Method for measuring size of genome of gesneriaceae plant

A technology of plant genome and Gesneriaceae, which is applied in the field of molecular genetics and evolution research, can solve the problems affecting the accuracy, the genome size measurement method is not clear, etc., and achieve the effect of simple operation and good repeatability

Active Publication Date: 2014-10-01
SOUTH CHINA BOTANICAL GARDEN CHINESE ACADEMY OF SCI
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Problems solved by technology

Therefore, the selection pressure of karst habitats may affect the accuracy of FCM to determine the genome size of plants in this region
Although this plant group plays an im...

Method used

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  • Method for measuring size of genome of gesneriaceae plant
  • Method for measuring size of genome of gesneriaceae plant
  • Method for measuring size of genome of gesneriaceae plant

Examples

Experimental program
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Embodiment 1

[0021] Taking Primula lycopersica as an example, collect 20 mg each of young leaves of Primula lycopersicum, tomato (Solanum lycopersicum cv. 'Stupicképolni rané') and rice (Oryza sativa ssp. japonica, 1C=0.43-0.45pg), With tomato as the main internal reference and rice as the second internal reference, the genome size of rice was re-calibrated according to the genome size of tomato; in a petri dish dripped with 1ml of nuclear extraction buffer LB01, the line leaves were quickly separated with a sharp blade. Chopped primula, tomato and rice, filtered through a 50 μm filter to obtain a suspension of cell nuclei, and kept on ice for later use; added RNA digestion enzyme to a final concentration of 50 μg ml -1 , digest for half an hour; add propidium iodide (PI) for staining, the final concentration is 100 μg ml -1 ; Light-proof staining, staining time is 40 minutes; adopt the flow cytometer (model CyFlow Space) of German Partec company to measure its genome, adopt 488nm laser ex...

Embodiment 2

[0023] This example is basically the same as Example 1, except that the materials are more than 100 species of the Gesneriaceae plant Primulus genus. After testing, the quality of the obtained results is all good, and the CV values ​​are all lower than 5%.

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Abstract

The invention discloses a method for measuring the size of the genome of a gesneriaceae plant. The method comprises the steps of taking fresh tender leaves of the gesneriaceae plant; with a tomato as a main internal reference and rice as a second internal reference, re-correcting the size of the genome of the rice by taking the size of the genome of the tomato as standard; according to the standard of adding 20 milligrams of tender leaves into 1ml of nuclear extraction buffering solution LB01, respectively adding the tender leaves of the target plant and the internal reference species into the nuclear extraction buffering solution LB01, chopping the tender leaves, performing filtration through a filtering net with the aperture of 50 microns to obtain a cell nucleus suspension, and putting the cell nucleus suspension on ice for later use; digesting RNA in the cell nucleus suspension through RNA digestive enzymes till the final concentration of the RNA digestive enzymes is 100 micrograms per ml, and then adding propidium iodide for fluorescence shading dyeing for 40 minutes; and measuring the size of the genome of the target plant through a flow cytometry. The method can measure the accurate sizes of the genomes of various gesneriaceae plants; the CV (Coefficient of Variation) is lower than 5 percent. The method is high in repetitiveness and easy to operate, and is suitable for gesneriaceae botanic research.

Description

Technical field: [0001] The invention belongs to the field of molecular genetics and evolution research, in particular to a method for measuring the genome size of Gesneriaceae plants. Background technique: [0002] Genome size (C-value) refers to the DNA content of haploid cell nuclei. Genome size variation is one of the important driving forces for speciation and evolution. More and more studies have shown that genome size plays an important role in many key biological research fields, such as ecology It has important theoretical and practical significance in the fields of biology, cytology, molecular biology, systematics and evolutionary biology. Theory predicts that the size of the genome is proportional to the complexity of the species, that is, the simpler the species, the smaller the genome; the more complex the species, the larger the genome. More and more studies have shown that even for species belonging to the same phylum, their genome size may vary greatly, whil...

Claims

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Application Information

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IPC IPC(8): G01N15/14
Inventor 王静刘娟康明
Owner SOUTH CHINA BOTANICAL GARDEN CHINESE ACADEMY OF SCI
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