Abstract
Cordyceps sinensis is an endoparasitic fungus widely used as a tonic and medicinal food in the practice of traditional Chinese medicine (TCM). In historical usage, Cordyceps specifically is referring to the species of C. sinensis. However, a number of closely related species are named themselves as Cordyceps, and they are sold commonly as C. sinensis. The substitutes and adulterants of C. sinensis are often introduced either intentionally or accidentally in the herbal market, which seriously affects the therapeutic effects or even leads to life-threatening poisoning. Here, we aim to identify Cordyceps by DNA sequencing technology. Two different DNA-based approaches were compared. The internal transcribed spacer (ITS) sequences and the random amplified polymorphic DNA (RAPD)-sequence characterized amplified region (SCAR) were developed here to authenticate different species of Cordyceps. Both approaches generally enabled discrimination of C. sinensis from others. The application of the two methods, supporting each other, increases the security of identification. For better reproducibility and faster analysis, the SCAR markers derived from the RAPD results provide a new method for quick authentication of Cordyceps.
Highlights
The term Cordyceps normally refers to the species of C. sinensis
According to the Chinese Pharmacopoeia 2015, C. sinensis is the only source of Cordyceps [3]
FigurFeig1u.rCe o1.rdCyocrdeypcseppsropdroudcuticotnionininCChhininaaaanndd iittss ccoolllleeccttiioonn. .(A(A) T) hTehgeegogeroagprhaipcahlicloaclaltoiocnastioofnms aojformajor Cord(CySco1er3dp–ySsc1pe6pr)oscdoplurloeccdttiueodcntiiosnintHesosit;ne(gsB;K()Bot)wntgwehlevelervbecaoclommmammrkeeerrtccsiiaawll essraaemmupspleeldessionof tfChC.iss.iensxienpnesenirsism(iSse1(n–SSt1.1–E2Sa)1cah2n)sdaaimntsdpaldietushlaateddrfuaonluttesrrants (S13–bSa1t6ch) ecso,lnle=ct4e.d in Hong Kong herbal markets were used in this experiment
Summary
Cordyceps has been used as medicine and a food supplement with an overwhelming list of pharmacological properties for hundreds of years in China. There are about 400 species in the Cordyceps genus in the world, mainly distributed in Eurasia, including Java, Sri Lanka, Tasmania, Japanese islands, China and Australia [1,2]. The term Cordyceps normally refers to the species of C. sinensis. According to the Chinese Pharmacopoeia 2015, C. sinensis is the only source of Cordyceps [3]. Cordyceps is rare and expensive, because it is wild and distributed at an elevation of 3000–5000 m, mainly in the provinces of Yunnan, Sichuan, Qinghai, Xizang and. GansMuoloecfulCesh20i1n5a, 2.0,Dpaugee–ptaogevery limited resources and huge market demand, Cordyceps is the most expensive herbal medicine in China, whose price is comparable to gold [4]. Tashpeeckt noof wsunchmcoenthtrodl iss tihne avuetrhifeicnaticoantionfgspheceirebsableimngatuesreidalsasbthye msoourrpcehomlaotgeirciaall., Tmheircerfoosrceo, pthiec and phytodcehvelmopicmael nmt oetfhqoudicskaarnedneoatsysumffietchioednst fionrtthe acuastheeonftiCcaotirodnyocfeCpos.rdInycdeeppsesnhdouenldt hfraovme gerenavtivroalnume.ental influencesThoer konroigwin omferthaowdsminatearuitahle, ntthiceatuinsgagheerobfalDmNaAtersieaqlsubeyncminogrpohrolmogoilceacl,umlaircrgoesncoeptiic manedthods can cpohmyptolcehmemenictatlhmeecthoondtsroalreonfoatustuhffeinciteinctaitniotnhe[5ca].seIonf Canordayttceempsp. tIntdoepenensduerentafrohmighenlyvirroenlimabelnetarlesult, two idnifflfueernecnets omreotrhigoidnsofhraawvembaeteerniald, tehveeulosapgeedofinDNaAutsheeqnuteincacitninggorCmoorldecyucleaprsg:enientitcemrneatlhotdrasncsacnribed sapmapcelcd(irIfioiTf(mefSIedT)prSelsen)remetsqgeeumiqneoetunntctehhenose(cdSecsaCosnhAnadatnRrvotd)ehl.etbohBefreeoaanrtnuhadtdnhoedemmvnoetemiltacohmapaoteipmdodlnispfiinl[eia5dfi]ra.eeupIdtnohppleayaonnmrtltiyaioccmtartupteiolmhnarigrppclthyCDtioocNarDdepAnypNscr(ueAoRprpAes(:rRPaiiaDAnhtt)Peie-gsDrhenilq)nay-ulseretdenrqleaciuanaeeblsicnlcnerhcgiabererecaswdhuctialsettrp,hraiatzcwceatedeorrhizigedh perceanmtapgliefioedf inretegrio-snpe(SciCfiAcRse).quBoetnhcemdeitvheordgsenacree, apnadrtibcuoltahrlayppaproparochpreisatheavine adlereaalidngy bweietnh sauchciegshsfully usedpfoerrcethnetaigdeenotfifiicnatteiro-snpoecfimficedsiecqiuneanlcpeladnitvseirngepnrceev, ioaunds stbuodthiesa[p6p–r8o]a.cBheyscohmavpearailnrgeatdhye obueetcnomes of twsuoccmesestfhulolydsu,setdheforRtAhePiDd-eSnCtifAicRatiomneothf omdedpicrionvalidpeladntsa inqupircekveiorusanstduduiesse[r6-–fr8i]e.nBdylycomtopoalrinfogr the authethneticoautticoonmoefs Cofotrwdoycmepetsh.ods, the RAPD-SCAR method provided a quicker and user-friendly tool for the authentication of Cordyceps
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