Effects of pH on growth and biochemical responses in Agarophyton vermiculophyllum under different temperature conditions

  • Palas Samanta
  • Sojin Jang
  • Sookkyung Shin
  • Jang Kyun KimEmail author


The effects of pH (6.2, 7.2, 8.2, 9.2, and 10.2) under rising temperature (30 °C vs 20 °C) on Agarophyton vermiculophyllum growth and bio-physiology were investigated. Results showed that A. vermiculophyllum exhibited lower growth rates under elevated temperature in all pH values. Chlorophyll a, carotenoid, and phycocyanin levels were significantly enhanced by temperature elevation (p < 0.05). Enhanced H2O2 production either at lower or higher pH values correlated with lipid peroxidation (LPO) levels under elevated temperature, which suggested oxidative stress development. Oxidative damage was more severe at elevated pH values, which is confirmed by higher reactive oxygen species (ROS) levels. Compared with ambient pH 8.2 value, lower pH values under elevated temperature lead to increase activities of superoxide dismutase (SOD), catalase (CAT), and glutathione S-transferase (GST), indicating that these enzymes played an important role to combat stress. However, decreased glutathione reductase (GR) and glutathione peroxidase (GPx) activities indicate least contribution for ROS scavenging at lower pH values. On contrary, SOD and CAT declined at elevated pH values compared with ambient pH, suggesting least contribution for ROS removal. Moreover, enhanced GR and GPx activities at elevated pH and temperature are not enough to scavenge ROS production. These data are consistent with higher H2O2 and LPO levels, and lower GST activities. Collectively, our results indicated that either pH fluctuations or elevated temperature displayed a disadvantageous influence on growth and bio-physiology of A. vermiculophyllum. Therefore, rising temperature alleviates adverse effects of seawater acidification, but it aggravates the negative effects of seawater alkalization on growth and bio-physiology of A. vermiculophyllum.


Agarophyton vermiculophyllum Rhodophyta Antioxidant enzymes pH Temperature Oxidative stress 


Author contribution

All authors have approved the final article and have participated in the research and/or article preparation.

Funding information

This study was supported by Basic Science Research Programs through the National Research Foundation of Korea (NRF) funded by the Ministry of Education (NRF-2017R1A6A1A06015181) and by the Ministry of Science and ICT (2019R1F1A1059663).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.


  1. Ali MB, Hahn EJ, Paek KY (2005) Effects of temperature on oxidative stress defense systems, lipid peroxidation and lipoxygenase activity in Phalaenopsis. Plant Physiol Biochem 43:213–223PubMedCrossRefPubMedCentralGoogle Scholar
  2. Asada K (1994) Production and action of active oxygen species in photosynthetic tissues. In: Foyer CH, Mullineauxs RH (eds) Causes of photooxidative stress and amelioration of defense system in plants. CRC Press, Boca Raton, pp 77–103Google Scholar
  3. Beer S, Eshel A (1985) Determining phycoerythrin and phycocyanin concentrations in aqueous crude extracts of red algae. Aust J Mar Freshw Res 36:785–792CrossRefGoogle Scholar
  4. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein using the principle of protein–dye binding. Anal Biochem 72:248–252PubMedPubMedCentralCrossRefGoogle Scholar
