关键词: 厚几丁质外壳, 微小节肢动物, 无损伤, 凭证标本

Abstract:

Objective In order to establish a method of obtaining high quality DNA templates from tiny thick-chitin arthropods without grinding or damaging morphology of the voucher specimens. Methods Tiny soil oribatid mites with thick-chitin were used as research materials. We used chitinase to digest the external chitin skeleton first,after the digestion,the individuals were treated with lysate. Amplification efficiency of the DNA barcodes were used as the indicator for DNA template quality. Working concentration and condition of chitinase were optimized. Morphology of the samples treated with or without the chitinase were compared. Results High quality DNA template could be obtained with the method we established,working concentration of chitinase was optimized to 1 mg/ml, and working time was optimized to 24 h. The morphology of oribatid mites treated with the chitinase has no obvious changes compared to the untreated ones, and could be used in the morphology species identification. Conclusion Chitinase is first applied to the digestion of chitin in the extraction of template DNA, high quality DNA could be obtained without damaging the morphological characteristics, after the extraction, the tiny arthropods could be mounted on the slides and kept as voucher specimen.

Key words: Thick chitin, Tiny arthropods, No damage, Voucher specimens