Macroalgal Endophytes from the Atlantic Coast of Canada: A Potential Source of Antibiotic Natural Products?
Abstract
:1. Introduction
2. Experimental Section
2.1. Algal Collection
2.2. Surface Sterilization of Algae and Culture Techniques
Algal Species | Sterilant Immersion Duration (Seconds) | |
---|---|---|
Bleach (5.25%) | Ethanol (70%) | |
Chondrus crispus | 5 | 10 |
Devaleraea ramentacea | 5 | 10 |
Mastocarpus stellatus | 5 | 15 |
Palmaria palmata | 0 | 15 |
Polysiphonia lanosa | 0 | 10 |
Porphyra purpurea | 0 | 10 |
Porphyra umbilicalis | 0 | 10 |
Ascophyllum nodosum | 0 | 15 |
Fucus spiralis | 0 | 15 |
Fucus vesiculosus | 0 | 15 |
Saccharina latissima | 5 | 15 |
Spongomorpha arcta | 0 | 20 |
Ulva intestinalis | 0 | 20 |
Ulva lactuca | 0 | 20 |
2.3. Isolation of Endophytes
2.4. Identification of Fungi
2.5. Preparation of Extracts
2.6. Antibacterial and Antifungal Activity Assay
2.7. Statistical Analyses
3. Results and Discussion
3.1. Isolation of Endophytic Fungi from Marine Algae
Seaweed Species | Total Algae Segments | Pieces with Endophytic Growth | Isolation Frequency (%) | Distinct Fungal Taxa |
---|---|---|---|---|
Chondrus crispus | 246 | 2 | 0.08 | 2 |
Devaleraea ramentacea | 108 | 46 | 43 | 13 |
Mastocarpus stellatus | 98 | 32 | 33 | 9 |
Palmaria palmata | 167 | 146 | 87 | 7 |
Polysiphonia lanosa | 67 | 12 | 18 | 5 |
Porphyra purpurea | 85 | 17 | 20 | 2 |
Porphyra umbilicalis | 217 | 156 | 72 | 3 |
Ascophyllum nodosum | 152 | 16 | 11 | 1 |
Fucus spiralis | 192 | 16 | 8 | 4 |
Fucus vesiculosus | 109 | 6 | 6 | 1 |
Saccharina latissima | 1138 | 95 | 8 | 13 |
Spongomorpha arcta | 184 | 49 | 27 | 11 |
Ulva intestinalis | 117 | 24 | 21 | 5 |
Ulva lactuca | 201 | 15 | 8 | 3 |
Total | 3081 | 632 | 26 | 79 |
Host species | Isolate | Species | Accession number |
---|---|---|---|
Chondrus crispus | AF8-053 | Aspergillus sp. I | KF572134 |
AF8-129 | Penicillium crustosum I | KF572146 | |
Devaleraea ramentacea | AF8-079 | Cladosporium sp. | KF572136 |
AF8-081 | Septate Pigmented I | - | |
AF8-083 | Septate Pigmented II | - | |
AF8-085 | Sterile Hyaline I | - | |
AF8-087 | Sterile Hyaline II | - | |
AF8-089 | Botrytis sp. I | - | |
AF8-091 | Penicillium decumbens I | KF572138 | |
AF8-093 | Septate Pigmented III | - | |
AF8-095 | Aureobasidium pullulans I | - | |
Devaleraea ramentacea | AF8-109 | Trametes versicolor | KF572133 |
AF8-111 | White Fluffy II | - | |
AF8-113 | Coniothyrium sp. | KF572141 | |
AF8-115 | Botrytis sp. II | - | |
Mastocarpus stellatus | AF8-055 | Penicillium sp. | - |
AF8-057 | Red Yeast I | - | |
AF8-059 | Septate Pigmented IV | - | |
AF8-061 | Red Yeast II | - | |
AF8-063 | Sterile Hyaline III | - | |
AF8-065 | Sterile Hyaline IV | - | |
AF8-067 | Coelomycete I | - | |
AF8-101 | Penicillium decumbens II | KF572139 | |
Palmaria palmata | AF8-069 | Sterile Hyaline V | - |
