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Rice grain gelatinization temperature main control gene ALK and its uses

A technology of gelatinization temperature and main effect gene, applied in the field of protein

Inactive Publication Date: 2005-01-26
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

[0006] The gelatinization temperature of rice, as an important index to evaluate and improve cooking and eating quality, has been reported in recent years, but only limited to phenotype, biochemistry and genetic positioning.

Method used

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  • Rice grain gelatinization temperature main control gene ALK and its uses
  • Rice grain gelatinization temperature main control gene ALK and its uses
  • Rice grain gelatinization temperature main control gene ALK and its uses

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] 1. Rice material

[0029] Rice (Oryza sativa ssp.) high gelatinization temperature indica variety “Shuangkezao” and low gelatinization temperature japonica variety “Cbao”.

[0030] 2. Analyze and target groups

[0031] The homozygous indica variety “Shuangkezao” was crossed with the japonica variety “Cbao”, F 1 A total of 7068 F 2 Individuals, and 1,352 individuals were selected as the positioning group. Take about 2 grams of young leaves from each plant at the seedling stage to extract DNA.

[0032] 3. Localization of ALK gene by SSR, STS, and CAPS markers

[0033] Genomic DNA for gene mapping was extracted from rice leaves using a modified CTAB method. About 100mg of rice leaves were taken, frozen in liquid nitrogen, ground into powder in a small mortar with a diameter of 5cm, transferred to a 1.5ml centrifuge tube to extract DNA, and the obtained DNA precipitate was dissolved in 100μl ultrapure water. Use 1 μl of DNA sample per SSR, STS or CAPS reaction.

[00...

Embodiment 2

[0043] 1. Cloning of ALK full-length gene sequence

[0044] Primers were designed according to the sequence of the early ALK gene of the indica rice bifamily, and high-fidelity PCR amplification and sequencing were carried out in three segments, the clones with completely correct sequences were selected, and they were connected into a 6.86kb gene by using the shared AgeI site and KpnI site fragment, which contains the full-length sequence of 1,766 bases upstream of the start codon ATG and 677 bases after the stop codon TGA.

[0045] 2. Construction of plant transformation vectors

[0046] Construction of ALK sense transformation vector. The cloned full-length ALK gene was inserted into the binary vector pCAMBIA1300 (CAMBIA Company, Australia) to obtain the plasmid pCAMBIA1300-Alk for transformation ( FIG. 4A ).

[0047] Construction of ALK antisense transformation vector. Using the total RNA of indica Shuangke early as a template, the 0.94kb fragment was amplified by RT-PCR...

Embodiment 3

[0049] Rice Genetic Transformation Experiment and Results

[0050] 1. Transformation acceptor material

[0051] Japonica rice: Cbao, Chunjiang 06 Indica rice: Shuangkezao, Minghui 63

[0052] 2. Agrobacterium strains and target genes

[0053] Agrobacterium strain: EHA105

[0054] Target gene: Alk (sense gene), Anti-Alk (antisense gene)

[0055] 3. Culture medium: commonly used medium for rice tissue culture

[0056] 4. Rice callus induction and subculture

[0057] The immature embryos of japonica rice "Cbao" and indica rice "Shuangkezao" were dehulled and sterilized, and inoculated into the medium for inducing callus. After 3 weeks of culture, the callus grew from the scutellum, and the vigorously growing, light yellow, relatively loose embryogenic callus was selected as the recipient for transformation.

[0058] 5. Agrobacterium infection and rice transformation

[0059] Cbao and Chunjiang 06 were transformed with the sense gene expression vector pCAMBIA1300-Alk; Shuan...

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Abstract

The invention provides a main effect gene controlling rice gelatinization point.The nucleotides sequence of the said gene is selected from(1) nucleotide sequence coding SEQ ID NO:2 amino acid sequence (2) sequence which can hybridize with sequence of (1) in strict condition, and at same time code rice gelatinization point controlling function nucleotides sequence. The invention also discloses a protein for controlling rice gelatinization point, plant expression vector containing inventive gene, method for cultivating plant. The method can make plant main storage organ gelatinization point change.

Description

Technical field: [0001] The invention relates to a main gene for controlling the gelatinization temperature of rice and the protein encoded by the gene. The invention also relates to a transgenic plant expression vector for controlling the gelatinization temperature of rice and a method for cultivating plants using the expression vector. The method can change the gelatinization temperature of the main storage organs of plant starch. Background technique: [0002] Gelatinization temperature is an important cooking quality of crops, especially rice. The gelatinization temperature is directly related to the structure of amylopectin in starch granules. The non-reducing ends of the amylopectin branch chains point to the surface of the granules and are arranged radially, forming a 9nm periodic semi-crystalline structure in which crystal layers and amorphous layers are alternately arranged [1] . The crystal layer is composed of adjacent A chains or B chains in the same branch cl...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H1/00A01H5/00C07K14/415C12N5/10C12N15/29C12N15/63
Inventor 李家洋钱前高振宇周奕华
Owner INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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