Application of a novel method PCR-ligase detection reaction for tracking predator—prey trophic links in insect-resistant GM rice ecosystem

Ecotoxicology - Tập 20 - Trang 2090-2100 - 2011
Kai Li1, Junce Tian2, Qinxi Wang1, Qiang Chen1, Mao Chen2, Huan Wang3, Yuxun Zhou1, Yufa Peng4, Junhua Xiao1, Gongyin Ye2,5
1Institute of Biological Sciences and Biotechnology, Donghua University, Shanghai, China
2State Key Laboratory of Rice Biology, Ministry of Agriculture Key Laboratory of Molecular Biology of Crop Pathogens and Insects, Institute of Insect Sciences, Zhejiang University, Hangzhou, China
3Shanghai Vocational and Technical College of Agriculture and Forestry, Shanghai, China
4State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing, China
5Institute of Insect Sciences, College of Agriculture and Biotechnology, Zhejiang University, Hangzhou, China

Tóm tắt

Insect-resistant genetically modified (IRGM) rice is on the verge of commercial release in China, however, its potential non-target effect on non-target insect natural enemies remains controversial. Tracking trophic interactions between predators and preys in IRGM rice ecosystem can provide new insights into better understanding of the ecological risks of IRGM rice. In the present study, a novel method based on ligase detection reaction (LDR), PCR-LDR was introduced to track 15 prey species in the gut of a predaceous spider Pirata subpiraticus, a dominant natural enemy in rice field. Our results indicated that PCR-LDR could provide high specificity and sensitivity in tracking prey-predator interactions in rice ecosystems. PCR-LDR could detect as little as 1,000th of DNA mixture. Reliable detection of DNA samples of prey species using PCR-LDR could be significantly affected by digestion time and prey species. In the analysis of 200 field-collected P. subpiraticus and 105 field-collected Tetragnatha maxillosa individuals using PCR-LDR, prey remains were identified in 78.3 and 74.3% of the individuals, respectively, from which significant predation differences between the two spider species were observed. Predation behavior of the spider species was not significantly different between Bt and non-Bt control rice lines. These results indicated that PCR-LDR can be used as an important tool for ecological studies, especially on the interactions between predators and preys in IRGM rice or other similar ecosystems.

