Protein related to rice ear sprouting period and encoding genes and uses thereof

A technology that encodes genes and heading dates is applied in the fields of application, genetic engineering, and plant genetic improvement to achieve the effects of expanding crop planting area, early maturity, and increasing crop yield

Inactive Publication Date: 2009-01-28
INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This patented technology allows us to modify certain genes on specific plants called ODDs (Oryza sativa L.) which are important components affecting their ability to reproduce during seed germination or flowering stages. By adjusting these genes we aimed at optimizing this process while also allowing earlier crops to be produced with optimal yields.

Problems solved by technology

This patents discusses different aspects related to improving rice cultivars such as drought resistance or increased grain yields due to factors including environmental stressors during their development stages. However, current methods involve modifying specific areas within these crop species through traditional approaches involving crossing over from wild ancestor populations towards domesticated ones, resulting in decreased productivity compared to native seedlings. There is thus a technical problem addressed in this patented text relating specifically to identifying rootage timing genes associated with rice pollinators called olives, particularly those involved in controlling later stage tending behavior.

Method used

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  • Protein related to rice ear sprouting period and encoding genes and uses thereof
  • Protein related to rice ear sprouting period and encoding genes and uses thereof
  • Protein related to rice ear sprouting period and encoding genes and uses thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Embodiment 1, the acquisition of OsDof12 and its coding gene

[0050] 1. Acquisition of the cDNA sequence of a protein related to rice heading stage

[0051] The rice variety Nipponbare was used as the experimental material, and total RNA was extracted from its leaves, which was reverse transcribed into cDNA. Using this cDNA as a template and using 5'-gtgcgaatgaaaagatttcaag-3' and 5'-gatctaaatattcaaatcattatatt-3' as primers, the cDNA sequence of a protein related to rice heading stage was amplified by PCR. The reaction mixture is as follows:

[0052] wxya 2 o

18.3μl

10×PCR buffer

2.5μl

dNTP Mixture (2.5mM)

2.5μl

Taq enzyme (5U / μl)

0.2μl

Primer (10mM)

0.25μl

Primer (10mM)

0.25μl

Template (reverse transcribed cDNA)

1μl

Total

25μl

[0053] PCR reaction conditions: pre-denaturation at 94°C for 4min; then denaturation at 94°C for 45S; annealing at 55°C for 45S; extension at 7...

Embodiment 2

[0062] Embodiment 2, Southern blot analysis

[0063] Genomic DNA of rice variety Nipponbare was extracted, digested with restriction endonucleases EcoR I and Hind III respectively and transferred to the membrane, using the OsDof12 full-length cDNA sequence obtained in the above example 1 from the 670th to 1378th deoxyribose nucleus at the 5' end The nucleotide fragments were used as probes for hybridization analysis. Hybridization results such as figure 2 As shown, where E is the result of Southern blot after EcoRI single-enzyme digestion, and H is the result of Southern blot after Hind III single-enzyme digestion. From figure 2 It can be seen that the hybridization signals of EcoR I and Hind III single-enzyme digestion are all single bands, indicating that OsDof12 exists in the form of a single copy in the rice genome.

Embodiment 3

[0064] Example 3. Analysis of subcellular localization and transcriptional activation domain of OsDof12

[0065] 1. Analysis of subcellular localization of OsDof12

[0066] As a result of analyzing the OsDof12 protein obtained in Example 1 above, no sequence similar to the reported nuclear localization signal was found. In order to analyze the localization of OsDof12 in cells, OsDof12 was digested with Nco I and the GFP-containing vector CaMV35S-sGFP(S65T)-nos3'(pUC18) (see reference Niwa Y , Hirano T, Yoshimoto K, Shimizu M, Kobayashi H.Non-invasivequantitative detection and applications of non-toxic, S65T-type green fluorescent proteinin living plants.Plant J.199918 (4): 445-463.) connected to get Recombinant expression vector 35S-OsDof12-GFP. After no frameshift was verified by sequencing, the recombinant expression vector 35S-OsDof12-GFP was transferred into onion epidermal cells with a gene gun, and the onion epidermal cells transformed with the empty vector CaMV35S-sGF...

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Abstract

The invention discloses a gene related to a heading date of rice as well as a coding protein and application thereof. The gene of OsDof12 related to the heading date of the rice is one of the following ribonucleotide sequences: 1) the sequence 1 in a sequence list; 2) the sequence 2 in the sequence list; 3) the ribonucleotide sequence of the protein sequence of the sequence 3 in a coding sequence list; and 4) a ribonucleotide sequence which has homology of more than 90 percent with the ribonucleotide sequence limited in the sequence 1 or the sequence 2 in the sequence list and codes the proteins with the same functions. The coding protein of the gene related to the heading date of the rice is an amino acid sequence which has the sequence 3 in the sequence list or the amino acid residue sequence of the sequence 3 which is substituted by one or a plurality of amino acid residues as well as deleted or added and has the protein derived from the sequence 3 with the same activity of the amino acid sequence of the sequence 3. The coding gene of OsDof12 related to the protein of the heading date of rice of the invention has important meanings on culturing the early plant variety, enlarging the crop planting area and improving the crop output.

Description

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Claims

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Application Information

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Owner INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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