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Rice final height-related protein, coding gene thereof and application thereof

An encoding gene and high correlation technology is applied to the rice plant height related protein and its encoding gene and application field, and can solve the problem that the composition of sugar groups and the type of glycosidic bonds cannot be inferred.

Active Publication Date: 2010-01-06
INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, it is not possible to infer the glycosyl composition and glycosidic bond type of the polysaccharide product from the amino acid sequence of a specific glycosyltransferase
The research on the function of cellulose synthase in vitro is still blank

Method used

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  • Rice final height-related protein, coding gene thereof and application thereof
  • Rice final height-related protein, coding gene thereof and application thereof
  • Rice final height-related protein, coding gene thereof and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1, Obtaining the full-length ORF of ND1 gene

[0037] 1. Rice narrow-leaf dwarf mutant nd1

[0038] a) Phenotype analysis of rice narrow-leaf dwarf mutant nd1

[0039] The seeds of the indica rice variety Zhongxian 3037 were irradiated with Co60 rays, and a dwarf and narrow-leaf mutant was screened, named nd1 (narrow leaf and dwarf 1). The basic characteristics of the mutant are: the plant is dwarfed, the leaves are narrowed and semi-involuted, and the number of tillers is increased. nd1 has mutant traits from the seedling stage, such as figure 1 As shown in A, the phenotype is more obvious after the plant matures, such as figure 1 Shown in B. After heading, the plant height of nd1 was about 60% of the wild-type plant height, and each leaf became narrower, shorter in length, and semi-involuted. The light energy utilization efficiency of the PSII system is an important indicator of plant senescence, and the light energy utilization efficiency of the plant le...

Embodiment 3

[0074] Embodiment 3, ND1 functional complementation experiment

[0075] 1) Construction of complementary expression vector pCsld4

[0076] BAC OsJNBa0027H05 (purchased from the National Gene Research Center of the Chinese Academy of Sciences) was digested with XbaI and HpaI to obtain a full-length DNA sequence containing 3005 bases upstream of the start codon ATG of ND1 and 3360 bases after the stop codon TGA. The long-sequence DNA fragment (10268bp) was cloned between the XbaI and SmaI recognition sites of pCAMBIA1300 (CAMBIA, Canberra, Australia), and the complementary expression vector pCsld4 was constructed. The constructed complementary vector pCsld4 was digested with BamHI, the coding region of ND1 gene was removed, and the 5'promoter region and 3'regulatory region of the gene were retained, thus the complementary control vector pCsld4T was constructed.

[0077] 2) ND1 function complementary

[0078] Complementary expression vector pCsld4 and complementary control vect...

Embodiment 4

[0080] Embodiment 4, cultivating the paddy rice that dwarf stalk and / or tiller number increase

[0081] 1) Construction of ND1 RNA interference vector

[0082] Total RNA was extracted from the leaves of rice Zhongxian 3037, and the first-strand cDNA was synthesized by reverse transcription using Oligo(dt)-18 as a primer and the extracted total RNA as a template. Using this cDNA as a template, primer 3(5'- ctgcag cattcacatgagagatcctc-3, the underline is the PstI recognition site) and primer 4 (5'- ctgcag agaagttgagcacgtggccg-3, the underline is the PstI recognition site), carry out the PCR amplification reaction, and the reaction conditions are as follows:

[0083] Reaction volume 50μl: template, 5μl (5ng); forward primer primer 3 and reverse primer primer 4, each final concentration 0.2μM; dNTP, each final concentration 200μM; LA Taq DNA polymerase, 2.5U; 10×Taq DNA polymerase Enzyme buffer, 5 μl; make up to 50 μl volume with double distilled water.

[0084] The reaction...

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Abstract

The invention discloses a rice final height-related protein, a coding gene thereof and application thereof. The protein is an a) protein or b) protein, wherein the a) protein is formed by an amino acid sequence expressed as the No.2 sequence in a sequence list; and the b) protein is formed by substituting and / or reducing and / or adding one or more amino acids to the amino acid sequence expressed as the No.2 sequence in sequence list, related to the final height of rice and derived from the a) protein. The coding gene of the protein particularly is a 1) gene, 2) gene or 3) gene: the 1) gene is a DNA molecule having a amino acid sequence expressed as the No.1 sequence in the sequence list; under strict conditions, the 2) gene can be crossed with the DNA sequence defined by the No.1 sequence in the sequence list and code the DNA molecule of the rice final height-related protein; and the 3) gene has over 90 percent homology with the 1) gene, and can code the DNA molecule of the rice final height-related protein. The rice final height-related protein and the coding gene thereof have important application values in molecular breeding work.

Description

technical field [0001] The invention relates to a rice plant height-related protein and its coding gene and application. Background technique [0002] The cell wall is a complex mainly composed of cellulose, hemicellulose, lignin, pectin, and protein (Delmer, 1999). The cell wall determines the size and shape of the cell, and provides mechanical support and elasticity for the cell, making the cell immune. attacked by pathogenic bacteria. Studies have shown that cellulose synthase (Ces) is a large class of glycosyltransferases responsible for the synthesis of cellulose in the cell wall. The first cloned plant cellulose synthase is CesA (Cellulose synthase A) of cotton (pear, 1996). It has been proved that CesA exists in the form of a gene family in plants, and multiple CesA genes are involved in the process of cellulose synthesis. work together. Protein structure analysis shows that a stretch of amino acids at the N-terminal of cellulose synthase CesA can form a special do...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/82C12N15/11C12N5/10C12N1/15C12N1/19C12N1/21A01H1/00A01H5/00
Inventor 程祝宽李明崔家骏唐丁
Owner INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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