In order to further solve the problem of non-targeted screening of plant components adulteration based on amplification technique in food, and to realize the high-throughput monitoring and evaluation of plant components adulteration, a non-targeted screening method was developed that based on amplification technique and using the common plant primers RbcL. The specificity, universality and sensitivity of this method were verified, and the sequencing results were analyzed by DNAMAN and the national center of biotechnology information database. Firstly, DNA were extracted from 27 plant species including adzuki bean, soybean, mung bean, sweet potato, potato, cassava, coix seed, wheat, corn, millet, rice, apricot, cashew nut, hazelnut, sesame, almond, walnut, peach, banana, pear, tomato, pumpkin, carrot, apple, peanut, pistachio and kidney bean. In addition, the second-generation sequencing analysis technology was used for high-throughput and non-directional screening of 4 kinds of spiked food samples. The sequencing results were consistent with product identification. The establishment of this method was of great significance to the identification and supervision for adulteration of plant-derived components and the protection of consumers′ rights and interests.
[1] 孙善峰, 黄晓宁, 韩北忠, 等.扩增子测序分析助力传统发酵食品微生物群落研究[J].中国酿造, 2019, 38(5):1-5.
SUN S F, HUANG X N, HAN B Z, et al.Amplicon sequencing analysis enhancing the understanding on microbial diversity of traditional Chinese fermented foods[J].China Brewing, 2019, 38(5):1-5.
[2] 刘伟, 张纪阳, 谢红卫.生物信息学[M].北京:电子工业出版社, 2014:5-15.
LIU W, ZHANG J Y, XIE H W.Bioinformations[M].Beijing:Publishing House of Electronics Industry, 2014:5-15.
[3] 颜亨梅, 钟文涛.动物捕食性天敌摄食分析方法的研究进展[J].生命科学研究, 2021, 25(1):1-8.
YAN H M, ZHONG W T.Research progress of feeding analysis methods for predatory animals[J].Life Science Research, 2021, 25(1):1-8.
[4] BUGLIONE M, MASELLI V, RIPPA D, et al. A pilot study on the application of DNA metabarcoding for non -invasive diet analysis in the Italian hare[J].Mammalian Biology, 2018, 88:31-42.
[5] JENSEN-VARGAS E, MARIZZI C.DNA barcoding for identification of consumer-relevant fungi sold in New York:A powerful tool for citizen scientists?[J].Foods(Basel, Switzerland), 2018, 7(6):87.
[6] XING B P, LIN H S, ZHANG Z L, et al.DNA barcoding for identification of fish species in the Taiwan Strait[J].PLoS One, 2018, 13(6):0198109.
[7] QASIM M, WANG B H, ZOU H S, et al.Phylogenetic relationship and genetic diversity of citrus psyllid populations from China and Pakistan and their associated Candidatus bacterium[J].Molecular Phylogenetics and Evolution, 2018, 126:173-180.
[8] HAMBÄCK P A, WEINGARTNER E, DALÉN L, et al.Spatial subsidies in spider diets vary with shoreline structure:Complementary evidence from molecular diet analysis and stable isotopes[J].Ecology and Evolution, 2016, 6(23):8 431-8 439.
[9] 白文明, 邢冉冉, 陈丽萍, 等.基于DNA条形码技术鉴别有毒鹅膏菌属物种[J].食品科学, 2021, 42(4):278-286.
BAI W M, XING R R, CHEN L P, et al.DNA barcoding for identification of toxic Amanita species[J].Food Science, 2021, 42(4):278-286.
[10] BENSON D A, CAVANAUGH M, CLARK K, et al.GenBank[J].Nucleic Acids Research, 2013, 41:D36-D42.
[11] COCHRANE G, ALDEBERT P, ALTHORPE N, et al.EMBL nucleotide sequence database:Developments in 2005[J].Nucleic Acids Research, 2006, 34(S1):D10-D15.
[12] MASHIMA J, KODAMA Y, KOSUGE T, et al.DNA data bank of Japan (DDBJ) progress report[J].Nucleic Acids Research, 2015, 44(D1):D51-D57.
