Study on the Ecolog ica l Rela tion sh ip Between a Red- tide Causa tive A lga and Three Stra in s of Bacter ia Isola ted from X iam en Harbor

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1 V o l. 22,N o. 12 A CTA ECOLO G ICA S IN ICA D ec., , 2, 1, 1, 1, 1 2, 3, (1. ; 2. ; 3., ) : [A lex and rium tam arense (L ebour) Balech, 1985 ], A. tam arense, 3 S5, S7, S10 A. tam arense :, A. tam arense, ;, 3 A. tam arense S10> S5> S7 : ; ; Study on the Ecolog ica l Rela tion sh ip Between a Red- tide Causa tive A lga and Three Stra in s of Bacter ia Isola ted from X iam en Harbor ZH EN G T ian2l ing 1, 2, T IAN Yun 1, SU J ian2q iang 1, L IAN Yu2W u 1, W AN G Yan2L i 1, HON G H ua2sheng 2, 3 (1. S chool of L if e S ciences, X iam en U niversity; 2. K ey L ab. : ( ) ; (D ) : ; : : (1955 ),,,, E2m ail: w shw zh@ jingxian. xm u. edu. cn f or M arine E nv ironm ental S ciencn of M inistry of E ducation; 3. E nv ironm ental S ciencn R esearch Centre, X iam en U niversity, X iam en , Ch ina). A cta Ecolog ica S in ica, 2002, 22 (12) : Abstract: T he physio logical and eco logical p roperties of A lex and rium tam arense, a k ind of red2tide causative alga, and the eco logical relationsh ip betw een th is alga and th ree strains of m arine bacteria iso lat2 ed from X iam en w estern sea area w ere exam ined in the contro lled eco logical conditionṡ A. grow th w ell and keep som e of its peculiarity w hen cultured under the labo rato ry conditionṡ tam arense can Its grow th curve is as sam e as that of the general m icroo rganism. Bacteria are no t found in the purely cultured A. tam arense cell. T h ree strains of Bacteria are iso lated from the sedim ents of X iam en H arbo ṙ A fter the mo r2 pho logical exam ination and the mo lecule2bio logical study, it can be identified that S5 p robably is a new specie of S p h ing om onas, S7 is B acillus m eg aterium and S10 is B acillus halm ap alus. T he relationsh ip be2 tw een th ree strains of bacteria S5, S7, S10 and the alga A. tam arense is studied by adding the bacteria s fil2 trate into the alga culture separately in the lag phase and exponential phase of the alga grow th. strains show different eco logical effecṫ alga s grow th and m ultip lication at the concentration of cellsgm l. T he th ree T he filtrate of bacteria S5 and S10 can also m arkedly inh ibit the S10 has the stronger inh ibiting a2 bility than that of S5. W h ile m iddle concentration ( cellsgml ) filtrate of S5 and S10 can m arkedly im 2 p rove the grow th and m ultip lication of the alga. S10 has the stronger p romo tion than that of S5 to the alga s grow th. T he inh ibition and p romo tion are bo th mo re rem arkable in the alga s lag phase than exponential phase. T he filtrate of S5 and S10 has no evident effect on the alga s grow th at the concentration of

2 cellsgm l. Bacterium S7 has no clear inh ibition o r p romo tion to the alga. bition and the po ssibility of red2tide contro l by bio logical m ethods w ere discussed. Key words: A lex and rium tam arense; m arine bacteria; eco logical relation sh ip : (2002) : Q 143 : A T he m echanism s of bacterial inh i2,,,,,,, 5000, ( ) 300, 40,,,,,,, ;, [1, ] A. tam arense, PSP ( ),,, A. tam arense PSP [2 1986, A. tam arense ] [3 ] [4 ; ],,, (A lex and rium tam arense), fg2, 22 1, 12L 12D, 3500lx cm 3 (S5 S7 S10) 113, 0. 1m l ( ), I2N a I 1m in 114 DA P I (Zeiss M icro scope, 100W HBO m ercury ligh t source; G365, FT 395, L P420), 10 ( ) : (cellgm l) = A S 1g(S 2 V ),A 10, S 1, S 2, V 115 (0122Λm ), 6m ldm F (N,

