Department of Botany , Don Bosco College , Tura 794002, Meghalaya , India.
AoB Plants. 2011;2011:plr012. doi: 10.1093/aobpla/plr012. Epub 2011 May 27.
Ilex khasiana is a rare and critically endangered holly endemic to the Khasi Hills of Meghalaya, India, and confined to a small number of pocket areas. In addition to conventional methods of propagation, endemic and threatened plants such as this could be more effectively multiplied and conserved using in vitro methods. Such techniques have the additional advantage of having a low impact on wild populations because they require a minimum of starting material. Our objective was to develop methodologies for the successful in vitro mass propagation of I. khasiana.
Seedlings were germinated in vitro under sterile conditions and nodal explants from these were transferred to Murashige and Skoog (MS) medium supplemented with 8.88 µM 6-benzyladenine and 4.64 µM kinetin.
This generated ∼10 shoots per explant. In a second approach, callus was obtained from seedling-derived leaf discs cultured on MS medium supplemented with 2,4-dichlorophenoxyacetic acid and 6-benzyladenine. Approximately 12 adventitious shoots per callus were regenerated from 83.33 % of the calli after transfer to MS medium supplemented with 6.63 µM 6-benzyladenine. The most effective treatment for inducing root formation on the shoots was transfer of shoots to half-strength MS medium with 9.84 µM indole-3-butyric acid. Regenerated plantlets with well-developed shoots and roots were hardened and transferred to open soil with 70 % survival after 4 weeks.
Both the methods described here are well suited for the mass multiplication of this critically endangered tree species.
Ilex khasiana 是一种罕见的、极危的冬青属植物，仅分布于印度梅加拉亚邦的卡西丘陵，且局限于少数几个小区域内。除了传统的繁殖方法外，这种特有种和受威胁的植物可以通过离体方法更有效地繁殖和保存。这些技术除了具有对野生种群影响较小的额外优势外，因为它们只需最少的起始材料。我们的目标是为 I. khasiana 的成功离体大规模繁殖开发方法。
在无菌条件下，对幼苗进行离体发芽，然后将这些幼苗的节间外植体转移到添加了 8.88µM 6-苄基腺嘌呤和 4.64µM 激动素的 Murashige 和 Skoog (MS) 培养基上。
每个外植体产生了约 10 个芽。在第二种方法中，从幼苗衍生的叶盘在添加了 2,4-二氯苯氧乙酸和 6-苄基腺嘌呤的 MS 培养基上获得愈伤组织。将约 12 个不定芽从 83.33%的愈伤组织再生到添加了 6.63µM 6-苄基腺嘌呤的 MS 培养基上。将芽转移到添加了 9.84µM 吲哚丁酸的半强度 MS 培养基上是诱导芽生根的最有效处理方法。再生的具有发育良好的芽和根的植物在 4 周后在开放土壤中进行硬化，成活率为 70%。
这里描述的两种方法都非常适合这种极危树种的大规模繁殖。
