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Detection of south Henan aromatic rice variety by Badh2 gene functional marker and application

A gene function and marker detection technology, applied in the field of genetic engineering, can solve the problems of inaccurate screening and cumbersome screening process, and achieve the effect of accurate and rapid detection and analysis

Inactive Publication Date: 2017-08-01
XINYANG NORMAL UNIVERSITY
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Problems solved by technology

[0005] In view of this, the molecular markers related to rice aroma genes in the present invention may be screened inaccurately in the genetic breeding process of aromatic rice, and the screening process is relatively cumbersome, which is not conducive to accurate and rapid detection and analysis of intermediate materials for aromatic rice breeding on a large scale. To solve the problem, the detection and application of a Badh2 gene functional marker of a fragrant rice variety in southern Henan is provided. The present invention uses 7 gene functional markers developed by the Badh2 gene to detect and analyze the fragrant rice varieties, aiming to clarify the fragrant rice resources. Variation types of aroma genes, and molecular marker-assisted selection of aromatic rice resources for molecular breeding of aromatic rice, laying the foundation for the cultivation of new varieties of high-quality aromatic rice

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  • Detection of south Henan aromatic rice variety by Badh2 gene functional marker and application
  • Detection of south Henan aromatic rice variety by Badh2 gene functional marker and application
  • Detection of south Henan aromatic rice variety by Badh2 gene functional marker and application

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Embodiment 112

[0027] Example 11.2 Gene function markers

[0028]The aroma gene Badh2 has multiple allelic forms in natural populations, and different alleles have different aromas. For the Badh2 gene, the 5'UTR region, the 2nd, 4th, 5th, 7th, 12th, 13th and There were insertions, deletions or single nucleotide variation sites in exon 14, and 7 functional markers of Badh2 gene were developed. Among them, the functional marker FME14 is the cleaved amplified polymorphic sequence marker (The cleaved amplified polymorphic sequence marker, CAPS), and the rest are ordinary PCR molecular markers. See Table 2 for details such as temperature. All the primers were synthesized by Nanjing GenScript Biotechnology Co., Ltd., and preliminary tests showed that the seven Badh2 gene functional markers were stable in the reaction system, and the results were clear and reliable.

[0029] Table 2 Badh2 gene functional marker information

[0030]

[0031]

Embodiment 2

[0032] Example 2 Detection of Badh2 Gene Functional Markers in Fragrant Rice Varieties in Southern Henan

[0033] 1) PCR amplification: DNA of the whole genome was extracted from the leaves of Henan fragrant rice variety and 4 control japonica rice by CTAB method.

[0034] PCR reaction system includes DNA template 2μL, 10×Buffer 2.5μL, 25mmol L -1 MgCl 2 1.5μL, 2.5mmol L -1 dNTP 2.0 μL, forward and reverse primers (12.5 μmol L -1 ) each 1.0μL, 5U·μL -1 Add 0.4 μl of Taq enzyme to 20 μl with sterile water. The reagents used were purchased from Shanghai Sangon Bioengineering Technology Co., Ltd.

[0035] The PCR reaction conditions were pre-denaturation at 94°C for 5 min, denaturation at 94°C for 30 s, annealing at 55°C to 61°C for 30 s, extension at 72°C for 30 s to 90 s, after 32 cycles of reaction, extension at 72°C for 10 min, and storage at 25°C.

[0036] 2) PCR product detection

[0037] The PCR product of functional marker FME14 was first reacted with BslI restrict...

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Abstract

The invention discloses detection of south Henan aromatic rice variety by a Badh2 gene functional marker and application, wherein a method for detecting the south Henan aromatic rice variety comprises the following steps: extracting DNA of a whole genome from leaves of the south Henan aromatic rice variety by adopting a CTAB (Hexadecyltrimethy Ammonium Bromide) method; performing PCR (Polymerase Chain Reaction) amplification by adopting the DNA extracted by using the Badh2 gene functional marker; performing electrophoresis on a PCR amplification product, and observing a result by using a gel-imaging system. The invention further discloses application of the Badh2 gene functional marker in the breeding of the aromatic rice variety. The invention authenticates the mutation type corresponding to the south Henan aromatic rice variety, and a foundation is laid for selecting and culturing high-quality new aromatic rice variety assisted by molecular markers.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and in particular relates to the detection and application of a Badh2 gene function marker of a fragrant rice variety in southern Henan. Background technique [0002] Rice (Oryza sativa L.) is one of the most important food crops in the world. More than half of the world's population and two-thirds of my country's population use rice as a staple food. With the improvement of living standards, people's demand for high-quality rice is increasing. However, the quality traits of rice are complex, mainly including nutritional quality, appearance quality, grinding quality, cooking and eating quality, etc. Among these qualities, consumers often pay more attention to the cooking and eating quality of rice. As a special type of cultivated rice, fragrant rice is regarded as a treasure in rice due to its strong aroma, fragrant rice and high nutritional value. Traditional fragrant rice varietie...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/686C12Q1/6895C12Q2600/13C12Q2600/156C12Q2565/125
Inventor 彭波孙艳芳袁红雨孔冬艳庞瑞华李金涛
Owner XINYANG NORMAL UNIVERSITY
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