Shekhawat Mahipal S, Kannan N, Manokari M, Ravindran C P
Biotechnology Laboratory, Department of Plant Science, M.G.G.A.C., Mahe, Pondicherry 673311, India.
Biotechnology Unit, K.M. Centre for Postgraduate Studies, Pondicherry 605 008, India.
J Genet Eng Biotechnol. 2015 Dec;13(2):209-214. doi: 10.1016/j.jgeb.2015.08.002. Epub 2015 Sep 9.
Methods were developed in the present investigation for cloning and large scale plant production of L. germplasm selected from the East-Coast region of South India. Nodal shoot segments were used as explants. The explants were dressed and surface sterilized with 0.1% (w/v) HgCl. Multiple shoots were induced (6.13 ± 0.22 shoots per explant) by proliferation of nodal shoot meristems on Murashige and Skoog (MS) semi-solid medium + 2.0 mg l 6-benzylaminopurine (BAP). The shoots of were further multiplied (16.45 ± 0.44 shoots per explant) on MS medium + 0.5 mg l each of BAP and Kinetin (Kin). The generated shoots were rooted on half-strength MS medium containing 2.5 mg l indole-3 butyric acid (IBA). By this method 67% shoots were rooted. About 97% shoots were rooted (8.33 ± 0.29 roots per shoot) when the cut ends of the shoots were treated with 300 mg l IBA for 5 min. The rooted plants were hardened and acclimatized in the greenhouse and successfully (100%) transplanted to the field.
本研究开发了从印度南部东海岸地区选出的罗勒种质克隆及大规模植株生产的方法。以带芽茎段作为外植体。外植体经修整后用0.1%（w/v）HgCl进行表面灭菌。在Murashige和Skoog（MS）半固体培养基 + 2.0 mg l 6-苄基腺嘌呤（BAP）上，通过带芽茎尖分生组织增殖诱导出多个芽（每个外植体6.13 ± 0.22个芽）。在MS培养基 + 0.5 mg l BAP和激动素（Kin）各一种的条件下，这些芽进一步增殖（每个外植体16.45 ± 0.44个芽）。所产生的芽在含有2.5 mg l吲哚-3-丁酸（IBA）的1/2强度MS培养基上生根。通过这种方法，67%的芽生根。当芽的切口端用300 mg l IBA处理5分钟时，约97%的芽生根（每个芽8.33 ± 0.29条根）。生根的植株在温室中进行炼苗和驯化，并成功（100%）移栽到田间。
