Jose Binoy, Satheeshkumar K, Seeni S
Plant Biotechnology Division, Tropical Botanic Garden and Research Institute, Palode, Thiruvananthapuram 695 562, India.
Pak J Biol Sci. 2007 Jan 15;10(2):349-55. doi: 10.3923/pjbs.2007.349.355.
A rapid clonal multiplication scheme comprising direct multiple shoot initiation and downsizing of the node with buds proliferated upon during subculture was developed for Plumbago rosea. Sixty five per cent of the nodes (approximately 2.0 cm) dissected out of young shoots from field grown plants and cultured in MS agar medium containing 3% sucrose and 15.4 microM BAP remained contamination free and responded at 95% rate with callusing at basal cut end and axillary bud break in 5 days followed by the formation of 2.41 +/- 0.14 shoots of 0.87 +/- 0.14 cm length in 3 weeks. Though differences in frequency and number of buds formed between nodes of 1-5 positions from the young shoots was negligible, the shoots emanated from the youngest node were shorter (0.92 +/- 0.19 cm) than those (2.3 +/- 0.50 cm) of the mature 5th node. Synergistic influence of BAP and auxins on caulogenesis was absent. Bud emergence in shorter (approximately 0.5 cm) nodes was delayed up to 3 weeks and extensive callus proliferation from the cut basal end overlapped the 8.2 +/- 0.37 axillary shoots/buds formed after 7 weeks. Reduction in the size (downsized) of the 2.0 cm node with buds to 1.0 cm by dissecting out the basal internodal segment having the callus and subculture of them (approximately 1.0 cm) with buds in contact with the medium for 3 weeks contributed to maximum multiplication of 42.1 +/- 5.40 shoot buds. Division of the shoot cluster and transfer of 2-3 shoots (0.5-1.5 cm) in a clump to MS basal liquid medium induced elongation of the shoots to 4.1 +/- 0.18 cm in 2 weeks. Shoots of 3.0-4.2 cm length were rooted within 3 weeks at 100% efficiency in vitro or ex vitro without hardening. In vitro rhizogenesis in presence of 0.49 microM IBA is recommended for enhanced rooting and high yield of commercially important tuberous roots during cultivation in the field.
为玫瑰紫茉莉开发了一种快速克隆繁殖方案，该方案包括直接诱导多芽形成以及在继代培养过程中对带有增殖芽的节段进行缩小处理。从田间种植的植株幼嫩枝条上切取的65%的节段（约2.0厘米），接种于含有3%蔗糖和15.4微摩尔/升苄氨基嘌呤（BAP）的MS琼脂培养基中，保持无污染，95%的节段有反应，基部切口端形成愈伤组织，腋芽在5天内萌发，3周后形成2.41±0.14个长度为0.87±0.14厘米的芽。尽管从幼嫩枝条第1至5节位的节段形成芽的频率和数量差异可忽略不计，但从最幼嫩节段发出的芽（0.92±0.19厘米）比成熟的第5节段发出的芽（2.3±0.50厘米）短。BAP和生长素对芽的形成没有协同作用。较短（约0.5厘米）节段的芽萌发延迟至3周，基部切口端大量愈伤组织增殖，与7周后形成的8.2±0.37个腋芽/侧枝重叠。通过切除带有愈伤组织的基部节间段，将带有芽的2.0厘米节段缩小至1.0厘米，并将其（约1.0厘米）带有芽且与培养基接触3周的节段进行继代培养，可使芽的增殖最大化，达到42.1±5.40个芽。将芽丛分割并将一丛中2 - 3个芽（0.5 - 1.5厘米）转移至MS基本液体培养基中，2周内可使芽伸长至4.1±0.18厘米。长度为3.0 - 4.2厘米的芽在3周内100%有效生根，无论是在试管内还是试管外，无需炼苗。建议在含有0.49微摩尔/升吲哚丁酸（IBA）的条件下进行试管内生根，以提高生根率，并在田间种植时提高具有商业重要性的块根的产量。