  5. Cade-Menun BJ, Paytan A (2010) Nutrient temperature and light stress alter phosphorus and carbon forms in culture-grown algae. Mar Chem 121:27–36CrossRefGoogle Scholar
  6. Caldeira K, Wickett ME (2003) Oceanography: anthropogenic carbon and ocean pH. Nature 425:365PubMedCrossRefPubMedCentralGoogle Scholar
  7. Cathcart R, Schwiers E, Ames BN (1983) Detection of picomole levels of hydroperoxides using a fluorescent dichlorofluorescein assay. Anal Biochem 134:111–116PubMedCrossRefPubMedCentralGoogle Scholar
  8. Chen B, Zou D, Zhu M, Yang Y (2017) Effects of CO2 levels and light intensities on growth and amino acid contents in red seaweed Gracilaria lemaneiformis. Aquac Res 48:2683–2690CrossRefGoogle Scholar
  9. Chen B, Zou D, Du H, Ji Z (2018) Carbon and nitrogen accumulation in the economic seaweed Gracilaria lemaneiformis affected by ocean acidification and increasing temperature. Aquaculture 482:176–182CrossRefGoogle Scholar
  10. Collen J, Davidson I (1999) Stress tolerance and reactive oxygen metabolism in the intertidal red seaweed Mastocarpus stellatus and Chondrus crispus. Plant Cell Environ 22:1143–1151CrossRefGoogle Scholar
  11. Corey P, Kim JK, Garbary DJ, Prithiviraj B, Duston J (2012) Bioremediation potential of Chondrus crispus (Basin Head) and Palmaria palmata: effect of temperature and high nitrate on nutrient removal. J Appl Phycol 24:441–448CrossRefGoogle Scholar
  12. D’Mello BR, Chemburkar MS (2018) Effect of temperature and pH variation on biomass and lipid production of Auxenochlorella pyrenoidosa. Res J Life Sci Bioinform Pharm Chem Sci 4:378–387Google Scholar
  13. Dhaka P, Singh GP (2018) Effect of pH on growth and biopigment accumulation of green alga Dunaliella salina. Int J Pharm Sci Res 9:271–276Google Scholar
  14. Dhindsa RS, Plumb-Dhindsa P, Thorpe TA (1981) Leaf senescence: correlated with increased levels of membrane permeability and lipid peroxidation, and decreased levels of superoxide dismutase and catalase. J Exp Bot 32:93–101CrossRefGoogle Scholar
  15. Dring MJ (2006) Stress resistance and disease resistance in seaweeds: the role of reactive oxygen metabolism. Adv Bot Res 43:175–207CrossRefGoogle Scholar
  16. Falkowski PG, Raven JA (1997) Aquatic photosynthesis. Blackwell, MaldenGoogle Scholar
  17. Gao K, Aruga Y, Asada K, Ishihara T, Akano T, Kiyohara M (1991) Enhanced growth of the red alga Porphyra yezoensis Ueda in high CO2 concentrations. J Appl Phycol 3:355–362CrossRefGoogle Scholar
  18. Gao K, Aruga Y, Asada K, Ishihara T, Akano T, Kiyohara M (1993) Calcification in the articulated coralline alga Corallina pilulifera, with special reference to the effect of elevated CO2 concentration. Mar Biol 117:129–132CrossRefGoogle Scholar
  19. Gerloff-Elias A, Spijkerman E, Pröschold T (2005) Effect of external pH on the growth, photosynthesis and photosynthetic electron transport of Chlamydomonas acidophila Negoro, isolated from an extremely acidic lake (pH 2.6). Plant Cell Environ 28:1218–1229CrossRefGoogle Scholar
  20. Gimmler H (2001) Acidophilic and acidotolerant algae. In: Rai LC, Gaur JP (eds) Algal adaptation to environmental stresses physiological, biochemical and molecular mechanisms. Springer, Heidelberg, pp 259–290CrossRefGoogle Scholar
  21. Gimmler H, Weis U (1999) Dunaliella acidophila: Life at pH 1.0. In: Avron M, Ben-Amotz A (eds) Dunaliella: physiology, biochemistry and biotechnology. CRC Press, Boca Raton, pp 99–133Google Scholar
