坛紫菜贝壳丝状体白斑病附生细菌及藻际细菌群落特征分析

doi:10.11924/j.issn.1000-6850.casb2022-0562

摘要/Abstract

摘要：

坛紫菜是最具经济价值的海产经济作物之一，贝壳丝状体白斑病严重影响坛紫菜育苗产业的健康发展。探究坛紫菜白斑病贝壳丝状体附生细菌群落和藻际细菌群落可为阐明白斑病的致病微生态机制提供一定的依据。本研究基于高通量测序技术分析了健康和患病的贝壳丝状体附生细菌、藻际细菌群落结构及对贝壳丝状体的影响。结果表明：白斑病贝壳丝状体上附生细菌丰度和多样性降低、藻际细菌丰度和多样性增加，健康池水样和患白斑病池水样的细菌群落结构有显著差异。健康池水样和患白斑病池水样中变形菌门Proteobacteria和拟杆菌门Bacteroidota为优势菌，水源水中蓝细菌门Cyanobacteria和变形菌门为优势菌；Nautella、Vibrio在患白斑病的贝壳丝状体水体中为优势菌；在白斑病贝壳丝状体附生细菌中土芽孢杆菌属Geobacillus丰度为18.25%，而健康贝壳丝状体仅为0.05%，差异极显著。培育池水体中高丰度的弧菌和贝壳丝状体上高丰度的土芽孢杆菌，都是坛紫菜贝壳丝状体白斑病的重要病因，在坛紫菜育苗期间藻际细菌及贝壳丝状体附生细菌群落结构的稳定是坛紫菜育苗的关键因素。

关键词: 坛紫菜, 白斑病, 细菌群落, 高通量测序, 贝壳丝状体

Abstract:

Porphyrahaitanensis is one of the most valuable marine economic crops. The shell conchocelis white spot disease has seriously affected the healthy development of Porphyrahaitanensis seedling industry. The study on the epiphytic bacterial community of shell conchocelis and the bacterial community in the phycosphere of Porphyrahaitanensis white spot disease can provide a basis for elucidating the pathogenic microecological mechanism of white spot disease. Based on high-throughput sequencing technology, this study analyzed the community structure of healthy and diseased shell conchocelis epiphytic bacteria and phycosphere bacteria and their effects on shell conchocelis. The results showed that the abundance and diversity of epiphytic bacteria on the conchocelis of white spot shellfish decreased, and the abundance and diversity of bacteria in the phycosphere increased. There were significant differences in the bacterial community structure between the healthy pool water and the pool water with white spot disease. Proteobacteria and bacteroidota were the dominant bacteria in water samples from healthy pools and pools suffering from white spot disease, Cyanobacteria and Proteobacteria were the dominant bacteria in water source; Nautella and Vibrio were dominant bacteria in the shell conchocelis body water with white spot disease. Among the epiphytic bacteria of white spot shell conchocelis, the abundance of Geobacillus was 18.25%, while that of healthy shell conchocelis was only 0.05%, with a highly significant difference. The high abundance of Vibrio in the water body of the cultivation pond and the high abundance of Geobacillus on the shell conchocelis are both important causes of Porphyrahaitanensis shell conchocelis white spot disease. The stability of the community structure of phycosphere bacteria and shell conchocelis epiphytic bacteria is the key factor in the seedling raising of Porphyrahaitanensis.

Key words: Porphyrahaitanensis, white spot disease, bacterial communities, high-throughput sequencing, shell conchocelis

缪凡, 刘鑫, 林岗, 陈燕婷, 刘燕飞, 郑怡. 坛紫菜贝壳丝状体白斑病附生细菌及藻际细菌群落特征分析[J]. 中国农学通报, 2023, 39(5): 154-164.

MIAO Fan, LIU Xin, LIN Gang, CHEN Yanting, LIU Yanfei, ZHENG Yi. Characteristics Analysis of Phycosphere Bacteria and Algal Bacterial Communities of Shell Conchocelis of Porphyrahaitanensis with White Spot Disease[J]. Chinese Agricultural Science Bulletin, 2023, 39(5): 154-164.

