Method for screening molecular markers associated with growth of spriodela polyrrhiza roots

A molecular marker, purple-backed duckweed technology, applied in biochemical equipment and methods, microbial determination/inspection, etc., to achieve the effect of fast detection and improved biological treatment efficiency

Active Publication Date: 2017-07-21
ZHEJIANG OCEAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The detection method of the invention has the advantages of rapidity, accuracy, sensitivity, etc., and can intuitively detect the genotypes of different individuals of the rusty spot, so as to quickly obtain the polymorphism map of the genetic variation of the functional gene microsatellite locus of the rusty spot, the microsatellite Markers can be applied to the fields of genetic variation and population genetic diversity analysis of rust spot moth, but there is still room for improvement in the detection speed of microsatellite markers

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  • Method for screening molecular markers associated with growth of spriodela polyrrhiza roots
  • Method for screening molecular markers associated with growth of spriodela polyrrhiza roots

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] The method for screening the growth-related molecular markers of Duckweed root includes clone culture, genomic DNA extraction, design of microsatellite primers, PCR amplification, and data statistics and analysis.

[0017] The composition and concentration of the nutrient solution for plant clone culture are: nitrogen 0.3~0.4g / L, phosphoric anhydride 0.1~0.15 g / L, potassium oxide 0.16~0.23 g / L, magnesium oxide 0.01~0.015 g / L and sulfur 0.012 ~0.02 g / L. The above-mentioned nutrient solution can comprehensively provide the macroelements and trace elements required for the growth of the purple-backed duckweed, the asexual reproduction speed is fast, the probability of gene mutation is low, and a large number of plants with the same gene can be obtained, and the sample size is sufficient.

[0018] The plant clones were cultured indoors in a light incubator for 50-70 days, the culture conditions were 22.5-23.5°C, the humidity was 73-83%, the light intensity was 1800-2200lux,...

Embodiment 2

[0023] The method for screening the growth-related molecular markers of Duckweed root includes clone culture, genomic DNA extraction, design of microsatellite primers, PCR amplification, and data statistics and analysis.

[0024] The composition and concentration of the nutrient solution for plant clone cultivation are: nitrogen 0.32 g / L, phosphoric anhydride 0.12 g / L, potassium oxide 0.19 g / L, magnesium oxide 0.012 g / L and sulfur 0.016 g / L. The above-mentioned nutrient solution can comprehensively provide the macroelements and trace elements required for the growth of the purple-backed duckweed, the asexual reproduction speed is fast, the probability of gene mutation is low, and a large number of plants with the same gene can be obtained, and the sample size is sufficient.

[0025] The plant clones were cultivated indoors for 60 days in a light incubator, the culture conditions were 23°C, humidity 78%, light intensity 2000lux, and light-dark time 16:8h. Under the above cultur...

Embodiment 3

[0030] The method for screening the growth-related molecular markers of Duckweed root includes clone culture, genomic DNA extraction, design of microsatellite primers, PCR amplification, and data statistics and analysis.

[0031] The composition and concentration of the nutrient solution for plant clone cultivation are: nitrogen 0.32g / L, phosphoric anhydride 0.12g / L, potassium oxide 0.2g / L, magnesium oxide 0.013g / L and sulfur 0.017g / L. The above-mentioned nutrient solution can comprehensively provide the macroelements and trace elements required for the growth of the purple-backed duckweed, the asexual reproduction speed is fast, the probability of gene mutation is low, and a large number of plants with the same gene can be obtained, and the sample size is sufficient.

[0032] The number of roots / piece, total root length / piece and average root length / piece were counted for each plant (leaves connected) in different culture lines.

[0033] Microsatellite primers were: Sp25: F: ...

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Abstract

The invention discloses a method for screening molecular markers associated with growth of spriodela polyrrhiza roots. The method comprises steps of culture of clones, extraction of genomic DNA, design of microsatellite primers, PCR amplification as well as data statistics and analysis. The invention has the beneficial effects that the method can be adopted to rapidly obtain a large number of DNA sequences of spriodela polyrrhiza marked by microsatellite primers, is fast in detection of microsatellite markers, and can rapidly obtain a polymorphic map showing high genetic variation of genetic marker gene loci Sp25, Sp30, Sp47 and Sp53 of spriodela polyrrhiza; the number of roots / piece, a total root length / piece and an average root length / piece of spriodela polyrrhiza are significantly associated with loci Sp25, Sp30, Sp47 and Sp53 (P is less than 0.01). The method helps understand influence of genotypic difference on the growth of spriodela polyrrhiza and purification ability of pollutants, and increases biological treatment efficiency of wastewater.

Description

technical field [0001] The invention relates to the technical field of DNA molecular markers, in particular to a method for screening molecular markers related to the growth of duckweed roots. Background technique [0002] Rich, highly heterozygous and good stability, co-dominant inheritance following Mendelian segregation law, easy PCR amplification, etc., have become widely used DNA markers in recent years, and are often used in family pedigree authentication and gene linkage analysis of biological resources , genetic map construction, germplasm identification, population genetic diversity and many other research fields. [0003] In the natural environment, Duckweed basically reproduces asexually, and when the environmental conditions are suitable, one generation can be cloned in 2 days, and the genetic diversity of the population is low. [0004] The prior art provides a variety of detection methods for satellite markers, for example, Chinese Invention Authorized Patent ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 徐娜娜徐嘉俊朱旭辉
Owner ZHEJIANG OCEAN UNIV
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