Study On Molecular Identification Of Common Poisonous And Confusable Edible Bulbous Plant Species Based On DNA Barcoding

Narcissus pseudonarcissus,Narcissus tazetta,Lycoris radiata,Zephyranthes candida,Tulipa gesneriana and Hyacinthus orienta.lis are common in China as ornamental plants,planted for urban greening and home landscaping.But these plants contain toxins and the morphological characteristics are similar to the edible species of bulbous plants,usually leading to accidental ingestion and food poisoning.How to identify the poisonous species of poisonous pulbs and confusable edible bulbs efficiently and accurately is very important for avoiding accidental ingestion and providing effective treatment of poisoning.DNA barcoding refers to a rapid,accurate and automated method for species identification,based on a short and standardized gene sequence.In this study,poisonous species of bulbous plants of Amaryllidaceae(Narcissus pseudonarcissus,Narcissus tazetta,Lycoris radiata,Zephyranthes Candida),poisonous species of bulbous plants of Liliaceae(Tulipa gesneriana,Hyacinthus orientalis)and edible plants of Liliaceae(Allium fistulosum,Allium tuberosum,Allium sativum,Allium cepa,Allium macrostemon,Allium chinense)were used to construct the table of classification of botany and database of DNA barcoding,and select the most suitable DNA barcoding for establishing the identification system with PCR-RFLP.The main results were as follows:1.Based on the morphological characteristics,the classification table of botany was listed for analyzing and distinguishing the morphological characteristics of Narcissus pseudonarcissus,Narcissus tazetta,Lycoris radiata,Zephyranthes candida,ulipa gesneriana,Hyacinthus orientalis,Allium fistulosum,Allium tuberosum,Allium sativum,Allium cepa,Allium macrostemon,Allium chinense.The morphological characteristics of the bulbs and leaves of poisonous species were found to be similar to those edible species,would be indistinguishable without further observing on the characteristics of the flowers.Therefore,it is easy to produce confusion in the case of samples deformed.It needs further analysis by different research methods,such as molecular markers.2.ITS2,matK,rbcL,psbA-trnH were selected as candidate DNA barcodes for the identification of poisonous species in Narcissus pseudonarcissus,Narcissus tazetta,Lycoris radiata,Zephyranthes candida,Tulipa gesneriana,Hyacinthus orientalis,Allium fistulosum,Allium tuberosum,Allium sativum,Allium cepa,Allium macrostemon,Allium chinense.Combining the distribution of K2P genetic distance,interspecies genetic distance difference and the identification efficiency of molecular phylogenetic tree,matK gene was selected as the optimal DNA barcode to identify poisonous species of bulbous plants.Simultaneously,it was found that the combination of matK and ITS2 had better identification accuracy.3.Six kinds of restriction enzyme Acc ?,Ava ?,ScrF ?,Bsm ?,Ban ?,and Bbs ?were selected to carry out the PCR-RFLP analysis of matK gene in Narcissus pseudonarcissus,Narcissus tazetta,Lycoris radiata,Zephyranthes candida,Tulipa gesneriana,Hyacinthus orientalis,Allium fistulosum,Allium tuberosum,Allium sativum,Allium cepa,Allium macrostemon and Allium chinense.The results showed that Acc ? could identify Lycoris and Liliaceae.Ban ? could distinguish Tulipa gesneriana,Hyacinthus orientalis from Allium fistulosum,Allium tuberosum,Allium fistulosum,Allium cepa and Allium macrostemon.Based on the identification of Acc? and Ban ?,the toxic species(Narcissus pseudonarcissus,Narcissus tazetta,Lycoris radiata,Zephyranthes candida,Tulipa gesneriana,Hyacinthus orientalis)in 12 bulbous plants could be identified.Combined with Ava ?,ScrF ? and Bsm ?,four kinds of toxic species of Lycoris could be fully identified.Bbs ? could simultaneously identify toxic species of Lycoris(Tulipa gesneriana and Hyacinthus orientalis).