1. phd project - sintef · 2017-12-13 · 1. phd project improve palmaria palmata hatchery...
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![Page 1: 1. PhD project - SINTEF · 2017-12-13 · 1. PhD project Improve Palmaria palmata hatchery techniques -from tetraspores to seedling linesPeter Schmedes, Mette M., Nielsen, Jens Kjerulf](https://reader034.vdocuments.net/reader034/viewer/2022042810/5f9a3f9a8c6cec60b7575cf6/html5/thumbnails/1.jpg)
1. PhD project
Improve Palmaria palmata hatchery techniques - from tetraspores to seedling lines
Peter Schmedes, Mette M., Nielsen, Jens Kjerulf PetersenPhD [email protected]
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DTU Aqua, Technical University of Denmark
Aqua
Experts in filter feeder cultivation and fisheries assessmentsDessimination Centre and Kitchen, Oyster Tours
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DTU Aqua, Technical University of Denmark
Palmaria in foodfound in high valued end-products
• Snack Food or dried Dulse (prices in Denmark up to 3-6 Euro/25g)
• Composition of polysaccharides, aminoacids and minerals (Mouritsen, 2013)
• Candidate as salmon feed supplement (Moroney et al 2014)
• Cultivation is not feasible with current cultivation techniques (Werner&Dring, 2011)
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DTU Aqua, Technical University of Denmark
In Nature Palmaria is often epi-phytic
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DTU Aqua, Technical University of Denmark
Cultivation Challenge: High input = relative low yield
1 long-line of seeded nets 130 kg FW (1.8 t FW)
2-3 kg (2-5 t FW)
(Werner & Dring, 2011)
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DTU Aqua, Technical University of Denmark
High percentage spore mortality at low agitation
2n/n Fertile tetraspores
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DTU Aqua, Technical University of Denmark
Hatchery steps based on life cycleof P. palmata
Tetrasporangia (fertility)
Can be found throughout the yearPeak season is Jan-April (in Irish waters)- 40.000 spores per gram tissue!
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DTU Aqua, Technical University of Denmark
My Objectives & Research Questions
Step 1. Prolong spore production. Can we induce fertility during summer or pre-natural occurence to expand the season?
Step 2. Can we optimise spore release?Release Experiment: What is the effect of spore release duration (time) and agitation?
Step 3. Investigate optimal use of spores.SporeUse1: Increase in seedling density by applying fertilisation step.
SporeUse2: Study surface preference of spores for settlement - effect of alginate coating and level of agitation.
SporeUse3:Q: Can we store, dispurse and get the spores to re-settle on substrate?Results on the effect of blending spores and rope type
50/50% ->100?
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DTU Aqua, Technical University of Denmark
1. Induction of spore formation 1. Attempt of pre-natural occurence
– (69 days from 25.10.2016 to 04.01.2017)– Exposure of treatment combinations
(= 16 treatments, n=5)
Two day lengthsLD = 12:16 (L:D)SD = 5:19 (L:D)40 PAR
F2-media100 % (882 uM)10 % (88,2 uM)
Binomial data (fertile/non-fertile) Proc Glimmix SAS modelClass of variable: Dates Replicate Fotoperiod Meristem Temp Nut DayFertility = Fotoperiod Meristem Temp Nut Day/ dist=binomial;randomreplicate*day ; /* code for each container
10 C5 C
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DTU Aqua, Technical University of Denmark
1. Induction of spore formation off-season (summer)
• Collection– Palmaria collected and start of experiment in early July– Additional tissue collected in late August
• Laboratory treatments: – Edge removal, dessication, temp cycles, red light, connection to basal
frond– Allantoin and Spermidine addition (metabolites) to medium
Sporangia has yet to be induced!
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DTU Aqua, Technical University of Denmark
Pre-treatment of triplicate plants(4 hours vs 24 h dehydration)
4h: Spore release at 10 °C for 98 hours with no agitation
Same amount of tissue
2. Spore Release Experiment
3 subsamples per replicate meanPre-treatment
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DTU Aqua, Technical University of Denmark
0
200
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600
800
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1200
1400
1600
1h/0 Lmin 3h/0 Lmin 1h/2.5 Lmin 3h/2.5 Lmin
Spo
re c
ount
mL-
1 g-
1
The effect of Time and Agitation of spore release yield
0
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600
800
1000
1200
1400
0 20 40 60 80 100
Spo
re c
ount
mL-
1
Time (h)
Accumulated spore release
n=3±SD
12 spores/ml/h
24 h dehydration show positive effect
No agitation- ~ 195 spores/ml/g/h
Higher release rate compared to 4 hours pre-treatment.
