PARK Mirye, PARK So Yun, HWANG Jinik, JUNG Seung Won, LEE Juyun, CHANG Man, LEE Taek-Kyun. Integration of the nuclease protection assay with sandwich hybridization (NPA-SH) for sensitive detection of Heterocapsa triquetra[J]. Acta Oceanologica Sinica, 2018, 37(5): 107-112. doi: 10.1007/s13131-018-1167-7
Citation: PARK Mirye, PARK So Yun, HWANG Jinik, JUNG Seung Won, LEE Juyun, CHANG Man, LEE Taek-Kyun. Integration of the nuclease protection assay with sandwich hybridization (NPA-SH) for sensitive detection of Heterocapsa triquetra[J]. Acta Oceanologica Sinica, 2018, 37(5): 107-112. doi: 10.1007/s13131-018-1167-7

Integration of the nuclease protection assay with sandwich hybridization (NPA-SH) for sensitive detection of Heterocapsa triquetra

doi: 10.1007/s13131-018-1167-7
  • Received Date: 2017-08-29
  • Rev Recd Date: 2017-12-11
  • Microalgae are photosynthetic microorganisms that function as primary producers in aquatic ecosystems. Some species of microalgae undergo rapid growth and cause harmful blooms in marine ecosystems. Heterocapsa triquetra is one of the most common bloom-forming species in estuarine and coastal waters worldwide. Although this species does not produce toxins, unlike some other Heterocapsa species, the high density of its blooms can cause significant ecological damage. We developed a H. triquetra species-specific nuclease protection assay sandwich hybridization (NPA-SH) probe that targets the large subunit of ribosomal RNA (LSU rRNA). We tested probe specificity and sensitivity with five other dinoflagellates that also cause red tides. Our assay detected H. triquetra at a concentration of 1.5×104 cells/mL, more sensitive than required for a red-tide guidance warning by the Korea Ministry of Oceans and Fisheries in 2015 (3.0×104 cells/mL). We also used the NPA-SH assay to monitor H. triquetra in the Tongyeong region of the southern sea area of Korea during 2014. This method could detect H. triquetra cells within 3 h. Our assay is useful for monitoring H. triquetra under field conditions.
  • loading
  • Antonella P, Luca G. 2013. The quantitative real-time PCR applications in the monitoring of marine harmful algal bloom (HAB) species. Environ Sci Pollut Res, 20(10):6851-6862
    Archambault M C, Bricelj V M, Grant J, et al. 2004. Effects of suspended and sedimented clays on juvenile hard clams, Mercenaria mercenaria, within the context of harmful algal bloom mitigation. Mar Biol, 144(3):553-565
    Ayers K, Rhodes L L, Tyrrell J, et al. 2005. International accreditation of sandwich hybridisation assay format DNA probes for micro-algae. New Zealand J Mar Freshw Res, 39(6):1225-1231
    Baek S H, Ki J S, Katano T, et al. 2011. Dense winter bloom of the dinoflagellate Heterocapsa triquetra below the thick surface ice of brackish Lake Shihwa, Korea. Phycol Res, 59(4):273-285
    Battocchi C, Totti C, Vila M, et al. 2010. Monitoring toxic microalgae Ostreopsis (dinoflagellate) species in coastal waters of the Mediterranean Sea using molecular PCR-based assay combined with light microscopy. Mar Pollut Bull, 60(7):1074-1084
    Cai Qingsong, Li Rongxiu, Zhen Yu, et al. 2006. Detection of two Prorocentrum species using sandwich hybridization integrated with nuclease protection assay. Harmful Algae, 5(3):300-309
    Chen Guofu, Liu Yang, Zhang Chunyun, et al. 2013. Development of rRNA-targeted probes for detection of Prorocentrum micans (Dinophyceae) using whole cell in situ hybridization. J Appl Phycol, 25(4):1077-1089
    Debelius B, Forja J M, DelValls Á, et al. 2009. Toxicity and bioaccumulation of copper and lead in five marine microalgae. Ecotoxicol Environ Saf, 72(5):1503-1513
    Diercks S, Medlin L K, Metfies K. 2008a. Colorimetric detection of the toxic dinoflagellate Alexandrium minutum using sandwich hybridization in a microtiter plate assay. Harmful Algae, 7(2):137-145
    Diercks S, Metfies K, Medlin L K. 2008b. Molecular probe sets for the detection of toxic algae for use in sandwich hybridization formats. J Plankton Res, 30(4):439-448
    Ebenezer V, Medlin L K, Ki J S. 2012. Molecular detection, quantification, and diversity evaluation of microalgae. Mar Biotechnol, 14(2):129-142
    Godhe A, Otta S K, Rehnstam-Holm A S, et al. 2001. Polymerase chain reaction in detection of Gymnodinium mikimotoi and Alexandrium minutum in field samples from southwest India. Mar Biotechnol, 3(2):152-162
