Ramseh M, Marx R, Mathan G, Pandian S Karutha
Department of Biotechnology, Alagappa University, Karaikudi, India.
J Environ Biol. 2009 May;30(3):441-4.
A reliable and reproducible protocol for contamination free plant recovery system from alginated encapsulated uninodal microcuttings of micropropagated Bacopa monnieri L. have been developed after storage at 18 degrees C for 45 days. Node segments excised from freshly micropropagated plants were encapsulated as single explant beads with 3.0% sodium alginate and 80 mM CaCl2 2 H2O. To find out the optimal concentration of fungicide bavistin for efficient plant recovery different concentrations of bavistin (1.0 - 15 mg l(-1)) were incorporated in to the encapsulation medium. 3.0mg l(-1) bavistin showed no reduction in plant conversion and generated maximum number of shoots (45.6 +/- 1.69) at high frequency with out any contamination after storage up to 45 days at 18 degrees C. At high concentrations (13 and 15 mg l(-1)), rupturing of calcium alginate coats after 8 - 9 days and gradual decline in the number of shoots indicates the toxic effect of bavistin on plant conversion. Encapsulated node cuttings stored up to 45 days regenerated shoots (5.2) and multiple shoots (45.6) in MS basal and hormone medium respectively. Maximum shoot length (8.2 +/- 0.37 cm) was observed from encapsulsted node cuttings incorporated with 3.0 mg l(-1) bavistin on MS basal medium. 90% of the recovered plantlets were hardened off and successfully established in soil.
已开发出一种可靠且可重复的方案，用于从微繁殖的假马齿苋经藻酸盐包封的单节微插穗中获得无污染的植物再生系统，该微插穗在18摄氏度下储存45天。从刚微繁殖的植株上切下的节段用3.0%的海藻酸钠和80 mM的CaCl₂·2H₂O作为单个外植体珠进行包封。为了找出高效植物再生所需的杀菌剂多菌灵的最佳浓度，将不同浓度的多菌灵（1.0 - 15 mg l⁻¹）加入到包封培养基中。3.0 mg l⁻¹的多菌灵在18摄氏度下储存长达45天后，对植物转化率没有降低，且高频产生最多数量的芽（45.6 ± 1.69）且无任何污染。在高浓度（13和15 mg l⁻¹）下，8 - 9天后藻酸钙包衣破裂且芽的数量逐渐减少，表明多菌灵对植物转化率有毒性作用。储存长达45天的包封节段分别在MS基本培养基和激素培养基中再生出芽（5.2）和多个芽（45.6）。在MS基本培养基上，观察到添加3.0 mg l⁻¹多菌灵的包封节段的最大芽长为（8.2 ± 0.37 cm）。90%的再生植株成功炼苗并在土壤中定植。
