Development of a PCR method for detection of <italic style="font-style: italic">Pseudoalteromonas marina</italic> associated with green spot disease in <italic style="font-style: italic">Pyropia yezoensis</italic>

Huichao YANG; Yongwei YAN; Jie LI; Lei TANG; Yunxiang MAO; Zhaolan MO

doi:10.1007/s00343-019-9045-5

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Development of a PCR method for detection of Pseudoalteromonas marina associated with green spot disease in Pyropia yezoensis

Ecology | Updated：2023-04-27

- Development of a PCR method for detection of Pseudoalteromonas marina associated with green spot disease in Pyropia yezoensis
- Development of a PCR method for detection of Pseudoalteromonas marina associated with green spot disease in Pyropia yezoensis
- 海洋湖沼学报（英文） 2020年38卷第1期页码：168-176
- Affiliations：
  
  1.Laboratory for Marine Fisheries Science and Food Production Processes, Qingdao National Laboratory for Marine Science and Technology, Key Laboratory of Maricultural Organism Disease Control, Ministry of Agriculture and Rural Affairs, Qingdao Key Laboratory of Mariculture, Epidemiology and Biosecurity, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China
  2.College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306, China
  3.Laboratory for Marine Biology and Biotechnology, Qingdao National Laboratory for Marine Science and Technology; Key Laboratory of Marine Genetics and Breeding(Ocean University of China), Ministry of Education, Qingdao 266003, China
- Author bio：
  
  MO Zhaolan, mozl@ysfri.ac.cn
- Funds：
  
  the China Agriculture Research System(CARS-50);the National High-Tech R & D Program of China(2012AA10A406);the National Science and Technology Infrastructure Platform Construction(2018DKA30470);the Aoshan Technology Innovation Program of Qingdao National Laboratory for Marine Science and Technology(2015ASKJ02);© Chinese Society for Oceanology and Limnology, Science Press and Springer-Verlag GmbH Germany, part of Springer Nature 2020
- DOI：10.1007/s00343-019-9045-5
  中图分类号：
- 收稿：2019-02-27，
  
  录用：2019-4-3，
  
  网络首发：2019-04-15，
  
  纸质出版：2020-01
- Accepted：
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Development of a PCR method for detection of Pseudoalteromonas marina associated with green spot disease in Pyropia yezoensis[J]. 海洋湖沼学报（英文）, 2020,38(1):168-176.

Huichao YANG, Yongwei YAN, Jie LI, et al. Development of a PCR method for detection of Pseudoalteromonas marina associated with green spot disease in Pyropia yezoensis[J]. Journal of Oceanology and Limnology, 2020, 38(1): 168-176.
Development of a PCR method for detection of Pseudoalteromonas marina associated with green spot disease in Pyropia yezoensis[J]. 海洋湖沼学报（英文）, 2020,38(1):168-176. DOI： 10.1007/s00343-019-9045-5.

Huichao YANG, Yongwei YAN, Jie LI, et al. Development of a PCR method for detection of Pseudoalteromonas marina associated with green spot disease in Pyropia yezoensis[J]. Journal of Oceanology and Limnology, 2020, 38(1): 168-176. DOI： 10.1007/s00343-019-9045-5.

摘要

Abstract

Pseudoalteromonas marina

is one of the potential pathogens that cause green spot disease (GSD) in

Pyropia yezoensis

. To prevent GSD from development and spread

an effective method to detect the pathogen at early GSD infection stages need to be established. In this research

PCR methods were established targeting the

dnaA

gene (encoding chromosome replication initiator protein) and the

dnaN

gene (encoding

sliding clamp of DNA polymerase Ⅲ protein) to detect

P. marina

with three primer pairs pws-

dnaA

2 (Forward

5'-ACCGCATTAACGAACTACTCGTG-3'; Reverse

5'-TGCCATTACCTACAGCATGG-3')

pcs-

dnaN

2 (Forward

5'-CTTACAACGTTATCAGCGGC-3'; Reverse

5'-GTTGAGTATTAAGTGATTGAGTAAGC-3') or pws-

dnaN

3 (Forward

5'-ACTTACAACGTTATCAGCGGC-3'; Reverse

5'-ACTGCTGTTTGAGTCTGCTAAC-3'). Three PCR methods corresponding to the three primer pairs sufficiently distinguished

