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Application of the rice glume sustained growth gene promoter nsgp

A promoter and gene technology, applied in genetic engineering, application, recombinant DNA technology, etc., can solve problems such as lemma protection

Active Publication Date: 2021-08-17
SOUTHWEST UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, ectopic expression of OsMADS1 / LHS1 can also cause lemmas (Wang et al.2016); ASP1 gene mutations can also lead to lemmas (Yoshida et al., 2012)

Method used

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  • Application of the rice glume sustained growth gene promoter nsgp
  • Application of the rice glume sustained growth gene promoter nsgp
  • Application of the rice glume sustained growth gene promoter nsgp

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] The cloning of embodiment 1 NSG promoter and the construction of GFP reporter gene expression vector

[0027] In this example, the expression pattern of the NSG promoter was detected by the reporter gene green fluorescent protein (GFP). A fusion expression vector (PNSG::NSG-GFP) with NSG gene promoter, gene coding frame sequence and GFP green fluorescent protein gene was constructed.

[0028] The specific construction process: first, the GFP sequence contained in the PA7 plasmid (purchased by the BioVector plasmid vector strain cell gene preservation center) was used as a template to amplify the GFP coding frame sequence (primers GFP-F and GFP-R, the sequences are shown in Table 1), And connected to the plant expression vector PCAMBIA1300 (BioVector plasmid carrier bacterial strain cell gene preservation center purchases), constructs the PCAMBIA1300-GFP intermediate vector, then amplifies the LOC_Os04g36650 gene promoter and coding frame sequence (comprising the NSG gen...

Embodiment 2

[0033] Embodiment 2 Acquisition and observation of transgenic plants

[0034]In this example, the fusion expression vector with the NSG gene's own promoter, gene coding frame sequence and GFP green fluorescent protein gene was sent to Wuhan Boyuan Biotechnology Co., Ltd. for genetic transformation experiments, and it was transformed into the nsg1-1 mutant Plants, obtained transgenic plant tissue culture seedlings, cultured them in a laboratory constant temperature light incubator until they were strong, transplanted them into the field, and observed their phenotypes after they matured. In this embodiment, a total of 20 transgenic plantlets were obtained. Among these plants, 4 plants still retained slightly elongated glumes or para-glucules. Among the remaining 8 transgenic plants, para-glucules, Huying and Yinghua all return to normal ( image 3 ). The results proved that these 8 transgenic plants can be used to detect the reporter gene, and can truly reflect the expression ...

Embodiment 3

[0035] Example 3 NSG1-GFP fusion protein expression observation

[0036] In this example, the transgenic plants whose phenotype returned to normal were selected, and the young inflorescences were taken at the booting stage, and the NSG-GFP fusion was observed under different magnifications of the OLYMPUS fluorescent macroscopic stereo microscope (MVX10) and the OLYMPUS laser confocal scanning microscope (FV100). Which parts or organs of the inflorescence the protein is expressed in. The young ears of 12 plants can see the expression of obvious green fluorescent signal, which proves that these plants are indeed transgenic plants that have been transferred into the pNSG-GFP fusion vector. The transgenic plants whose phenotype returned to normal were used as materials, and the GFP signal was observed by ordinary fluorescence microscope and laser confocal fluorescence microscope to explore the expression pattern of NSG gene. Ordinary fluorescence microscope observation revealed t...

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Abstract

The invention belongs to the technical field of molecular biology, and in particular relates to the application of NSG promoter of rice glume sustainable growth gene. The invention provides the use of the NSG promoter of the rice glume sustainable growth gene in rice breeding. The NSG gene promoter NSGP provided by the present invention can be specifically expressed in the early inflorescences, subgliats and glumes of rice, and can be used to study and improve the development of plant glumes. The invention provides a new option for improving the breeding of gramineous grain crops.

Description

technical field [0001] The invention relates to the technical field of rice development, in particular to the application of NSGP promoter of rice glume sustainable growth gene. Background technique [0002] Rice is one of the most important food crops in the world, and it is also an ideal model crop for the study of monocotyledonous plant developmental biology. The yield of rice is composed of three factors: the number of panicles per unit area, the number of solid grains per panicle, and the weight of thousand grains. The number of grains per panicle is one of the important factors in the yield of gramineous grain crops such as rice. It is determined by the number of spikelets per panicle and the seed setting rate. However, the seed setting rate is easily affected by environmental factors and it is difficult to improve it. Therefore, increasing the number of grains per panicle is the consensus of domestic and foreign researchers engaged in research on rice breeding. The n...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/113C12N15/82A01H5/02A01H6/46
CPCC07K14/415C12N15/8231C12N15/8261C12N15/827
Inventor 李云峰庄慧何光华张婷桑贤春杨正林
Owner SOUTHWEST UNIV
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