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Kev txhim kho ntawm phenylethanoid glycosides ntau lawm los ntawm fungal elicitor nyob rau hauv cell suspension kab lis kev cai ntawm Cistanche deserticola

Cui-tao Lu¹, ∗ & Xing-guo Mei²

Ntsiab lus Cistanche deserticola, fungal elicitor,phenylethanoid glycoside, cog cell culture

Abstract
Thaum, nyob rau 15th hnub ntawm kev loj hlob, ib tug elicitor los ntawmFusarium solanitau ntxiv ntawm 40 mg l^-1 rauCistanche deserticolacell suspension cultures, cov ntsiab lus ntawm echinacoside, acteoside, thiab tag nrhophenylethanoid glycosides(PeGs) nyob rau hauv cov kab lis kev cai tag nrho tau nce ntxiv tom ntej 27 d los ntawm ntau dua 100 feem pua ​​​​rau 15 mg g^-1qhuav wt, 9 mg g -1 qhuav wt thiab 57 mg g^-1 dry wt, ua. Qhov kawg biomass (1.3 mg qhuav wt ml^-1) tsis cuam tshuam.

Cistanche deserticola

Taw qhia

Cistanche deserticolaMa. (Orobanchaceae), ib hom tshuaj ntsuab ntawm Tuam Tshoj yog dav siv nyob rau hauv cov tshuaj ib txwm siv rau ntau yam kev kho mob xws li siv raws li ib tug sedative los yog ib tug tiv thaiv kab mob intensifier (Hsu et al. 1995, Chang li al. 1995).Phenylethanoid glycosides(PeGs) yog cov bioactive metabolites loj hauvCistanche deserticola(Xiong li al. 2000) thiab muaj ntau yam xws li echinacoside, acteoside, cisacteoside, isoacteoside, 2-acetylacteoside, cistanoside A, thiab osmanthuside B (Wang li al. 2000). Nrog rau kev xav tau ntau ntxiv rau cov khoom lag luam no thiab kev poob sai sai hauvCistanche deserticolacov nroj tsuag, kab lis kev cai ntawm tes yuav yog ib qho kev ntxim nyiam lwm txoj kev tsim khoom. Ntau tus elicitors (tshwj xeeb tshaj yog fungal elicitors) tuaj yeem txhim kho metabolite ntau lawm hauv cov nroj tsuag kab lis kev cai (Wang li al. 2001). Li et al. (1998) pom tias cov fungal elicitor ntawm Fusarium solani cais los ntawm cov hauv paus hniav ntawm ob qho tib si.Cistanche deserticolathiab nws tus tswv, Halaxylon ammodendron, tuaj yeem txhim kho cov amino acid ntsiab lus hauv calli ntawmCistanche deserticolaTab sis kev hloov pauv ntawm PeGs cov ntsiab lus hauv kab callus no tsis tau hais hauv lawv txoj kev kawm. Nyob rau hauv daim ntawv no, cov teebmeem ntawm ib tug elicitor los ntawm F. solani ntawm biosynthesis ntawm tag nrho cov PeGs thiab ob lub ntsiab bioactive PeGs (echinacoside thiab acteoside), thiab ntawm kev hloov ntawm cell loj hlob tau kawm nyob rau hauv lub ncua kev kawm ntawv kab lis kev cai ntawmCistanche deserticola.

Cov ntaub ntawv thiab cov txheej txheem

Cog kab lis kev cai
Callus yog induced los ntawm nplai ntawmCistanche deserticolathiab khaws cia ntawm Gamborg's B5 nruab nrab (1968) ntxiv nrog 0.5 mg indole-3-acetic acid l^-1, 2 mg Kinetin l^-1,thiab 1 g casein hydrolyzate l^-1. Cov pH raug hloov mus rau 6 ua ntej kom tsis muaj menyuam. Tag nrho cov kev sim tau ua nyob rau hauv 250 ml co flasks, txhua tus muaj 100 ml nruab nrab ntawm ib tug rotary shaker (120 rpm thiab 25 degree) nyob rau hauv qhov tsaus ntuj. Lub inoculum yog hais txog 100 g tshiab wt l^-1.

cov ntsiab lus zoo ntawm cistanche deserticola--phenylethanoid glycoside

Kev npaj ntawm fungal elicitor

Kev sib cais ntawm cov kab mob fungus, Fusarium solani, thiab kev npaj ntawm fungal elicitor tau ua raws li tau piav qhia yav dhau los (Li et al. 1998, Yu et al. 2001). Ntau tus nqi (carbohydrate sib npaug) ntawm fungal elicitor tau ntxiv rau cov kab lis kev cai ntawm lub khob cawv rau hnub tim 15 d (tag nrho cov kab lis kev cai yog 42 d).

