Method for improving starch quality in rice

A starch and quality technology, applied in the field of plant genetic engineering in molecular biology, can solve problems such as the reduction of mature mRNA, affect the amylose content, and reduce the accumulation of GBSS, so as to achieve short improvement cycles, promote human health, and improve quality. Effect

Inactive Publication Date: 2018-06-22
袁米农业科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Among them, compared with Wxa, because the base at the 5' end of the first intron cleavage site is mutated from G to T, the cleavage efficiency i

Method used

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  • Method for improving starch quality in rice
  • Method for improving starch quality in rice
  • Method for improving starch quality in rice

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] Embodiment 1: the construction of recombinant vector and the establishment of transformed Agrobacterium:

[0064] (1), the plasmid Puc18-Glb with the glutenin GluB-1 promoter (Glb) (according to EMBL sequence X5433314 carries out PCR amplification and clones on the Puc18 vector (Takara company product), the method refers to Lee et al, 2001, Constitutive and seed-specific expression of a maize lysine) was digested with EcoRI and Hind III, separated and recovered the target fragment, and connected with pCAMBIA1300 (Cambia company product) after double digestion with EcoRI and Hind III to form an intermediate vector with T4 ligase, Then the cDNA fragment of SBE1 gene (for the sequence see figure 1 ) was digested with KpnI and SacI, separated and recovered the target fragment, connected with the intermediate vector with KpnI and SacI and connected with T4 ligase to form a recombinant vector, the above restriction enzymes (EcoRI, Hind III, KpnI and SacI) and T4 ligase were ...

Embodiment 2

[0068] Example 2: Agrobacterium-mediated genetic transformation:

[0069] (1), induction:

[0070] The mature rice variety Wuyujing No. 7 seeds were shelled, then treated with 70% ethanol by volume for 1min, and the surface of the seeds with 2% sodium hypochlorite (NaClO) was sterilized for 15min; the seeds were washed 4-5 times with sterilized water; The seeds were placed on the japonica rice induction medium; the inoculated medium was placed in a dark place for 4 weeks at a temperature of 26°C.

[0071] (2), Succession:

[0072] Select bright yellow, compact and relatively dry embryogenic calli, put them on the subculture medium of japonica rice in the dark, and cultivate them at 26°C for 2-3 weeks.

[0073] (3), pre-cultivation:

[0074] Select compact and relatively dry embryogenic calli, put them on the japonica rice pre-medium in the dark, and cultivate them for 4-5 days at 26°C.

[0075] (4), Agrobacterium culture:

[0076] Pre-cultivate Agrobacterium strain S1 on ...

Embodiment 3

[0088] DNA was extracted from leaves of transgenic plants, and polymerase chain reaction (PCR) was performed. PCR program: 94°C pre-denaturation for 5 minutes; 33 cycles (94°C denaturation for 1 minute; 55°C annealing for 1 minute; 72°C extension for 2 minutes, 72°C extension for 7 minutes; ).

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Abstract

The invention discloses a method for improving starch quality in rice. The method comprises the following steps: A, constructing a recombinant vector; B, establishing transformed agrobacterium; C, obtaining an independent transformed plant; D, performing PCR amplification; E, selecting a homozygous single plant; and F, performing phenotypic selection on the selected homozygous single plant. The step further comprises A, recombinant carrier construction and transformed agrobacterium establishment: enabling cDNA of an SBE1 gene and a rice grain specific glutelin GluB-1 promoter to be in enzyme-cut connection with an expression vector pCAMBIA1300 so as to construct the recombinant carrier. The method disclosed by the invention adopts a transgenic method to improve the rice grain quality, is short in improvement cycle and high in efficiency and can be directly applied to the improvement practice of other crop qualities.

Description

technical field [0001] The invention relates to the technical field of molecular biology plant genetic engineering, in particular to a method for improving the quality of rice starch. Background technique [0002] Rice is one of the most important food crops in the world, and about 1 / 2 of the population in the world takes it as staple food. Rice breeding in my country has long focused on increasing yield, while ignoring the improvement and improvement of quality (Huang Fasong et al., 1998). With the development of society, industry and agriculture have higher and higher requirements for rice quality, and quality breeding has become a very important aspect of rice breeding. [0003] The main component of rice endosperm is starch, so the content and properties of starch directly affect the quality of rice. Starch is stored in semicrystalline plastid amyloplasts composed of highly ordered glucose polymers. [0004] According to its structure, starch can be divided into amylo...

Claims

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Application Information

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IPC IPC(8): C12N15/82C12N15/54A01H5/10A01H6/46
CPCC12N9/107C12N15/8205C12N15/8234C12N15/8245C12Y204/01018
Inventor 张国栋刘佳音张佩邹丹丹吴洁芳王晶罗碧米铁柱
Owner 袁米农业科技有限公司
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