  22. Gorman L, Kraemer GP, Yarish C, Boo SM, Kim JK (2017) The effects of temperature on the growth and nitrogen content of Gracilaria vermiculophylla and Gracilaria tikvahiae from LIS, USA. Algae 32:57–66CrossRefGoogle Scholar
  23. Guedes AC, Amaro HM, Pereira RD, Malcata FX (2011) Effects of temperature and pH on growth and antioxidant content of the microalga Scenedesmus obliquus. Biotechnol Prog 27:1218–1224PubMedCrossRefPubMedCentralGoogle Scholar
  24. Habig WH, Pabst MJ, Jakoby WB (1974) Glutathione S-transferases. The first enzymatic step in mercapturic acid formation. J Biol Chem 249:7130–7139PubMedPubMedCentralGoogle Scholar
  25. Häder DP, Williamson CE, Wängberg SÅ, Rautio M, Rose KC, Gao K, Helbling EW, Sinha RP, Worrest R (2015) Effects of UV radiation on aquatic ecosystems and interactions with other environmental factors. Photochem Photobiol Sci 14:108–126PubMedCrossRefPubMedCentralGoogle Scholar
  26. Halliwell B, Gutteridge JMC (1989) Protection against oxidants in biological systems: the super oxide theory of oxygen toxicity. In: Halliwell B, Gutteridge JMC (eds) Free radicals in biology and medicine. Clarendon Press, Oxford, pp 86–123Google Scholar
  27. Heath RL, Packer L (1968) Photo peroxidation in isolated chloroplasts I. Kinetics and stoichiometry of fatty acid peroxidation. Arch Biochem Biophys 125:189–198PubMedCrossRefPubMedCentralGoogle Scholar
  28. Howland RJM, Tappin AD, Uncles RJ, Plummer DH, Bloomer NJ (2000) Distributions and seasonal variability of pH and alkalinity in the Tweed Estuary, UK. Sci Total Environ 251-252:125–138PubMedCrossRefPubMedCentralGoogle Scholar
  29. Jiang H, Zou D, Lou W, Deng Y, Zeng X (2018) Effects of seawater acidification and alkalization on the farmed seaweed, Pyropia haitanensis (Bangiales, Rhodophyta), grown under different irradiance conditions. Algal Res 31:413–420CrossRefGoogle Scholar
  30. Khalil ZI, Asker MMS, El-Sayed S, Kobbia IA (2010) Effect of pH on growth and biochemical responses of Dunaliella bardawil and Chlorella ellipsoidea. World J Microbiol Biotechnol 26:1225–1231PubMedCrossRefPubMedCentralGoogle Scholar
  31. Kim JK, Yarish C (2014) Development of a sustainable land-based Gracilaria cultivation system. Algae 29:217–225CrossRefGoogle Scholar
  32. Kim JK, Kraemer GP, Neefus CD, Chung IK, Yarish C (2007) Effects of temperature and ammonium on growth, pigment production and nitrogen uptake by four species of Porphyra (Bangiales, Rhodophyta) native to the New England coast. J Appl Phycol 19:431–440CrossRefGoogle Scholar
  33. Kim JK, Kraemer GP, Yarish C (2014) Field scale evaluation of seaweed aquaculture as a nutrient bioextraction strategy in Long Island Sound and the Bronx River Estuary. Aquaculture 433:148–156CrossRefGoogle Scholar
  34. Kim JK, Pereira R, Yarish C (2016) Tolerances to hypo-osmotic and temperature stresses in native and invasive Gracilaria species. Phycologia 55:257–264CrossRefGoogle Scholar
  35. Kim JK, Yarish C, Hwang EK, Park MS, Kim YD (2017) Seaweed aquaculture: cultivation technologies, challenges and its ecosystem services. Algae 32:1–13CrossRefGoogle Scholar
  36. Koch M, Bowes G, Ross C, Zhang XH (2013) Climate change and ocean acidification effects on seagrasses and marine macroalgae. Glob Chang Biol 19:103–132PubMedCrossRefPubMedCentralGoogle Scholar
