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Method of judging rapid growth period of cyanobacteria based on cell division ftsZ gene expression quantity

A technology of gene expression and cell division, which is applied in the field of water environment management and cyanobacteria bloom control, can solve the problems such as the identification method of rapid growth period of no cyanobacteria, and achieve the effect of rapid and high-throughput detection

Active Publication Date: 2018-10-16
NANJING INST OF GEOGRAPHY & LIMNOLOGY
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Problems solved by technology

Patent document CN104388544A discloses a method for judging the limit of overwintering cyanobacteria overwintering recovery period by using the expression level of pseudovacuum gene (gvpC), which has played a certain role in the control of lake cyanobacteria, but there is no relevant method for judging the rapid growth period of cyanobacteria.

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  • Method of judging rapid growth period of cyanobacteria based on cell division ftsZ gene expression quantity
  • Method of judging rapid growth period of cyanobacteria based on cell division ftsZ gene expression quantity
  • Method of judging rapid growth period of cyanobacteria based on cell division ftsZ gene expression quantity

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Embodiment Construction

[0025] The technical solution of the present invention will be further described in detail below in conjunction with the accompanying drawings and embodiments, taking Taihu Lake as an example.

[0026] Method of the present invention comprises the steps:

[0027] (1) Algae samples from the complete water column were collected at different sampling points in Taihu Lake, mixed and filtered, concentrated and quickly transferred to RNase-free cryopreservation tubes, placed in liquid nitrogen, brought back to the laboratory, and stored at -70°C. The sampling time is from March to June 2017. The RNA of each sample was extracted, and the RNA was reverse-transcribed to obtain Microcystis cDNA.

[0028] RNA extraction steps: transfer the GF / C filter membrane stored in low temperature to Lysing Matrix B (Lysing Matrix B, glass bead diameter 0.1 mm), add 500 μL Buffer RLT, and place it in the Fast Prep-24 sample processing instrument. Set the parameters of the FastPrep-24 sample proces...

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Abstract

The invention provides a method of judging rapid growth period of cyanobacteria based on cell division ftsZ gene expression quantity. In-situ sampling is performed on a lake; samples are subjected toRNA extraction before reverse transcription; ftsZ and 16SrRNA are subjected to primer amplification before reverse transcription is performed to obtain microcystis cDNA; fluorescence quantitative PCR(polymerase chain reaction) amplification is performed on ftsZ and 16SrRNA genes of each sample; with 16SrRNA as a housekeeping gene and ftsZ as a target gene, relative expression quantity of ftsZ gene in unit cells is deducted; microcystis separated in situ from the lake are subjected to indoor culture experiment, a regression equation of microcystis growth rate mu and ftsZ gene relative expression quantity is established; rapid growth period of microcystis is determined; meanwhile, the ftsZ gene relative expression quantity is used as a threshold of window for the rapid growth period of cyanobacteria; the period greater than the threshold is determined as the rapid growth period of lake cyanobacteria. The method herein can provide rapid high-throughput detection, detection results of multiple samples are attained within hours, and the rapid growth period of cyanobacteria can be judged accurately.

Description

technical field [0001] The invention relates to a method for discriminating the rapid growth period of cyanobacteria based on the expression of cell division ftsZ gene, belongs to the field of water environment treatment and cyanobacteria bloom control, and relates to eutrophication treatment and ecological restoration technologies of large shallow lakes and reservoirs. Background technique [0002] The problem of eutrophication of lakes in our country is becoming more and more serious, and cyanobacteria blooms occur frequently, which has seriously affected the development of society and the drinking water problem in some areas. Cyanobacteria blooms in lakes often have Microcystis as the dominant population. Existing studies have shown that the formation process of algal blooms can be divided into four stages: sinking and overwintering, recovery and rising, rapid growth, and algal bloom accumulation. It can be seen that the rapid growth period before the formation and accum...

Claims

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Application Information

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IPC IPC(8): C12Q1/6851C12Q1/04G06F19/18G06F17/18C12R1/89
CPCC12Q1/6851G06F17/18G16B20/00C12Q2531/113C12Q2563/107
Inventor 史小丽阳振于洋张民陈开宁
Owner NANJING INST OF GEOGRAPHY & LIMNOLOGY
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