P. marina

from 22 bacterial species

thus resulting in detection limits of 4 to 4×10

CFU cells or 2.37×10

to 2.37×10

fg of

P. marina

DNA per PCR reaction. In an artificial infection experiment of

P. yezoensis

infected with

P. marina

all established PCRs successfully detected

P. marina

at early GSD infection stages. The results show that the established PCRs are specific and sensitive

and are potential for applications in early diagnosis of GSD in

Pyropia

关键词

Keywords

references

S Arasaki . Studies on red rot of Porphyra tenera . Nippon Suisan Gakkaishi , 1947 . 13 ( 3 ): 74 - 90 . DOI: 10.2331/suisan.13.74 http://doi.org/10.2331/suisan.13.74 .

J Cao , S C Wang , C W Yao , Z Xu , X M Xu . Hypolipidemic effect of porphyran extracted from Pyropia yezoensis in ICR mice with high fatty diet . Journal of Applied Phycology , 2016 . 28 ( 2 ): 1315 - 1322 . DOI: 10.1007/s10811-015-0637-y http://doi.org/10.1007/s10811-015-0637-y .

H Y Ding , J H Ma . Simultaneous infection by red rot and chytrid diseases in Porphyra yezoensis Ueda . Journal of Applied Phycology , 2005 . 17 ( 1 ): 51 - 56 . DOI: 10.1007/s10811-005-5523-6 http://doi.org/10.1007/s10811-005-5523-6 .

H Y Ding . Study on development process of green-spot disease in Porphyra yezoensis . Journal of Anhui Agricultural Sciences , 2008 . 36 ( 11 ): 4626 - 4628 . DOI: 10.3969/j.issn.0517-6611.2008.11.085 http://doi.org/10.3969/j.issn.0517-6611.2008.11.085 .

S Egan , M Gardiner . Microbial dysbiosis: rethinking disease in marine ecosystems . Frontiers in Microbiology , 2016 . 7 991 DOI: 10.3389/fmicb.2016.00991 http://doi.org/10.3389/fmicb.2016.00991 .

S Z Feng , H Y Xu , Y Q Song , H P Zhang , Z H Sun . Phylogenetic analysis of closely related Leuconostoc mesenteroides species based on dnaA and rpoB gene homologues . China Dairy Industry , 2016 . 44 ( 2 ): 8 - 11 . DOI: 10.3969/j.issn.1001-2230.2016.02.002 http://doi.org/10.3969/j.issn.1001-2230.2016.02.002 .

Y Fujita , B Zenitani . Studies on pathogenic Pythium of laver red rot in Ariake sea farm-Ⅱ: experimental conditions and nutritional requirements for growth.Experimental conditions and nutritional requirements for growth . Nippon Suisan Gakkaishi , 1977 . 43 ( 1 ): 89 - 95 . DOI: 10.2331/suisan.43.89 http://doi.org/10.2331/suisan.43.89 .

Fujita Y. 1990. Diseases of cultivated Porphyra in Japan. In : Akatsuka I ed. Introduction to Applied Phycology. SPB Academic Publishing, The Hague. p.177–190.

C M M Gachon , T Sime-Ngando , M Strittmatter , A Chambouvet , G H Kim . Algal diseases: spotlight on a black box . Trends in Plant Science , 2010 . 15 ( 11 ): 633 - 640 . DOI: 10.1016/j.tplants.2010.08.005 http://doi.org/10.1016/j.tplants.2010.08.005 .

