Molecular markers for identifying PHYB wild type and mutant of rice phytochrome gene

Abstract

The invention discloses molecular markers for identifying the PHYB wild type and mutant of a rice phytochrome gene. The nucleotide sequences of the markers related to phyB wild type site detection and phyB mutant site detection are shown as Seq No.1-2 respectively. A primer PHYBF4 / PHYBR1 is used for carrying out PCR amplification, and an amplification product is subjected to BamH I digestion and electrophoretic analysis, wherein if the PCR product can not be digested by BamH I and is a 175bp DNA fragment, the PCR product is the wild type phyB, and if the PCR product can be digested by BamH I, and the digestion products are a 25bp DNA fragment and a 151bp DNA fragment, the PCR product is the mutant phyB1. The molecular markers have the advantages that amplification is easy and fast, and cost is low. The molecular markers can be used for effectively, rapidly and reliably identifying the phyB allelotype, and a powerful tool is provided for subsequent application of the phyB mutant in breeding population screening.

Description

technical field [0001] The invention relates to a molecular marker for distinguishing rice phytochrome gene PHYB wild type and mutant, and belongs to the field of plant biotechnology. Background technique [0002] Higher plants use photoreceptors such as phytochromes, cryptochromes, and phototropins to sense changes in light quality, light intensity, and photoperiod in their growth environment, and regulate their own growth and development. , to adapt to the surrounding environment as much as possible. Among them, phytochrome mainly senses red light (R) and far-red light (FR). The phytochrome gene family in rice includes three members: PHYA, PHYB and PHYC. Among them, the PHYB gene is located on the short arm of the third chromosome of rice and is a single copy. Takano et al. (2005) obtained multiple phyB mutants through gamma ray-induced mutation screening, which were named phyB1-phyB5. Among them, the phyB1 mutant inserts a base C at 1644 bp inside the PHYB gene, there...

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Problems solved by technology

Although Takano et al. (2005) developed a method to identify phyB1 mutations in the article Distinct and Cooperative Functions of Phytochromes A, B, and C in the Control of Deetiolation and Flowering in Rice (The Plant Cell, Vol. However, the amplified fragment of this marker is relatively large (1Kb) and has a high GC content. It needs to be amplified with a high GC PCR buffer, and the PCR amplification time is long, and it is used for restriction enzyme digestion. Dicer NlaIV is a rare restriction endonuclease, which is difficult to purchase and expensive (3.49 yuan / U, NEB Beijing), which increases the cost of identification and is not suitable for large-scale identification; there are often phenomena such as out of stock and out of stock , seriously affecting the smooth progress of the appraisal work

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