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Method for detecting growth rate of skeletonema costatum

A growth rate and rib technology, applied in the detection field of planktonic diatoms, can solve the problem that basic physiological and ecological parameters such as planktonic algae on-site growth rate cannot be measured, and achieve the effect of improving specificity

Inactive Publication Date: 2010-01-20
OCEAN UNIV OF CHINA
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Problems solved by technology

[0015] The technical problem to be solved by the present invention is to provide a technical method for accurately estimating the on-site growth rate of Scutellaria costarum, which is used for estimating the productivity of planktonic algae, studying the change of planktonic algae populations, constructing ecological dynamic models and even predicting red tides, overcoming existing The defect that basic physiological and ecological parameters such as the on-site growth rate of a single species of planktonic algae cannot be measured in the technology

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  • Method for detecting growth rate of skeletonema costatum
  • Method for detecting growth rate of skeletonema costatum
  • Method for detecting growth rate of skeletonema costatum

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Embodiment Construction

[0055] In the embodiment of the present invention, the culture process of Skeletonemacostatum is: Skeletonemacostatum (Skeletonemacostatum) is cultivated to logarithmic growth phase with F / 2 medium in the laboratory, culture conditions: light / dark cycle is 12h / 12h , the light intensity is 4000Lux, and the culture temperature is 22-25°C.

[0056] Collect 5 parts of Skeletalum costanum (cell density is 4.1×107 cells / mL) cultured for one week about 3.8×10 7 cells (the weight of the algae concentrate is about 150mg). Use the following RNA extraction methods to extract:

[0057] (1) Collect algae cells and grind them into fine powder with a mortar under the condition of continuously adding liquid nitrogen;

[0058] (2) Add the grinding liquid to 1mL TRIZOL reagent, and homogenize at room temperature for 5min;

[0059] (3) Add 200 μL of chloroform vigorously, shake for 15 seconds, and incubate at room temperature for 3 minutes;

[0060] (4) Centrifuge at 12000rpm for 15min at 4°...

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Abstract

The present invention discloses a method to test the growth rate of the skeletonema costatum, which includes the procedures as follows: a. the extraction of RNA of skeletonema costatum, b. separation and cloning of partial sequences of the PCNA gene of the skeletonema costatum, c. cloning of partial sequences of Cytb gene of skeletonema costatum, d. a standard real-time quantitative PCR curve used to test genes of PCNA and Cytb of skeletonema costatum is developed, based on the gene sequences of both PCNA and Cytb, e. the growth rate of the skeletonema costatum is tested based on the expression of quantity (REQ) of gene PCNA of skeletonema costatum, with PCNA as the target gene and the Cytb as the housekeeping gene. The present invention provides a technical method for accurate estimation on the growth rate on the spot of the skeletonema costatum, which can be used to estimate the productivity of the floating algae, study the variance of the alga species, construct the zoological dynamic model and even forecast the red tide.

Description

technical field [0001] The invention relates to a method for detecting planktonic diatoms, in particular to a method for detecting the growth rate of Skeletalum costatum by real-time fluorescent quantitative PCR. Background technique [0002] Scutellaria mesocostalis is a typical representative of wide-temperature and wide-salt species. It is widely distributed, from the Arctic to the equator, from high-salt waters in open seas to coastal low-salt water masses, and even in semi-permanent waters, but it is most common in the coast. Sclerostena mesoscarcinus forms dominant species in late summer and early autumn, and causes red tides. When red tides occur, the cell density is 4.3×10 3 ~7.2×10 5 mL -1 (Wang et al., 1991). The density of Scutellaria costatum in the critical state of red tide in the Yangtze Estuary is 7×106 m-3, accounting for about 96% of the total phytoplankton (Hong et al., 1992). In my country's Pearl River Estuary in the South China Sea, the Jiulong Rive...

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12R1/89
Inventor 米铁柱何闪英赵丽媛孙静于志刚甄毓
Owner OCEAN UNIV OF CHINA
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