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Gene OsPRO related to content of fragrance of fragrant rice and application of encoding protein of gene OsPRO

A technology related to protein and aroma, applied in the fields of application, genetic engineering, plant genetic improvement, etc.

Inactive Publication Date: 2014-09-10
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, there is no research report on the synthesis process and mechanism of proline to 2-AP

Method used

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  • Gene OsPRO related to content of fragrance of fragrant rice and application of encoding protein of gene OsPRO
  • Gene OsPRO related to content of fragrance of fragrant rice and application of encoding protein of gene OsPRO
  • Gene OsPRO related to content of fragrance of fragrant rice and application of encoding protein of gene OsPRO

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0065] Example 1 Fragrant rice OSPRO Gene acquisition

[0066] 1. Primer design

[0067] According to NCBI (http: / / www.ncbi.nlm.nih.gov / ) about OSPRO Primers were designed for the cDNA sequences of homologous genes, and the primer sequences were as follows:

[0068] OsPRO-F (as shown in SEQ ID NO: 3, the underline is a restriction endonuclease Hin dIII recognition site)

[0069] 5’-ACTTGG AAGCTT CAGGCACAAGTCCGACCTTC-3';

[0070] OsPRO-R (as shown in SEQ ID NO: 4, the underline is a restriction endonuclease Mlu I recognition site)

[0071] 5'-AATATC ACGCGT TTACCGGAGCAGCTGTCTGT-3'.

[0072] 2. PCR amplification

[0073] Using primers OsPRO-F1 and OsPRO-R1 to amplify cDNA from 2-week-old seedling leaves of japonica rice variety Yunjingyou 14 as a template OSPRO Gene. PCR amplification is a routine method.

[0074] After the reaction, the PCR amplification product was subjected to 1% agarose gel electrophoresis, and a DNA fragment of about 1428bp was recovered a...

Embodiment 2

[0075] Example 2 Genetic transformation to identify the function of the target gene

[0076] In this example, by constructing overexpression OSPRO Genetic transgenic rice, study its PRO activity and the change of aroma (2-AP) content in rice, to explore OSPRO Effects of genes on aroma content in aromatic rice and its mechanism.

[0077] 1. Construction of expression vector

[0078] Will OSPRO The gene was cloned into the multiple cloning site of the plant overexpression vector pCAMBIA1380 (purchased from CAMBIA Company, Australia) Hin dIII and Mlu Between the I restriction site, obtain containing OSPRO Plant expression vectors for genes.

[0079] (1) Construction of expression vector

[0080] OsPRO-F and OsPRO-R primers respectively introduced Hind III and Mlu I two enzyme cutting sites, there is also a single on the constitutive vector pCAMBIA1380 Hind III and Mlu I enzyme cutting site, therefore using primers OsPRO-F and OsPRO-R, using the cDNA of japonica...

Embodiment 3

[0126] Example 3 OSPRO Gene Affects the Synthesis Mechanism of Aroma Substance 2-AP in Aromatic Rice

[0127] 1. After extensive research, the inventor has explored OSPRO The specific mechanism of genes affecting the aroma content of fragrant rice, as shown in the attached Figure 4 As shown, among them, OSPRO The OsPRO protein (proline oxidase) encoded by the gene plays a crucial role. Under the action of proline oxidase, proline is converted into pyrroline-5-carboxylic acid, which is the conversion of proline into The first step of 2-AP is also a necessary step.

[0128] 2. The inventor passed the OSPRO Gene RNAi silencing, construction of transgenic rice

[0129] (1) Vector construction

[0130] Primers were designed according to the sequence of SEQ ID NO: 1, and the primer sequences are as follows:

[0131] OsPRO-RNAi-F (as shown in SEQ ID NO: 7, the underline is a restriction endonuclease Bam H I recognition site)

[0132] 5'-ATAGTA GGATCC GACGAAGCTGCCATGGAA...

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Abstract

The invention belongs to the technical field of plant genetic engineering and discloses a gene OsPRO related to content of fragrance of fragrant rice and application of an encoding protein of the gene OsPRO. The cDNA nucleotide sequence of the gene OsPRO is shown in SEQ ID NO:1; and an amino acid sequence of the encoding protein of the gene OsPRO is shown in SEQ ID NO:2. The application refers to application of the gene OsPRO and the encoding protein thereof in improvement of the content of fragrance of fragrant rice. The content of fragrance of fragrant rice in which the gene is introduced is obviously increased, a mechanism for increasing the content of fragrance of fragrant rice through the gene OsPRO is clear, the rice gene OsPRO can be inserted into a multiple cloning site of a plant expression vector to prepare a recombinant expression vector, and a transgenic plant is further constructed. The gene OsPRO has great guide application values for cultivating highly flavored type fragrant rice varieties during production and controlling the content of fragrance of fragrant rice through a transgenic method.

Description

Technical field [0001] The invention is the field of plant genetic engineering technology.More specifically, involving a gene related to the aroma content of fragrant rice OSPRO And its application of protein. Background technique [0002] Xiang rice is usually called fragrant rice. It is a treasure in rice. The incense rice is rich in various trace elements, amino acids and protein.In particular, the eight types of amino acids are rich in content, showing excellent nutritional quality.The high -quality brand rice with high sales price and good rice quality on the market today is mixed with fragrant rice. Fragrant rice not only has edible significance, but also has high economic value (You Qingru and Huang Tingxu. Research and breeding use.Mai Technology, 2003, 20 (3): 30-33).Although the sales price of Xiangmi is more than twice as high as conventional high -quality rice, the consumption of the world's fragrant rice is still increasing year by year, which has attracted great att...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/53C12N9/06C12N15/84A01H5/00
Inventor 田华唐湘如潘圣刚段美洋肖立中钟克友
Owner SOUTH CHINA AGRI UNIV
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