(A) Oocyte, (B) Spermary

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(A) Oocyte, (B) Spermary 25 Years of PICES: Celebrating the Past, Imagining the Future November 2-13, 2016 Session 1: Multiple factors affecting the dynamics of shallow-water corals and other organisms < Jeju Island > < Korea map > Riman North-Korea < Jeju University > Tushima Yellow-Sea Kuroshio Contents 01 Introduction & Objectives 02 Material and method 03 Result 04 Summary 05 Discussion 01 Introduction & Objective Global warming -> SST(Surface Sea temperature) has been increased – 0.41 for the past 30yrs / 1.29 increase since 1968 in Korea Several℃ coral species have migrated℃ towards higher-latitude (Yamano et al. 2011) Increased subtropical species in marine ecosystem of Jeju Island < Seaweed based > < Coral based > In this study, annual reproductive cycle and reproductive strategy of two Screlactininan corals, Alveopora japonica and Oulastrea crispata, were analyzed 01 Introduction & Objective Alveopora japonica kingdom: Animallia phylum: Cnidaria class: Anthozoa order: Scleractinia family: Acroporidae genus: Alveopora <Alveopora japonica> <Community of coral> In shallow rocky foreshores, usually nested among algae and soft corals Hemispherical shape, small and less than 40 mm in diameter A hermaphroditic brooding coral with oocytes and spermaries developing on separate mesenteries of the polyp (Harii et al., 2001) 01 Introduction & Objective Oulastrea crispata kingdom: Animallia phylum: Cnidaria class: Anthozoa order: Scleractinia family: Acroporidae genus: Oulastrea <Oulstrea crispata> <Community of coral> In murky water and at low tide, colonies size about 10-15cm A hermaphroditic coral with oocytes and spermaries developing on same mesenteries of the polyp (Lam, 2000) A broadcast spawner and planula brooder (Lam, 2000) 02 Material and method • Sampling period : January ~ December in 2015 • Location : North (Geumneung) and South (Bomok), 5-10m depth • Target species : Alveoprora japonica & Oulastrea crispata • Sampling frequency : Over than 5 colonies for each specie Monthly 02 Material and method . Standard Histological work for coral research Sample collection Directly fixation with 10% formalin solution 10% HCl, 20% citric acid with 50% formic Decalcification acid solution Embedding paraffin blocks Tissue processor machine and then heated paraffin embedding machine Sectioning & Staining Tissue was Sectioned in 6~7µm and stained followed by H&E staining method Identifying organisms of corals and sexual Observation gonads, image analysis 02 Material and method . Image analysis by “Image J” 1 2 3 2 1 3 90° 1. Maximum diameter, (um) 2. Perpendicular diameter, (um) 3. Surface are (um2) 02 Material and method . Planulae observation Water tank Collecting larvae (with 200µm mesh) (200µm mesh) Light 2% glutaraldehyde 50, 70, 90, 95, 100% Ethanol for dehydration ZEISS Supra 55VP, Korea Basic Science Institute 30, 50, 70, 100% Isoamyl acetate for substitution Vacuum dry, Water storage coating <Flow-out aquarium> <SEM picture process> 03 Result (SST of sampling sites) North : 24.01 30 South : 25.02 ℃ 25 ℃ 20 15 North South 10 5 0 M A M J J A S O N D . SST(Surface Sea Temperature) : 13 ~ 25.02℃ 03 Result (A. japonica & O. crispata Oogenesis) Alveopora japonica Oulastrea crispata A B E F O O C D O G O H N N O O . (A) Stage 1(Empty stage) in mesoglea(Scale bar=50µm). (B) Stage 2, early development of oocyte(Scale bar=50µm). (C) Stage 3, developing oocyte(Scale bar=100µm). (D) Stage 4, mature oocyte(Scale bar=100µm). (E) Stage 1(Empty stage) in mesoglea in January(Scale bar=50µm). (F) Stage 2, early development of oocyte(Scale bar=50µm). (G) Stage 3, late development of oocyte(Scale bar=50µm). (H) Stage 4, Mature oocyte(Scale bar=100µm). N: nucleus, O: oocyte. 