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21 August 2007 Influence of cytokinin levels on in vitro propagation of shy suckering chrysanthemum “Arka Swarna” and activation of endophytic bacteria
Bindu Panicker, Pious Thomas, Tolety Janakiram, Rangarajan Venugopalan, Sathyanarayana B. Narayanappa
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Abstract

In an effort to develop a sustainable protocol for the micropropagation of a shy suckering elite chrysanthemum cv. Arka Swarna (yellow pompon type), in vitro cultures were established using surface-sterilized nodal microcuttings (1–1.5 cm) from polyhouse-grown plants on MS medium containing 3% sucrose, 0.25% phytagel, and 5 μM benzyl adenine (BA) or kinetin. Microbial contamination in the range of 6–24% was encountered during the first in vitro passage. Apparently clean cultures after one passage on MS basal medium were transferred to medium with BA or kinetin (0, 1, 5, 10, or 20 μM) in culture bottles, and were monitored for eight in vitro passages (1 mo. each) for growth and microbial contamination. Plant growth regulator (PGR)-free medium was the best for sustainable micropropagation over successive in vitro passages yielding a single shoot from cultured microcuttings. Higher cytokinin levels inhibited rooting and induced one or more shorter shoots with close nodes resulting in low propagation rates. All apparently clean stocks revealed covert endophytic bacteria during tissue-indexing using bacteriological media. Three distinct bacterial morphotypes were isolated from such stocks, identified based on 16S rRNA gene sequence analysis as different morphotypes of Curtobacterium citreum. The endophytes tended to show obvious growth on chrysanthemum culture medium with increase in cytokinin levels (5–20 μM), but such growth was not noticed in inoculations on MS medium without plants. Sustainable micropropagation of cv. Arka Swarna for more than 2 yr with the resident endophytic bacteria in covert form was realized on PGR-free MS medium giving a net propagation rate of three to four times over a subculture cycle of 2–3 wk.

Bindu Panicker, Pious Thomas, Tolety Janakiram, Rangarajan Venugopalan, and Sathyanarayana B. Narayanappa "Influence of cytokinin levels on in vitro propagation of shy suckering chrysanthemum “Arka Swarna” and activation of endophytic bacteria," In Vitro Cellular and Developmental Biology - Plant 43(6), 614-622, (21 August 2007). https://doi.org/10.1007/s11627-007-9061-6
Received: 28 December 2006; Accepted: 20 June 2007; Published: 21 August 2007
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KEYWORDS
16S rRNA gene
Dendranthema grandiflora
Endophytic actinobacteria
Microbial contamination
Plant tissue culture
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