Biomass accumulation thiab viability assay
Cells los ntawm ib lub khob cawv yog ntxuav nrog dej distilled thiab khov-dried mus rau qhov hnyav tas li. Lub qhuav wt tau txiav txim siab li no. Cell viability (feem pua ​​​​ntawm cov hlwb siv tau hauv cov neeg ntawm tes) tau txiav txim los ntawm kev txo qis ntawm 2,3, 5-triphenyl tetrazolium chloride (Towill & Mazur 1975).

phenylethanoid glycosidentawmcistanche deserticola

Kev txiav txim siab ntawm cov ntsiab lus ntawm echinacoside, acteoside, thiab tag nrhophenylethanoid glycosides(PeGs)
Extraction ntawm PeGs tau ua raws li txoj kev ntawm Du & Liu (1993). Khov-qhuavCistanche deserticolahlwb tau muab rho tawm nrog 70 feem pua ​​(v/v) ethanol. Cov ethanol feem yog concentrated, degreased nrog dej-saturated ethyl acetate, thiab ces leached nrog dej-saturated n-butanol. N-butanol feem yog concentrated, yaj nyob rau hauv dej, loaded rau hauv ib tug macropore resin kem (AB-8, tshuaj cov khoom ntawm Nankai University, Tianjin, Tuam Tshoj), thiab eluted nrog dej thiab methanol, successively. Methanol feem raug sau thiab tag nrho cov ntsiab lus PeGs tau txiav txim los ntawm kev siv echinacoside raws li tus qauv ntawm 334 nm. Echinacoside thiab acteoside tau txiav txim siab los ntawm kev siv Nova Pak C18 kem (250 × 4.6 mm) hauv HPLC system (dej) nrog lub ntsuas UV (334 nm).

Cov txiaj ntsig

Cov nyhuv ntawm fungal elicitor ntawm biomass tsub zuj zuj thiab tag nrho PeGs biosynthesis Los ntawm fungal elicitor ntawm cell biomass thiab tag nrho PeGs tsub zuj zuj nyob ntawm qhov ntau npaum li cas ntxiv (carbohydrate equivalents). Raws li pom nyob rau hauv daim duab 1, qis koob tshuaj elicitor tau zoo siab rau qhov qhuav wt ntawmCistanche deserticolahlwb. Lub ncov yog nrog tus elicitor ntawm 20 mg l^-1(muab 1.76 mg qhuav wt ml^-1).
Tag nrho PeGs tsub zuj zuj zuj zus tau zoo dua los ntawm cov fungal elicitor ntawm 20 mg l.^-1ib 80mg l^-1. Elicitor ntawm 40 mg l-1 tau txais txiaj ntsig siab tshaj plaws (57 mg g^-1), uas yog ntau tshaj ob zaug uas tau txais nyob rau hauv lub non-elicitor tswj kab lis kev cai. Qhov kev sim ntxiv siv cov fungal elicitor ntawm 40 mg1^-1.

Fig. 1. Qhuav cell hnyav thiab tag nrho PeGs tsub zuj zuj nyob rau hauv ncua kev kawm ntawv cell kab lis kev cai ntawmCistanche deserticolanyob rau hauv teb rau ntau yam tshuaj ntawm fungal elicitor. Ib tug fungal elicitor tau ntxiv rau qhov nruab nrab ntawm 15th d. Cov qauv coj mus kuaj tom qab 27 d. ▲, Thaum kawg qhuav cell hnyav; ◆, tag nrho cov ntsiab lus ntawm PeG. Cov txiaj ntsig yog txhais tau tias cov txiaj ntsig triplicate thiab cov khoom yuam kev sawv cev rau cov qauv sib txawv.

