Method for increasing amylose content of plants

A technology for amylose content and plants, applied in the fields of botany and molecular biology, can solve the problems of difficulty in meeting the urgent needs of the starch processing industry and time-consuming problems

Inactive Publication Date: 2014-09-03
SHANGHAI INST OF BIOLOGICAL SCI CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Relying on traditional breeding to change the quality of starch takes a long time, requires a lot of manpower and material resources, and is difficult to meet the urgent needs of the starch processing industry

Method used

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  • Method for increasing amylose content of plants
  • Method for increasing amylose content of plants
  • Method for increasing amylose content of plants

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Experimental program
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Effect test

preparation example Construction

[0101] The present invention has no particular limitation on the preparation method of the polynucleotide fragment, including but not limited to: chemical synthesis method, in vitro transcription method and so on. It should be understood that those skilled in the art can prepare the polynucleotide fragments in various ways.

[0102] construct

[0103]According to the polynucleotide sequence provided by the present invention, it is possible to design a polynucleotide construct that can be processed by the plant to interfere with the gene expression of GBSSI, SBE I or SBE II after being introduced into the plant, thereby affecting GBSSI, SBE I or SBE II gene expression to regulate starch composition in plants. Therefore, the present invention provides an isolated or artificially constructed polynucleotide construct, which can be transcribed by plant cells and expressed as the small molecule RNA in the cells. As a preferred mode of the present invention, the polynucleotide cons...

Embodiment 1

[0215] Embodiment 1, the detection of sweet potato regeneration plant

[0216] 1. Rooting test

[0217] Randomly select 379 strains from the regenerated plants and carry out the rooting test on the medium containing hygromycin, the results show that 92.35% (350) strains can take root and grow normally on the SBM containing 10mg / L hygromycin , when the concentration of hygromycin was increased to 20mg / L for more stringent selection, 349 of these lines could still take root and survive within a week. In view of the fact that high concentrations of hygromycin have a certain inhibitory effect on the growth of transgenic plants, showing that the growth of the plants is slowed down, it is a cheap, convenient, fast and effective way to eliminate false positive strains by screening with 10mg / L hygromycin, and at the same time, it can carry out transgenic The expansion of strains provides enough materials for subsequent experiments.

[0218] 2. PCR detection

[0219] A pair of speci...

Embodiment 2

[0229] Embodiment 2, the quality determination of transgenic novel starch

[0230] 1. Amylose content

[0231] With reference to the standard method for determining the amylose content of rice, the amylose content of the existing sweet potato and common crop starches that have not been transgenic by the inventor was determined, as shown in Table 6. The results showed that the average amylose content of the five sweet potato varieties used for genetic transformation ranged from 28% to 31%.

[0232]Restricted by the limitations and accuracy of the absorbance method, when the amylose content in rice is less than 5%, it can only be qualitatively explained that the amylose content is very low and cannot be accurately measured. The measured value of the amylose content of glutinous rice starch in Table 5 also confirmed this conclusion. Therefore can be defined as novel waxy sweet potato when the sweet potato starch amylose content that records is lower than 5%, the amylose content...

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Abstract

The invention relates to a method for increasing the amylose content of plants. The invention provides enzymes, namely granule-bound starch synthase I (GBSS i), starch branching enzyme I (SBE I) and/or starch branching enzyme II (SBE II) which are applicable to genetic engineering operation in a starch synthesis path and are capable of remarkably changing starch composition of the plants. The invention also provides a novel method capable of favorably adjusting starch in plants, which comprises the following step of reducing the expression of the enzymes to obtain a series of transgenic plants which are different in amylose and amylopectin contents and proportions. The invention also provides a construct or carrier for adjusting the starch composition of plants.

Description

[0001] The present invention is a divisional application of the invention patent application whose application number is CN201110401231.X. technical field [0002] The invention belongs to the fields of botany and molecular biology; more specifically, the invention relates to a method for regulating the content of amylose and amylopectin in plants. Background technique [0003] Starch is the storage material of higher plants, algae and some microbial cells. It is a colorless and odorless white powder and widely exists in various plants. The properties of starch are closely related to its structure. Starch with different amylose content has huge differences in properties, which directly affects its industrial application. In the distribution of starch, it is especially rich in plant seeds (such as rice, wheat, corn) and roots and tubers (such as cassava, sweet potato, potato), which can be used as raw materials for industrial starch production. For example, corn, sweet potato...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/87C12N15/82C12N15/84A01H5/00C12N15/113
Inventor 张鹏杨俊赵姗姗
Owner SHANGHAI INST OF BIOLOGICAL SCI CHINESE ACAD OF SCI
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