T Hall . BioEdit: an important software for molecular biology . GERF Bulletin of Biosciences , 2011 . 2 ( 1 ): 60 - 61 . http://d.old.wanfangdata.com.cn/NSTLQK/NSTL_QKJJ04776284/ http://d.old.wanfangdata.com.cn/NSTLQK/NSTL_QKJJ04776284/ , .

X J Han , Y X Mao , J Li , G Y Li , C Li , L Liu , Z L Mo . Identification and pathogenecity of a novel pathogen causing green rot disease in Porphyra haitanensis . Journal of Fisheries of China , 2015 . 39 ( 11 ): 1721 - 1729 . DOI: 10.11964/jfc.20150209720 http://doi.org/10.11964/jfc.20150209720 .

J L Kerwin , L M Johnson , H C Whisler , A R Tuininga . Infection and morphogenesis of Pythium marinum in species of Porphyra and other red algae . Canadian Journal of Botany , 1992 . 70 ( 5 ): 1017 - 1024 . DOI: 10.1139/b92-126 http://doi.org/10.1139/b92-126 .

G H Kim , T A Klochkova , D J Lee , S H Im . Chloroplast virus causes green-spot disease in cultivated Pyropia of Korea . Algal Research , 2016 . 17 293 - 299 . DOI: 10.1016/j.algal.2016.05.023 http://doi.org/10.1016/j.algal.2016.05.023 .

G H Kim , K H Moon , J Y Kim , J Shim , T A Klochkova . A revaluation of algal diseases in Korean Pyropia ( Porphyra ) sea farms and their economic impact . Algae , 2014 . 29 ( 4 ): 249 - 265 . DOI: 10.4490/algae.2014.29.4.249 http://doi.org/10.4490/algae.2014.29.4.249 .

T A Klochkova , J B Shim , M S Hwang , G H Kim . Hostparasite interactions and host species susceptibility of the marine oomycete parasite, Olpidiopsis sp., from Korea that infects red algae. . Journal of Applied Phycology , 2012 . 24 ( 1 ): 135 - 144 . DOI: 10.1007/s10811-011-9661-8 http://doi.org/10.1007/s10811-011-9661-8 .

V Kumar , E Zozaya-Valdes , S Kjelleberg , T Thomas , S Egan . Multiple opportunistic pathogens can cause a bleaching disease in the red seaweed Delisea pulchra . Environmental Microbiology , 2016 . 18 ( 11 ): 3962 - 3975 . DOI: 10.1111/1462-2920.13403 http://doi.org/10.1111/1462-2920.13403 .

M S Kwak , T A Klochkova , S Jeong , G H Kim . Olpidiopsis porphyrae var.koreanae, an endemic endoparasite infecting cultivated Pyropia yezoensis in Korea . Journal of Applied Phycology , 2017 . 29 ( 4 ): 2003 - 2012 . DOI: 10.1007/s10811-017-1109-3 http://doi.org/10.1007/s10811-017-1109-3 .

P Y Lai . Investigation and countermeasure of rot disease of Porphyra haitanensis in Fuding City in autumn 2008 . Modern Fisheries Information , 2009 . 24 ( 7 ): 6 - 9 . DOI: 10.3969/j.issn.1004-8340.2009.07.002 http://doi.org/10.3969/j.issn.1004-8340.2009.07.002 .

S J Lee , B Y Jee , M H Son , S R Lee . Infection and cox2 sequence of Pythium chondricola (Oomycetes) causing red rot disease in Pyropia yezoensis (Rhodophyta) in Korea . Algae , 2017 . 32 ( 2 ): 155 - 160 . DOI: 10.4490/algae.2017.32.5.16 http://doi.org/10.4490/algae.2017.32.5.16 .

S J Lee , S W Park , J H Lee , Y S Kim . Diseases of the cultivated Porphyra at seocheon area . Journal of Fish Pathology , 2012 . 25 ( 3 ): 249 - 256 . DOI: 10.7847/jfp.2012.25.3.249 http://doi.org/10.7847/jfp.2012.25.3.249 .