图/表 15

参考文献 46

[1]	朱文嘉, 王联珠, 郭莹莹, 等. 我国紫菜产业现状及质量控制[J]. 食品安全质量检测学报, 2018, 9(13):3353-3358.
[2]	陈必链, 林跃鑫, 黄键. 坛紫菜的营养评价[J]. 中国海洋药物, 2001(2):51-53.
[3]	邓茜莹, 李缓缓, 曾荣急, 等. 紫菜发酵提取物的体外降血脂功能分析[J]. 食品与发酵工业, 2019, 45(12):109-113.
[4]	白露, 范晓丹, 张学武. 坛紫菜木瓜蛋白酶水解肽的抗肿瘤活性研究[J]. 食品工业科技, 2016, 37(9):352-356,362.
[5]	邱小明. 响应面优化法提取坛紫菜多酚及抗氧化研究[J]. 闽南师范大学学报(自然科学版), 2019, 32(2):81-89.
[6]	张全斌, 李大新, 于鹏展, 等. 坛紫菜多糖对脾细胞活性的影响[J]. 中国海洋药物, 2003(6):14-18.
[7]	GACHON C M M, SIME-NGANDO T, STRITTMATTER M, et al. Algal diseases: spotlight on a black box[J]. Trends in plant science, 2010, 15(11):633-640. doi: 10.1016/j.tplants.2010.08.005 pmid: 20833575
[8]	刘一萌, 马家海, 文茜. 福建坛紫菜赤腐病的病程及病原鉴定[J]. 福建农林大学学报(自然科学版), 2013, 42(1):18-22.
[9]	韩晓娟, 茅云翔, 李杰, 等. 一株引起坛紫菜绿斑病病原的分离鉴定及致病性研究[J]. 水产学报, 2015, 39(11):1721-1729.
[10]	GUAN X, LI J, ZHE Z, et al. Characterizing the microbial culprit of white spot disease of the conchocelis stage of Porphyrayezoensis (Bangiales,Rhodophyta)[J]. Journal of applied phycology, 2013, 25(5):1341-1348 doi: 10.1007/s10811-013-9976-8 URL
[11]	贲秀林. 条斑紫菜丝状体培育的病害及防治[J]. 水产养殖, 2007(1):43-44.
[12]	徐梦雅, 杨锐, 刘棋琴, 等. Vibrio mediterranei 117-T6引发的坛紫菜黄斑病的初步研究[J]. 水产学报, 2020, 44(4):661-671.
[13]	李金波. 条斑紫菜丝状体白斑病病原真菌的分离及鉴定[D]. 武汉: 华中农业大学, 2006.
[14]	MITCHELL B R. Chemotactic and growth responses of marine bacteria toalgal extracellular products[J]. The biological bulletin, 1972, 143:265-277. doi: 10.2307/1540052 URL
[15]	刘莹璐, 陈从立, 付亮, 等. 藻际环境中微生物胞间通讯行为及作用[J]. 微生物学报, 2022, 62(1):33-46.
[16]	孙墨可, 孙晓文, 田娟, 等. 综述高通量转录组测序技术在生物中的应用[J]. 南方农机, 2018, 49(19):16-17.
[17]	李晓霞. 坛紫菜养殖病烂的原因探析及其对策[J]. 中国水产, 2009(4):57-58.
[18]	沈梅丽. 温州坛紫菜养殖周期中的藻际微生物多样性[D]. 宁波: 宁波大学, 2013.
[19]	李芸, 刘发龙, 王巧晗, 等. 藻际微环境中藻体与微生物相互作用[J]. 河北渔业, 2019(3):43-48.
[20]	盖珊珊. 条斑紫菜丝状体黄斑病菌群特征及其与水体环境关系的研究[D]. 大连: 辽宁师范大学, 2017.
[21]	XIE X, HE Z, HU X, et al. Large-scale seaweed cultivation diverges water and sediment microbial communities in the coast of Nan'ao Island, South China Sea[J]. Science of the total environment, 2017, 15:598:97-108.
[22]	RAVINDRA PAL, SINGH, et al. Seaweed-microbial interactions: key functions of seaweed-associated bacteria[J]. Fems microbiology ecology, 2014, 88(2):213-230. doi: 10.1111/1574-6941.12297 pmid: 24512602
[23]	KLOCHKOVA T A, SHIM J B, HWANG M S, et al. Host-parasite interactions and host species susceptibility of the marine oomycete parasite, Olpidiopsis sp. from Koreathat infects red algae[J]. Journal of applied phycology, 2012, 24(1):135-144. doi: 10.1007/s10811-011-9661-8 URL
[24]	GOECKE F, LABES A, WIESE J, et al. Chemical interactions between marine macroalgae and bacteria[J]. Marine ecology progress, 2010, 409(6):267-300. doi: 10.3354/meps08607 URL
[25]	BLOUIN N A, BRODIE J A, GROSSMAN A C, et al. Porphyra: a marine crop shaped by stress[J]. Trends in plant science, 2011, 16(1):29-37. doi: 10.1016/j.tplants.2010.10.004 pmid: 21067966
[26]	张孟. 铜绿微囊藻对水体微生物群落结构与功能的影响[D]. 杭州: 浙江工业大学, 2019.
[27]	MARTINEZ J N, PADILLA P. Isolation and characterization of agar-digesting Vibrio species from the rotten thallus of Gracilariopsisheteroclada Zhang et Xia[J]. Marine environmental research, 2016, 119:156-160. doi: 10.1016/j.marenvres.2016.05.023 URL