  37. Kumar M, Kumari P, Gupta V, Reddy CRK, Jha B (2010) Biochemical responses of red alga Gracilaria corticata (Gracilariales, Rhodophyta) to salinity induced oxidative stress. J Exp Mar Biol Ecol 391:27–34CrossRefGoogle Scholar
  38. Kwon SY, Jeong YJ, Lee HS, Kim JS, Cho KY, Allen RD, Kwak SS (2002) Enhanced tolerances of transgenic tobacco plants expressing both superoxide dismutase and ascorbate peroxidase in chloroplasts against methyl viologen-mediated oxidative stress. Plant Cell Environ 25:873–882CrossRefGoogle Scholar
  39. Lee DH, Kim YS, Lee CB (2001) The inductive responses of the antioxidant enzymes by salt stress in the rice (Oryza sativa L.). J Plant Physiol 158:737–745CrossRefGoogle Scholar
  40. Lesser MP (2006) Oxidative stress in marine environments: biochemistry and physiological ecology. Annu Rev Physiol 68:253–278PubMedCrossRefPubMedCentralGoogle Scholar
  41. Lichtenthaler H, Wellburn A (1983) Determinations of total carotenoids and chlorophylls a and b of leaf extracts in different solvents. Biochem Soc Trans 11:591–592CrossRefGoogle Scholar
  42. Lignell Å, Pedersén M (1989) Effects of pH and inorganic carbon concentration on growth of Gracilaria secundata. Br Phycol J 24:83–89CrossRefGoogle Scholar
  43. Liu BH, Lee YK (2000) Secondary carotenoids formation by the green alga Chlorococcum sp. J Appl Phycol 12:301–307CrossRefGoogle Scholar
  44. Liu CX, Zou DH (2015) Do increased temperature and CO2 levels affect the growth, photosynthesis, and respiration of the marine macroalga Pyropia haitanensis (Rhodophyta)? An experimental study. Hydrobiologia 745:285–296CrossRefGoogle Scholar
  45. Loggini B, Scartazza A, Brugnoli E, Navari-Izzo F (1999) Antioxidative defense system, pigment composition and photosynthetic efficiency in two wheat cultivars subjected to drought. Plant Physiol 119:1091–1100PubMedPubMedCentralCrossRefGoogle Scholar
  46. Lu IF, Sung MS, Lee TM (2006) Salinity stress and hydrogen peroxide regulation of antioxidant defense system in Ulva fasciata. Mar Biol 150:1–15CrossRefGoogle Scholar
  47. Luo MB, Liu F (2011) Salinity-induced oxidative stress and regulation of antioxidant defense system in the marine macroalga Ulva prolifera. J Exp Mar Biol Ecol 409:223–228CrossRefGoogle Scholar
  48. Machalek KM, Davison IR, Falkowski PG (1996) Thermal acclimation and photoacclimation of photosynthesis in the brown alga Laminaria saccharina. Plant Cell Environ 19:1005–1016CrossRefGoogle Scholar
  49. Maharana D, Das PB, Verlecar XN, Pise NM, Gauns MU (2015) Oxidative stress tolerance in intertidal red seaweed Hypnea musciformis (Wulfen) in relation to environmental components. Environ Sci Pollut Res 22:18741–18749CrossRefGoogle Scholar
  50. Middelboe AL, Hansen PJ (2007) Direct effects of pH and inorganic carbon on macroalgal photosynthesis and growth. Mar Biol Res 3:134–144CrossRefGoogle Scholar
  51. Misra HP, Fridovich I (1972) The role of superoxide anion in the autoxidation of epinephrine and a simple assay for superoxide dismutase. J Biol Chem 247:3170–3175PubMedPubMedCentralGoogle Scholar
  52. Mohsen AF, Hasr AH, Metwalli AM (1973) Effect of temperature variations on growth, reproduction, amino acid synthesis, fat and sugar content in Ulva fasciata Delile plants. Hydrobiologia 43:451–460CrossRefGoogle Scholar
  53. Nejrup LB, Staehr PA, Thomsen MS (2013) Temperature- and light-dependent growth and metabolism of the invasive red algae Gracilaria vermiculophylla – a comparison with two native macroalgae. Eur J Phycol 48:295–308CrossRefGoogle Scholar
