三裂叶野葛毛状根的生长及其培养基营养物质的消耗变化

Relationship Between Growth and Medium Nutrient Consumption During the Culture of Pueraria phaseoloides Hairy Roots

研究了发根农杆菌(Agrobacterium rhizogenes)ATCC15834遗传转化产生的三裂叶野葛(Pueraria phaseoloides)毛状根在液体培养过程中生长及其部分营养物质消耗的关系.结果表明:三裂叶野葛毛状根液体培养0～4d内处于生长迟滞期、8～16d为快速生长期、16d后进入生长平台期.培养基的PO43-、硝态氮和铵态氮在毛状根液体培养过程中被逐渐吸收和消耗,培养16d时培养基中的PO43-被消耗殆尽,其浓度仅为培养基起始PO43-浓度的0.26%;培养基的铵态氮和硝态氮则在培养20d时才消耗殆尽;而培养基中的Ca2+浓度在培养过程中逐渐降低,但在培养20d时仍未被完全消耗,其浓度约为起始浓度的30.5%.培养基的pH值随培养时间的延长而不断降低,培养20d后pH值由5.62降低到4.09;而毛状根的颜色也随培养基pH值的降低和培养时间的延长逐渐由白色变成浅黄色和浅褐色.该结果为今后设计合适的培养基以开展野葛毛状根的大规模液体培养来生产葛根素提供了可能性.

Relationship between nutrient consumption in the medium and growth of Pueraria phaseoloides hairy roots induced by Agrobacterium rhizogenes ATCC15834 were investigated during liquid culture. The results showed that hairy roots grew very slowly in 0 -4 days, very fast from 8 to 16 days. During liquid culture, PO4^3-, NO3^- and NH4^＋ in the medium were gradually absorbed and utilized with time. PO4^3- of the medium was used up at day 16 of the culture, the content of which was 0. 26% of the initial amount. While the NO3^- and NH4^＋ in the medium was almost used up at day 20. Ca^2 ＋ of the medium was gradually absorbed and utilized during liquid culture and it was not used up at day 20, the content of which was still 30.5% of the initial amount. It was also observed that the pH value of the medium decreased gradually from 5.62 at day 0 to 4.09 at day 20 of the culture and the color of the hairy roots cultured in liquid medium turned gradually from white into light yellow and finally partly brown with time and decrease in pH value. The results presented here had provided the possibilities on how to prepare optimum medium for large scale cultivation and production of puerarin from P. phaseoloides hairy roots.