Removing external DNA contamination from arthropod predators destined for molecular gut‐content analysis

Molecular Ecology Resources - Tập 12 Số 3 - Trang 464-469 - 2012
Matthew H. Greenstone1, Donald C. Weber1, Thomas A. Coudron2, Mark E. Payton3, Jing Hu1
1United States Department of Agriculture, Agricultural Research Service, Invasive Insect Biocontrol and Behavior Laboratory, Beltsville, MD 20705, USA.
2United States Department of Agriculture, Agricultural Research Service, Biological Control of Insects Research Laboratory, Columbia, MO 65203, USA
3Department of Statistics, 301 MSCS Building, Oklahoma State University, Stillwater, OK 74078, USA

Tóm tắt

AbstractEcological research requires large samples for statistical validity, typically hundreds or thousands of individuals, which are most efficiently gathered by mass‐collecting techniques. For the study of interspecific interactions, molecular gut‐content analysis enables detection of arthropod predation with minimal disruption of community interactions. Field experiments have demonstrated that standard mass‐collection methods, such as sweep netting, vacuum sampling and foliage beating, sometimes lead to contamination of predators with nontarget DNA, thereby compromising resultant gut‐content data. We deliberately contaminated immature Coleomegilla maculata and Podisus maculiventris that had been fed larvae of Leptinotarsa decemlineata by topically applying homogenate of the alternate prey Leptinotarsa juncta. We then attempted to remove contaminating DNA by washing in ethanol or bleach. A 40‐min wash with end‐over‐end rotation in 80% EtOH did not reliably reduce external DNA contamination. Identical treatment with 2.5% commercial bleach removed most externally contaminating DNA without affecting the detectability of the target prey DNA in the gut. Use of this bleaching protocol, perhaps with minor modifications tailored to different predator–prey systems, should reliably eliminate external DNA contamination, thereby alleviating concerns about this possible source of cross‐contamination for mass‐collected arthropod predators destined for molecular gut‐content analysis.

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Tài liệu tham khảo

Benton AH, 1981, Observations on the spring and summer behavior of the 12‐spotted ladybird beetle, Coleomegilla maculata (DeGeer) (Coleoptera: Coccinellidae), Journal of the New York Entomological Society, 89, 102

10.1111/j.1755-0998.2010.02857.x

10.1046/j.1461-9563.2003.00181.x

10.1007/s00442-008-1095-x

10.1111/j.1365-294X.2005.02628.x

10.1017/S000748530700497X

10.1111/j.1755-0998.2010.02922.x

10.1093/ee/23.1.193

10.1080/09583150500086474

10.1093/ee/37.4.990

Harwood JD, 2008, Recent Advances in Insect Physiology, Toxicology and Molecular Biology, 41

10.1111/j.1365-294X.2004.02331.x

10.1111/j.1365-294X.2007.03482.x

10.1093/ee/20.3.841

Heimpel GE, 1992, A survey of arthropod predators of Leptinotarsa decemlineata (Say) in Delaware potato fields, Journal of Agricultural Entomology, 9, 137

Hemenway R, 1967, Ground beetles of the genus Lebia Latreille in Arkansas (Coleoptera: Carabidae): ecology and geographical distribution, Arkansas Academy of Science Proceedings, 21, 15

10.1006/bcon.1997.0506

10.1007/s00114-009-0641-7

10.1111/j.1365-294X.2007.03238.x

10.1111/j.1365-294X.2010.04602.x

10.1111/j.1365-294X.2007.03613.x

10.1017/S0007485311000575

10.1016/j.baae.2007.08.012

10.1520/JFS15532J

10.1016/j.jinsphys.2009.11.020

10.1890/08-1882.1

10.1111/j.1365-2311.1992.tb01171.x

10.1653/0015-4040(2008)91[294:MSOEIF]2.0.CO;2

Prince AM, 1992, PCR: How to kill unwanted DNA, BioTechniques, 12, 358

10.1016/j.soilbio.2010.07.007

Ruberson JR, 1998, Predators of budworm/bollworm eggs in cotton: an immunological study, Proceedings, Beltwide Cotton Conference, 1095

10.1111/j.1365-294X.2004.02216.x

10.1023/A:1014577114915

10.1111/j.1365-294X.2004.02216.x

10.1046/j.1365-294X.2002.01471.x

10.1016/j.baae.2009.10.006

10.1673/1536-2442(2006)6[1:PPAHSO]2.0.CO;2

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Molecular Ecology Resources

Tập 12 Số 3

464-469

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