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Title: <i style="">In vitro </i>cloning of apple (<i style="">Malus domestica </i>Borkh) employing forced shoot tip cultures of M9 rootstock
Authors: Dalal, M Amin
Das, B
Sharma, A K
Mir, M Amin
Sounduri, Amarjeet Singh
Keywords: apple
dwarfing rootstocks
forced explants
in vitro propagation
Issue Date: Oct-2006
Publisher: CSIR
Series/Report no.: Int. Cl.<sup>8</sup> A01H4/00
Abstract: For micropropagation of adult apple <i style="">(Malus domestica </i>Borkh) rootstock M9, explants were harvested from pre-chilled (4±3°C) dormant cuttings forced in growth chamber. Primary explants were established by using a slightly modified version of culture initiation procedure developed earlier. Multiple shoots, raised by axillary branching of surviving explants, were obtained from established cultures in two cultural pathways: (i) repeat transfer of primary explants on the MS supplemented with BAP (2.22 µ<i style="">M</i>), IBA (0.49 µ<i style="">M</i>) and Kn (2.2 µ<i style="">M</i>), and (ii) repeat subculture of harvested microshoots and basal portion of sectored proliferating clumps on the same but suitably PGR amended medium, in which cytokinins concentration was reduced by half. A 4-fold shoot multiplication was achieved during each sub culture of 3±1 week’s duration that repeatedly produced crop of proliferated shoots without loss of vigour. Nodal segments (5-15 mm), obtained from <i style="">in vitro </i>raised microshoots were also used to initiate a new cycle of proliferating cultures. Isolated cloned microshoots (15-20 mm) with apical bud were cultured on MS basal medium supplemented with IBA (14.70 µ<i style="">M</i>) for 8±3 d to initiate root. The microshoots were re-implanted in PGR free half strength basal MS medium with full complement of organics for 4±1 week for root development. The transferred<i style=""> in vitro </i>hardened plantlets to polyvinyl cups or polybags, under carefully controlled descending RH regime of 95% to 70±5% over a period of 5±1 week, resulted in 80% <i style="">ex vitro </i>survival. The present protocol highlighted a novel strategy of micropropagation of apple rootstock M9 using three-step culture initiation procedure of forced primary explants.
Description: 543-550
ISSN: 0975-0967 (Online); 0972-5849 (Print)
Appears in Collections:IJBT Vol.05(4) [October 2006]

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