  54. Paglia DE, Valentine WN (1967) Studies on the quantitative and qualitative characterization of erythrocyte glutathione peroxidase. J Lab Clin Med 70:158–169Google Scholar
  55. Park JH, Kim JK, Kong JA, Depuydt S, Brown MT, Han T (2017) Implications of rising temperatures for gametophyte performance of two kelp species from Arctic waters. Bot Mar 60:39–48CrossRefGoogle Scholar
  56. Pereira M, Bartolomé MC, Sánchez-fortún S (2013) Influence of pH on the survival of Dictyosphaerium chlorelloides populations living in aquatic environments highly contaminated with chromium. Ecotoxicol Environ Saf 98:82–87PubMedCrossRefPubMedCentralGoogle Scholar
  57. Raikar SV, Iima M, Fujita Y (2001) Effect of temperature, salinity and light intensity on the growth of Gracilaria spp. (Gracilariales, Rhodophyta) from Japan, Malaysia and India. Indian J Mar Sci 30:98–104Google Scholar
  58. Reddy AR, Chaitanya KV, Vivekanandan M (2004) Drought-induced response of photosynthesis and antioxidant metabolism in higher plants. J Plant Physiol 161:1189–1202CrossRefGoogle Scholar
  59. Rocha CMR, Sousa AMM, Kim JK, Magalhães JMCS, Yarish C, do Pilar Gonçalves M (2019) Characterization of agar from Gracilaria tikvahiae cultivated for nutrient bioextraction in open water farms. Food Hydrocoll 89:260–271CrossRefGoogle Scholar
  60. Ross C, Van Alstyne KL (2007) Intraspecific variation in stress-induced hydrogen peroxide scavenging by the Ulvoid macroalga Ulva lactuca. J Phycol 43:466–474CrossRefGoogle Scholar
  61. Rueness J (2005) Life history and molecular sequences of Gracilaria vermiculophylla (Gracilariales, Rhodophyta), a new introduction to European waters. Phycologia 44:120–128CrossRefGoogle Scholar
  62. Samanta P, Shin SK, Jang S, Kim JK (2019a) Comparative assessment of salinity tolerance based on physiological and biochemical performances in Ulva australis and Pyropia yezoensis. Algal Res 42:101590CrossRefGoogle Scholar
  63. Samanta P, Shin SK, Jang S, Song YC, Oh S, Kim JK (2019b) Stable carbon and nitrogen isotopic characterization and tracing nutrient sources of Ulva blooms around Jeju coastal areas. Environ Pollut 245:113033PubMedCrossRefPubMedCentralGoogle Scholar
  64. Sampath-Wiley P, Neefus CD, Jahnke LS (2008) Seasonal effects of sun exposure and emersion on intertidal seaweed physiology: fluctuations in antioxidant contents, photosynthetic pigments and photosynthetic efficiency in the red alga Porphyra umbilicalis Kützing (Rhodophyta, Bangiales). J Exp Mar Biol Ecol 361:83–91CrossRefGoogle Scholar
  65. Sang M, Wang M, Liu J, Zhang C, Li A (2012) Effects of temperature, salinity, light intensity, and pH on the eicosapentaenoic acid production of Pinguiococcus pyrenoidosus. J Ocean Univ China 11:181–186CrossRefGoogle Scholar
  66. Sergiev I, Alxieva V, Karanov E (1997) Effect of spermone, atrazine and combination between them on some endogenous protective systems and stress markers in plants. Comp. Rend. Acad. Bulg. Sci., 51: 121-124.Google Scholar
  67. Stæhr PA, Wernberg T (2009) Physiological responses of Ecklonia radiata (Laminariales) to a latitudinal gradient in ocean temperature. J Phycol 45:91–99PubMedCrossRefPubMedCentralGoogle Scholar
  68. Sung MS, Hsu YT, Wu TM, Lee TM (2009) Hypersalinity and hydrogen peroxide upregulation of gene expression of antioxidant enzymes in Ulva fasciata against oxidative stress. Mar Biotechnol 11:199–209PubMedCrossRefPubMedCentralGoogle Scholar