    Hyka P, Lickova S, Pribyl P, et al. 2013. Flow cytometry for the development of biotechnological processes with microalgae. Biotechnol Adv, 31(1):2-16
    Jedlicki A, Fernández G, Astorga M, et al. 2012. Molecular detection and species identification of Alexandrium (Dinophyceae) causing harmful algal blooms along the Chilean coastline. AoB Plants, 2012:pls033
    Jiang Liying, Ilag L L. 2014. Detection of endogenous BMAA in dinoflagellate (Heterocapsa triquetra) hints at evolutionary conservation and environmental concern. PubRaw Sci, 1(2):1-8
    Ki J S, Han M S. 2006. A low-density oligonucleotide array study for parallel detection of harmful algal species using hybridization of consensus PCR products of LSU rDNA D2 domain. Biosens Bioelectron, 21(9):1812-1821
    Lee J Y, Han M S. 2007. Change of blooming pattern and population dynamics of phytoplankton in Masan bay, Korea. Journal of the Korean Society of Oceanography, 12(3):147-158
    Lee C K, Lee O H, Lee S G. 2005. Impacts of temperature, salinity and irradiance on the growth of ten harmful algal bloom-forming microalgae isolated in Korean coastal waters. Journal of the Korean Society of Oceanography, 10(1):79-91
    Lee C, Limand W. 2006. Variation of harmful algal blooms in Masan-Chinhae Bay. ScienceAsia, 32(S1):51-56
    Litaker R W, Tester P A, Duke C S, et al. 2002a. Seasonal niche strategy of the bloom-forming dinoflagellate Heterocapsa triquetra. Mar Ecol Prog Ser, 232:45-62
    Litaker R W, Warner V E, Rhyne C, et al. 2002b. Effect of diel and interday variations in light on the cell division pattern and in situ growth rates of the bloom-forming dinoflagellate Heterocapsa triquetra. Mar Ecol Prog Ser, 232:63-74
    Lu Songhui, Hodgkiss I J. 2004. Harmful algal bloom causative collected from Hong Kong waters. Hydrobiologia, 512(1-3):231-238
    Morel F M M, Price N M. 2003. The biogeochemical cycles of trace metals in the oceans. Science, 300(5621):944-947
    Naito K, Matsui M, Imai I. 2005. Ability of marine eukaryotic red tide microalgae to utilize insoluble iron. Harmful Algae, 4(6):1021-1032
    Park J, Jeong H J, Yoo Y D, et al. 2013. Mixotrophic dinoflagellate red tides in Korean waters:distribution and ecophysiology. Harmful Algae, 30(S1):S28-S40
    Priyadarshani I, Rath B. 2012. Commercial and industrial applications of micro algae-a review. J Algal Biomass Utln, 3(4):89-100
    Spolaore P, Joannis-Cassan C, Duran E, et al. 2006. Commercial applications of microalgae. J Biosci Bioeng, 101(2):87-96
    Suh S S, Park M, Hwang J, et al. 2016. Detection of the dinoflagellate, Cochlodinium polykrikoides, that forms algal blooms using sandwich hybridization integrated with nuclease protection assay. Biotechnol Lett, 38(1):57-63
    Tas S. 2015. A prolonged red tide of Heterocapsa triquetra (Ehrenberg) F. Stein (Dinophyceae) and phytoplankton succession in a eutrophic estuary in Turkey. Mediterr Mar Sci, 16(3):621-627
    Tyrrell J V, Connell L B, Scholin C A. 2002. Monitoring for Heterosigma akashiwo using a sandwich hybridization assay. Harmful Algae, 1(2):205-214
    Venugopalan C, Kapoor H C. 1997. Single step isolation of plant RNA. Phytochemistry, 46(8):1303-1305
    Xin Zeyu, Yu Zhigang, Wang Tanchun, et al. 2005. Identification and quantification of the toxic dinoflagellate Gymnodinium sp. with competitive enzyme-linked immunosorbent assay (cELISA). Harmful Algae, 4(2):297-307
    Zhen Yu, Mi Tiezhu, Yu Zhigang. 2008. Detection of Phaeocystis globosa using sandwich hybridization integrated with nuclease protection assay (NPA-SH). J Environ Sci, 20(12):1481-1486
    Zhen Yu, Mi Tiezhu, Yu Zhigang. 2009. Detection of several harmful algal species by sandwich hybridization integrated with a nuclease protection assay. Harmful Algae, 8(5):651-657
    Zhen Yu, Yu Zhigang, Cai Qingsong, et al. 2007. Detection of two diatoms using sandwich hybridization integrated with nuclease protection assay (NPA-SH). Hydrobiologia, 575(1):1-11
  • 加载中

Catalog

    通讯作者: 陈斌, bchen63@163.com
    • 1. 

      沈阳化工大学材料科学与工程学院 沈阳 110142

    1. 本站搜索
    2. 百度学术搜索
    3. 万方数据库搜索
    4. CNKI搜索

    Article Metrics

    Article views (676) PDF downloads(462) Cited by()
    Proportional views
    Related

    /

    DownLoad:  Full-Size Img  PowerPoint
    Return
    Return