Detection method for biological community structures of ocean brown tide

Abstract

The invention discloses a detection method for the biological community structures of an ocean brown tide. According to the detection method, miniature organism 18S rDNA in a water area environment is detected by virtue of a metagenomics high-throughput sequencing technology, the obtained OUTs and species annotations are combined, and then diversity, uniformity, abundance and dominance index analysis is carried out on the OUTs, thus judging the dominance degrees of organisms in the ocean brown tide; statistics and judgment for the dominance evolvement rules of dominant species in brown tide outbreaks are promoted. The embodiment of the invention designs and verifies primer sequences designed with regard to the organisms in the brown tide, and a verification for the actual brown tide outbreak cases proves that the method disclosed by the invention is capable of authenticating the biological community structures of the ocean brown tide in ocean water, providing a technical support for brown tide monitoring operation, and providing a technical support for avoiding environment risks for healthy mariculture.

Description

technical field [0001] The invention relates to the technical field of detection of marine pico-eukaryotic species, in particular to a detection method of marine brown tide biological community structure. Background technique [0002] Research overview of brown tide at home and abroad: brown tide is a harmful algal bloom caused by microscopic algae, which can also be understood as a red tide of picoplankton, which is a brown tide. , The duration is longer. American research institutions named this harmful algal bloom "brown tide" to distinguish it from the general "red tide", and it was adopted by relevant international agencies (Cosper EM, et al., 1990). In recent years, brown tides have occurred frequently in the Bohai Sea of ​​China, and the main species of disaster is Aureococcus anophagefferens, which belongs to microalgae. This microalgae is extremely harmful to the aquaculture industry, causing shellfish to stagnate or even die. China is the third country to experi...

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Problems solved by technology

[0003] Traditionally, the identification and detection of plankton mainly rely on the microscope to observe the morphological differences for classification. However, this method has certain limitations in practical applications: first, plankton, especially micro and pico plankton, are small in size, Difficult to observe, difficult to operate, conventional microscopic examination cannot achieve species identification; secondly, this method relies on experienced taxonomic researchers, requires solid taxonomic skills, and is time-consuming and laborious, not suitable for large-scale sample research; Finally, the morphological differences of many plankton are not obvious, and it is difficult to distinguish and easy to cause human error

Because brown tide species are pico organisms smaller than 3 μm, it is impossible to identify its species by general methods such as microscope identification. There are many pico-organisms that may have brown tides, and DNA sequencing technology is used to detect the 18S rDNA of pico-organisms in the target waters, and the obtained DNA fragment sequences are input into GenBank for retrieval and comparison, and the Latin and Chinese names of corresponding species are searched. , and then can identify the species of brown tide organisms, there is no proprietary primer sequence designed for brown tide organisms, and no 18S rDNA sequencing comparison method is used in the analysis of brown tide biological community structure. The present invention uses this While expanding the method, specially designed and verified the primer sequences designed for brown tide organisms, and used the probability of clone sequencing to count the parameters such as the dominance degree, community diversity index, and evenness index of various pico organisms

[0012] 2010.11.24 published invention patent "a detection method of phytoplankton community structure" (application number: 201010184810) is a detection method using denaturing gradient gel electrophoresis (DGGE) technology to study the detection method of phytoplankton community structure, but this method cannot identify specific Species, can only count parameters such as biodiversity based on electrophoretic strips, but cannot know the name of dominant species and its degree of dominance, etc.

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