AF8-071 | Hypoxylon sp. | KF572135 | |
AF8-073 | Penicillium decumbens III | KF572149 | |
AF8-075 | Penicillium chrysogenum I | KF572137 | |
AF8-077 | Helicomyces sp. | - | |
AF8-103 | Penicillium crustosum II | KF572147 | |
AF8-105 | Botrytis sp. III | - | |
AF8-107 | Sterile Hyaline VI | - | |
AF8-123 | Sterile Hyaline VII | - | |
Polysiphonia lanosa | AF8-097 | Sterile Hyaline VIII | - |
AF8-099 | Aureobasidium pullulans II | KF572140 | |
AF8-117 | Sterile Hyaline IX | - | |
AF8-119 | Botryotinia fuckeliana | KF572148 | |
AF8-121 | Sterile Hyaline X | - | |
Porphyra purpurea | AF8-125 | Sterile Hyaline XI | - |
AF8-127 | Sterile Hyaline XII | - | |
Porphyra umbilicalis | AF8-049 | Sterile Hyaline XIII | - |
AF8-051 | Sterile Hyaline XIV | - | |
AF8-131 | Aspergillus sydowii | KF572145 | |
Ascophyllum nodosum | AF1-141B | Aspergillus sp. II | - |
Fucus spiralis | AF1-021A | Black Hyaline I | - |
AF1-021B | Septate Pigmented V | - | |
AF1-021D | Pigmented Hyaline I | - | |
AF1-021G | Sterile Hyaline XV | - | |
Fucus vesiculosus | AF2-059A | Sterile Hyaline XXI | - |
Saccharina latissima | AF1-029A2 | White Hyaline IV | - |
AF1-029B2 | Black Hyaline II | - | |
AF1-073C | Penicillium chrysogenum II | KF572144 | |
AF1-073D | Aspergillus sp. III | - | |
AF1-073E | White Hyaline I | - | |
AF1-073G | Sterile Beige I | - | |
AF1-073N | Sterile Beige II | - | |
AF2-055B | Penicillium soppii I | KF572151 | |
AF2-055F | Sterile Hyaline XVI | - | |
AF2-055G | White Hyaline II | - | |
AF2-055O | Penicillium chrysogenum III | KF572150 | |
AF2-055P | Pigmented Hyaline IV(Red) | - | |
Spongomorpha arcta | AF1-037C2 | Sterile Hyaline XVII | - |
AF1-037F | Sterile Beige III | - | |
AF1-153A | Pigmented Hyaline VII | - | |
AF1-153B | Sterile Hyaline XVIII | - | |
AF1-153C | White Hyaline III | - | |
AF1-153D | Penicillium spinulosum | KF572143 | |
AF1-153H | Black Hyaline III | - | |
AF1-153J | Pigmented Hyaline VI | - | |
AF1-153L | Pigmented Hyaline V | - | |
AF1-153M | Penicillium sp. | KF572152 | |
AF1-153N | Penicillium soppii II | - | |
Ulva intestinalis | AF2-063C | Sterile Hyaline XIX | - |
AF2-063E | Septate Pigmented VIII | - | |
AF2-063F | Pigmented Hyaline VIII | - | |
AF2-063G | Aspergillus sp. V | - | |
AF2-063H | Septate Pigmented VI | - | |
Ulva lactuca | AF1-033A2 | Septate Pigmented VII | - |
AF1-033B | Sterile Hyaline XX | - | |
AF1-033C | Penicillium chrysogenum IV | KF572142 |
3.2. Antimicrobial Screening Results of Algal-Derived Fungal Endophytes
Fungal Taxa | Host Alga | Extract 1 Source | Inhibition (%) 2 | ||
---|---|---|---|---|---|
SA 3 | PA 3 | CA 3 | |||
Aspergillus sp. I | C. crispus | Mycelia | - | 18 ± 1 | - |
Aspergillus sp. I | C. crispus | Media | 28 ± 3 | - | 15 ± 3 |
Penicillium crustosum I | C. crispus | Media | 43 ± 1 | - | - |