Lub sijhawm kawm ntawm cell loj hlob thiab PeGs biosynthesis
Daim duab 2 qhia txog lub sij hawm kawm ntawm cell viability thiab PeGs (echinacoside thiab acteoside) biosynthesis ntawm suspension cells tom qab kab lis kev cai raug 40 mg l.^-1fungal elicitor nyob rau hauv 15th d. Tom qab 27 d ntawm kab lis kev cai, echinacoside thiab acteoside, ob lub ntsiab bioactive PeGs hauvCistanche deserticola, ob leeg nce considerably. Nyob rau 39th d qhov siab tshaj plaws tsub zuj zuj ntawm ob qho tib si tiav, ncav cuag 14.7 mg g.^-1qhuav wt 8,9 mg g^-1 dry wt, ua. Qhov sib txawv tseem ceeb hauv cell viability ntawm elicitor-kho kab lis kev cai thiab kev tswj tsis tau pom.

Daim duab 2. Cell viability thiab echinacoside thiab acteoside biosynthesis nyob rau hauv kev ncua cell kab lis kev cai ntawmCistanche deserticolatom qab kev kho mob los ntawm 40 mg l^-1tus kab mob fungal. fungal elicitor tau ntxiv rau qhov nruab nrab ntawm 15th d.◆, Echinacoside-kho nrog elicitor; ◇, echinacoside-tswj; ●, acteoside-kho nrog elicitor; ○,

acteoside-tswj; ▲, cell viability-kho nrog elicitor; △, cell viability-control. Cov txiaj ntsig yog txhais tau tias cov txiaj ntsig triplicate thiab cov khoom yuam kev sawv cev rau cov qauv sib txawv.

Kev sib tham

Ntau yam khoom tau los ntawm cov kab mob (cell phab ntsa fragments, polysaccharides, oligosaccharides, glycoproteins, thiab lwm yam) tau siv los ua ntau yam fungal elicitors nyob rau hauv theem nrab metabolite ntau lawm nrog ntau hom nroj tsuag cell. Hauv daim ntawv no, homogenate (tsuas yog tsim los ntawm polysaccharides) los ntawm cov kab mob fungus Fusarium solani cais los ntawm cov hauv paus hauv paus.Cistanche deserticolathiab Halaxylon ammodendron tau pom tias yog ib qho elicitor zoo rau PeGs biosynthesis hauv kab lis kev cai ntawm tes.Cistanche deserticola. Kev siv cov elicitor no los txhawb PeGs synthesis hauv hlwb tsis yog qhov tseem ceeb xwb tab sis tau nthuav tawm sai sai tom qab elicitation. Qhov kev txiav txim siab no tau pom zoo nrog qee qhov kev tshawb fawb yav dhau los ntawm cov kab lis kev cai cog qoob loo (Wang li al. 2001).

Hauv kev kawm dhau los ntawm Li et al. (1998), thaum ib tug fungal elicitor los ntawm F. solani tau ntxiv rau cov kab lis kev cai nruab nrab, callus induced los ntawm cov noob ntawmCistanche deserticolatuaj yeem ciaj sia nyob rau hauv lub sijhawm ntev subculture, thiab cov iridoids ntau lawm tuaj yeem txhim kho. Nws qhia tau hais tias cov kab lis kev cai tuaj yeem tsim cov metabolites theem nrab ntawm cov kab mob tiv thaiv thaum tawm tsam los ntawm cov kab mob. Txawm hais tias iridoids tsis tau txiav txim siab hauv peb qhov kev sim, peb tau pom tias polysaccharides los ntawm F. solani tuaj yeem siv los ua cov kab mob fungal zoo hauv kev tsim cov PeGs nrog.Cistanche deserticolahlwb.

cov ntsiab lus zoo ntawm cistanche deserticola--phenylethanoid glycoside

Los ntawm: ' Txhim kho ntawmphenylethanoid glycosidesntau lawm los ntawm ib tug fungal elicitor nyob rau hauv cell suspension kab lis kev cai ntawmCistanche deserticola' by Cui-tao Lu¹, ∗ & Xing-Guo Mei²

----Biotechnology Letters 25: 1437–1439, 2003. © 2003 Kluwer Academic Publishers. Luam tawm hauv Netherlands.

Cov ntaub ntawv

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