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A biosensor and sensing method for visually displaying the degree of CD stress on plant roots

A biosensing and plant root system technology, applied in botany equipment and methods, biochemical equipment and methods, plant regeneration, etc., can solve the problems of poor visualization results, measurement results errors, poor repeatability, etc., and achieve easy operation , beautiful results, quick and easy method

Inactive Publication Date: 2020-07-24
NANJING AGRICULTURAL UNIVERSITY
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AI Technical Summary

Problems solved by technology

Visually observe whether the rice root system is affected by cadmium stress. The traditional method is to measure the elongation of rice roots under different concentrations of Cd stress, which is time-consuming and laborious, and the elongation of rice roots is affected by external environmental factors (temperature, pH, nutrition, etc.) etc.) have a large impact, resulting in large errors in measurement results and poor repeatability
In addition, the results of the root elongation experiment are mainly presented in the form of data, and the visualization results are poor

Method used

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  • A biosensor and sensing method for visually displaying the degree of CD stress on plant roots
  • A biosensor and sensing method for visually displaying the degree of CD stress on plant roots
  • A biosensor and sensing method for visually displaying the degree of CD stress on plant roots

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Embodiment 1

[0046] (1) The specific steps for the development of a biosensor (root system of OsNAC300p-GUS transgenic rice) that quickly and visually display the degree of Cd stress in plant roots are as follows:

[0047] 1. Extract rice genomic DNA (see method one), detect template DNA quality (OD 260 / 280 =1.80) and concentration (C=860ng / μl);

[0048] 2. Use OsNAC300 promoter-specific primers (Table 1) to amplify the OsNAC300p fragment (see method 2);

[0049] 3. Introduce the OsNAC300p fragment into the pDONR vector by BP reaction (see method 3), and transform Escherichia coli (see method 5);

[0050] 4. Analyze the correctness of the OsNAC300p sequence by first-generation sequencing (sequence table seqNo3, OsNAC300p sequence);

[0051] 5. Integrate the OsNAC300p fragment into pGWB3 (Nakagawa et al., 2007) by LR reaction (see method 4); and transform Escherichia coli to obtain positive colonies, shake the bacteria and extract the plasmid pGWB3-OsNAC300p (see method 6).

[0052] 6. Tra...

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Abstract

The invention relates to a biosensor and sensing method for visually displaying the Cd stress degree of plant roots, and belongs to the technical field of biology. The promoter sequence of an NAC300 gene is connected to a GUS reporter gene, the obtained gene is introduced to a paddy rice genome, the NAC300 gene promoter OsNAC300p senses the Cd stress degree of the external environment, and regulates the expression of downstream GUS reporter gene, and the external Cd stress degree of plant roots is determined according to the shade of GUS dyeing. The biosensor and method are convenient, can identify the Cd stress condition of paddy rice roots in a relatively short time, and no expensive equipment is required. Moreover, the color variation degree is used to display the Cd stress degree of plant roots. The visual manner meets the requirement of a modern research report. Therefore, the biosensor and method have relatively good popularization prospects with the respect relative to business product development and scientific research.

Description

technical field [0001] The invention relates to a biosensor and a sensing method for visually displaying the Cd stress degree of plant roots, belonging to the field of biotechnology. Background technique [0002] Cadmium (Cd) is one of the most toxic heavy metal elements in the environment. Once absorbed by plants and enters the food chain, it will accumulate in the human body, which is extremely harmful to health. According to Xinhua News Agency, the Guangdong Food Safety Commission recently announced the results of random inspections of rice in the Guangzhou market in 2013, showing that 40% of the samples had Cd exceeding the standard. In 2011, the State Council approved the "Twelfth Five-Year Plan" for Comprehensive Prevention and Control of Heavy Metal Pollution, which reflects the country's great importance to the prevention and control of heavy metals. In order to control and alleviate the accumulation of Cd in the edible parts of plants and reduce the potential threa...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/82A01H5/10A01H4/00G01N21/78
CPCA01H4/00C12N15/8205C12N15/8212G01N21/78
Inventor 郑录庆胡淑宝沈振国
Owner NANJING AGRICULTURAL UNIVERSITY
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