Cladosporium sp. | D. ramentacea | Mycelia | 36 ± 1 | 9 ± 1 | - |
Septate Pigmented I | D. ramentacea | Mycelia | - | - | 17 ± 2 |
Sterile Hyaline I | D. ramentacea | Mycelia | - | 20 ± 1 | - |
Sterile Hyaline I | D. ramentacea | Media | 29 ± 2 | - | - |
Botrytis sp. I | D. ramentacea | Mycelia | 94 ± 1 | - | - |
P. decumbens I | D. ramentacea | Mycelia | 36 ± 4 | - | 98 ± 1 |
P. decumbens I | D. ramentacea | Media | - | - | 99 ± 1 |
Septate Pigmented III | D. ramentacea | Mycelia | 47 ± 4 | - | - |
Aureobasidium pullulans I | D. ramentacea | Media | 82 ± 4 | 51 ± 2 | 13 ± 2 |
Trametes versicolor | D. ramentacea | Mycelia | - | 17 ± 2 | - |
White Fluffy II | D. ramentacea | Media | - | - | 12 ± 1 |
Coniothyrium sp. | D. ramentacea | Mycelia | 31 ± 1 | - | - |
Coniothyrium sp. | D. ramentacea | Media | 25 ± 2 | 28 ± 2 | - |
Botrytis sp. II | D. ramentacea | Mycelia | - | - | 14 ± 1 |
Botrytis sp. II | D. ramentacea | Media | 97 ± 1 | - | - |
Red Yeast I | M. stellatus | Mycelia | 31 ± 2 | - | - |
Septate Pigmented IV | M. stellatus | Mycelia | - | 16 ± 3 | - |
Septate Pigmented IV | M. stellatus | Media | 23 ± 1 | - | 70 ± 3 |
P. decumbens II | M. stellatus | Media | - | - | 12 ± 1 |
Sterile Hyaline V | P. palmata | Mycelia | - | 11 ± 1 | 15 ± 4 |
Sterile Hyaline V | P. palmata | Media | - | - | 16 ± 1 |
Hypoxylon sp. | P. palmata | Mycelia | - | 13 ± 3 | 30 ± 3 |
Hypoxylon sp. | P. palmata | Media | - | - | 18 ± 2 |
P. decumbens III | P. palmata | Mycelia | - | - | 16 ± 5 |
P. chrysogenum I | P. palmata | Media | 58 ± 3 | - | - |
Helicomyces sp. | P. palmata | Mycelia | 21 ± 1 | - | - |
Helicomyces sp. | P. palmata | Media | - | - | 21 ± 5 |
P. crustosum II | P. palmata | Mycelia | 82 ± 5 | - | 22 ± 3 |
P. crustosum II | P. palmata | Media | 39 ± 2 | - | - |
Botrytis sp. III | P. palmata | Media | 92 ± 1 | - | - |
Sterile Hyaline VI | P. palmata | Mycelia | - | - | 13 ± 2 |
Sterile Hyaline VI | P. palmata | Media | 19 ± 6 | - | - |
Sterile Hyaline VII | P. palmata | Media | 90 ± 2 | - | 24 ± 2 |
Aureobasidium pullulans II | P. lanosa | Mycelia | 49 ± 1 | 34 ± 3 | - |
Sterile Hyaline IX | P. lanosa | Media | - | - | 40 ± 5 |
Botryotinia fuckeliana | P. lanosa | Mycelia | 96 ± 1 | - | 30 ± 2 |
Botryotinia fuckeliana | P. lanosa | Media | 41 ± 2 | - | - |
Sterile Hyaline X | P. lanosa | Mycelia | - | - | 20 ± 11 |
Sterile Hyaline XI | P. purpurea | Mycelia | 22 ± 1 | - | - |
Sterile Hyaline XI | P. purpurea | Media | 34 ± 9 | - | - |
Sterile Hyaline XII | P. purpurea | Mycelia | 50 ± 1 | - | - |
Sterile Hyaline XIII | P. umbilicalis | Media | - | - | 33 ± 1 |
Sterile Hyaline XIV | P. umbilicalis | Mycelia | - | 14 ± 1 | 27 ± 6 |
Sterile Hyaline XIV | P. umbilicalis | Media | 22 ± 5 | - | - |
Aspergillus sydowii | P. umbilicalis | Media | 51 ± 3 | - | - |
Aspergillus sp. II | A. nodosum | Mycelia | 66 ± 4 | - | - |
