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The RNAi fragment of Msl1 and its application in enhancing the sensitivity of plants to cadmium stress

A sensitive and fragmented technology, applied in the field of RNAi, can solve the problems of MSL1 gene function without any research, achieve good application value, repair soil cadmium pollution, and improve sensitivity

Active Publication Date: 2021-10-19
CHINA JILIANG UNIV
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  • Claims
  • Application Information

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Problems solved by technology

However, the function of MSL1 gene in rice development and resistance has not been studied

Method used

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  • The RNAi fragment of Msl1 and its application in enhancing the sensitivity of plants to cadmium stress
  • The RNAi fragment of Msl1 and its application in enhancing the sensitivity of plants to cadmium stress
  • The RNAi fragment of Msl1 and its application in enhancing the sensitivity of plants to cadmium stress

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Embodiment one, research material and method

[0027] 1. Experimental materials and growth conditions

[0028] Wild-type rice (Oryza sativa, Zhonghua 11, referred to as ZH11) was used as the control group, and MSL1 RNAi transgenic rice positive lines (i4; i8; i11) were used as the experimental group. Seeds of wild-type and transgenic strains were treated in the dark at 28°C for 3 days, and after germination, they were sown on the grid and placed in an incubator at 30°C, 13 hours of light, 22°C, 11 hours of darkness, and a relative humidity of 70%. Concentration of Kimura B solution for hydroponic pre-cultivation. After 14 days, they were replaced with complete nutrient solution for cultivation. The solution was changed every three days. After being cultured for 8 weeks and treated with 60 μmol / L CdCl2 for 14 days, observe and compare the phenotypes of RNAi transgenic lines (i4; i8; i11) and wild-type ZH11 rice plants before and after cadmium treatment, and conduct re...

Embodiment 2

[0073] Embodiment two, experimental result:

[0074] 1. Analysis of expression characteristics of MSL1 gene:

[0075] Such as Figure 4 As shown, the spatiotemporal specific expression of MSL1 gene was detected by GUS staining of MSL1-GUS transgenic rice. It was found that MSL1 was highly expressed in anthers and nodes. Detection of MSL1 gene expression in rice treated with cadmium by real-time quantitative PCR. The 4-week-old rice seedlings were treated with 10 μM CdCl2 for 1-24 hours, and the expression level of MSL1 in rice roots did not change significantly. After 24h of cadmium stress, the expression of MSL1 in rice leaves increased significantly.

[0076] 2. Construction of MSL1 RNAi vector and transgenic rice:

[0077] A 250bp fragment of MSL1RNAi was amplified by PCR, and the amplified product was subjected to 1% agarose gel electrophoresis. KpnⅠ / SacⅠ double enzyme digestion of the cloning vector pMD19-MSL1RNi plasmid obtained fragments with the same sequence siz...

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Abstract

The present invention provides an RNAi fragment whose sequence is shown in SEQ ID NO:5. The invention also provides a vector containing the RNAi fragment. The RNAi fragment or carrier of the present invention can be used to improve the sensitivity of plants to cadmium stress, cultivate transgenic plants with increased sensitivity to cadmium stress, regulate plant cadmium stress, regulate plant absorption and transport of cadmium, and be used for soil cadmium pollution restoration, wherein The plant can be rice.

Description

technical field [0001] The invention relates to the technical field of RNAi, in particular to an RNAi fragment of MSL1 and its application in improving plant cadmium stress sensitivity. Background technique [0002] Rice (Oryza sativa L.) is the most important crop in the world, and more than half of the world's population uses rice as a staple food. Due to rapid industrialization and industrial pollution caused by industrial activities, extensive application of heavy metal cadmium-containing fertilizers or pesticides, mining and human activities, a large amount of cadmium is released into soil, air and water. In Asian countries including China, up to 50% of cadmium intake comes from rice and its products. Once heavy metals enter the soil-crop system, it is difficult to get rid of them, so the problem of cadmium pollution in rice has attracted great attention from relevant departments in various countries around the world. Cadmium not only poisons rice and reduces its yiel...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/113C12N15/82A01H5/00A01H6/46B09C1/10
CPCB09C1/105C12N15/1137C12N15/8218C12N15/8271C12N2310/14
Inventor 丁艳菲朱诚孙骏威丁丽红王飞娟蒋晗江琼
Owner CHINA JILIANG UNIV
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