Increased freezing stress tolerance of Nicotiana tabacum L. cv. Bright Yellow-2 cell cultures with the medium addition of Ascophyllum nodosum (L.) Le Jolis extract

  • Mahbobeh Zamani-BabgohariEmail author
  • Alan T. Critchley
  • Jeff Norrie
  • Balakrishnan Prithiviraj
Plant Tissue Culture


The study of bioactive components of the brown alga Ascophyllum nodosum (L.) Le Jolis has shown that they promote growth and increase productivity of plants. However, a standardized model system providing consistent responses to such bioactive components has yet to be established. Given that freezing stress, especially in northern climates, reduces plant growth and productivity, a technique was developed to protect plant cells under freezing stress using the natural products made from the abundant A. nodosum as a cell culture supplement. In this study, a homogenous cell culture system of Nicotiana tabacum L. cultivar Bright Yellow-2 (BY-2) suspension cells was used to investigate the bioactivity and protection level of this alga extract under freezing temperatures, and BY-2 cell growth, physiology, and molecular changes were measured in the presence or absence of the extract. The results indicated that A. nodosum extract significantly improved BY-2 cell survival after exposure to freezing temperatures. Inclusion of alga extract also improved cell growth, membrane stability, and nuclear integrity, and reduced cell death of cold-stressed BY-2 suspension cultures. It was concluded that A. nodosum extract influenced cellular and molecular regulation and triggered mechanisms, such as osmolyte accumulation and antioxidant activity, to combat freezing stress in BY-2 suspension cells.


Freezing stress Ascophyllum nodosum (L.) Le Jolis extract Suspension culture Growth Nicotiana tabacum L. cv. Bright Yellow-2 



The authors would like to thank the government of Nova Scotia for providing through a Nova Scotia Research and Innovation Graduate Scholarship. The authors also thank Dalhousie University for providing Entrance and Graduate Scholarships, Natural Sciences and Engineering Research Council of Canada (NSERC) for research funding, as well as Acadian Seaplants Ltd. team for supporting this project. Sincere thanks are expressed to Drs. Dhriti Battacharyya, Tudor Borza, Pushp Sheel Shukla, and Sridhar Ravichandran for their consistent help throughout the study.

Authors’ contributions

BP designed and supervised the research. MZ conducted experiments and wrote the manuscript. AC and JN contributed in providing resources, supporting the research, and advising throughout the experiments. All authors read and approved the manuscript.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

11627_2019_9972_MOESM1_ESM.docx (17 kb)
ESM 1 (DOCX 17 kb)


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Copyright information

© The Society for In Vitro Biology 2019

Authors and Affiliations

  1. 1.Department of Plant, Food and Environmental Sciences, Agricultural CampusDalhousie UniversityTruroCanada
  2. 2.Acadian Seaplants LimitedDartmouthCanada

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