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Method for improving content of protein and combined lysine in wheat seeds

A wheat seed and protein technology, applied in botany equipment and methods, horticultural methods, angiosperms/flowering plants, etc., can solve the problems of lysine deficiency and affecting the nutritional value of grains, etc.

Inactive Publication Date: 2010-09-01
JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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Problems solved by technology

[0016] In view of the lack of lysine in wheat seeds, which affects the nutritional value of its seeds, a method for increasing the content of bound lysine and protein in wheat seeds through genetic engineering is provided

Method used

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  • Method for improving content of protein and combined lysine in wheat seeds
  • Method for improving content of protein and combined lysine in wheat seeds
  • Method for improving content of protein and combined lysine in wheat seeds

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Embodiment 1

[0069] A method for improving the content of protein and bound lysine in wheat seeds, which comprises the following steps:

[0070] A. Construction of plasmid pUC18-PCN

[0071] Using pBI2301 as template, 5'-GCC CTGCAG CGTTCAAACATTTGGCAATAAAG-3' (with PstI restriction site) and 5'-GCC AAGCTT CCCGATCTAGTAACATAGATGAC-3' (containing HindIII restriction site) is primer amplification Nos3' sequence, wherein said Nos3' sequence is a known nucleic acid sequence ( http: / / www.ncbi.nlm.nih.gov / ; GenBank: AF485783), 1% agarose electrophoresis, the PCR product is purified and recovered, with PUC18 as the basic plasmid ( figure 2 ), using restriction endonucleases HindIII and PstI to carry out complete double digestion of Nos3' fragment and PUC18 respectively, 1% agarose electrophoresis, and recovering the enzyme-cut fragments, and using T 4 DNA ligase ligated overnight at 16°C (20ul reaction system containing 1×T 4 DNA ligase buffer, PUC18 fragment 0.03pmol, Nos3'fragment 0.3pmol,...

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Abstract

The invention relates to the field of wheat breeding, in particular to a method for improving content of protein and combined lysine in wheat seeds, which guides high chili lysine protein gene (cflr) into conventional wheat by adopting a genetic engineering method and expresses the gene to improve the content of the protein and the combined lysine in the wheat seeds. The method is characterized by comprising the following steps of: constructing an expression vector pAHC25-Cf1r of the high chili lysine protein gene (cflr); guiding the pAHC25-Cf1r into an acceptor wheat embryo cell through a gene gun to perform callus induction and plant regeneration on the wheat embryo cell so as to obtain a T5-generation transgenic homozygous strain; and detecting cflr gene expression abundance in progeny seeds of the T5-generation transgenic homozygous strain, measuring the content of the protein and the combined lysine in the seeds, and screening the strains of which the content of the protein and the combined lysine in the seeds is remarkably improved. Compared with the conventional germ plasm breeding method, the method has the advantages of short breeding cycle, obvious improvement effect and little influence on important agronomic traits.

Description

technical field [0001] The invention relates to the field of wheat breeding. The capsicum high lysine protein gene (cflr) is introduced into conventional wheat and expressed by using a genetic engineering method, so as to increase the content of total protein and bound lysine in wheat seeds. technical background [0002] Wheat is one of the most important food crops in the world. Its grain products not only provide people with calories, but also are an important source of plant protein for people. They can be processed into steamed buns, noodles, bread, biscuits, pastries and other foods. However, the lack of lysine, an essential amino acid in wheat seed protein, affects the complete utilization of its seed protein. The average content of lysine in wheat seed storage protein is only about 0.44% (Duan Min et al., 1997), while the ideal content of lysine announced by the International Food and Agriculture Organization (FAO) is 5.5%, the lysine content of wheat seed protein It...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82A01H4/00A01H5/00
Inventor 孙晓波房瑞马鸿翔余桂红周淼平张旭许玲
Owner JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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