Selection of a marker gene to construct a reference library for wetland plants, and the application of metabarcoding to analyze the diet of wintering herbivorous waterbirds

• Published in: PeerJ
• Main Authors: Yang, Yuzhan, Zhan, Aibin, Cao, Lei, Meng, Fanjuan, Xu, Wenbin
• Other Authors: National Basic Research Program of China, National Natural Science Foundation of China, State Key Laboratory of Urban and Regional Ecology, Chinese Academy of Sciences
• Format: Article in Journal/Newspaper
• Language: English
• Published: PeerJ 2016
• Subjects: Anser fabalis
• Online Access: http://dx.doi.org/10.7717/peerj.2345; https://peerj.com/articles/2345.pdf; https://peerj.com/articles/2345.xml; https://peerj.com/articles/2345.html

Food availability and diet selection are important factors influencing the abundance and distribution of wild waterbirds. In order to better understand changes in waterbird population, it is essential to figure out what they feed on. However, analyzing their diet could be difficult and inefficient using traditional methods such as microhistologic observation. Here, we addressed this gap of knowledge by investigating the diet of greater white-fronted goose Anser albifrons and bean goose Anser fabalis , which are obligate herbivores wintering in China, mostly in the Middle and Lower Yangtze River floodplain. First, we selected a suitable and high-resolution marker gene for wetland plants that these geese would consume during the wintering period. Eight candidate genes were included: rbc L, rpo C1, rpo B, mat K, trn H- psb A, trn L (UAA), atp F -atp H, and psb K -psb I. The selection was performed via analysis of representative sequences from NCBI and comparison of amplification efficiency and resolution power of plant samples collected from the wintering area. The trn L gene was chosen at last with c/h primers, and a local plant reference library was constructed with this gene. Then, utilizing DNA metabarcoding, we discovered 15 food items in total from the feces of these birds. Of the 15 unique dietary sequences, 10 could be identified at specie level. As for greater white-fronted goose, 73% of sequences belonged to Poaceae spp., and 26% belonged to Carex spp. In contrast, almost all sequences of bean goose belonged to Carex spp. (99%). Using the same samples, microhistology provided consistent food composition with metabarcoding results for greater white-fronted goose, while 13% of Poaceae was recovered for bean goose. In addition, two other taxa were discovered only through microhistologic analysis. Although most of the identified taxa matched relatively well between the two methods, DNA metabarcoding gave taxonomically more detailed information. Discrepancies were likely due to biased PCR amplification in ...