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A protein related to rice yield and its coding gene and application

A rice and gene technology, applied in the field of genetic engineering, can solve the problems that have not existed, and achieve the effect of increasing the yield and reducing the area of ​​arable land.

Active Publication Date: 2021-05-11
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Kapoor et al. (2008) conducted a preliminary bioinformatics analysis on the rice AGO gene family from the perspectives of gene structure, chromosome location, and phylogenetic relationships among members of the gene family, and found that OsAGO2 was affected by low temperature, high salinity, drought, etc. The expression is up-regulated under abiotic stress, and it is speculated that OsAGO2 may have a stress-resistant function (Kapoor et al, 2008), but there is no research on its function in improving rice yield

Method used

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  • A protein related to rice yield and its coding gene and application
  • A protein related to rice yield and its coding gene and application
  • A protein related to rice yield and its coding gene and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] The cloning of embodiment 1 OsAGO2 gene, the construction of overexpression vector

[0042] Take the seedling leaf parts of rice variety ZH11, use TriZolReagent (Invitrogen Company, its article number: 15596026) to extract the total RNA of the leaves, and use a UV spectrophotometer to detect the purity and concentration of the extracted total RNA, and take 1 μg of total RNA as the initial For the reverse transcription reaction, the reverse transcriptase used is AMV (TAKARA Company), and the steps of the reverse transcription reaction refer to the instructions for use of the reverse transcriptase. Using the reverse transcription product as a template, primers are designed according to the cDNA sequence of the gene provided by NCBI (http: / / www.ncbi.nlm.nih.gov / ), and the primer sequence is as follows: (3-3475bp)

[0043] SEQ ID NO: 3

[0044] AGO2F:aaaaACTAGTaatcgaaacgtcgttctgatcg

[0045] SEQ ID NO: 4

[0046] AGO2R:aaaaACTAGTgctgtcgtcagatgaagaac

[0047] PCR amplifi...

Embodiment 2

[0053] Example 2 The AGO2 overexpression vector was transferred into the callus of normal japonica rice variety Zhonghua 11 by using the genetic transformation method mediated by Agrobacterium EHA105.

[0054] The transformation method is described in the following literature: Zhou Lingyan, Jiang Dagang, Wu Hao et al., Establishment of a rice transformation system based on TAC vector. Acta Genetica Sinica, 2005, 32: 514-518. After screening, pre-differentiation, and differentiation, 5 transformed plants of the T0 generation were obtained. The T0 generation was detected by PCR and quantitative PCR, and the positive transformed plants were identified from the DNA level and the RNA level.

[0055] image 3 , after PCR identification, all were positive, and the PCR primer sequences were as follows:

[0056] SEQ ID NO: 7

[0057] Primer 1: 5'-GACAGCGTCTCCGACCTGAT-3';

[0058] SEQ ID NO: 8

[0059] Primer 2: 5'-CATCGCCTCGCTCCAGTCAAT-3';

[0060] The PCR amplification conditions...

Embodiment 3

[0062] Example 3. Identification of Yield Traits of OsAGO2, Os04g0615700 Overexpression Transgenic Plants

[0063] The transgenic plants identified as positive by PCR were cultivated to maturity, and the plant morphology in different periods was observed. The results were as follows: Figures 5a-5e As shown, the obtained transgenic plants of the three homozygous lines (OsAGO2-1, OsAGO2-2, OsAGO2-3) all showed a phenotype of significantly increased grain number per panicle. Compared with the control plants, the transformed plants had obvious changes in the following four yield traits: increased 100-grain weight; increased grain number in the main panicle; increased grain length (Fig. 5d); and increased yield per plant. Figure 6 Shown are 2 transgenic OsAGO2 plants and 1 control plant.

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Abstract

The invention belongs to the field of genetic engineering and relates to the application of rice OsAGO2 and Os04g0615700 genes in improving rice yield. The present invention constructs OsAGO2, Os04g0615700 gene overexpression vectors and transforms them into rice, which can increase the yield of rice from many aspects such as 100-grain weight, number of grains in the main panicle, and grain length, which is of great significance in production practice.

Description

technical field [0001] The invention belongs to the field of genetic engineering and relates to the application of rice OsAGO2 and Os04g0615700 genes in improving rice yield. Background technique [0002] Rice is an important food crop in the world, as the staple food of half of the world's population, and also a model plant for the study of monocots. Increasing rice yield is one of the important ways to maintain world economic stability and promote the development of human society (Hao W and Lin H X., 2010). The agronomic traits composed of the number of effective panicles, the number of grains per panicle and the thousand-grain weight are the three major factors that determine the yield of rice, and are also the key direction of rice breeding and improvement. At this stage, with the increase of population and the decrease of arable land, increasing rice production is still an urgent problem to be solved in the future (Zuo J R and Li J Y., 2014). [0003] High-yield breed...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/82C12N15/11A01H5/10A01H6/46
CPCC07K14/415C12N15/8261
Inventor 庄楚雄郑少燕
Owner SOUTH CHINA AGRI UNIV
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