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Rice OsABCC9 gene and application thereof in cadmium transport

A gene and rice technology, applied in the field of rice OsABCC9 gene and its application in transporting cadmium

Active Publication Date: 2020-12-29
GUANGXI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, no new rice ABC transporter family genes have been found to be cadmium-tolerant in the existing reports

Method used

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  • Rice OsABCC9 gene and application thereof in cadmium transport
  • Rice OsABCC9 gene and application thereof in cadmium transport
  • Rice OsABCC9 gene and application thereof in cadmium transport

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] 1. Plant Material and Growth Conditions

[0029] The materials used in this example are wild-type Nipponbare rice and two OsABCC9 mutant plants.

[0030] Soak the rice seeds in tap water, put them in an incubator at 28°C for 2 days to germinate in the dark, put the seeds on a strainer, and place them in a medium containing 0.5mM CaCl 2 In a 4L plastic container of deionized water, culture was continued for 2 days at 28°C. After the root grows to 2-3cm, it is used for subsequent related experiments.

[0031] 2. Obtaining the complete sequence of the OsABCC9 gene and constructing the phylogenetic tree

[0032] In order to obtain the CDS sequence of OsABCC9, total RNA was extracted from rice roots using the Trizol kit (Lifetechnologies) according to the product instructions. Take 1 μg of total RNA and use the Hiscript II Q RT SuperMix Kit (Vazyme) synthesis kit to synthesize the first cDNA. The obtained cDNA was used as a template to obtain the complete sequence of OsA...

Embodiment 2

[0045] 1. Using the method of PEG-mediated transformation, the plasmid GFP-OsABCC9 or the GFP empty vector obtained in Example 1 were co-transformed with the tonoplast marker plasmid AtTPK1 in rice protoplast cells, and after incubation at room temperature, confocal laser A scanning microscope (TCS SP8; Leica) was used to take pictures, and the results were as follows: Figure 5 shown.

[0046] Depend on Figure 5 As shown in A-D, the GFP empty vector (green fluorescence) localizes in the cytoplasm and nucleus, while the tonoplast marker AtTPK1 (red fluorescence) localizes only on the tonoplast. However, by Figure 5 As shown in E-H, the colocalization results of GFP-OsABCC9 and AtTPK1 revealed that the green fluorescence and red fluorescence can be fused, indicating that OsABCC9 is localized on the tonoplast membrane.

[0047] 2. Phenotype analysis and Cd content analysis of OsABCC9 mutant plants

[0048] In order to study the tolerance of OsABCC9 mutant plants to Cd, 15-...

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Abstract

The invention provides a rice OsABCC9 gene and application thereof, and belongs to the technical field of plant genetic engineering. The nucleotide sequence of the rice OsABCC9 gene is as shown in SEQID No. 1. The rice OsABCC9 gene provided by the invention has the effect of transporting cadmium, and the protein encoded by the rice OsABCC9 gene plays an important role in the cadmium detoxification process of plants. The expression of the OsABCC9 gene in rice roots is highly induced by Cd. Subcellular localization analysis shows that the OsABCC9 is localized on a vacuolar membrane. Plant experiment analysis shows that the OsABCC9 gene has the effect of transporting Cd. The OsABCC9 gene disclosed by the invention can be used as a cadmium transport protein and plays an important role in thecadmium detoxification process of rice.

Description

technical field [0001] The invention belongs to the technical field of plant genetic engineering, and in particular relates to a rice OsABCC9 gene and its application in transporting cadmium. Background technique [0002] Cadmium (Cd) is one of the metal elements that is highly toxic to animals and plants. Excess cadmium in soil can be absorbed by plant roots, thereby inhibiting plant growth and reducing crop yield. Moreover, the cadmium absorbed by the root can be transferred from the root to the aerial part, and then accumulate in the fruit and grain, and enter the human body through dietary intake. Long-term intake of toxic cadmium can cause renal insufficiency, fractures, cancer, etc. many diseases. Therefore, elucidating the transport process and accumulation mechanism of Cd in plants will help to produce and develop low-Cd crops, thereby reducing or reducing the intake of Cd by humans. [0003] As a non-essential element, Cd mainly enters the root cells of plants th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C12N15/82A01H5/00A01H6/46
CPCC07K14/415C12N15/8271
Inventor 夏继星付珊黄晶晶杨广哲
Owner GUANGXI UNIV