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Application of CIPK2 to improvement of mercury resistance/tolerance ability of rice

A kind of use and tolerance technology, applied in the field of plant genetic engineering, can solve the problems of heavy metal stress and other problems, achieve good application prospects, promote growth and development, resist adversity stress or increase yield

Inactive Publication Date: 2018-07-31
LANZHOU UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Currently, the known genes involved in abiotic stresses such as salt, osmotic pressure, and desiccation include AtCIPK6, NtCIPK2, and HbCIPK2, etc., but they have nothing to do with heavy metal stress
[0006] The currently known functions of genes AtCIPK2 and HsCIPK2 are to increase resistance to stresses such as salt, temperature, drought and osmosis, but have nothing to do with heavy metal stress

Method used

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  • Application of CIPK2 to improvement of mercury resistance/tolerance ability of rice
  • Application of CIPK2 to improvement of mercury resistance/tolerance ability of rice
  • Application of CIPK2 to improvement of mercury resistance/tolerance ability of rice

Examples

Experimental program
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Effect test

Embodiment 1

[0067] Embodiment 1, the cultivation of Arabidopsis thaliana, Qinghai-Tibet Plateau annual wild barley and rice

[0068] Arabidopsis wild-type (Col-0) seeds were surface-sterilized (disinfected with 70% ethanol for 10 minutes, then sterilized with 10% NaClO for 30 minutes, and finally rinsed with water for 8 times), cold-treated at 4°C for 3 days in the dark, and placed in 0.5 ×Murashige&Skoog (MS) agar plate (1.5% agar and 1% sucrose, W / V, pH 5.6) cultured. Seedlings were grown in artificial climate culture room (day and night temperature 22 / 20°C, 16 / 8 hour photoperiod, light intensity 80 μmol m -2 the s -1 ), the agar plate was placed vertically.

[0069] The annual wild barley X74 (Hordeum spontaneum C.Koch) and rice Nipponbare (Oryza sativa L.ssp.japonica) seeds on the Qinghai-Tibet Plateau were surface-sterilized with 70% ethanol for 10 minutes, followed by 10% NaClO for 30 minutes, and finally rinsed with water for 8 times . In the dark at 25°C, sterile barley seeds ...

Embodiment 2

[0070] Cloning of embodiment 2, Arabidopsis AtCIPK2 and wild barley HsCIPK2 gene

[0071] The full-length cDNAs of ATCIPK2 (AAF86506; AT5G07070) and OsCIPK2 (ACD7694) were used as probes in the full-length cDNA library of barley (Hordeum vulgare L.; http: / / earth.lab.nig.ac.jp / ~dclust / cgi- bin / barley_pub / ) to search for homologous genes. Based on the highly homologous cDNA sequences to ATCIPK2 and OsCIPK2, the full-length coding sequence (CDS) of HSCIPK2 was amplified by reverse transcription (RT)-PCR. The total RNA of annual wild barley X74 on the Qinghai-Tibet Plateau was isolated with RNeasy Plant Mini Kit (QiAGEN), and then the first-strand cDNA was synthesized with SuperScript III First-Strand Synthesis System (Invitrogen). The amino acid alignment of CIPK2 was analyzed by bioinformatics using Lasergene software (MegAlign). Alignment analysis of amino acid sequences revealed that, similar to OsCIPK2 and ATCIPK2, HSCIPK2 contains two CIPK-specific functional domains, n...

Embodiment 3

[0077] Embodiment 3, the cultivation of AtCIPK2 and HsCIPK2 gene overexpression transgenic Arabidopsis

[0078] The original recipient material of Arabidopsis thaliana transgene was wild type Arabidopsis (Col-0), and the wild type Arabidopsis Col-0 was introduced into wild type Arabidopsis thaliana Col-0 by inflorescence dipping method mediated by Agrobacterium. Please refer to the published master's degree thesis "Functional Analysis of Arabidopsis Clathrin Light Chain" (Zhejiang Normal University, 2012.4).

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Abstract

The invention discloses usage of a CBL interacting protein kinase gene. The CBL interacting protein kinase gene is used for constructing a transgenic plant which has mercury stress resistance / tolerance. The gene is an Arabidopsis thaliana gene AtCIPK2 for constructing transgenic Arabidopsis thaliana with mercury stress resistance / tolerance; or the gene is a Tibet Plateau annual wild barley (Hordeum spontaneum C.Koch) gene HsCIPK2 for constructing the transgenic Arabidopsis thaliana with mercury stress resistance / tolerance; or the gene is used for constructing transgene rice with mercury stressresistance / tolerance.

Description

technical field [0001] The invention belongs to the field of plant genetic engineering. Specifically, the present invention relates to the application of CIPK2 in improving the resistance / tolerance of mercury in rice. Background technique [0002] In order to adapt to the changing living environment, plants must respond to various biotic and abiotic stresses, thus establishing a series of regulatory mechanisms during evolution, such as sensing and decoding various stress signals, signal transduction and expression of stress-related genes. During plant growth and development, Ca 2+ Signaling is a central regulator of the physiological response of plant cells to stress (Dodd et al., 2010). Ca 2+ The change of the signal is first sensed and decoded by the calcium sensor, and then the calcium sensor interacting protein transmits the signal to the downstream, thereby activating the expression of downstream early response genes, and finally causing various physiological respons...

Claims

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Application Information

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IPC IPC(8): C12N15/82C12N15/54A01H5/00A01H6/20A01H6/46
CPCC12N9/1205C12N15/8271
Inventor 潘建伟潘伟槐郑仲仲沈金秋严旭
Owner LANZHOU UNIVERSITY
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