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Gene related to rice leaf color variation, premature senility and stress tolerance, protein coded by gene and application of protein

A technology of rice and genes, applied in the field of protein, achieves the effect of broad application prospects

Active Publication Date: 2020-11-06
CHINA NAT RICE RES INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] There are few studies on NLRC3 in plants, and there are only a few reports in legumes. For example, in pea (pisum sativum L.), the combination of two symbiotic genes NOD3 and NOD5 promotes nodulation and active nitrogen fixation in plants. and high yield, but there is no relevant report on rice

Method used

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  • Gene related to rice leaf color variation, premature senility and stress tolerance, protein coded by gene and application of protein
  • Gene related to rice leaf color variation, premature senility and stress tolerance, protein coded by gene and application of protein
  • Gene related to rice leaf color variation, premature senility and stress tolerance, protein coded by gene and application of protein

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0071] 1. Rice material:

[0072] Rice (Oryza sativa L.) mutants ypd1-1 and ypd1-2, the original wild material is the japonica rice variety "Nipponbare (NPB)".

[0073] Rice seedling yellow dwarf late stage senescence mutants ypd1-1 and ypd1-2 are from mutations produced by Nipponbare EMS mutagenesis (such as figure 1 shown), the mutant was obtained in Zhejiang Province, China. The agronomic traits compared with the control varieties are as follows:

[0074]

[0075] ** represents a very significant difference between the mutant and the wild type, P<0.01.

[0076] The mutagenesis method is specifically as follows: soak rice seeds with 1.5% EMS for 8 hours, rinse them with clean water and germinate normally; select the senescence mutants ypd1-1 and ypd1-2 in the seedling stage and the later stage of yellow dwarfing in the M1 generation, and use them after multiple generations of stable inheritance. for population construction and gene mapping.

[0077] In the ypd1-1 and ...

Embodiment 2

[0100] Plant Transformation:

[0101] Using the genome of the japonica rice variety "Nipponbare" as a template, primers were designed according to the target gene:

[0102] F1-5'-gctcggtacccggggatcc TTTGAGACCGAGTACAGAGAAGTATGT-3'

[0103] R1-5'-TAGCGACATAGCAGTCCAACTT-3';

[0104] F2-5'-TTGGAGCTGCTATGTCGCTAGG-3'

[0105] R2-5'-CTAGAGTAAATAGACAAAGTACATGTCAGTTAGT-3';

[0106] F3-5'-ACTTTGTCTATTTTACTCTAGTGGCAACTATG-3'

[0107] R3-5'-aggtcgactctagaggatccCAACCAATGACACTTAAAAAGTAAAAAAA-3'.

[0108] The PCR amplification system is: 50 μL PCR reaction system: template DNA 2 μL; 2×PCR buffer 25 μL; 2rnmoldNTP (Roche) 10 μL; KODFX (TOYOBO) enzyme 1 μL; 10 μM PrimerF 3 μL; 10 μM PrimerR 3 μL; ddH 2 O 6 μL; PCR amplification conditions: pre-denaturation at 94°C for 2 min; denaturation at 98°C for 10 s, annealing at 60°C for 30 s, extension at 68°C for 4 min; a total of 35 cycles; extension at 68°C for 10 min, and incubation at 15°C.

[0109] Remarks: The genome of the japonica rice va...

Embodiment 3

[0115] OsYPD1 genome structure and cDNA analysis:

[0116]According to the prediction in Rice Genome Annotation Project (RGAP) (https: / / rapdb.dna.affrc.go.jp / viewer / gbrowse / irgsp1 / ?name=chr01%3A1..50000), OsYPD1 (LOC_Os06g13050) It encodes a peroxidase family protein with 18 exons and 17 introns, the full length of gDNA is 9862bp, and the full length of cDNA is 2532bp. In order to further explore the function of the target gene, we designed two pairs of primers with different combinations (including the sequences available in the sequencing primers) for the amplification of the predicted full-length or partial fragments of cDNA. The newly designed primer sequences are: AnYPD1-GFP1F / R: cttctgcaggagctcggtaccATGGACACTCACCTCGTCGTCG; CTGCACAACATCGTCACCAGAC; AnYPD1-GFP2F / R: AGGACAGTCATATGGAGGGTGG; full-length, so we suspect that the predicted cDNA sequence in RGAP may not be the real coding sequence of OsYPD1. In order to verify this speculation, we used the predicted cDNA sequenc...

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Abstract

The invention relates to plant genetic engineering, and discloses a gene related to rice leaf color variation, premature senility and stress tolerance, a protein coded by the gene and application of the protein. Nucleotide sequences of the gene are shown as SEQ ID No: 1 and SEQ ID No: 2. The rice OsYPD1 gene is cloned by utilizing a map-based cloning technology, and the function of the gene is identified by utilizing a transgenic complementation experiment; and meanwhile, the influence on rice aging and stress tolerance is studied by utilizing a gene coding product, and the product can be usedfor improving the stress tolerance of rice.

Description

technical field [0001] The invention relates to plant genetic engineering, in particular to a gene related to rice leaf color variation, premature aging and stress tolerance, the protein encoded by the gene and its application. Background technique [0002] Pattern recognition receptors (PRR), such as TLR (Toll-like receptor), NLR (nucleotide-binding domain, leucine-rich repeat containing), CLR (C-type lectin receptor), RLR (RIG-I-like receptor) is the host An important part of the immune system, among which NOD (nucleotide oligomerization domain)-like receptors NLRs are pattern recognition receptors related to immunity and inflammation caused by pathogen-associated molecular patterns (PAMPs) and damage / danger-associated molecular patterns (DAMPs). NLRs were classified according to their domain structure and phylogenetic relationship into NLRA / CIITA, NLRB / NAIP, NLRC, NLRP and NLRX1 subfamilies using standard nomenclature. NLRC is one of the largest and most important groups...

Claims

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Application Information

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IPC IPC(8): C12N15/29C07K14/415C12N15/82A01H5/00A01H6/46
CPCC07K14/415C12N15/8261C12N15/8273
Inventor 朱丽钱前陈冬东王小琦曾大力张光恒胡江陈光沈兰张强任德勇郭龙彪董国军高振宇
Owner CHINA NAT RICE RES INST
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