3 12 : 2065 N 2 ) (A R ) ( 5 ) 015h,, 850 ( E x = 430nm, Em = 670nm ) : (a), 015h, 1 fg2 (b) F a Table 1 The ingredien ts of improved fg2 culture medium for algae (c) 1 10%, 30s Compounds Amount ( ) F b, 1000m l ( ) Sto re seaw ater Ch l2a : 1. M ajo r nutrient ZnSO 4 7H 2O m g Ch l2a = FD R g(r 1) (F b F a) V 1gV 2 N ano 3 75m g CoC l2 6H 2O 0. 01m g, FD, FD = (Λg gm l); N ah 2PO 4 H 2O 5m g M nc l2 4H 2O 0. 18m g R, R = 5; V 1 (m l); V 2 N am oo 4 2H 2O m g (m l); F 2. a, F T race elem ent b 3. V itam in N a2ed TA 2H 2O 4. 36m g B m g FeC l2 6H 2O 3. 15m g B Λg 116 CuSO 4 5H 2O 0. 01m g B io tin 0. 1Λg DNA 16S rdna PCR, (10mmo lgl T rishc l, 10mmo lgl N ac l),, 1Λl 500Λl, ΛL, 16S rdna PCR, : 16S+ 5 A GA CT T T GA TCCT GGCTCA G3, 1486R 5 CGGCTA CCT T GT TA CGA CT TC3, 16S rdna 5 3 PCR 94 2 m in, 94 1 m in, 55 1m in, 72 2 m in, 30, 1%, S rdna PCR Q ia Q uick PCR Purification K it, 377 DNA (AB I PR ISM ), 16S+, 1486R BLA ST, Pc2 gene610,,, Phylip315 DNA dist, N eighbo r ,, 1 10cm 2216E 2d,, 14, 3 S5, S7, S10, S5 (S p h ing om onas),, S7 B acillus m eg aterium ( 2 ), B acillus m eg aterium ; S10, B acillus halm ap alus 16S rdna 2% 212 A. tam arense 20, 100 cellgm l,, 8d, d,,,, 10d,,,, A. tam arense, 213 A. tam arense

4 A. tam arense A. tam arense,, 1 6 : cellgm l, , m l 10m l, S cellg m l, S cellgm l 1, S5,,, , A. tam arense S ,, S S5 2 S7 F ig. 1 T he effect on alga cell density (cellgm l) w hen F ig. 2 T he effect on alga cell density (cellgm l) w hen the filtrate of bacteria S5 is added to A. tam arense cul2 the filtrate of bacteria S7 is added to A. tam arense cul2 ture during the lag phase ture during the lag phase 1. Tem arense contro l; 2. Tem arense+ S5; 2. Tem arense 1. Tem arense contro l; 2. Tem arense+ S7; 3. Tem arense + S ; 4. T em arense+ S S ; 4. T em arense+ S , S7 S5, S7 A. tam arense, S S7 10-2, S7 A. tam arense 3, S10 S5 S10 S5 3 S10 4 S5 F ig. 3 T he effect on alga cell density (cellgm l) w hen F ig. 4 T he effect on alga ch lo rophyll content ( 10-3 the filtrate of bacteria S10 is added to A. tam arense cul2 Λggm l) w hen the filtrate of bacteria S5 is added to ture during the lag phase A. tam arense culture during the lag phase

5 12 : 2067 S10,, S S S S5, S10 A. tam arense, A. tam arense Ch l2a 4 6, A + S5 A + S10 Ch l2a, S10,, S5 S10, S5 S10, S S Ch l2a S S Ch l2a, S10 5 S7 F ig. 5 T he effect on alga ch lo rophyll content ( S10 Λggm l) w hen the filtrate of bacteria S7 is added to F ig. 6 T he effect on alga ch lo rophyll content ( 10-3 A. tam arenseculture during the lag phase Λggm l) w hen the filtrate of bacteria S10 is added to A. tam arenseculture during the lag phase 5, S7, S7 Ch l2a, Ch l2a,,, A. tam arense, 8,, S5 8 S7 F ig. 7 T he effect on alga cell density (cellgm l) w hen the filtrate of bacteria S5 is added to A. tam arense cul2 ture during the exponential phase F ig. 8 T he effect on alga cell density (cellgm l) w hen the filtrate of bacteria S7 is added to A. tam arense cul2 ture during the exponential phase S7, 8, S7,

