Method for measuring size of genome of gesneriaceae plant

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...

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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 important role in the study of karst habitat adaptation and resource conservation in my country, the most suitable method for determining its genome size has not yet been clarified.

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