Tài liệu tham khảo

Admassu B, Juen A, Traugott M (2006) Earthworm primers for DNA-based gut content analysis and their cross-reactivity in a multi-species system. Soil Biol Biochem 38:1308–1315 Agustí N, De Vicente MC, Gabarra R (1999) Development of sequence amplified characterized region (SCAR) markers of Helicoverpa armigera: a new polymerase chain reaction-based technique for predator gut analysis. Mol Ecol 8:1467–1474 Agustí N, Shayler SP, Harwood JD, Vaughan IP, Sunderland KD, Symondson WOC (2003) Collembola as alternative prey sustaining spiders in arable ecosystems: prey detection within predators using molecular markers. Mol Ecol 12:3467–3475 Alvarez-Alfageme F, Ferry N, Castanera P, Ortego F, Gatehouse AMR (2008) Prey mediated effects of Bt maize on fitness and digestive physiology of the red spider mite predator Stethorus punctillum Weise (Coleoptera: Coccinellidae). Transgenic Res 17:943–954 Asahida T, Yamashita Y, Kobayashi T (1997) Identification of consumed stone flounder, Kareius bicoloratus (Basilewsky), from the stomach contents of sand shrimp, Crangon affinis (De Haan) using mitochondrial DNA analysis. J Exp Mar Biol Ecol 217:153–163 Chapman EG, Romero SA, Harwood JD (2010) Maximizing collection and minimizing risk: does vacuum suction sampling increase the likelihood for misinterpretation of food web connections? Mol Ecol Res 10:1023–1033 Chen M, Zhao JZ, Ye GY, Fu Q, Shelton AM (2006) Impact of insect-resistant transgenic rice on target insect pests and non-target arthropods in China. Insect Sci 13:409–420 Chen M, Liu Z, Ye G, Shen Z, Hu C, Peng Y, Altosaar I, Shelton AM (2007) Impacts of transgenic cry1Ab rice on non-target planthoppers and their main predator Cyrtorhinus lividipennis (Hemiptera: Miridae) – A case study of the compatibility of Bt rice with biological control. Biol Control 42:242–250 Chen M, Zhao JZ, Collins HL, Earle ED, Cao J, Shelton AM (2008) A critical assessment of the effects of Bt transgenic plants on parasitoids. PLoS ONE 3:e2284 Chen M, Ye GY, Liu ZC, Fang Q, Hu C, Peng YF, Shelton AM (2009) Analysis of Cry1Ab toxin bioaccumulation in a food chain of Bt rice, an herbivore and a predator. Ecotoxicology 18:230–238 Cohen M, Chen M, Bentur JS, Heong KL, Ye GY (2008) Bt rice in Asia: potential benefits, impact, and sustainability. In: Romeis J, Shelton AM, Kennedy GG (eds) Integration of insect-resistant GM crops within IPM programs. Springer, Dordrecht, pp 223–248 Desneux N, Bernal JS (2010) Genetically modified crops deserve greater ecotoxicological scrutiny. Ecotoxicology 19:1642–1644 Desneux N, Ramirez-Romero R, Bokonon-Ganta AH, Bernal JS (2010) Attraction of the parasitoid Cotesia marginiventris to host frass is affected by transgenic maize. Ecotoxicology 19:1183–1192 Ferry N, Mulligan EA, Majerus MEN, Gatehouse AMR (2007) Bitrophic and tritrophic effects of Bt Cry3A transgenic potato on beneficial, non-target, beetles. Transgenic Res 16:795–812 Folmer O, Black M, Hoeh W, Vrijenhoek R, Lutz R (1994) DNA primers for amplification of mitochondrial cytochrome c oxidase subunit I from diverse metazoan invertebrates. Mol Mar Biol Biotech 3:294–299 Harper GL, King RA, Dodd CS, Harwood JD, Glen DM, Bruford MW, Symondson WO (2005) Rapid screening of invertebrate predators for multiple prey DNA targets. Mol Ecol 14:819–827 Harwood JD (2008) Are sweep net sampling and pitfall trapping compatible with molecular analysis of predation? Environ Entomol 37:990–995 Harwood JD, Desneux N, Yoo HJS, Rowley DL, Greenstone MH, Obrycki JJ, O’Neil RJ (2007) Tracking the role of alternative prey in soybean aphid predation by Orius insidiosus: a molecular approach. Mol Ecol 16:4390–4400 Juen A, Traugott M (2006) Amplification facilitators and multiplex PCR: tools to overcome PCR-inhibition in DNA-gut-content analysis of soil-living invertebrates. Soil Biol Biochem 38:1872–1879 Kagata H, Ohgushi T (2006) Bottom-up trophic cascades and material transfer in terrestrial food webs. Ecol Res 21:26–34 King RA, Read DS, Traugott M, Ttrugott M, Symondson WOC (2008) Molecular analysis of predation: a review of best practice for DNA-based approaches. Mol Ecol 17:947–963 Kuuska AK, Cassel-Lundhagena A, Kvarnhedenb A et al (2008) Tracking aphid predation by lycosid spiders in spring-sown cereals using PCR-based gut-content analysis. Basic Appl Ecol 9:718–725 Li K, Chen B, Zhou