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Molecular analysis of physiological responses to changes in nitrogen in a marine macroalga, Porphyra yezoensis (Rhodophyta)

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Abstract

The rhodophyte seaweed Porphyra yezoensis, known more commonly world-wide as “nori”, is an important commercial crop in Japan. Cultivation of nori in Japan is often affected by outbreaks of “iroochi”, a discoloration of the thalli due to a decrease in inorganic nutrients in the culture area that in turn decreases the amount of photosynthetic pigments in the thalli. Treating thalli with inorganic nitrogen can reverse iroochi. In this paper, we report on the characterization of three P. yezoensis genes, a nitrate transporter (PyNRT2) and two urea transporters (PyUT1 and PyUT2), which may be involved in reversing iroochi. The predicted length of the PyNRT2 protein was 479 amino acids (AA), while PyUT1 and PyUT2 were 740 and 680 AA, respectively. PyNRT2 was more similar to NRT2 from a chromophyte than to NRTs from Chlamydomonas and higher plants. The two P. yezoensis UTs had 56% AA identity to each other, and showed the closest relationship to higher plant and yeast DUR3 proteins which formed a subfamily of the sodium-solute symporter protein family. Hydrophobicity plots of the AA sequences showed that the PyNRT2, PyUT1, and PyUT2 included 12, 15, and 16 transmembrane domains, respectively. Southern blot analysis indicated that the P. yezoensis genome has a single NRT2-encoding gene and at least four UT-encoding genes. Expression analysis of PyNRT2 and PyUT genes showed that the messenger RNA level of the PyNRT2 gene reached a maximum after 48 h in the nitrate starvation condition and was then restored to the constitutive level, while expression of the PyUT genes was induced in proportion to treatment times in the nitrate starvation condition. These results suggest that the PyNRT2 and PyUT are responsible for the high-affinity nitrate/urea transport systems that operate under low external nitrate concentrations.

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Abbreviations

AA:

amino acid(s)

DIN:

dissolved inorganic nitrogen

EST:

expressed sequence tag

NNP:

nitrate-nitrite porter

NRT:

nitrate transporter

nt:

nucleotide(s)

ORF:

open reading frame

PCR:

polymerase chain reaction

PES:

Provasoli’s enriched seawater

RACE:

rapid amplification of cDNA ends

ss:

single-stranded

SSS:

sodium-solute symporter

TM:

transmembrane domain

UT:

urea transporter

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Acknowledgments

This study was supported by the National Research Institute of Fisheries Science, Fisheries Research Agency, Japan, and was funded in part by Grant-in-Aids from the Fisheries Agency, Government of Japan and the Japan Society for the Promotion of Science. We thank Dr. M. Iwabuchi and Dr. T. Fukunaga of the Fukuoka Fisheries and Marine Technology Research Center, Japan, for supplying P. yezoensis strain FA-89 and for help with its culture. We also thank Dr. M. Kobayashi and M. Yoshikawa of National Research Institute of Fisheries Science, Japan, for help with the experiments.

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Authors and Affiliations

  1. Laboratory of Marine Biochemistry, Graduate School of Bioresources, Mie University, 1577 Kurima-machiya, Tsu, Mie, 514-8507, Japan

    M. Kakinuma, D. A. Coury, C. Nakamoto & H. Amano

  2. Fisheries Research Division, Mie Prefectural Science and Technology Promotion Center, 1-6277-4 Shiroko, Suzuka, Mie, 510-0243, Japan

    K. Sakaguchi

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  1. M. Kakinuma
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  2. D. A. Coury
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  3. C. Nakamoto
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  4. K. Sakaguchi
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  5. H. Amano
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Correspondence to M. Kakinuma.

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Kakinuma, M., Coury, D.A., Nakamoto, C. et al. Molecular analysis of physiological responses to changes in nitrogen in a marine macroalga, Porphyra yezoensis (Rhodophyta). Cell Biol Toxicol 24, 629–639 (2008). https://doi.org/10.1007/s10565-007-9053-7

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