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Biotization of in vitro calli and embryogenic calli of oil palm (Elaeis guineensis Jacq.) with diazotrophic bacteria Herbaspirillum seropedicae (Z78)

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A continuous discovery of novel practices for in vitro plant culture is essential to promote better plant growth. An alternative approach to the conventional micropropagation practice could be achieved through biotization by supplying selected diazotrophs to the plant materials. Due to this considerable potential, an artificial symbiosis was created between the Herbaspirillum seropedicae strain Z78 (ATCC 35893) and in vitro oil palm (Elaeis guineensis Jacq.). This experiment was carried out with the aim of studying the effects of H. seropedicae inoculation on growth of oil palm calli and embryogenic calli. A total of 4 treatments were tested for calli and embryogenic calli of oil palm on modified Murashige–Skoog medium, namely: T1: 100 % nitrogen source + 100 % 2,4D; T2: 25 % nitrogen source + 10 % 2,4D; T3: Z78 sonicated cell + 25 % nitrogen source + 10 % 2,4D and T4: Z78 pellet cell + 25 % nitrogen source + 10 % 2,4D. The ability of the inoculum to produce phytohormone indole-3-acetic acid was detected by ultra-performance liquid chromatography. Scanning electron microscopy and transmission electron microscopy showed successful artificial symbiosis of calli and embryogenic calli-bacterium. This study has demonstrated that Z78 pellet cell and sonicated cell have the potential to promote growth of in vitro calli and embryogenic calli of oil palm under symbiosis conditions.

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2, 4 Dichlorophenoxyacetic acid


Acetylene reduction assay


Biological nitrogen fixation


Embryogenic calli


Indole-3-acetic acid


Murashige–Skoog medium




Herbaspirillum seropedicae strain Z78 (ATCC 35893)


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The experiment was supported by Grant 1001/PBIOLOGI/815088. Authors are thankful to the School of Biological Sciences, Universiti Sains Malaysia, Penang, Malaysia for research facilities; and the Advanced Biotechnology and Breeding Centre (ABBC) of Malaysian Palm Oil Board (MPOB), Bangi, Selangor, Malaysia for the oil palm planting materials. We also thank Dr Samuel Allen, Agricultural Research Scientist, New Mexico State University, for his review of this manuscript.

Authors’ contributions

SL Lim performed this experiment, analyzed the data and prepared the article; I Zamzuri provided the plant materials and consultation for the experimental design, outcomes, plant histological study and revised the article; S Subramaniam was involved in experimental design, provided plant tissue culture opinions and revised the article; and HG Amir concieved this experiment, provided consultation in the overall experiment and data analysis and amended the article. All authors have read and approved the manuscript.

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Correspondence to Hamzah Ghazali Amir.

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Lim, S., Subramaniam, S., Zamzuri, I. et al. Biotization of in vitro calli and embryogenic calli of oil palm (Elaeis guineensis Jacq.) with diazotrophic bacteria Herbaspirillum seropedicae (Z78). Plant Cell Tiss Organ Cult 127, 251–262 (2016).

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  • Artificial symbiosis
  • Oil palm
  • Herbaspirillum seropedicae
  • In vitro culture