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TRANSCRIPT
1Karenia mikimotoi
Karenia 13
Karenia
LAMP LAMP
LAMP
RNA NASBA
226 5 ictyocha
ictyocha
ictyochaichtyocha
LAMP NASBAKarenia LAMP
K. mikimotoi NASBA
271
2
1) LAMPictyocha
124
ictyocha26 5 ictyocha octonaria ictyocha
icicitus glo osus 9000 rpm, 5TE 5 Chele PBS DNA DNArDNA ITS1 LSU D1-D2 PCR
GenBank ictyocha . glo osus
26 5 . octonaria LAMP. octonaria Ooc1KSK Primer E plorer
er.3 FUJIITSU Solutions LAMPLAMP
Karenia digitataK. digitata Karlodinium
26 K. digitataGF/F
apata et al. (2000)HPLC
K. digitata LAMPK. digitata LAMP
27 K. digitataTE
5 Chele PBS DNA LAMP2) NASBA
K. mikimotoi NASBAK. mikimotoi 3.69 104 cells mL-1 1.5 mL 10,000 rpm 3 4
-80 RNeasyPlusE tragen 50 L
E tragenNASBA 5 L
5,540 554 55.4 0.55 0.05 0.005 0.0005 cellsK. mikimotoi NASBA
NASBA1.5 mL 65 5 41
NASBA 60 NASBA3 15
NASBA
125
NASBA RNA 65NASBA 65
NASBA 65 K. mikimotoi
E tragen
3 mK. mikimotoi 23 cells mL-1 1 mL
-80 300 L TEE tragen
1 1 2 1 2 2 33 3 10
L NASBA 11.5 cells1.15 cells 2
NASBANASBA RNA RNA
RNA DNANASBA RNA
DNARNA DNA
RNARNA
DNAK. mikimotoi
NASBA
K. mikimotoi 5.35 104 cells mL-1 2 mL 3,000 rpm 5-80 20 0, 1, 3, 6, 24, 48, 72
-80 E tragen 10 LNASBA
NASBAPCR
K. mikimotoiK. mikimotoi 1.32 104 cells mL-1 0.1 mL 1.5
mL -802015 10 26 3 m 10 L
126
Chaetoceros spp. keletonema spp.4 25
. octonaria NASBA. octonaria rRNA ITS rRNA LSU D1D2
NASBADNA PCR DNA
31 LAMP
ictyocha1 .
octonaria . i ula . speculum1 ITS1-5.8s-ITS2 ITS
rDNA LSU D1-D2 LSU ITS. i ula . octonaria . speculum
. glo osus NIWA1008 . i ula2
. i ula 2
spine2
. octonaria . speculum. octonaria . speculum GenBank
. octonaria NIWA1026
. octonaria. speculum. speculum
26 5 . octonaria LAMPLSU ITS ITS
6 LAMP . octonaria. speculum ITS
LAMP . octonariaNIWA1026 . speculum GenBank
. octonaria Ooc3mi KSKDNA 220 cells L-1 LAMP
64 62 63 3
127
0.022 cells L-1 2 cells 100L 4
23.5 22.968.1 62.2 30
70 .octonaria LAMP
. octonaria . speculum
K. digitataKarenia cf. digitata
ventral pore K digitata sulcus tube-shaped structure 5 K. digitata Keto-19 -butanoylo yfuco anthin-like, 4-keto-19 -butanoylo yfuco anthin, Keto-he anoylo yfuco anthin-like,
4-keto-19 -he anoylo yfuco anthin 6 apata et al. (2012)6 Karenia
Karlodinium K. digitata KarlodiniumK. digitata Karlodinium
K. digitata LAMPK. digitata
2 6 7 10 2023 K. digitata
LAMP K. digitata
LAMPK. digitata
2 NASBAK. mikimotoi NASBA
RNeasyPlus E tragen E tragen0.55 cells
7 E tragen 0.05 cells3 E tragen 1
cell E tragen NASBA
NASBANASBA 65 41 41
8 65
128
0.05 cells55.4 cells
65NASBA
E tragen
NASBA K. mikimotoi
923 cells K. mikimotoi
NASBANASBA
20 K. mikimotoi 72 NASBA10 PCR 24
NASBA
72 NASBA RNA
11 PCR 1NASBA 2 RNA PCR
2 DNA RNA
PCR NASBA2 DNA
NASBA 3K. mikimotoi NASBA
. octonaria NASBA7 DNA ITS rRNA LSU D1-D2PCR 12
RNA NASBA
HPLC
129
K. digitata
3apata M, Rodriuez F, Garrido JL. Separation of chlorophylls and carotenoids from marine
phytoplankton: a new HPLC method using a reversed phase C8 column and pyridine-containing mobile phases. Mar Ecol Prog Ser 195, 29-45, 2000.
apata M, Fraga S, Rodr guez F, Garrido JL. Pigment based chloroplast types in Dinoflagellates. Mar Ecol Prog Ser 465, 33 52, 2012.
1 ictyocha icicitus glo osus ictyocha glo osa
130
1. rDNA ITS1-5.8s-ITS2 ictyocha/ n 1000, 50
2 2015 Karenia digitata LAMP
131
0
10
20
30
40
50
60
70
2.2E+1 2.2E+0 2.2E 1 2.2E 2 2.2E 3
Reactio
ntim
e(m
in.)
Concentration (cells l 1)
w/o Loop w/ Loop
051015202530354045
60 61 62 63 64 65
Reactio
ntim
e(m
in.)
Temperature ( C)
w/o loop w/ loop
3. ictyocha octonariaLAMP
4. ictyocha octonariaLAMP
5. Karenia digitata Kdi01TKTA: apical groove tube-shaped stracture B:C: vesicle
2. ictyocha i ula (a, b) ictyocha octonaria (c, d) a: b: c: d: . i ula
4 spine
132
6 Karenia digitata Kalrodinium vene icum Karenia mikimotoi
A: K. digitata Kdi03TKT B: K. digitataKdi USK C: K. vene icum B1 D: K. mikimotoi
Km69-9 AX14: Chlorophyll C3, 8: Divinyl Protochlorophyllide a, 9: Chlorophyll C2,
13: Keto-19 -butanoylo yfuco anthin-like, 14: 4-keto-19 -butanoylo yfuco anthin, 16: 19 -butanoylo yfuco anthin, 19: Fuco anthin, 22: Keto-he anoylo yfuco anthin-like, 23: 4-keto-19 -he anoylo yfuco anthin, 26: 19 -he anoylo yfuco anthin, 28: Diadino anthin.
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7 NASBA Karenia mikimotoi
8 NASBA 65 Karenia mikimotoi
134
9 E tragen
1 2 2 22 3 3 3
1 11.5 cells2 1.2 cells
10 NASBA20 Karenia mikimotoi
PCRN
11 NASBA25 Karenia mikimotoi
PCR N
12 ictyocha octonaria NASBA
DNA PCR1: Oct D2 F1, R1, 2:
Oct D2 R1, R2, 3: Oct ITS1 F1, R1, 4: Oct ITS1 F1, Oct ITS2 R1, 5: Oct ITS2 F1, Oct ITS2 R1, 6: F41, R5, 7: F41, R61, 8: Negative control
135