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A molecular approach to species identification of Chenopodiaceae pollen grains in surface soil¹

Laboratory of Quantitative Vegetation Ecology, Institute of Botany, Chinese Academy of Sciences, 20 Nanxincun, Xiangshan, Haidian District, Beijing 100093, P.R. China; School of Engineering and Applied Science, University of Pennsylvania, Philadelphia 19104 USA

ABSTRACT

Pollen identification and classification are important not only for palynologists, but also for systematists and ecologists. Because palynological methods for the identification of pollen in surface soil until now could resolve at best to the generic level, we have developed a molecular approach to species-level identification of Chenopodiaceae pollen in surface soils. Surface soil samples were collected in the central area of Junggar Desert Basin, Xinjiang, China. Fresh leaves of 19 Chenopodiaceae species were sampled for DNA sequencing, establishing a database of internal transcribed spacer (ITS) regions of nuclear ribosomal DNA for Chenopodiaceae. Individual chenopod pollen grains in a soil sample were separated from the soil and the ITS1 region of each pollen grain was amplified using nested PCR and sequenced. By comparing the amplified ITS1 sequences to those in the Chenopodiaceous database, we identified the pollen in the soil samples to the level of species. The new method provides a technical reference for species identification of soil surface pollen for other families. This work is necessary for further efforts to interpret the relationship of surface soil pollen to vegetation characteristics. It also has significant potential for enhancing the ability to identify pollen in clinical airborne allergen or criminological studies.

Key Words: Chenopodiaceae • internal transcribed spacer sequence • Junggar Desert Basin • modern pollen • pollen identification