[13] 张稚兰, 邢炳鹏.小球藻属DNA条形码的鉴定研究[J].海洋开发与管理, 2021, 38(2):69-76.
ZHANG Z L, XING B P.Assessment of candidate DNA sequences for barcoding of the genus Chlorellla[J].Ocean Development and Management, 2021, 38(2):69-76.
[14] LI X L, HONG M H.Aqueous extract of Dendrobium officinale confers neuroprotection against hypoxic-ischemic brain damage in neonatal rats[J].The Kaohsiung Journal of Medical Sciences, 2020, 36(1):43-53.
[15] JIN W T, JIN X H, SCHUITEMAN A, et al.Molecular systematics of subtribe Orchidinae and Asian taxa of Habenariinae (Orchideae, Orchidaceae) based on plastid matK, rbcL and nuclear ITS[J].Molecular Phylogenetics and Evolution, 2014, 77:41-53.
[16] 杨荣武. 分子生物学[M].南京:南京大学出版社, 2017:636-637.
YANG R W.Molecular Biology[M].Nanjing:Nanjing University Press, 2017:636-637.
[17] 姚丹丹, 陈佐, 朱军, 等.基于coxⅠ、ITS和rbcL基因序列的马尾藻系统进化分析[J].分子植物育种, 2020,20(4):1 153-1 162.
YAO D D, CHEN Z, ZHU J, et al.Genetic evolution analysis of Sargassum based on coxⅠ, ITS and rbcL gene sequences[J].Molecular Plant Breeding, 2020,20(4):1 153-1 162.
[18] FAWLEY M W, FAWLEY K P, CAHOON A B.Finding needles in a haystack-Extensive diversity in the eustigmatophyceae revealed by community metabarcode analysis targeting the rbcL gene using lineage-directed primers[J].Journal of Phycology, 2021, 57(5):1 636-1 647.
[19] 陈玉花, 撒切尔, 肖田梅, 等.蒙药蜀葵花HPLC指纹图谱及rbcL序列分子鉴定研究[J].中草药, 2020, 51(21):5 607-5 612.
CHEN Y H, SA Q E, XIAO T M, et al.Fingerprint and molecular identification of rbcL sequence of Mongolian medicine Althaeae Roseae Flos by HPLC[J].Chinese Traditional and Herbal Drugs, 2020, 51(21):5 607-5 612.
[20] 林晓霞, 鹿炎, 李会丽, 等.基于matK和rbcL序列的石斛属植物亲缘关系研究[J].生物学杂志, 2021, 38(1):46-50.
LIN X X, LU Y, LI H L, et al.Genetic relationship analysis of Dendrobium plants based on chloroplast matK and rbcL gene[J].Journal of Biology, 2021, 38(1):46-50.
[21] 赵琳娜, 胡凤月, 吴孝槐, 等.用于转基因检测的番木瓜基因组DNA提取方法的比较[J].现代食品科技, 2010, 26(2):188-191.
ZHAO L N, HU F Y, WU X H, et al.Study on DNA extraction from papaya for detection of genetically modified organism[J].Modern Food Science and Technology, 2010, 26(2):188-191.
[22] LANDEWEERT R, LEEFLANG P, KUYPER T W, et al.Molecular identification of ectomycorrhizal mycelium in soil horizons[J].Applied Environmental Microbiology, 2003, 69(1):327-333.
[23] 夏梦雷, 杨帆, 陆锴, 等.高通量测序技术在传统发酵食品微生物群落中的应用研究[J].中国酿造, 2021, 40(5):1-7.
XIA M L, YANG F, LU K, et al.Advances in the application of high-throughput sequencing technology in traditional fermented foods[J].China Brewing, 2021, 40(5):1-7.
[24] 翟清燕, 任易婕, 郑世超, 等.三种植物源性成分阳性质粒的构建及检测方法的验证[J].食品与发酵工业, 2020, 46(17):237-241.
ZHAI Q Y, REN Y J, ZHENG S C, et al.Verification of detection method and construction of plasmid molecule for three plant-derived components[J].Food and Fermentation Industries, 2020, 46(17):237-241.