Aspergillus sp. II | A. nodosum | Media | 85 ± 1 | - | 15 ± 3 |
Black Hyaline I | F. spiralis | Media | 41 ± 2 | - | - |
Septate Pigmented V | F. spiralis | Mycelia | 39 ± 3 | 8 ± 1 | - |
Septate Pigmented V | F. spiralis | Media | 27 ± 3 | - | - |
Pigmented Hyaline I | F. spiralis | Mycelia | 53 ± 4 | - | - |
White Hyaline IV | S. latissima | Media | - | - | 30 ± 1 |
Black Hyaline II | S. latissima | Mycelia | - | - | 13 ± 5 |
P. chrysogenum II | S. latissima | Mycelia | 25 ± 4 | - | - |
P. chrysogenum II | S. latissima | Media | 31 ± 4 | - | - |
Aspergillus sp. III | S. latissima | Mycelia | 25 ± 1 | - | - |
Sterile Beige II | S. latissima | Media | - | - | 14 ± 3 |
White Hyaline II | S. latissima | Mycelia | 28 ± 3 | - | 27 ± 5 |
White Hyaline II | S. latissima | Media | 21 ± 1 | - | 18 ± 9 |
Pigmented Hyaline IV(Red) | S. latissima | Mycelia | - | 12 ± 1 | - |
Pigmented Hyaline IV(Red) | S. latissima | Media | - | - | 17 ± 1 |
Sterile Hyaline XVII | S. arcta | Mycelia | - | - | 13 ± 1 |
Pigmented Hyaline VII | S. arcta | Media | - | 15 ± 3 | - |
White Hyaline III | S. arcta | Mycelia | 27 ± 7 | - | - |
White Hyaline III | S. arcta | Media | - | - | 13 ± 3 |
Penicillium spinulosum | S. arcta | Mycelia | 65 ± 4 | - | - |
Penicillium spinulosum | S. arcta | Media | 37 ± 1 | - | - |
Pigmented Hyaline V | S. arcta | Media | 87 ± 3 | - | 82 ± 1 |
Septate Pigmented VIII | S. arcta | Mycelia | - | 17 ± 1 | - |
Septate Pigmented VIII | S. arcta | Media | 36 ± 2 | - | - |
Penicillium soppii II | S. arcta | Media | - | 9 ± 3 | - |
Sterile Hyaline XIX | U. intestinalis | Media | - | 9 ± 2 | 22 ± 2 |
Septate Pigmented VI | U. intestinalis | Mycelia | - | - | 13 ± 2 |
Septate Pigmented VII | U. lactuca | Mycelia | - | - | 18 ± 2 |
Sterile Hyaline XX | U. lactuca | Mycelia | - | 24 ± 3 | - |
4. Conclusions
Acknowledgments
Conflicts of Interest
References
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Flewelling, A.J.; Ellsworth, K.T.; Sanford, J.; Forward, E.; Johnson, J.A.; Gray, C.A. Macroalgal Endophytes from the Atlantic Coast of Canada: A Potential Source of Antibiotic Natural Products? Microorganisms 2013, 1, 175-187. https://doi.org/10.3390/microorganisms1010175
Flewelling AJ, Ellsworth KT, Sanford J, Forward E, Johnson JA, Gray CA. Macroalgal Endophytes from the Atlantic Coast of Canada: A Potential Source of Antibiotic Natural Products? Microorganisms. 2013; 1(1):175-187. https://doi.org/10.3390/microorganisms1010175
Chicago/Turabian StyleFlewelling, Andrew J., Katelyn T. Ellsworth, Joseph Sanford, Erica Forward, John A. Johnson, and Christopher A. Gray. 2013. "Macroalgal Endophytes from the Atlantic Coast of Canada: A Potential Source of Antibiotic Natural Products?" Microorganisms 1, no. 1: 175-187. https://doi.org/10.3390/microorganisms1010175