Detection of Phaeocystis globosa using sandwich hybridization integrated with nuclease protectio
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J ou rn al of En viro nm ental S cienc es 2 0(2 00 8) 1 48 1– 14 86
Detection of Phaeocystis globosa using sandwich hybridization integrated with nuclease protection assay (NPA-SH)
Байду номын сангаас
Introduction
Phae ocy stis globosa S che rff el is a ge nus of the fa mily P rym nesiophyta ( or Ha ptophyta ) with e urytherm al a nd e uryha line c har acte ristic s ( Pe ng et al., 2005) . The c ell sha pe of P. globosa is sphe rica l, a nd the y often f orm c olonie s, w hic h a re covere d with soft colloid outside. The re is abunda nt polysa cc haride in the algae . W hen the m icr oalgae m ultiply m assive ly and form the har mf ul algae bloom ( HAB ), they will ca use serious problem s to the a quatic or ganisms a nd the envir onme nt. F irst, the algae c ontaining colloid and polysac cha ride stick to the fi sh gills a nd aff e ct the brea th and fee ding of fi sh, w hic h lea ds to the de ath of fi sh be ca use of hypoxia or su ff oc ation. Se cond, the e norm ous m icroa lgae bioma ss proba bly lea ds to dissolve d oxyge n defi cit in the wa ter body. Thir d, P. globosa has be en c onfi rm e d to produc e som e toxin, such as hem olytic substanc es, which ar e a mixture of liposa cc haride s, to da m age fi sh gills (Qi e t al ., 2002). Ther efor e, the HAB of P. globosa c an da ma ge the wate r e nvironm ent a nd e ve n lead to fi sh de ath in la rge a re as in se rious condition. This dam a ge is mor e ser ious to ca ge c ulture and pra wn br ee ding. In China, P. globosa is m a inly distr ibuted in the c oastal water s of Fujian and Gua ngdong Pr ovinc e. F rom O ctober in 1997 to Fe brua ry in 1998, the HA B of P. globosa br oke out for the first tim e in the coa sta l wate rs
A bstra ct P ha eocys tis glob os a Sch er ffel is on e of th e co mmon harmfu l alg ae s pecies in coas tal w aters of th e s ou th eas tern Ch in a. In this s tu dy, s and wich h yb rid izatio n integ rated with nu cleas e pro tection ass ay (N PA-SH ) was us ed to q ualitativ ely an d q uan titatively d etect P. g lo bo sa . Resu lts sh ow ed th at this meth od had g oo d app licability and v alidity in an alyzing th e s amples fro m labo rato ry cultu res an d from fields . Th e linear regres sion eq uatio n fo r P. glob os a was ob tained , and the low est d etectio n n umber o f cells was 1.8 × 1 04 cells . Statis tics s ho wed that there w as no d is tinct di fference b etw een th e res ults of detecting the microalg ae by N PA-SH and tradition al micro sco py. T his techn iq ue has go od reliab ility, accuracy, an d can g ive a remark ably high samp le pro cess in g rate. Sand wich hy bridization in tegrated w ith nu cleas e p rotection as say w ill prov id e an e ffi cien t alternative to microsco pic metho d for mon itoring an d inv estigatin g th e b lo om o f P. glob os a. Key wo rds: harmfu l algae bloo ms; Ph aeo cystis glob os a Sch er ffel; sand wich hy bridizatio n in tegrated with nu cleas e pro tectio n as say (NPA-SH )
Z HE N Yu 1 ,2 , M I Tiezhu 1 ,2 , YU Zhigang 2,∗
1. Key Laborat ory of M arine E nv ironm ent and E c ology, Mi nistry of E ducat ion, O ce an Uni ve rsity of China, Q ingdao 266100, Chi na. E -mai l: zheny u@ouc.e du.c n 2. Key L aboratory of Mar ine C hem istry The ory and Te chnology, M inist ry of Educ ation, O c ean U niv ersit y of C hina, Q ingdao 266100, China Re c eived 4 J anua ry 2008; revi sed 19 F ebruary 2008; a cc epte d 12 M arc h 2008