Y, Huang R, Liang Y, Wang Q, Xiao Z, Xiao J (2009) Multiplex quantification of 16S rDNA of predominant bacteria group within human fecal samples by polymerase chain reaction-ligase detection reaction (PCR-LDR). J Microbiol Method 76:289–294 Liu ZC, Ye GY, Hu C, Datta SK (2002) Effects of Bt transgenic rice on population dynamics of main non-target insect pests and dominant spider species in rice paddies. Acta Phytophyl Sin 29:138–144 Liu ZC, Ye GY, Fu Q, Zhang ZT (2003a) Indirect impact assessment of transgenic rice with cry1Ab gene on predations by the wolf spider, Pirata subpiraticus. Chin J Rice Sci 17:175–178 Liu ZC, Ye GY, Hu C, Datta SK (2003b) Impact of transgenic indica rice with a fused gene of cry1Ab/cry1Ac on the rice paddy arthropod community. Acta Entomol Sin 46:454–465 Liu ZC, Ye GY, Hu C (2004) Effects of Bacillus thuringiensis transgenic rice and chemical insecticides on arthropod communities in paddy-fields. Chin J Appl Ecol 15:2309–2314 Lövei GL, Andow DA, Arpaia S (2009) Transgenic insecticidal crops and natural enemies: a detailed review of laboratory studies. Environ Entomol 38:293–306 Ma J, Li D, Keller M, Schmidt O, Feng X (2005) A DNA marker to identify predation of Plutella xylostella (Lep., Plutellidae) by Nabis kinbergii (Hem., Nabidae) and Lycosa sp (Aranaea, Lycosidae). J Appl Entomol 129:330–335 Mathews RW, Mathews JR (1970) Malaise trap studies of flying insects in a New York mesic forest. I. Ordinal composition and seasonal abundance. NY Entomol Soc 78:52–59 Monzó C, Sabater-Muñoz B, Urbaneja A, Castañera P (2009) Tracking medfly predation by the wolf spider, Pardosa cribata Simon, in citrus orchards using PCR-based gut-content analysis. B Entomol Res 100:145–152 Ramirez-Romero R, Bernal JS, Chaufaux J, Kaiser L (2007) Impact assessment of Bt-maize on a moth parasitoid, Cotesia marginiventris (Hymenoptera : Braconidae), via host exposure to purified Cry1Ab protein or Bt-plants. Crop Protection 26:953–962 Romeis J, Meissle M, Bigler F (2006) Transgenic crops expressing Bacillus thuringiensis toxins and biological control. Nat Biotechnol 24:63–71 Romeis J, Bartsch D, Bigler F, Candolfi MP, Gielkens MM, Hartley SE, Hellmich RL, Huesing JE, Jepson PC, Layton R, Quemada H, Raybould A, Rose RI, Schiemann J, Sears MK, Shelton AM, Sweet J, Vaituzis Z, Wolt JD (2008) Assessment of risk of insect-resistant transgenic crops to nontarget arthropods. Nat Biotechnol 26:203–208 Sanvido O, Romeis J, Bigler F (2009) An approach for post-market monitoring of potential environmental effects of Bt-maize expressing Cry1Ab on natural enemies. J Appl Entomol 133:236–248 Sheppard SK, Harwood JD (2005) Advances in molecular ecology: tracking trophic links through predator-prey food-webs. Funct Ecol 19:751–762 Simon C, Frati F, Beckenbach A, Crespi B, Liu H, Flook P (1994) Evolution, weighting, and phylogenetic utility of mitochondrial gene sequences and a compilation of conserved polymerase chain reaction primers. Ann Entomol Soc Am 87:651–701 Symondson WOC, Glen DM, Ives AR, Langdon CJ, Wiltshire CW (2002) Dynamics of the relationship between a generalist predator and slugs over five years. Ecology 83:137–147 Thompson JD, Gibson TJ, Plewniak F, Jeanmougin F, Higgins DG (1997) The CLUSTAL_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Res 25:4876–4882 Wu JH, Liu WT (2007) Quantitative multiplexing analysis of PCR-amplified ribosomal RNA genes by hierarchical oligonucleotide primer extension reaction. Nucleic Acids Res 35:e82 Wu JC, Lu ZQ, Yang JS, Su ZL (1993) Habitat niche and predation effect of natural enemies of insect pests in paddy field. Acta Entomol Sin 36:323–331 Ye GY, Shu QY, Yao HW, Cui HR, Cheng XY, Hu C, Xia YW, Gao MW, Altosaar I (2001) Field evaluation of resistance of transgenic rice containing a synthetic cry1Ab gene from Bacillus thuringiensis Berliner to two stem borers. J Econ Entomol 94:270–276 Ye GY, Yao HW, Shu QY, Cheng X, Hu C, Xia YW, Gao MW, Altosaar I (2003) High levels of stable resistance in transgenic rice with a synthetic cry1Ab gene from Bacillus thuringiensis Berliner to rice leaffolder, Cnaphalocrocis medinalis (Guenée) under field conditions. Crop Prot 22:171–178 Zhang GF, Lu ZC, Wan FH, Lovei GL (2007) Real-time PCR quantification of Bemisia tabaci (Homoptera: Aleyrodidae) B-biotype remains in predator guts. Mol Ecol Notes 7:947–954