Protein related to plant tillering number and coding gene and application thereof

A technology for encoding genes and plants, applied in the direction of plant gene improvement, application, plant peptides, etc., can solve the problems of little knowledge

Inactive Publication Date: 2010-07-07
INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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Problems solved by technology

[0004] In the model plant Arabidopsis in the past two decades, the synthesis and signal transduction pathways of brassinosteroids have been studied relatively clearly (Gendron, J.M. and Wang, Z.Y. (2007) Multiple mechanisms modulate brassinosteroid signaling.Curr Opin Plant Biol, 10 , 436-441.; Li, J.and Jin, H.(2007) Regulation of brassinosteroid signaling. Trends Plant Sci, 12, 37-41.), however, in the important monocot model plant rice, little is known about

Method used

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  • Protein related to plant tillering number and coding gene and application thereof
  • Protein related to plant tillering number and coding gene and application thereof
  • Protein related to plant tillering number and coding gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Embodiment 1, the acquisition of the protein DLT related to the number of plant tillers and its coding gene

[0046] 1. Obtaining mutants and their phenotypes

[0047] A dwarf mutant was found from a rice mutant library (from the Institute of Genetics and Developmental Biology, Chinese Academy of Sciences). This mutant not only has a semi-dwarf plant height but also has a significantly reduced number of tillers. The phenotype of the mutant is as follows figure 1 shown. Wherein, WT is wild-type rice, and dlt is the mutant rice obtained from the above screening. The plant height of this mutant rice is about 60% of the wild-type rice Zhonghua 11, the number of tillers is about half of the wild-type rice, and the plant type is compact, the leaves and spikes are erect, the plant fertility is reduced, and the ratio of inverted two stem nodes Significantly down-regulated; microscopic section observation found that the cell length of the mutant rice became smaller and the cel...

Embodiment 2

[0052] Example 2, DLT gene is involved in the signal transduction of brassinosteroid

[0053] 1. Comparison of the sensitivity of mutant rice and wild-type rice to brassinosteroids

[0054] According to literature Hong, Z., Ueguchi-Tanaka, M., Umemura, K., Uozu, S., Fujioka, S., Takatsuto, S., Yoshida, S., Ashikari, M., Kitano, H. and Matsuoka , M. (2003) A ricebrassinosteroid-deficient mutant, ebisu dwarf (d2), is caused by a loss of function of a new member of cytochrome P450.Plant Cell, 15, 2900-2910. The method of using the microdrop method to test the wild type Sensitivity of leaf angles of rice and mutant rice screened in Example 1 above to brassinosteroids. After germination of the wild-type rice Zhonghua 11 (from the Institute of Genetics and Developmental Biology, Chinese Academy of Sciences) and the mutant rice screened in Example 1 above at 30°C for two days, seeds with the same germination state were selected and sown in water. , after three days of growth (10 ho...

Embodiment 3

[0066] Embodiment 3, the rice tiller number of trans DLT gene increases

[0067] 1. Construction of expression vector containing DLT gene

[0068] Using the PCR method, using 5'-CCATGGATGTTGGCGGGTTGCTCGTTCTCGT-3' as the forward primer and 5'-AGATCTGATGTTGGCGGGTTGCTCGTTCTCGT-3' as the reverse primer, the open reading frame sequence of the DLT gene was amplified from the rice genomic DNA, namely the sequence listing Bases 771-2621 from the 5' end of sequence 1. Restriction enzymes NcoI and BglII were added to the ends of the forward and reverse primers respectively. After the PCR product was recovered, it was ligated into the pMD18-T vector (purchased from TAKARA Company), and after sequencing to ensure correctness, it was double-digested with NcoI and BglII, and the digested product was ligated into the binary expression vector pCAMBIA1302 (purchased from Cambia, Australia, http: / / www.cambia.org / daisy / cambia / home.html), the recombinant expression vector pCAMBIA1302-DLT contai...

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Abstract

The invention discloses protein relevant to brassinolide signal conduction and a coding gene and the application thereof; the protein is the protein in the 1) or 2); 1) the protein is formed according to an amino acid sequence shown by a sequence 2 in a sequence table; 2) the amino acid residue sequence of the sequence 2 in a sequence table is substituted and / or lost and / or added by one or a plurality of the amino acid residues and is derived by 1) relevant to the plant tillering number. In the invention, DLT gene is cloned by utilizing a rice half-stuntedness little-tillering mutant, and molecule complementation verification is carried out; the result shows that DLT participates in the brassinolide signal conduction process to control the height, tillering and other phenotypes of the rice; after the DLT gene is transferred to the rice, the fact that the tillering number of the transgenosis rice with high DLT gene expression level is obviously increased; the invention has high potential application value on molecular breeding and improving crop production by utilizing genetic engineering measures to regulate plant hormone level.

Description

technical field [0001] The invention relates to a protein related to plant tiller number, its coding gene and application. Background technique [0002] During the first green revolution in the last century, breeding experts bred semi-dwarf rice and wheat varieties, which greatly improved the lodging resistance of crops, successfully resolved the contradiction between high yield and lodging, and thus increased world grain production by about 2.5 Times, made a great contribution to solving the world food problem. However, the molecular nature of the genes responsible for this revolution—the synthesis of the plant hormone gibberellin and the signaling genes SD1 and RHT1—was unclear at the time. Today, we have been able to purposefully explore and discover genes that can affect crop yields, so that they can be used by genetic engineering methods, so that crop yields can be "revolutionarily" improved again. [0003] Brassinolide is an important class of phytohormones involved ...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N15/63C12N15/82C12N5/10C12N1/15C12N1/19C12N1/21C12N15/11A01H5/00
CPCC12N15/8298C07K14/415
Inventor 储成才童红宁金芸刘文波李峰方军朱立煌
Owner INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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