  69. Takahashi T, Sutherland SC, Chipman DW, Goddard JG, Newberger T, Sweeney C (2014) Climatological distributions of pH, pCO2, total CO2, alkalinity, and CaCO3 saturation in the global surface ocean. Carbon Dioxide Information Analysis Center, Oak Ridge National Laboratory, U.S. Department of Energy, Oak RidgeGoogle Scholar
  70. Tammam AA, Fakhry EM, El-Sheekh M (2011) Effect of salt stress on antioxidant system and the metabolism of the reactive oxygen species in Dunaliella salina and Dunaliella tertiolecta. Afr J Biotechnol 10:3795–3808Google Scholar
  71. Taraldsvik M, Myklestad S (2000) The effect of pH on growth rate, biochemical composition and extracellular carbohydrate production of the marine diatom Skeletonema costatum. Eur J Phycol 35:189–194CrossRefGoogle Scholar
  72. Vardi A, Berman-Frank I, Rozenberg T, Hadas O, Kaplan A, Levine A (1999) Programmed cell death of the dinoflagellate Peridinium gatunense is mediated by CO2 limitation and oxidative stress. Curr Biol 9:1061–1064PubMedCrossRefPubMedCentralGoogle Scholar
  73. Wu H, Shin SK, Jang SJ, Yarish C, Kim JK (2018) Growth and nutrient bioextraction of Gracilaria chorda, G. vermiculophylla, Ulva prolifera and U. compressa under hypo- and hyper-osmotic conditions. Algae 33:329–340CrossRefGoogle Scholar
  74. Xu Z, Zou D, Gao K, Li M (2011) Effects of temperature, irradiance level and nitrate concentration on the uptake of inorganic phosphorus in Gracilaria lemaneiformis (Rhodophyta). J Fish China 35:1023–1029CrossRefGoogle Scholar
  75. Xu K, Chen H, Wang W, Xu Y, Ji D, Chen C, Xie C (2017) Responses of photosynthesis and CO2 concentrating mechanisms of marine crop Pyropia haitanensis thalli to large pH variations at different time scales. Algal Res 28:200–210CrossRefGoogle Scholar
  76. Yang Y, Fei X, Song J, Hu H, Wang G, Chung IK (2006) Growth of Gracilaria lemaneiformis under different cultivation conditions and its effects on nutrient removal in Chinese coastal waters. Aquaculture 254:248–255CrossRefGoogle Scholar
  77. Yang Y, Chai Z, Wang Q, Chen W, He Z, Jiang S (2015) Cultivation of seaweed Gracilaria in Chinese coastal waters and its contribution to environmental improvements. Algal Res 9:236–244CrossRefGoogle Scholar
  78. Young AJ, Frank HA (1996) Energy transfer reactions involving carotenoids: quenching of chlorophyll fluorescence. J Photochem Photobiol B 36:3–15PubMedCrossRefPubMedCentralGoogle Scholar
  79. Zhao YK, Zhang WS, Wang YN (2008) Research progress in physiology and molecular biology of plant responses to high pH. Chin J Eco-Agric 16:783–787Google Scholar
  80. Zou D, Gao K (2013) Thermal acclimation of respiration and photosynthesis in the marine macroalga Gracilaria lemaneiformis (Gracilariales, Rhodophyta). J Phycol 49:61–68PubMedCrossRefPubMedCentralGoogle Scholar
  81. Zou D, Liu S, Du H, Xu J (2012) Growth and photosynthesis in seedlings of Hizikia fusiformis (Harvey) Okamura (Sargassaceae, Phaeophyta) cultured at two different temperatures. J Appl Phycol 24:1321–1327CrossRefGoogle Scholar

Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  1. 1.Department of Marine ScienceIncheon National UniversityIncheonRepublic of Korea
  2. 2.Research Institute of Basic SciencesIncheon National UniversityIncheonRepublic of Korea
  3. 3.System Toxicology Research CenterKorea Institute of ToxicologyDaejeonRepublic of Korea

Personalised recommendations