6 ,,,, S7,, S7 A. tam arense S10 S5 9, S10, S10, S10, S10, A. tam arense 10 12,, 9 S10 10 S5 F ig. 9 T he effect on alga cell density (cellgm l) w hen F ig. 10 T he effect on alga ch lo rophyll content ( 10-3 the filtrate of bacteria S10 is added to A. tam arense cul2 Λggm l) w hen the filtrate of bacteria S5 is added to A. ture during the exponential phase tam arense culture during the lag phase 11 S7 12 S10 F ig. 11 T he effect on alga ch lo rophyll content ( 10-3 Λggm l) w hen the filtrate of bacteria S7 is added to A. tam arense culture during the lag phase F ig. 12 T he effect on alga ch lo rophyll content ( 10-3 Λggm l)w hen the filtrate of bacteria S10 is added to A. tam arense culture during the lag phase Fukam i K, F lavobacterium sp. G. m ik im otoi, Chattonella, H eterosig ina akash iw o S keletonem a costatum Gym od inium nag asak iense [5 ] 3 A. tam arense,, S5 S10 A. tam arense S10 S7 A. tam arense

7 12 : 2069, , P seud om onas [6 ], B acillus [7 ], [5 B d ellov ibrio, F lavobacterium sp. S ap rosp ira sp. ], S10 B acillus halm ap alus P seud om onas aerug inosa, [8 ], 12 [9 P seud om onas stu tz eri ], Chattonella antiqua Im ai I H iro sh im a 4 A lterom onas C. antiqua [10 ] Ish io 1993 V ibrio alg oinf estus, (D G I), C. antiqua, P. stu tz eri D G I, (4 3 [11 ),, ],,,, [12, 13 ], A. tam arense,,,, [14, ],,,,, 312,,, A. tam arense,,,, :,,,, A. tam arense,,,a. tam arense,, 28,A. tam arense 1 2d ,,,, ,, ,, ;,

8 , [ 1 ] Zheng T L ( ), Zhung T C ( ), CaiL Z ( ). T he ro le of m icrobes in bio rem ediation of m arine po l2 luted environm enṫ J ou rnal of X iam en U niversity (in Ch inese) ( ), 2001, 40 (2) : [ 2 ] Su H M. A toxic dinoflagellate first reco rded in T aiw an. P roc. 1st lnṫ Symp. on Red T ides, N YgE lsevire, [ 3 ] Q i Y Z ( ), Q ian F ( ). Study on the taxonom y of several species of dinoflagellate in D aya Bay. O ceanolo2 g ia E T L im nolog ia S inica (in Ch inese) ( ), 1994, 25 (2) : [ 4 ] L in Y S ( ). Toxic dinoflagellate A lex and rium tam arense a causative red2tide algae in sh rimp s pond in X i2 am en area. J ou rnal of O ceanog rap hy in T aiw an S trait (in Ch inese), 1996, 15 (1) : [ 5 ] Fukam i K. Iso lation and p roperties of a bacterium inh ibiting the grow th of Gym nodinium nagasak iense. N ipp on S u isan Gakkaish i, 1992, 58 (6) : [ 6 ] D akham a A. Iso lation and identification of antialgal substances p roduced by P seudomonas aerugino sa. J. A pp l. P hy col., 1993, (5) : [ 7 ] Reim R L. T he characterization of a Bacillus capable of blue2green bactericidal activity. Can. J. M icrobiol., 1974, 20: [ 8 ] M iddelboem, Sondergard M, L etarte Y, et al. A ttached and free2living bacteria: P roducion and po lym er hydro ly2 sis during a diatom bloom. M icrobial E cology, 1995, 29: [ 9 ] D akham a A. Stim ulato ry and inh ibito ry effects of P seudomonas on the grow th of algae. Can. T ech. R ep. F ish. A quaṫ S ci., 1989, 1714: [ 10 ] Im ai I, Ish ia Y, Sakaguch i K, et al. A lgicidal m arine bacteria iso lated from no rthern H iro sh im a Bay. J ap an. F isheries S cience, 1995, 61 (4) : [ 11 ] Sh insaku H ayash ida, et al. Iso lation of anti2algal P seudomonas stutzeri strains and their lethal activity fo r Chat2 tonella antiqua A gric. B iol. Chem., 1991, 55 (3) : [ 12 ] L in F D ( ), Zhang C ( ), Zou J Z ( ). Ro le of bacteria in the gow th p rocess of phytop lank ton ( ). M arine S cience(in Ch inese) ( ), 1996, (6) : [ 13 ] L ian Y W ( ),W ang Y L ( ), Zheng T L ( ). A dvance in the research on interaction betw een red tide alage and bacteria ( ). M arine S cience ( in Ch inese) ( ), 1999, (1) : [ 14 ] Caio la M G & Peilegrini S. L ysis of M icrocystis aerugino sa by Bdellovibrio2like bacteria. J. P hcol., 1984, (20) :

The Effects of Physica l and Chem ica l Cond ition s on Form ing M ycelia l Pellet of P hanerochaete ch rysosp orium and B iosorption of L ead

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