Abstract

Twenty-two sacred lotus (Nelumbo nucifera), 46 taros (Colocasia esculenta) and 10 arrowheads (Sagittaria trifolia) were used as materials and combined with EST-SSR (expressed sequence tag-simple sequence repeats) primers developed by our laboratory. Core primers were screened from a large number of primers that were able to distinguish all materials with a high frequency of polymorphisms. Six pairs, twenty pairs and three pairs of core primers were screened from sacred lotus, taro, and arrowhead, respectively. The SSR fingerprints of these three important aquatic vegetables, producing 17-, 87- and 14-bit binary molecular identity cards, respectively, were separately determined by using the core primers. Since there were few core primers of sacred lotus and arrowhead, 3 and 9 primer pairs with higher polymorphic information content (PIC), respectively, were selected as candidate primers. These core and candidate primers were used to identify the purities of No.36 space lotus, Shandong 8502 taro and Wuhan arrowhead, which were 93.3% (84/90), 98.9% (89/90) and 100.0% (90/90), respectively. The fingerprints, displayed as binary molecular identification cards of three important aquatic vegetables, were obtained, and their purity was successfully determined with EST-SSR labeling technology. Phylogenetic trees were also constructed to analyze the genetic diversity of 22 sacred lotus, 46 taros and 10 arrowheads. This study classifies and identifies germplasm resources and is an important reference to test the authenticity and variety purity of other aquatic vegetables in the future.

Highlights

  • The authenticity of the variety and quality and purity of seeds are extremely significant for crop and vegetable production[15]

  • DNA fingerprints are generally constructed by three methods: the characteristic locus method, the single primer method and the core primer combination method, the most efficient of which is the core primer combination method[4,19,20,21,22]

  • Identifying the the core primers of sacred lotus, taro and arrowhead is a key step for SSR fingerprint construction and purity identification, and it is a prerequisite for the commercial application of SSR fingerprints

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Summary

Introduction

The authenticity of the variety and quality and purity of seeds are extremely significant for crop and vegetable production[15]. Identification by SSR markers is accurate, can be accomplished within a short time, is low-cost, and is immune to environmental and self factors. Due to the limited research on molecular markers, most aquatic vegetables do not have established molecular marker fingerprints or DNA fingerprints constructed from EST-SSR markers. There is no way to identify the authenticity of varieties or the quality and purity of seeds by molecular markers. The purpose of this study is to construct DNA fingerprints for aquatic vegetables (sacred lotus, taro and arrowhead) by using a large number of EST-SSR molecular markers developed in our laboratory and to identify the purity of these aquatic vegetables. We aim to provide theoretical knowledge for the construction of DNA fingerprints and purity identification of other aquatic vegetables

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