Effects Of Precursors And Elicitors On Growth And Alkaloids Accumulation In Shoots Of Lycoris Radiate

In this article, the difference of alkaloid content in different tissues of L.radiatawas studied, the conditions of liquid culture of shoots of L.radiata were optimizedand the liquid-shake culture method was established; the effects of precursorcompounds and elicitors on growth and alkalids accumulation of shoots of L.radiatawere studied; the effects of adding time of precursor compounds and elicitors ongrowth and alkalids accumulation of shoots of L.radiata were studied. The followingresults were obtained:1. Analysis of alkaloids in L.radiata. tissues. The results showed that thecontents of three alkaloids in different tissues of L.radiata were inconsistent.Thecontent of lycorine from high to low was: flower> bulb> scape> leaf> root>shoot> root from tissue culture> callus; the content of lycoramine from high to lowwas: flower> bulb> leaf> scape> root>shoot> root from tissue culture> callus; thecontent of galanthamine from high to low was: flower> leaf> root> shoot> bulb>scape> root from tissue culture> callus. the galanthamine content of shoot was230.197μg g-1DW,Just below the flower,leaf and root,but higher than callus, and was21.79times of callus. Flower was not suitable for tissue culture due to its floweringperiod is shorter;shoots were easy to induce, easy culture and not limited by seasonchanged compared with leaf,root,and callus. Therefore shoot was the most suitableexperimental material for the study of alkaloid synthesis. There were5alkaloidsidentificated by GC-MS from the bulbs of L.radiata, respectively Lycorine,lycoramine, galantamine, tazettine, pluviine.2.The liquid culture of shoots of L.radiata. The results showed that the form ofmedium,hormone proportion,concentration of sucrose and illumination time had ainfluence on growth and accumulation of lycorine, lycoramine and galantamine ofshoots.The best culture conditions were: liquid culture,6-BA and NAA concentrationswere2mg L^-1and0.2mg L^-1, the concentration of sucrose was40g L^-1, illuminationtime was16h d-1.3. Effects of precursor compounds on shoots of L.radiata.The results showed that the effect of tyramine on growth and alkaloid content ofshoots was not consistent. High concentrations of tyramine made for the accumulationof Amaryllidaceae alkaloids,however low concentrations of tyramine could promotethe growth of shoots;the best concentration of tyramine was250μmol L^-1. The results showed that the effects of protocatechualdehyde on growth andalkaloid content of shoots were consistent.The addition of low concentrations ofprotocatechualdehyde could promote the growth as well as the accumulation ofAmaryllidaceae alkaloids of shoot; the best concentration of protocatechualdehydewas50μmol L^-1.4. Effects of elicitors on shoots of L.radiata. The results showed that lowconcentrations of salicylic acid could promote the growth as well as the accumulationof Amaryllidaceae alkaloids of shoot.The best concentration of salicylic acid ongrowth of shoots was50μmol L^-1,however the best concentration of salicylic acid onalkaloids accumulations was10μmol L^-1. The results showed that the GI and ADB ofMeJA group were lower than the control group.The addition of MeJA≤200μmol L^-1can promote the accumulation of lycorine, lycoramine, galanthamine. The productionof lycorine lower than control group and and there is no obvious difference betweencontrol and MeJA group in the production of lycoramine and galanthamine. Theconcentration of MeJA was100μmol L^-1combined the effects on growth withalkaloids content.5. Effects of elicitors and precursors on shoots of L.radiata.The results showed that the addition of salicylic acid and tyramine andprotocatechualdehyde could promote the growth as well as the accumulation ofAmaryllidaceae alkaloids of shoot.When the concentration of tyramine andprotocatechualdehyde was150μmol·L^-1, the GI and ADB of shoots were119%and115%of the control;The concentrations of lycorine, lycoramine and galanthaminewere the highest, which were302%,320%,388%of the control.The results showed that the effects on growth and alkaloids contents weredifferent between adding MeJA alone and adding MeJA, tyramine andprotocatechualdehyde together.The addition of MeJA, tyramine andprotocatechualdehyde together promoted the growth and the accumulations of drymatter of shoots; however the contents of lycorine, lycoramine and galanthamine werelower than the control group. And the inhibition of alkaloid synthesis was strongerthan the promotion of shoots growth. Therefore, the addition of MeJA and tyramineand protocatechualdehyde together cannot promote the accumulation ofAmaryllidaceae alkaloids.6. Time effects of elicitors and precursorsThe results showed that adding tyramine at cultured at12th d the GI and ADB of shoots were largest,but the contents of lycorine and galanthamine were highest whichadding tyramine at9th d.Combine time effects of elicitors and precursors on growthand alkaloids content,adding tyramine at9th d could promote the accumulation oflycorine and galanthamine,however at12th d could promote the accumulation oflycoramine.The results showed that adding salicylic acid at12th d the GI and ADB of shootswere largest,but the contents of lycorine and galanthamine were highest which addingsalicylic acid at9th d.The results showed that the GI and ADB of shoots were largest that addingsalicylic acid and tyramine and protocatechualdehyde together at12th d after culturedand the contents of lycorine and galanthamine were highest,while the content oflycoramine were similar to the content of the9th d...