2. Release Experiment
Pre-treatment(4 h dehydration)
Spore release at 10 °C for 98 hours with no agitation
Same amount of tissue (n=3)
2 hours
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DTU Aqua, Technical University of Denmark
0
200
400
600
800
1000
1200
1400
1600
1800
1h/0 Lmin 3h/0 Lmin 1h/2.5 Lmin 3h/2.5 Lmin
Num
ber
of r
elea
sed
spor
es
The effect of Time and Agitation of spore release yield
0
200
400
600
800
1000
1200
1400
0 20 40 60 80 100
Spo
re c
ount
mL-
1
Time (h)
Accumulated spore release
n=3±SD
12 spores/ml/h
24 h dehydration + agitation show positive effect
No agitation- ~ 195 spores/ml/g/h
Higher release rate compared to 4 hours pre-treatment.
With agitation- Higher concentration- Higher release rate
~ 202 spores/ml/g/h
2. Release Experiment
Pre-treatment(4 h dehydration)
Spore release at 10 °C for 98 hours with no agitation
Same amount of tissue (n=3)
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DTU Aqua, Technical University of Denmark
3. Applying a fertilization step(SporeUse1)
12 h drying
15 mL
Air
+: addition of 10 mL- : addition of 10 mL water
x 6
500 mL
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DTU Aqua, Technical University of Denmark
3. Result: Applying fertilization step
(n=6) ±SE-2
0
2
4
6
8
10
12
14
Day 11 Day 18 Day 29 Day 40Cou
nt (
# p
er 1
0 cm
rop
e)
Number of seedlings
+-
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DTU Aqua, Technical University of Denmark
4. Spore settlement(SporeUse2)
• Preference study• Assess effect of alginate coating and agitation
- Clean rope- Coated rope• Spores in excess• N=3
Day-1
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DTU Aqua, Technical University of Denmark
• Preference study• Assess effect of alginate coating and agitation
- Clean rope- Coated rope• Spores in excess• N=3
Day-1 Day20 Day41 Day70
4. Spore settlement(SporeUse2)
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DTU Aqua, Technical University of Denmark
4. Results: Spore settlementAlginate coating + agitation
0
20
40
60
80
100
120
140
160
Sse
edlin
g co
unt
(per
10
cmro
pe)
Number of seedlings dependent on agitation
0 Lmin + Alg
0.5Lmin + Alg
2.5Lmin +Alg
0 Lmin
0.5Lmin
2.5Lmin
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DTU Aqua, Technical University of Denmark
4. Spore settlement(SporeUse3)Can we store, dispurse and re-settle spores to substrate?
30 daysLow light5 °C
1 minute
15 mL
Air agitation0.5 Lmin-1
2.5 Lmin-1
Ropes5 mm Danline (PV)2 mm Kuralon
+
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DTU Aqua, Technical University of Denmark
4. Results: Spore settlement(SporeUse3)
Danline
Kuralon
Settlement: 3 daysAttachment 14 days
0
10
20
30
40
50
60
70
80
Day3 Day14
Spo
re c
ount
(pe
r 10
cm
rop
e)
Settlement and attachment of Tetraspores
Kuralon (2.5Lmin)
Danline(2.5Lmin)
Danline (0.5Lmin)
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DTU Aqua, Technical University of Denmark
4. Results: Spore settlement(SporeUse3)
0
20
40
60
80
100
120
Day3 Day14
Spo
re c
ount
(pe
r 10
cm
rop
e)
Settlement and attachment of Tetraspores
Kuralon (2.5Lmin)
Danline(2.5Lmin)
Danline (0.5Lmin)Danline
Kuralon
Settlement: 3 daysAttachment 14 days
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DTU Aqua, Technical University of Denmark
4. Results: Spore settlement(SporeUse3)
Danline
Kuralon
Settlement: 3 daysAttachment 14 days
0
50
100
150
200
250
Day 3 Day14
See
dlin
g co
unt
(per
10
cm r
ope)
Settlement and attachment of seedlings
Kuralon (2.5Lmin)
Danline(2.5Lmin)
Danline (0.5Lmin)
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Findings and suggestions for improved P. palmata hatchery
• Time duration and agitation increase spore release and agitation resulted in higher seedling density
• Blending the spore release provide dispersal of spores and seedlingsKuralon rope show higher density compared to Danline rope
• Fertilization step; adding male gamets increase numbers of seedlings• No clear effect of Alginate pretreatment
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Thanks for listening!
facebook.com/skaldyrcenterwww.skaldyrcenter.dk/www.skaldyrcenter.aqua.dtu.dk/
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DTU Aqua, Technical University of Denmark
Fertility (Jan-April)
Spore release
Germination1:1
Photo: Constanza C.
Seedlings
Biomass
1 season0.3-0.9 kg/m
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DTU Aqua, Technical University of Denmark