03 Result (A. japonica & O. crispata Spermagenesis) Alveopora japonica Oulastrea crispata A B E F mf mf C D G H S S . (A) Stage 1(Empty stage)(Scale bar=100µm) (B) Stage 2, Primordial spermary(Scale bar=50µm) (C) Stage 3, Developing spermary(Scale bar=50µm) (D) Stage 4, Mature spermary(Scale bar=50µm) . (E) Stage 1 in mesoglea(Scale bar=100µm) (F) Stage 2, Primordial spermary(Scale bar=100µm) (G) Stage 3, Developing speramry(Scale bar=50µm) (H) Stage 4, Mature spermary(Scale bar=200µm) mf: mesenterial filament, S: spermary 03 Result . Frequency of reproductive stages of A. japonica from North, (A) Oocyte, (B) Spermary Early Empty Late Ripe Spawning Spawning <A> <B> 100% 100% 90% 90% 80% 80% 70% 70% 60% 60% 50% 50% 40% 40% Frequency(%) 30% 30% 20% 20% 10% 10% 0% 0% Jan Feb Mar Apr May Jun Jul Aug Sep Oct Nov Dec Jan Feb Mar Apr May Jun Jul Aug Sep Oct Nov Dec 2015 2015 03 Result . Frequency of reproductive stages of A. japonica from South, (A) Oocyte, (B) Spermary Early Empty Late Ripe Spawning Spawning <A> <B> 100% 100% 90% 90% 80% 80% 70% 70% 60% 60% 50% 50% 40% 40% Frequency(%) 30% 30% 20% 20% 10% 10% 0% 0% Jan Feb Mar Apr May Jun Jul Aug Sep Oct Nov Dec Jan Feb Mar Apr May Jun Jul Aug Sep Oct Nov Dec 2015 2015 03 Result . Frequency of reproductive stages of O. crispata from South, (A) Oocyte, (B) Spermary Early Empty Late Ripe Spawning Spawning <A> <B> 100% 100% 90% 90% 80% 80% 70% 70% 60% 60% 50% 50% 40% 40% Frequency(%) 30% 30% 20% 20% 10% 10% 0% 0% Jan Feb Mar Apr May Jun Jul Aug Sep Oct Nov Dec Jan Feb Mar Apr May Jun Jul Aug Sep Oct Nov Dec 2015 2015 03 Result (North, GMD & SA of A. japonica ) 200 180 160 140 120 Spawning period 100 GMD(um) 80 60 40 20 0 30000 1 2 3 4 5 6 7 8 9 10 11 12 25000 20000 15000 10000 Surface (um^2)area 5000 0 1 2 3 4 5 6 7 8 9 10 11 12 03 Result (South, GMD & SA of A. japonica ) 450 400 350 300 250 200 Spawning period GMD (um) 150 100 50 0 160000 1 2 3 4 5 6 7 8 9 10 11 12 140000 120000 2) 100000 80000 60000 Surface (um^are 40000 20000 0 1 2 3 4 5 6 7 8 9 10 11 12 03 Result (South, GMD & SA of O. crispata ) 100 90 80 70 60 50 Spawning period GMD (um) 40 30 20 10 0 7000 1 2 3 4 5 6 7 8 9 10 11 12 6000 5000 4000 3000 Surface (um^2)are 2000 1000 0 1 2 3 4 5 6 7 8 9 10 11 12 03 Result . Planulae of A. japonica (South) <A> <B> • From July and Aug A. japonica samples 637 μm • Size : About 600~700µm for each <C> <D> • Ability to mobile (Swimming) • Symbiodiniums which 498.1 μm facilitate photosynthesis were symbiodinium found at endotherm <A>,<B>, Pictures by digital optical microscope, (Scale bar= 100µm, 200µm), <C>,<D> SEM picture of Planulae,(Scale bar= 20µm, 10µm) 04 Summary Slightly difference in maturation period between A. japonica from North and South <In the North> Oocytes: March ~ August Spermaries: May ~ July <In the South> Oocytes: April ~ August Spemaries: May ~ August, September Planulae of A. japonica were collected from July to August (at South site) The average size - 600µm, possessed motility <For O. crispata> Oocytes: May ~ September Spermaries : May ~ October 05 Discussion A. japonica and O. crispata are a hermaphroditic brooding coral with oocytes and spermaries developing on separate mesenteries of the polyps Seasonal patterns of gametogenesis were shown from those species Gonial mitosis occurring in late January The major spawning of those two species appeared to followed a period of rising the water temperature in September in Jeju Island More research on physiology and ecology of those species are required in various area Long-term environmental monitoring is also necessary to prepare for climate changes ありがとうございます。 谢谢您。 Merci. ًﺷﻛﺭﺍ Благодарю вас La ringrazio! .
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