Ou Shaoyun, Shi Heping, Tsang Eric Pokeung
Guangdong Provincial Key Laboratory of Biotechnology for Plant Development, College of Life Science, South China Normal University, Guangzhou 510631, China.
Sheng Wu Gong Cheng Xue Bao. 2010 Mar;26(3):378-85.
To study the possibilities for improvement of the ornamental character and production of secondary metabolites by using Wedelia trilobata hairy roots, we investigated the induction of W. trilobata L. hairy roots and its consumption changes of carbon resource, nitrogen resource, phosphate and calcium in the medium during liquid culture. The results showed that hairy roots could be incited from the cut edges of leaf explants 7 days after inoculation with Agrobacterium rhizogenes ATCC15834 and could have an autonomous growth on the medium without phytohormones. The PCR amplification showed that rol genes of Ri plasmid of A. rhizogenes was integrated and expressed into the genome of transformed hairy roots. The hairy root line grew very slowly in 0-7 days, very fast from 7 to 21 days. During the liquid culture of hairy roots, sucrose, NO3(-)-N, PO4(3-) and Ca2+ in the medium could be gradually absorbed and utilized with time. The content of NO3(-)-N in the medium was 5.8% of the initial amount at day 7, while sucrose content was about 50% of the initial amount. At day 35, the NO3(-)-N and sucrose content in the medium was 1.82% and 3.39% of the initial amount, respectively. In combination with Ca2+ consumption, PO4(3-) of the medium was rapidly absorbed and utilized. At day 7, the content of PO4(3-) in the spent medium was only 1.76% of the initial amount; but even at day 35, the content of Ca2+ in the spent medium was still 61.3% of the initial amount. The results presented here had provided the possibilities on improvement the ornamental character and how to prepare optimum medium for large scale cultivation and production of secondary metabolites from W. trilobata L. hairy roots.
为了研究利用南美蟛蜞菊毛状根改善观赏性状和生产次生代谢产物的可能性，我们研究了南美蟛蜞菊毛状根的诱导及其在液体培养过程中对培养基中碳源、氮源、磷酸盐和钙的消耗变化。结果表明，接种发根农杆菌ATCC15834 7天后，叶外植体的切口边缘可诱导出毛状根，且在无植物激素的培养基上能自主生长。PCR扩增表明，发根农杆菌Ri质粒的rol基因已整合并表达到转化毛状根的基因组中。毛状根系在0 - 7天生长非常缓慢，7 - 21天生长非常迅速。在毛状根液体培养过程中，培养基中的蔗糖、NO3(-)-N、PO4(3-)和Ca2+可随时间逐渐被吸收利用。培养7天时，培养基中NO3(-)-N的含量为初始量的5.8%，而蔗糖含量约为初始量的50%。培养35天时，培养基中NO3(-)-N和蔗糖含量分别为初始量的1.82%和3.39%。与Ca2+消耗相结合，培养基中的PO4(3-)被迅速吸收利用。培养7天时，消耗后的培养基中PO4(3-)的含量仅为初始量的1.76%；但即使在培养35天时，消耗后的培养基中Ca2+的含量仍为初始量的61.3%。本文的结果为改善南美蟛蜞菊的观赏性状以及如何制备用于大规模培养和生产南美蟛蜞菊毛状根次生代谢产物的最佳培养基提供了可能性。