Li J, Mou Z J, Yang H C, Mao Y X, Yan Y W, Mo Z L. 2018. Isolation and identification the pathogen of Pyropia yezoensis green spot disease. Progress in FisheryScience , published online First, August 2018. https: //doi.org/10.19663/j.issn2095-9869.20180710002. (in Chinese)

Y M Liu , J H Ma , Q Wen . Simultaneous infection by red rot and olpidiops disease in laver Porphyra haitanensis . Journal of Dalian Ocean University , 2012 . 27 ( 6 ): 546 - 550 . DOI: 10.3969/j.issn.2095-1388.2012.06.011 http://doi.org/10.3969/j.issn.2095-1388.2012.06.011 .

Q Q Lu , W Zhou , J Y Zhu , B L Yan , J Z Ni , L E Yang . The history, status quo and development trend of Pyropia yezoensis industry of China . Marine Economy in China , 2018 . ( 3 ): 3 - 11 . .

J H Ma . A preliminary study on the red rot disease of Porphyra yezoensis . Journal of Shanghai Fisheries University , 1996 . 5 ( 1 ): 1 - 7 . http://en.cnki.com.cn/article_en/cjfdtotal-ssdb199601000.htm http://en.cnki.com.cn/article_en/cjfdtotal-ssdb199601000.htm , .

M Mohkam , N Nezafat , A Berenjian , M A Mobasher , Y Ghasemi . Identification of Bacillus probiotics isolated from soil rhizosphere using 16S rRNA, recA, rpoB gene sequencing and RAPD-PCR . Probiotics and Antimicrobial Proteins , 2016 . 8 ( 1 ): 8 - 18 . DOI: doi.org/10.1007/s12602-016-9208-z http://doi.org/doi.org/10.1007/s12602-016-9208-z .

I Mokrousov , A Vyazovaya , V Zhuravlev , T Otten , J Millet , W W Jiao , A D Shen , N Rastogi , B Vishnevsky , O Narvskaya . Real-time PCR assay for rapid detection of epidemiologically and clinically significant Mycobacterium tuberculosis Beijing genotype isolates . Journal of Clinical Microbiology , 2014 . 52 ( 5 ): 1691 - 1693 . DOI: 10.1128/JCM.03193-13 http://doi.org/10.1128/JCM.03193-13 .

Z J Mou . Etiological Study on Pyropia/Porphyra yezoensis and Screening of Agarase-Producing Bacteria , : Qingdao Ocean University of China , 2012 . p.33 - 35 . .

Y Nakao , T Onohara , T Matsubara , Y Fujita , B Zenitani . Bacteriological studies on diseases of cultured Laver—Ⅰ.Green spot rotting-like deterioration of laver frond by bacteria, in vitro . Nippon Suisan Gakkaishi , 1972 . 38 ( 6 ): 561 - 564 . DOI: 10.2331/suisan.38.561 http://doi.org/10.2331/suisan.38.561 .

C S Park , M Kakinuma , H Amano . Forecasting infections of the red rot disease on Porphyra yezoensis Ueda (Rhodophyta) cultivation farms . Journal of Applied Phycology , 2006 . 18 ( 3-5 ): 295 - 299 . DOI: 10.1007/s10811-006-9031-0 http://doi.org/10.1007/s10811-006-9031-0 .

L Provasoli . Media and prospects for the cultivation of marine algae. In: Watanabe A, Hattori A eds. Cultures and Collections of Algae , : Tokyo Japan Society of Plant Physiology , 1968 . p.63 - 75 . .

Y Saito , T Matsusato , K Yoshikawa . On the symptoms of "green spot" and "crape" in nori ( Porphyra ) culture . Bulletin of Nansei National Fisheries Research Institute , 1972 . 5 1 - 9 . .

