Development of an efficient tissue culture protocol for callus formation and plant regeneration of wetland species <Emphasis Type="Italic">Juncus effusus</Emphasis> L. |
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Authors: | L Xu U Najeeb R Raziuddin W Q Shen J Y Shou G X Tang W J Zhou |
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Institution: | (1) Institute of Crop Science, Zhejiang University, Hangzhou, 310029, China;(2) The University of Nottingham Ningbo, Ningbo, 315100, China;(3) Present address: Zhuji Municipal Agro-Tech Extension Center, Zhuji, 311800, China;(4) Present address: Department of Plant Breeding and Genetics, NWFP Agricultural University, Peshawar, Pakistan; |
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Abstract: | Wetland species mat rush (Juncus effusus L.) is an important economic plant, but no information is available regarding plant regeneration, callus induction, and its
proliferation from in vitro seed grown plantlets. The present study investigates the effects of growth regulator combinations and medium innovation on
tissue culture system of five mat rush varieties. Addition of N6-benzyladenine (BA) and 2,4-dichlorophenoxyacetic acid (2,4-D)
in Murashige and Skoog (MS) medium showed significantly positive effect on callus proliferation, plant regeneration, and its
multiplication compared to the medium devoid of BA. The highest callus induction frequency (80.95%, 90.48%, 75.40%, 70.83%,
and 83.33%) was observed in MS medium containing 0.5 mg L−1 (2.2 μM) BA in Yinlin-1, Nonglin-4, Gangshan, Taicao, and Taiwan green, respectively. Various growth regulator combinations
with successive subculture (medium replacement) were found essential to develop organogenic calluses and to regenerate shoots.
The combination of 0.1 mg L−1 BA (0.4 μM) and 2 mg L−1 2,4-D (9.0 μM) in MS medium was found best for callus proliferation for all the varieties under trial. The plant regeneration
required two steps involving successive medium replacements as well as optimal hormonal balances. Successful plant regeneration
(over 70%) was observed only by transferring the organogenic callus from regeneration medium I MS medium containing 0.5 mg
L−1 BA (2. μM) and 1.0 mg L−1 kinetin (KT; 4.6 μM)] to the regeneration medium II MS medium containing 0.5 mg L−1 BA (2.2 μM), 1.0 mg L−1 KT (4.6 μM) and 3.0 mg L−1 indoleacetic acid (IAA; 17.1 μM)]. Our results confirmed the importance of the ratio of auxin (IAA) to cytokinin (BA and
KT) in the manipulation of shoot regeneration in J. effusus L. The maximum plant survival frequency and multiplication rates (90.97% and 5.40 and 94.23% and 8.25) were recorded in the
presence of 0.5 mg L−1 BA (2.2 μM) in the 1/2 MS multiplication medium for the varieties of Nonglin-4 and Taicao, respectively. About 100% survival
rate was also observed for all the varieties in soil conditions. The efficient plant regeneration system developed here will
be helpful for rapid micropropagation and further genetic improvement in J. effusus L. |
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