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RNAi fragment of MSL1 and application of RNAi fragment to increase of sensibility of plants to cadmium stress

A sensitive and fragmented technology, applied in the field of RNAi, which can solve the problem that the function of MSL1 gene has not been studied.

Active Publication Date: 2019-07-19
CHINA JILIANG UNIV
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  • Abstract
  • Description
  • 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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  • RNAi fragment of MSL1 and application of RNAi fragment to increase of sensibility of plants to cadmium stress
  • RNAi fragment of MSL1 and application of RNAi fragment to increase of sensibility of plants to cadmium stress
  • RNAi fragment of MSL1 and application of RNAi fragment to increase of sensibility of plants to cadmium stress

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1. Research materials and methods

[0027] 1. Experimental Materials and Growth Conditions

[0028] Wild-type rice (Oryza sativa, Zhonghua 11, ZH11 for short) was used as the control group, and MSL1RNAi transgenic rice positive lines (i4; i8; i11) were used as the experimental group. Seeds of wild-type and transgenic lines were treated in the dark at 28°C for 3 days, and after germination were sown on grids and placed in an incubator at 30°C, 13 hours light, 22°C, 11 hours dark, 70% relative humidity, with half Concentration of Kimura B solution for hydroponic preculture. After 14 days, it was replaced with a complete nutrient solution for cultivation. The solution was changed every three days. The phenotypes of RNAi transgenic lines (i4; i8; i11) and wild-type ZH11 rice plants before and after cadmium treatment were observed and compared, and related physiological and molecular analysis .

[0029] 2. Analysis of gene expression characteristics

[0030] 1)...

Embodiment 2

[0073] Embodiment two, experimental result:

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

[0075] like 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. The expression of MSL1 gene in rice after cadmium treatment was detected by real-time quantitative PCR. 4-week-old rice seedlings were treated with 10 μM CdCl2 for 1-24 h, and there was no significant change in the expression of MSL1 in rice roots. The expression of MSL1 in rice leaves increased significantly after 24 h of cadmium stress.

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

[0077] A 250bp fragment of MSL1 RNAi was amplified by PCR, and the amplified product was subjected to 1% agarose gel electrophoresis. KpnI / SacI double digestion of the cloning vector pMD19-MSL1RNi plasmid yielded fragments with the same sequence size. ...

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Abstract

The invention provides an RNAi fragment. The sequence of the RNAi fragment is as shown in SEQID NO:5. The invention also provides a carrier containing the RNAi fragment. The RNAi fragment or the carrier disclosed by the invention can be used for increasing the sensibility of plants to cadmium stress, breeding transgenic plants of which the sensibility to cadmium stress is increased, adjusting andcontrolling the cadmium stress of the plants, adjusting and controlling cadmium absorption and transfer of the plants, and repairing soil subjected to cadmium pollution, wherein the plants can be oryza sativa.

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 the sensitivity of plants to cadmium stress. Background technique [0002] Rice (Oryza sativa L.) is the most important crop in the world and is the staple food for more than half of the world's population. Due to rapid industrialization and industrial pollution brought about 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 bodies. 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 eliminate 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 a...

Claims

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Application Information

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Patent Type & Authority Applications(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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