[28]	FUJITA Y. Diseases of cultivated Porphyra in Japan[J]. in Introduction to Applied Phycology, ed. I. Akatsuka (The Hague: SPB Academic Publishing), 177-190.
[29]	MAHMUD Z H, NEOGI S B, KASSU A, et al. Seaweeds as a reservoir for diverse Vibrio parahaemolyticus populations in Japan[J]. International Journal of food microbiology, 2007, 118(1):92-96. doi: 10.1016/j.ijfoodmicro.2007.05.009 URL
[30]	HUDSON J, GARDINER M, DESHPANDE N, et al. Transcriptional response of Nautellaitalica R11 towards its macroalgal host uncovers new mechanisms of host-pathogen interaction[J]. Molecular ecology, 2017, 27(8):1820-1832. doi: 10.1111/mec.14448 URL
[31]	FERNANDES N., CASE R. J., LONGFORD S. R, et al. Genomes and virulence factors of novel bacterial pathogens causing bleaching disease in the marine red alga deliseapulchra[J]. PloS One, 2011, 6(12):e27387. doi: 10.1371/journal.pone.0027387 URL
[32]	梁珊珊. 坛紫菜丝状体和海带配子体的藻际细菌群落结构分析[D]. 福州: 福建师范大学, 2019.
[33]	WANG W, WU L, XU K, et al. The cultivation of Pyropiahaitanensis has important impacts on the seawater microbial community[J]. Journal of applied phycology, 2020, 32(4):2561-2573. doi: 10.1007/s10811-020-02068-6 URL
[34]	MIRANDA L N, HUTCHISON K, GROSSMAN A R, et al. Diversity and abundance of the bacterial community of the red macroalgaporphyraumbilicalis: Did bacterial farmers produce macroalgae?[J]. PloS One, 2013, 8(3):e58269. doi: 10.1371/journal.pone.0058269 URL
[35]	MATSUO, YOSHIHIDE, IMAGAWA, et al. Isolation of an algal morphogenesis inducer from a marine bacterium[J]. Science, 2005, 307(5715):1598. pmid: 15761147
[36]	LACHNIT T, M BLÜMEL, IMHOFF J F, et al. Speciﬁc epibacterial communities on macroalgae: Phylogeny matters more than habitat[J]. Aquatic biology, 2009, 5:181-186. doi: 10.3354/ab00149 URL
[37]	王璐璐. 土芽孢杆菌TS3-9胞外多糖的初步研究[D]. 杭州: 浙江工商大学, 2013.
[38]	KNOLL A H, BAULD J. The evolution of ecological tolerance in prokaryotes[J]. Transactions of the Royal Society of Edinburgh Earth Sciences, 1989, 80(3-4):209-223.
[39]	DR ZEIGLER. The Geobacillus paradox: why is a thermophilic bacterial genus so prevalent on a mesophilic planet?[J]. Microbiology-(UK), 2014, 160(1-11). doi: 10.1099/mic.0.071696-0 URL
[40]	NATCHEVA M, BESCHKOV V. Denitrification Performance of a Culture of Thermophilic Aerobic Bacteria NBIMCC 3729[J]. Chemical & biochemical engineering quarterly, 2003, 17(2):123-129.
[41]	FUJII K, TOMINAGA Y, OKUNAKA J, et al. Microbial and genomic characterization of Geobacillusthermodenitrificans OS27,a marine thermophile that degrades diverse raw seaweeds[J]. Applied microbiology and biotechnology, 2018, 102(11):4901-4913. doi: 10.1007/s00253-018-8958-9 URL
[42]	杨慧超. 条斑紫菜(Pyropiayezoensis)的病害调查及其绿斑病病原的PCR检测方法[D]. 上海: 上海海洋大学, 2019.
[43]	吴月红, 许学伟. 赤杆菌科微生物分类研究进展[J]. 微生物学通报, 2016, 43(5):1082-1094.
[44]	熊玉琴, 杨锐, 杨华田, 等. 藻际芽孢杆菌对坛紫菜生理的影响[J]. 生物学杂志, 2018, 35(1):37-41.
[45]	XIONG Y Q, YANG R, SUN X X, et al. Effect of the epiphytic bacterium & ITBacillus & ITsp WPySW2 on the metabolism of & ITPyropiahaitanensis&IT[J]. J ApplPhycol, 2018, 30(2):1225-1237.
[46]	MORIARTY D J. Control of luminous vibrio species in penaeid aquaculture ponds[J]. Aquaculture, 1998, 164(1-4):351-358. doi: 10.1016/S0044-8486(98)00199-9 URL

