三角褐指藻藻液细菌的分离鉴定及其对藻细胞生长的影响
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摘要
为了解藻液分离细菌的种类及其对微藻生长的影响,本研究采用梯度稀释涂布法从三角褐指藻培养液分离得到3株细菌,基于16S rDNA基因序列分析,鉴定3株细菌P1、P2和P3分别为Loktanella vestfoldensis,Erythrobacter citreus和Aquamicrobium sp。将3株分离菌分别以10~4、10~6和10~8cfu/mL3个浓度梯度加入三角褐指藻培养液中比较其对微藻生长的影响。结果表明,P1和P3菌株在中浓度促进微藻生长,但高浓度却存在不同程度的抑制效应,而P2菌株在本实验浓度范围内均表现出促进作用,且促进作用随浓度升高而加强。这说明P2菌株可作为促进三角褐指藻生长的候选菌株构建有利微藻生长的藻菌共培养体系。
In order to understand the species of bacteria in the phycosphere and their effects on the growth of microalgae, three bacteriastrains were isolated from phaeodactylum tricornutum by means of gradient dilution coating. Based on 16 S rDNA gene sequence analysis, three strains belong to Loktanella vestfoldensis, Erythrobacter citreus and Aquamicrobiumsp. Besides, we studied the effect of different concentrations(10~4, 10~6 and 10~8 cfu/mL) of bacteria on the growth of phaeodactylum tricornutum by co-culture. The results showed that: P1 and P3 strains showed promoting effect at medium concentration and inhibiting effect at high concentration. While P2 strain still showed promoting effect in the experimental range of concentration, and the promoting effect was enhanced with increasing concentration. This shows that, P2 strain can be used as a candidate strain to promote the growth of phaeodactylum tricornutum, and will help to build a co-culture system of bacteria and microalgae for promoting the growth of microalgae.
In order to understand the species of bacteria in the phycosphere and their effects on the growth of microalgae, three bacteriastrains were isolated from phaeodactylum tricornutum by means of gradient dilution coating. Based on 16 S rDNA gene sequence analysis, three strains belong to Loktanella vestfoldensis, Erythrobacter citreus and Aquamicrobiumsp. Besides, we studied the effect of different concentrations(10~4, 10~6 and 10~8 cfu/mL) of bacteria on the growth of phaeodactylum tricornutum by co-culture. The results showed that: P1 and P3 strains showed promoting effect at medium concentration and inhibiting effect at high concentration. While P2 strain still showed promoting effect in the experimental range of concentration, and the promoting effect was enhanced with increasing concentration. This shows that, P2 strain can be used as a candidate strain to promote the growth of phaeodactylum tricornutum, and will help to build a co-culture system of bacteria and microalgae for promoting the growth of microalgae.
引文
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[2] 高亚辉,荆红梅,黄德强. 海洋微藻胞外产物研究进展[J]. 海洋科学,2002,26(3):35-38.
[3] Ding Y, Miao J, Wang Q, Zheng Z, Li G, Jian J, Wu Z. Purification and characterization of a psychrophilic glutathione reductase from Antarcticice microalgae Chlamydomonas sp. Strain ICE-L[J]. Polar Biol, 2007,31(1): 23-30.
[4] Grossart H P, Levold F, Allgaier M,et al. Marine diatom species harbour distinct bacterial communities[J]. Environ Microbiol, 2005,7(6):860-873.
[5] Grossart H P, Czub G, Simon M. Specific interactions of planktonic algae and bacteria:consequences for aggregation processes and organic matter cycling in the sea[J].EnvironMicrobiol, 2006, 8(6):1074-1084.
[6] 王少沛,曹煜成,李卓佳,等.水生环境中细菌与微藻的相互关系及其实际应用[J].南方水产, 2008(1): 76-80.
[7] Govindaraju K,Kiruthiga V,Kumar V G, et al. Extracellular synthesis of silvernanoparticles by a marine alga, Sargassum wightii Grevilli and their antibacterial effects[J]. JNanosci Nanotechnol, 2009, 9(9): 5497-501.
[8] Wang X,Li Z,Su J, et al. Lysis of a red-tide causing alga, Alexandrium tamarense, caused bybacteria from its phycosphere[J]. Biol. Control, 2010,52(2): 123-130.
[9] Jiang H M, Gao K S. Effects of lowering temperature during culture on theproductionof polyunsaturated fatty acids in the marine diatom Phaeodactylum tricornutum[J].Journal of Phycology, 2004, 40: 651-654.
[10] Fajardo A R, Cerdan L E, Medina A R, et al. Lipid extraction from the microalgaPhaeodactylum tricornutum[J]. Eur J Lipid Sci, 2007, 109(2): 120-126.
[11] 侯和胜,任晓咏,佟少明. 低温胁迫对三角褐指藻生长和生理生化特性的影响[J]. 辽宁师范大学学报(自然科学版),2011,(01):89-92.
[12] 符茹,黄长干,王海英. 营养条件对三角褐指藻生长和油脂积累的影响[J]. 湖北农业科学,2011,(11):2292-2294+2304.
[13] Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S. MEGA5: molecular evolutionary genetics analysisusing maximum likelihood, evolutionary distance, andmaximum parsimony methods[J]. Mol Biol Evol, 2011,28:2731–2739
[14] 刘建康. 高级水生物学[M].北京:科学出版社,1999:63-72.
[15] 乔洪金,刘相全,马晶晶,王际英,张利民. 5 株海洋微藻伴生细菌的分离鉴定与功能分析[J]. 安徽农业科学,2012(29):14421-14424.
[16] Schafer H,Abbas B,Witte H.Genetic diversity of ‘satellite’ bacteria present in cultures of marine diatoms[J].FEMS Microbiol Ecol,2002,42: 25-35
[17] 黄志涛,宋协法,李勋,万荣,董登攀,石蓉蓉,翟介明. 基于高通量测序的石斑鱼循环水养殖生物滤池微生物群落分析[J]. 农业工程学报,2016,S1:242-247.
[18] Gonzalez J M,Kiene R P,Moran M A.Transformation of sulfurcompounds by an abundant lineage of marine bacteria in the alpha-subclass of the class Proteobacteria[J].Appl Environ Microbiol,1999,65:3810-3819.
[19] 李筠,周宏霞,刘佳琳,张跃峰,陈吉祥,张晓华. 青岛近岸特征环境中海洋异养细菌的分布规律及其分子鉴定[J]. 中国海洋大学学报(自然科学版),2006,06:965-970.
[20] Lechner U, Baumbach R, Becker D, Kitunen V, Auling G,Salkinoja-Salonen M. Degradation of 4-chloro-2-methylphenol by an activated sludge isolate and its taxonomic description[J]. Biodegradation,1995, 6:83-92
[21] Hyun MJ,Kim JM,Jeon CO. Aquamicrobium aestuariisp. nov., a marine bacterium isolated from a tidal flat[J]. Int JSyst Evol Microbiol,2013,63:4012–4017
[22] Haaber J,Middelboe M.Viral lysis of Phaeocystis pouchetii:implications for algal population dynamics and heterotrophic C, Nand P cycling[J]. The ISME Journal, 2009,3(4): 430-441.
[23] Rodrigues R, Williams P. Inorganic nitrogen assimilation bypicoplankton and whole plankton in a coastal ecosystem[J]. LimnolOceanogr, 2002, 47(6): 1608-1616
[24] 徐金森,郑天凌,郭清华,等.两种海洋细菌对赤潮藻的细胞生物量的影响研究[J]. 海洋科学,2002,26(12) : 57-60.