Maximising Production of Palmaria Palmata (Linnaeus) Weber & Mohr, 1805
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FINAL REPORT TO MASTS under Visiting Fellowship Scheme 2015 Dr J Craig Sanderson Kai Ho, Tasmanian Sea Vegetables, Associate University Tasmania. 27 July 2015 - 23 October 2015 Maximising Production of Palmaria palmata (Linnaeus) Weber & Mohr, 1805 Scottish Association Marine Sciences Supported by: Mara Seaweed Loch Fyne Oysters Deakin University and NAFC Marine Centre University of the Highlands and the Islands. Acknowledgements Thanks to Dr Adam Hughes for driving this project from the beginning and overall support while in Scotland. We were grateful to have received funding through MASTS which enabled the project. Dr Phillip Kerrison and the technical staff at SAMS facilitated the seaweed culture making it possible to do more than anticipated within the set time frame. Dr Claire Gachon and Yacine Badis provided inspiration for future research. Dr Liz Cook facilitated off site support. Helpful discussions were made possible through Phycomorph at the meeting in Porto and we were very grateful for Dr Lesley Mc Evoy’s support in the Sheltlands. The final laboratory work could not have been completed without the very capable SAMS intern Sandra Yeregui Osa. ABSTRACT The aim of this project was to investigate maximising spore release and settlement success for the commercial red algal species Palmaria palmata. Difficulties were encountered in obtaining fertile material to complete the original experimental design so the direction was altered to incorporate investigating mass production of Palmaria palmata through vegetative propagation. This involved obtaining plantlets and tetraspores from fragments and cell aggregates of meristematic and sub-meristimatic tissue. This fulfiled the initial aim of the original project through enabling mass production of Palmaria palmata and has the added bonus of not requiring a source of fertile material meaning it would be possible to conduct at any time of the year. Success in these areas will facilitate the beginning of a future industry supplying Palmaria for edible markets and may lead to opportunities for culturing other algae for more varied uses in the commercial sector. Vegetative propagation of Palmaria palmata plants was initially sourced through cutting meristems into small pieces approx. 1 x 1 mm. Macerated tissue was then trialled and then very small (100µm) sections of meristematic tissue generated using a microtome. Growth of plant pieces of all sizes was a consistent 10% per day. Culturing of segments of meristems from Osmundia sp and Chondrus crispus for comparison resulted in growth rates of 2 and 3% per day respectively. Freezing of Palmaria plants was shown to kill all plant tissue except the meristems. Use of a freeze microtome for generating plant fragments from meristems may be worth following up. This method has the added advantage that freezing kills off all associated biota, sterilising the fragments. The production of new plantlets for Palmaria occurred principally from the meristematic tissue. As part of this project, preliminary work was conducted on generating adventive material from plant tissues other than the meristem using auxins and other shoot producing compounds. Results were confounded by plant infections. Further work needs to be conducted with an emphasis on maintaining axenic culture conditions. Best growth was obtained using commercially available seaweed extract plant growth promoters. Tips grown from bladelets of Palmaria blades in a commercial aquaculture farm grew from 450 to 58000mm2 over a period of 56 days equivalent to 8.7% per day. This indicates potential growth to full size from spores or very small plant pieces within six months when grown in tanks on land. Reports of growth rates elsewhere for Palmaria palmata of 20+% indicate a potential growth period to full size plants within 2 months. From tetraspores that were released from fertile plants collected as part of this project, and from Palmaria plants maintained in tanks as part of this project, an infecting oomycete was found that has only recently been identified and yet to be reported in the literature. This may have particular relevance for any industry based on cultivating Palmaria from spores as crashes of cultures of Palmaria have been common place and this fungus may be the cause. The find expands our knowledge in this area. An endophyte of Palmaria palmata was also identified: Acrochaetium secundatum which is a very common contaminant on blades of Palmaria. This identification expanded the range of this ecologically significant microorganism. ABSTRACT INTRODUCTION PART 1 Investigation to maximise spore release and settlement success for the commercial red algal species Palmaria palmata (Linnaeus) Weber & Mohr, 1805 PART 2 Mass culture of Palmaria palmata from meristematic tissue. PART 3 Growth and adventive tissue accelerating compounds. REFERENCES Appendix 1 Identification of meristematic tissue through freezing. Appendix 2 Growth of Palmaria fronds from excised adventitious shoots (tips). Appendix 3 Palmaria Oomycete infection Appendix 4 Palmaria Endophyte/epiphyte Appendix 5 Phycomorph Workshop Porto, Portugal September 2015. Appendix 6 Shetlands visit, September 2015. Appendix 7 MASTS, Marine Alliance for Science and Technology for Scotland meeting October 2015. INTRODUCTION In many developed countries significant proportions of the population are obese or overweight. There is compelling evidence from literature that seaweeds should become more common in global diets. Seaweeds are high in protein, dietary fibre, long-chain omega-3 fatty acids and a suite of vitamins, minerals and antioxidants. Regular consumption can reduce obesity and associated illnesses, such as diabetes and heart disease, whilst potentially enhancing the health of societies now and for future generations. Worldwide seaweed production via aquaculture has doubled in the past 10 years and now accounts for a total annual harvest of 23.8 Mt valued at over US$6 billion globally. Other than culture Porphyra, which is the most valuable edible seaweed species in the world, culture of red seaweeds is secondary to the easier to culture brown seaweeds, despite red seaweeds generally being of higher nutritional value. Demand for red seaweeds currently exceeds supply. Red seaweeds are also the source of both unique chemical compounds and various gelling agents which are widely used as food additives. The red alga Palmaria palmata (L.) O. Kuntze is a conspicuous vegetation component in the low intertidal and upper subtidal zones along both sides of the North Atlantic Ocean, from the Arctic to cold-temperate regions. The alga has been harvested and used for centuries in the coastal areas of Northern Europe and North America. Pressure on wild stocks has encouraged investigation into mass culture methods which has met some success for tank and rope cultivation. The original aim of this project was to further exploit the current knowledge of Palmaria life history traits to provide mass spore release on cue at any time of the year. A number of substrates for on-growing Palmaria were to be investigated. Problems were encountered sourcing tetrasporic plants for this investigation and the project brief was expanded to investigate mass production of Palmaria palmata through vegetative propagation as outlined in a relatively recent publication by Titlyanov et al. (2006) entitled ‘New methods of obtaining plantlets and tetraspores from fragments and cell aggregates of meristematic and sub-meristimatic tissue of the red alga Palmaria palmata’. This fulfils the initial aim of the original proposed project through enabling mass production of Palmaria. It also has the added bonus of not requiring a source of fertile material and can be conducted at any time of the year. Success in these areas will facilitate the beginning of a future industry supplying Palmaria for edible markets and may lead to other opportunities for culturing red algae for more varied uses in the commercial sector. This proposed project fits within the MASTS ‘Productive Seas’ research theme. Aquaculture is pivotal to the rural economy of some areas within Scotland and is likely to expand into the production of other non-food products and services through biofuels, marine biotechnology and genomics. Research into growing Palmaria complements aquaculture production of seaweeds for biofuels and is a nutrient extractive aquaculture operation as distinct from fin fish aquaculture. PART 1 Investigation to maximise spore release and settlement success for the commercial red algal species Palmaria palmata (Linnaeus) Weber & Mohr, 1805 Palmaria palmata fronds are usually fertile in the first six months of the year although this can vary depending on geographic location. In the vicinity of Oban, fronds may become fertile as early as October-November (SAMS personnel pers com). The seasonality of reproduction did not agree with the timing of this project (August-October). Pang and Luning 2006 have induced tetraspory in Palmaria palmata under conditions of short day (8:16 dark: light) and low temperature: 10oC within 5 weeks. This was trialled as part of the project but there were difficulties associated with maintaining these conditions. Inducing tetraspory is also dependent on having tetrasporic plants and as tetrasporic plants have the same appearance the same as male plants, there is the possibility that all plants collected were males. Inducing tetraspory was not successful. Careful examination of field populations of P. palmata at Ellenabeich adjacent to Easdale