样品	温度/℃	盐度/‰	pH	溶解氧/(mg/L)
患白斑病池水样	29.3	33.5	8.63	7.3
健康池水样	28.6	33.3	8.76	7.1
水源水	29.9	33.8	8.8	7.2

样品	温度/℃	盐度/‰	pH	溶解氧/(mg/L)
患白斑病池水样	29.3	33.5	8.63	7.3
健康池水样	28.6	33.3	8.76	7.1
水源水	29.9	33.8	8.8	7.2

样品编号	样品性质	有效序列数	OUT数	覆盖度
PT11	患白斑病池水样	70438	1191	0.98
PT12	患白斑病池水样	117649	1487	0.98
PT13	患白斑病池水样	170417	1641	0.98
PT21	健康池水样	78881	854	0.99
PT22	健康池水样	116235	783	0.99
PT23	健康池水样	35574	767	0.99
PT31	水源水	91794	1209	0.98
PT32	水源水	85952	914	0.99
PT33	水源水	42813	1269	0.98
PT41	白斑病贝壳丝状体	56141	420	0.99
PT42	白斑病贝壳丝状体	91462	726	0.99
PT43	白斑病贝壳丝状体	91400	193	1
PT51	健康贝壳丝状体	276019	1049	0.99
PT52	健康贝壳丝状体	68964	1808	0.98
PT53	健康贝壳丝状体	136253	1755	0.98

样品编号	样品性质	有效序列数	OUT数	覆盖度
PT11	患白斑病池水样	70438	1191	0.98
PT12	患白斑病池水样	117649	1487	0.98
PT13	患白斑病池水样	170417	1641	0.98
PT21	健康池水样	78881	854	0.99
PT22	健康池水样	116235	783	0.99
PT23	健康池水样	35574	767	0.99
PT31	水源水	91794	1209	0.98
PT32	水源水	85952	914	0.99
PT33	水源水	42813	1269	0.98
PT41	白斑病贝壳丝状体	56141	420	0.99
PT42	白斑病贝壳丝状体	91462	726	0.99
PT43	白斑病贝壳丝状体	91400	193	1
PT51	健康贝壳丝状体	276019	1049	0.99
PT52	健康贝壳丝状体	68964	1808	0.98
PT53	健康贝壳丝状体	136253	1755	0.98

样品名称	Chao1指数	Shannon指数	Simpson指数	覆盖度
患白斑病池水样(PT1)	2095.68±238.2^a	6.49±0.76^a	0.95±0.04^a	0.98
健康池水样(PT2)	1241.89±71.33^b	3.87±0.37^b	0.69±0.04^b	0.99
水源水(PT3)	1540.22±295.26^b	6.07±0.32^a	0.94±0.01^a	0.99