Detection of Phaeocystis globosa using sandwich hybridization integrated with nuclease protection assay (NPA-SH)
Байду номын сангаас
Introduction
Phae ocy stis globosa S che rff el is a ge nus of the fa mily P rym nesiophyta ( or Ha ptophyta ) with e urytherm al a nd e uryha line c har acte ristic s ( Pe ng et al., 2005) . The c ell sha pe of P. globosa is sphe rica l, a nd the y often f orm c olonie s, w hic h a re covere d with soft colloid outside. The re is abunda nt polysa cc haride in the algae . W hen the m icr oalgae m ultiply m assive ly and form the har mf ul algae bloom ( HAB ), they will ca use serious problem s to the a quatic or ganisms a nd the envir onme nt. F irst, the algae c ontaining colloid and polysac cha ride stick to the fi sh gills a nd aff e ct the brea th and fee ding of fi sh, w hic h lea ds to the de ath of fi sh be ca use of hypoxia or su ff oc ation. Se cond, the e norm ous m icroa lgae bioma ss proba bly lea ds to dissolve d oxyge n defi cit in the wa ter body. Thir d, P. globosa has be en c onfi rm e d to produc e som e toxin, such as hem olytic substanc es, which ar e a mixture of liposa cc haride s, to da m age fi sh gills (Qi e t al ., 2002). Ther efor e, the HAB of P. globosa c an da ma ge the wate r e nvironm ent a nd e ve n lead to fi sh de ath in la rge a re as in se rious condition. This dam a ge is mor e ser ious to ca ge c ulture and pra wn br ee ding. In China, P. globosa is m a inly distr ibuted in the c oastal water s of Fujian and Gua ngdong Pr ovinc e. F rom O ctober in 1997 to Fe brua ry in 1998, the HA B of P. globosa br oke out for the first tim e in the coa sta l wate rs
A bstra ct P ha eocys tis glob os a Sch er ffel is on e of th e co mmon harmfu l alg ae s pecies in coas tal w aters of th e s ou th eas tern Ch in a. In this s tu dy, s and wich h yb rid izatio n integ rated with nu cleas e pro tection ass ay (N PA-SH ) was us ed to q ualitativ ely an d q uan titatively d etect P. g lo bo sa . Resu lts sh ow ed th at this meth od had g oo d app licability and v alidity in an alyzing th e s amples fro m labo rato ry cultu res an d from fields . Th e linear regres sion eq uatio n fo r P. glob os a was ob tained , and the low est d etectio n n umber o f cells was 1.8 × 1 04 cells . Statis tics s ho wed that there w as no d is tinct di fference b etw een th e res ults of detecting the microalg ae by N PA-SH and tradition al micro sco py. T his techn iq ue has go od reliab ility, accuracy, an d can g ive a remark ably high samp le pro cess in g rate. Sand wich hy bridization in tegrated w ith nu cleas e p rotection as say w ill prov id e an e ffi cien t alternative to microsco pic metho d for mon itoring an d inv estigatin g th e b lo om o f P. glob os a. Key wo rds: harmfu l algae bloo ms; Ph aeo cystis glob os a Sch er ffel; sand wich hy bridizatio n in tegrated with nu cleas e pro tectio n as say (NPA-SH )
Z HE N Yu 1 ,2 , M I Tiezhu 1 ,2 , YU Zhigang 2,∗
1. Key Laborat ory of M arine E nv ironm ent and E c ology, Mi nistry of E ducat ion, O ce an Uni ve rsity of China, Q ingdao 266100, Chi na. E -mai l: zheny u@ouc.e du.c n 2. Key L aboratory of Mar ine C hem istry The ory and Te chnology, M inist ry of Educ ation, O c ean U niv ersit y of C hina, Q ingdao 266100, China Re c eived 4 J anua ry 2008; revi sed 19 F ebruary 2008; a cc epte d 12 M arc h 2008
