Nair L G, Seeni S
Plant Biotechnology Division, Tropical Botanic Garden and Research Institute, Pacha-Palode, Thiruvananthapuram, India.
Indian J Exp Biol. 2001 Jul;39(7):697-704.
Nodes, shoot tips, internodes and leaf bases (approximately 1.0 cm) excised from young vines of the flowering woody climber, Celastrus paniculatus WilId. sub. sp. paniculatus (Celastraceae) were cultured in Murashige and Skoog (MS) medium containing agar (0.6%), sucrose (3%) and varied concentrations of 6-benzyl aminopurine (BAP) and kinetin. All the explant types were regenerative and maximum number (3.6) and frequency (94%) of axillary shoot formation of (5.08 cm long) was recorded in the nodes cultured in BAP (1 mg L(-1)) after 6 weeks. Combinations of BAP (1 mg L(-1)) and indole-3-acetic acid/l-naphthalene acetic acid (0.01-1 mg L(-1); IAA/NAA) tested with nodes induced formation of less number (3 and 2.2) of shoots at same frequency (94%). All the explant types viz. node, shoot tip, internode and leaf base of in vitro derived shoots responded earlier and better in lower concentrations of BAP (0.5-2 mg L(-1)) with formation of 8, 3.1, 6.4 and 1.8 shoots respectively during the same period. In spite of the advanced and increased caulogenic responses, differences in cytokinin requirements between different explants observed during culture initiation still persisted with the nodes, shoot tips, internodes and petiole segments responding best at 0.5, 1 and 2 mg L(-1) BAP, respectively. The repeated reculture up to 10 cycles of the nodes from the shoot cultures each at 6-week intervals enabled multiplication and stocking of shoots without decline. Rooting of 3-7 cm shoot cuttings was induced in half-strength MS liquid medium containing IAA (1 mg L(-1)) with formation of 7.25 roots of 2.41 cm length within 6 weeks. Rooted plants were established at 84-96% rate in community pots without hardening, the least value (84%) being obtained with NAA- induced thick and calloid rooted plants. Four month old community potted plants were reintroduced into native forest habitats at 95% efficiency and 8 months after restoration, the plants were uniform in morphological, growth, cytological and peroxidase and esterase isozyme characteristics.
从开花木质藤本植物南蛇藤（Celastrus paniculatus WilId. sub. sp. paniculatus，卫矛科）的幼嫩藤蔓上切取节点、茎尖、节间和叶基部（约1.0厘米），接种于含有琼脂（0.6%）、蔗糖（3%）以及不同浓度6-苄基腺嘌呤（BAP）和激动素的Murashige和Skoog（MS）培养基中。所有外植体类型都具有再生能力，6周后，在添加1毫克/升BAP的培养基中培养的节点上，腋芽形成的数量最多（3.6个），频率最高（94%），形成的腋芽长度为5.08厘米。用节点测试BAP（1毫克/升）与吲哚-3-乙酸/1-萘乙酸（0.01 - 1毫克/升；IAA/NAA）的组合，在相同频率（94%）下诱导形成的芽数量较少（3个和2.2个）。所有外植体类型，即体外衍生芽的节点、茎尖、节间和叶基部，在较低浓度的BAP（0.5 - 2毫克/升）下反应更早且更好，在此期间分别形成8个、3.1个、6.4个和1.8个芽。尽管生根反应有所提前且增强，但在培养起始阶段观察到的不同外植体对细胞分裂素需求的差异仍然存在，节点、茎尖、节间和叶柄切段分别在0.5毫克/升、1毫克/升和2毫克/升BAP时反应最佳。将茎尖培养的节点每隔6周重复继代培养多达10个周期，能够实现芽的增殖和保存且不会衰退。在含有1毫克/升IAA的1/2强度MS液体培养基中诱导3 - 7厘米长的茎段生根，6周内形成7.25条长度为2.41厘米的根。未经炼苗的生根植株在营养钵中的定植率为84% - 96%，用NAA诱导形成粗壮且愈伤状根的植株定植率最低（84%）。4个月大的营养钵植株以95%的效率重新引入原生森林栖息地，恢复8个月后，这些植株在形态、生长、细胞学以及过氧化物酶和酯酶同工酶特征方面表现一致。
