Sharma Urvashi, Mohan J S S
Department of Biosciences, Sardar Patel University, Vallabh Vidyanagar 388 120, India.
Indian J Exp Biol. 2006 Jun;44(6):499-505.
Reduction of vitrification in in vitro raised shoots derived from shoot bases and immature floral buds along with inflorescence axis used as explants of C. borivilianum, a rare medicinal herb is described. Shoot multiplication was obtained on MS medium with 2 mg l(-1) benzylaminopurine (BAP) + 0.1 mg l(-1) indole-3-butyric acid (IBA) and MS medium with 2 mg l(-1) kinetin (Kin) + 0.1 mg l(-1) 2,4-dichlorophenoxy acetic acid (2,4-D) from shoot bases and inflorescence axis respectively. Best multiplication rates were obtained from both the explants on MS medium with 2 mg l(-1) BAP. Vitrification of shoots in cultures appeared during the multiplication stage. Culture bottles with aerated caps reduced the vitrification to 80%. Reduction of BAP concentration from 2 mg l(-1) to zero during subsequent subcultures also minimized vitrification. Use of 0.5-2 mg l(-1) Kin produced healthy shoots when compared to BAP. In vitro raised shoots rooted on Knop salts containing iron and vitamins of MS medium, 2 mg l(-1) IBA and 0.1% activated charcoal. About 80% plantlets survived upon soil transfer. Scanning electron microscopic and image analyzer studies reveal the morphological structural differences between the leaves of normal and vitrified plantlets.
描述了在体外培养源自茎基部、未成熟花芽以及用作珍稀药用植物波棱瓜外植体的花序轴的芽时,玻璃化现象的减少情况。在添加2 mg l(-1)苄氨基嘌呤(BAP)+ 0.1 mg l(-1)吲哚-3-丁酸(IBA)的MS培养基上以及分别在添加2 mg l(-1)激动素(Kin)+ 0.1 mg l(-1) 2,4-二氯苯氧乙酸(2,4-D)的MS培养基上,从茎基部和花序轴获得了芽增殖。在添加2 mg l(-1) BAP的MS培养基上,两种外植体均获得了最佳增殖率。培养过程中芽的玻璃化现象出现在增殖阶段。带有通气帽的培养瓶将玻璃化现象减少到了80%。在后续继代培养中将BAP浓度从2 mg l(-1)降至零也使玻璃化现象最小化。与BAP相比,使用0.5 - 2 mg l(-1) Kin可产生健康的芽。体外培养的芽在含有MS培养基铁和维生素的Knop盐、2 mg l(-1) IBA和0.1%活性炭的培养基上生根。约80%的植株在移栽到土壤后存活。扫描电子显微镜和图像分析仪研究揭示了正常植株和玻璃化植株叶片之间的形态结构差异。