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New method for formulation of full waxy starch wheat germplasm

A technology for waxy wheat and wheat, applied in the field of cultivating whole waxy wheat and obtaining new germplasm of whole waxy wheat, which can solve problems such as difficult commercial application and poor agronomic traits

Inactive Publication Date: 2012-10-24
INST OF CEREAL & OIL CROPS HEBEI ACAD OF AGRI & FORESTRY SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Waxy or part of waxy wheat starch can improve the quality of noodle processing, but the waxy wheat in my country is all bred through the aggregation of the three deletion sites of GBSSⅠ, which is poor in agronomic properties and cannot be directly used for noodle processing. Difficult for widespread commercial application

Method used

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  • New method for formulation of full waxy starch wheat germplasm
  • New method for formulation of full waxy starch wheat germplasm
  • New method for formulation of full waxy starch wheat germplasm

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Example 1. Obtaining of transgenic wheat and detection of its Wx protein expression

[0048] 1. Construction of RNAi expression vector pCAMBIA3301-GBSS

[0049] Firstly, the pCAMBIA3301 vector was modified as follows, pTCK303 vector was digested with KpnI and SpeI, and the digested product was purified and recovered by 1% agarose gel electrophoresis. Ligate a 478bp rice intron sequence excised by enzymes to the pMD19-T vector (Takara (Dalian) Company, D102A), name the resulting recombinant plasmid pMD19-T-intron, and transform it into DH5α Competent cells, pick positive clones and send them to the company for sequence determination. After the sequencing results were correct, the pMD19-T-intron and pCAMBIA3301 vectors were digested with KpnI and SpeI, and the rice intron sequence was connected to the multiple cloning site between the CaMV35S promoter and the NOS terminator of the pCAMBIA3301 vector. details as follows:

[0050] Using the GBSSⅠ cDNA (GenBank: Y16340.1)...

Embodiment 2

[0075] Embodiment 2, the starch quality characteristic detection of transgenic wheat progeny

[0076] 1. Determination of grain starch and protein content of transgenic wheat offspring

[0077] T obtained in Example 1 3 The seeds of the transgenic wheat Wax-1-3-1 were used as experimental materials, and the starch quality characteristics were tested. The specific operation is as follows: the grain starch content is determined according to the polarimeter method of GB / T1568521995. The content of amylose in wheat grains was determined by dual-wavelength spectrophotometry, the main wavelength for the determination of amylose content was 580nm, and the reference wavelength was 500nm. The total nitrogen content of the grain was measured by an automatic azotometer (Kjeldahl method) made in Switzerland, and the total nitrogen content of the grain was multiplied by 5.7 to be the protein content of the grain. Simultaneously with wild-type Konon 199 seeds and T 3 Transgenic wheat tr...

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Abstract

The invention discloses a method for cultivation of full waxy wheat. The method for cultivation of full waxy wheat provided in the invention includes the following step of: inhibiting the expression of GBSS (granule-bound starch synthase) I gene in target wheat to obtain the full waxy wheat. The inhibition of expression of the GBSS I gene in the target wheat is realized by transferring a DNA fragment as shown in formula "SEQforward-X-SEQbackward" into the target wheat. Specifically, the SEQforward is the top 288 nucleotides in sequence 1 of a sequence table; the SEQbackward is in reverse complement with the SEQforward; the X is a spacer sequence between the SEQforward and the SEQbackward, and in sequences, the X is not complementary with and the SEQforward and the SEQbackward. Experiments prove that the wheat GBSS I gene can be silenced through a transgene silencing technology (RNAi) to obtain a GBSS I gene expression-inhibiting full waxy wheat new germplasm, and the transgenic material has excellent comprehensive agronomic characters, thus being able to be directly used for selective breeding of wheat low-amylose new strains (varieties) or be used as a patent material of breeding.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a method for cultivating full-waxy wheat, in particular to a method for silencing the wheat granule-bound starch synthase gene (GBSSⅠ) through the transgene silencing technique (RNAi), thereby obtaining a new variety of full-waxy wheat qualitative method. Background technique [0002] The genetic improvement of wheat starch quality is called "biotechnology to turn starch into gold" abroad, and waxy wheat is one of the research hotspots. [0003] The grain starch of whole waxy wheat does not contain amylose or contains very little (usually less than 1%). The starch viscosity characteristics of its flour are very different from those of ordinary wheat. It is a new wheat material and can be widely used in food It has great development and application value in industries such as processing, specialty food development, processing raw materials related to starch, papermaking, textiles, and ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82A01H5/00A01H1/02C12N15/113C12N15/63C12N1/21C12N5/10
Inventor 李辉刘茜胡梦芸张颖君胡亚萍
Owner INST OF CEREAL & OIL CROPS HEBEI ACAD OF AGRI & FORESTRY SCI
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