R A Sandaa , L Brunvold , T Magnesen , Ø Bergh . Monitoring the opportunistic bacteria Pseudoalteromonas sp.LT-13 in a great scallop, Pecten maximus hatchery . Aquaculture , 2008 . 276 ( 1-4 ): 14 - 21 . DOI: 10.1016/j.aquaculture.2008.02.007 http://doi.org/10.1016/j.aquaculture.2008.02.007 .

T Sawabe , H Makino , M Tatsumi , K Nakano , K Tajima , M M Iqbal , I Yumoto , Y Ezura , R Christen . Pseudoalteromonas bacteriolytica sp.nov., a marine bacterium that is the causative agent of red spot disease of Laminaria japonica . International Journal of Systematic and Evolutionary Bacteriology , 1998 . 48 ( 3 ): 769 - 774 . DOI: doi.org/10.1099/00207713-48-3-769 http://doi.org/doi.org/10.1099/00207713-48-3-769 .

S Sekimoto , T A Klochkova , J A West , G W Beakes , D Honda . Olpidiopsis bostrychiae sp.nov.: an endoparasitic oomycete that infects Bostrychia and other red algae (Rhodophyta) . Phycologia , 2009 . 48 ( 6 ): 460 - 472 . DOI: 10.2216/08-11.1 http://doi.org/10.2216/08-11.1 .

T L Skovhus , N B Ramsing , C Holmström , S Kjelleberg , I Dahllöf . Real-time quantitative PCR for assessment of abundance of Pseudoalteromonas species in marine samples . Applied and Environmental Microbiology , 2004 . 70 ( 4 ): 2373 - 2382 . DOI: 10.1128/AEM.70.4.2373-2382 http://doi.org/10.1128/AEM.70.4.2373-2382 .

M Sunairi , H Tsuchiya , T Tsuchiya , Y Omura , Y Koyanagi , M Ozawa , N Iwabuchi , H Murooka , M Nakajima . Isolation of a bacterium that causes Anaaki disease of the red algae Porphyra yezoensis . Journal of Applied Bacteriology , 1995 . 79 ( 2 ): 225 - 229 . DOI: 10.1111/j.1365-2672.1995.tb00939.x http://doi.org/10.1111/j.1365-2672.1995.tb00939.x .

S Suto , O Umebayashi . On the "perforating disease" in nori ( Porphyra ) culture . Nippon Suisan Gakkaishi , 1954 . 19 ( 12 ): 1176 - 1178 . DOI: 10.2331/suisan.19.1176 http://doi.org/10.2331/suisan.19.1176 .

Y N Wang , N S Feng , Q Li , J Ding , Y Y Zhan , Y Q Chang . Isolation and characterization of bacteria associated with a syndrome disease of sea urchin Strongylocentrotus intermedius in North China . Aquaculture Research , 2013 . 44 ( 5 ): 691 - 700 . DOI: 10.1111/j.1365-2109.2011.03073.x http://doi.org/10.1111/j.1365-2109.2011.03073.x .

S Wei , R Chelliah , B J Park , J H Park , F Forghani , Y S Park , M S Cho , D S Park , D H Oh . Molecular discrimination of Bacillus cereus group species in foods (lettuce, spinach, and kimbap) using quantitative realtime PCR targeting groEL and gyrB . Microbial Pathogenesis , 2018 . 115 312 - 320 . DOI: 10.1016/j.micpath.2017.12.079 http://doi.org/10.1016/j.micpath.2017.12.079 .

Y Yan , J H Ma , P Xu , Q H Sun , H Q Wang . Pseudoalteromonas citrea , the causative agent of green-spot disease of Porphyrae yezoensis . Journal of Fishery Sciences of China , 2002 . 9 ( 4 ): 353 - 358 . DOI: 10.3321/j.issn:1005-8737.2002.04.014 http://doi.org/10.3321/j.issn:1005-8737.2002.04.014 .

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Development of a PCR method for detection of Pseudoalteromonas marina associated with green spot disease in Pyropia yezoensis

DOI：10.1007/s00343-019-9045-5

摘要

Abstract

关键词

Keywords

references