A method for molecular marker identification of high nitrogen use efficiency genes in rice

Abstract

The invention discloses a molecular marker identification method of a paddy high nitrogen use efficiency gene, and belongs to the technical field of bioengineering. NRT1.1B is a high nitrogen use efficiency gene. According to the molecular marker identification method, a molecular marker composed of four primers is designed based on mononucleotide difference of wild type NRT1.1B gene with mutant type nrt1.1b gene, and is added into a same tube PCR reaction system so as to realize PCR amplification of DNA of different paddy plants; if characteristic strips 368bp and 243bp are obtained via amplification at the same time, the DNA is a homozygote containing NRT1.1B gene; if characteristic strips 368bp and 177bp are obtained via amplification at the same time, the DNA is a homozygote containing nrt1.1b gene, and if characteristic strips 368bp, 243bp, and 177bp are obtained via amplification at the same time, the DNA is a heterozygote containing NRT1.1B gene and nrt1.1b gene. The molecular marker identification method is rapid, convenient, and accurate in gene type detection of NRT1.1B gene; operation is simple; cost is low; the molecular marker identification method is extremely suitable for target gene identification of large-scale paddy materials; and important reference is provided for paddy resource gene identification and molecular marker-associated selective breeding.

Description

technical field [0001] The invention relates to a molecular marker identification method of rice high nitrogen utilization efficiency gene (NRT1.1B), which belongs to the field of biotechnology engineering and is specially used for rice resource identification and marker-assisted breeding of rice high nitrogen utilization efficiency NRT1.1B gene. Background technique [0002] Rice is one of the most important grain crops in my country. The national rice planting area accounts for about 30% of the grain crop area, and the output is close to half of the total grain output (Zhang Jiayu, Jilin Agricultural University, 2008), which provides more than 60% of my country's population. rations. According to botanical classification, rice varieties grown in my country belong to Asian cultivated rice. There are two subspecies of Asian cultivated rice, namely the indica subspecies and the japonica subspecies (Yang Youxin et al., Science Bulletin, 2009, (15): 2212-2218), and they all sho...

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Problems solved by technology

[0004] The technical problem of the present invention: the present invention aims at identifying the NRT1.1B gene by sequencing, which is expensive, complicated and time-consuming. The difference between the existing bases T and C, combined with the principle of tera-primer ARMS-PCR primer design, developed four specific primers for the gene and amplified rice genomic DNA in the same tube PCR reaction to accurately and quickly identify the NRT1.1B gene It provides an important reference for resource screening and identification of corresponding target genes and molecular marker-assisted selection breeding

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