用 [J]. 华东昆虫学报, 2005, 14(2):109−114. HUANG K H, GUO Q X, YU Y, et al. The application of molecular marker methods in entomology studies [J]. Entomological Journal of East China, 2005, 14(2): 109−114.(in Chinese)
[9]	MUN J H, KIM J S, SONG Y H, et al. Molecular genetic diagnosis of four fruit fly species (Tephritidae) [J]. Journal of Asia-Pacific Entomology, 2000, 3(2): 89−94. doi: 10.1016/S1226-8615(08)60061-8
[10]	吴佳教, 胡学难, 赵菊鹏, 等. 9种检疫性实蝇PCR-RFLP快速鉴定研究 [J]. 植物检疫, 2005, 19(1):2−6. doi: 10.3969/j.issn.1005-2755.2005.01.002 WU J J, HU X N, ZHAO J P, et al. Rapid identification among 9 species of quarantine fruit flies(Diptera: Tephritidae)by PCR-RFLP [J]. Plant Quarantine, 2005, 19(1): 2−6.(in Chinese) doi: 10.3969/j.issn.1005-2755.2005.01.002
[11]	张亮, 张智英. 云南六种实蝇的RAPD快速鉴定 [J]. 应用生态学报, 2007, 18(5):1163−1166. doi: 10.3321/j.issn:1001-9332.2007.05.038 ZHANG L, ZHANG Z Y. Random amplified polymorphic DNA identification of six Bactrocera (Diptera: Tephritidae) species in Yunnan Province of Southwest China [J]. Chinese Journal of Applied Ecology, 2007, 18(5): 1163−1166.(in Chinese) doi: 10.3321/j.issn:1001-9332.2007.05.038
[12]	林丽莉, 吴佳教, 曾玲, 等. 二种大实蝇的PCR-RFLP快速鉴定方法 [J]. 昆虫知识, 2007, 44(4):588−592. LIN L L, WU J J, ZENG L, et al. Rapid identification of two species of Tetradacus by PCR-RFLP [J]. Chinese Bulletin of Entomology, 2007, 44(4): 588−592.(in Chinese)
[13]	CHUA T H, CHONG Y V, LIM S H. Species determination of Malaysian Bactrocera pests using PCR-RFLP analyses (Diptera: Tephritidae) [J]. Pest Management Science, 2010, 66(4): 379−384. doi: 10.1002/ps.1886
[14]	KAKOULI-DUARTE T, CASEY D G, BURNELL A M. Development of a diagnostic DNA probe for the fruit flies Ceratitis capitata and Ceratitis rosa (Diptera: Tephritidae) using amplified fragment-length polymorphism [J]. Journal of Economic Entomology, 2001, 94(4): 989−997. doi: 10.1603/0022-0493-94.4.989
[15]	刘慎思, 张桂芬, 万方浩. 基于mtDNA COI基因的离腹寡毛实蝇属常见种DNA条形码识别和系统发育分析 [J]. 昆虫学报, 2014, 57(3):343−355. LIU S S, ZHANG G F, WAN F H. DNA barcoding and phylogenetic analysis of common species of the genus Bactrocera (Diptera: Tephritidae) based on mtDNA COI gene sequences [J]. Acta Entomologica Sinica, 2014, 57(3): 343−355.(in Chinese)
[16]	LUNT D H, ZHANG D X, SZYMURA J M, et al. The insect cytochrome oxidase I gene: Evolutionary patterns and conserved primers for phylogenetic studies [J]. Insect Molecular Biology, 1996, 5(3): 153−165. doi: 10.1111/j.1365-2583.1996.tb00049.x
[17]	JAMNONGLUK W, BAIMAI V, KITTAYAPONG P. Molecular phylogeny of tephritid fruit flies in the Bactrocera tau complex using the mitochondrial COI sequences [J]. Genome, 2003, 46(1): 112−118. doi: 10.1139/g02-113
[18]	JAMNONGLUK W, BAIMAI V, KITTAYAPONG P. Molecular evolution of tephritid fruit flies in the genus Bactrocera based on the cytochrome oxidase I gene [J]. Genetica, 2003, 119(1): 19−25. doi: 10.1023/A:1024481032579
[19]	王芙蓉, 吴薇, 荣德福. 分子生物学技术在检疫性昆虫鉴定中的应用前景 [J]. 安徽农业科学, 2008, 36(8):3149−3151, 3162. doi: 10.3969/j.issn.0517-6611.2008.08.051 WANG F R, WU W, RONG D F. Application prospect of molecular biological techniques in identification of quarantine insect [J]. Journal of Anhui Agricultural Sciences, 2008, 36(8): 3149−3151, 3162.(in Chinese) doi: 10.3969/j.issn.0517-6611.2008.08.051
[20]	黄振, 陈韶萍, 谢婧, 等. 应用种特异性PCR技术快速鉴定辣椒实蝇 [J]. 昆虫学报, 2015, 58(4):460−466. HUANG Z, CHEN S P, XIE J, et al. Rapid identification of Bactrocera latifrons (Diptera: Tephritidae) by using species-specific PCR technique [J]. Acta Entomologica Sinica, 2015, 58(4): 460−466.(in Chinese)