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Function and application of transcription inhibitor LIP1 for regulating and controlling rice yield

A yield and rice technology, applied in application, plant products, genetic engineering, etc., can solve problems such as unclear molecular mechanism and affecting the formation of rice yield

Inactive Publication Date: 2021-05-04
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In summary, LAX1 affects the formation of rice yield by regulating the formation of panicle branches
[0004] LAX1 plays an important role in regulating the formation of panicle branches and affecting yield, but the molecular mechanism of LAX1 function is still unclear

Method used

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  • Function and application of transcription inhibitor LIP1 for regulating and controlling rice yield
  • Function and application of transcription inhibitor LIP1 for regulating and controlling rice yield
  • Function and application of transcription inhibitor LIP1 for regulating and controlling rice yield

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Example 1: Identification of the LIP1 gene

[0044] 1. LIP1 is the interacting protein of LAX1

[0045] The present invention uses yeast two-hybrid technology to identify a protein interacting with LAX1, which is named LAX1Interacting Protein1 (abbreviated as LIP1). The present invention constructs five yeast vectors, namely pGBK-LAX1 (1-159aa), pGBK-LAX1 (1-40aa), pGBK-LAX1 (31-99aa), pGBK-LAX1 (△bHLH) and pGAD-LIP1. Among them, pGBK-LAX1(1-159aa), pGBK-LAX1(1-40aa), pGBK-LAX1(31-99aa); pGAD-LIP1 four yeast vectors are constructed as follows: design primers BK-LAX1-1F / BK- LAX1-159R, BK-LAX1-1F / BK-LAX1-40R, BK-LAX1-31F / BK-LAX1-99R; AD-LIP1-F / AD-LIP1-R (see Table 1 for primer sequences), wild type "Zhonghua No. 11" variety (conventional rice variety cultivated by the Institute of Crop Science, Chinese Academy of Agricultural Sciences, referred to as ZH11) cDNA of the young panicle (2mm-5mm) was used as a template to amplify the target fragment, and TA cloned it into pE...

Embodiment 2

[0055] Example 2: LIP1 regulates panicle branching and spikelet formation in rice

[0056] 1. Overexpression of LIP1 leads to reduction of lateral spikelets

[0057] In order to analyze the function of LIP1 gene in the development of panicle lateral meristem, genetic analysis was carried out by creating materials with overexpression of LIP1. Design primers LIP1(FL-cDNA)-OX-F / LIP1(FL-cDNA)-OX-R (see Table 2 for the primer sequence), use the ZH11 young ear (2mm-5mm) cDNA as a template to amplify the full length of LIP1 cDNA fragment, TA cloned into pEASY-T3 (see Figure 11 , purchased from Beijing Quanshijin Biotechnology Co., Ltd.) and verified by sequencing; pEASY-T3 with the correct sequence was digested with KpnI (purchased from Treasure Bioengineering Dalian Co., Ltd.) to obtain the exogenous fragment of LIP1 (FL-cDNA). T4 ligase was connected to the overexpression vector pU1301 (see Figure 18 ), name the correct vector as pU1301-LIP1 (see Figure 19 ), and electropora...

Embodiment 3

[0067] Example 3: LIP1 regulates panicle lateral meristem formation by limiting the active region of LAX1

[0068] 1. LIP1 inhibits the transcriptional activity of LAX1

[0069] In order to explore the molecular regulation mechanism of LIP1 on LAX1, the present invention carried out an in vivo transcription activity experiment for analysis. Specifically: design primers GFP-Protein-F / GFP-Protein-R (see Table 3 for the primer sequence) and LAX1-Protein-F / LAX1-Protein-R (see Table 3 for the primer sequence), use the GFP gene and the LAX1 gene The CDS was used as a template to amplify the target fragment, and TA cloned into pEASY-T3 (see Figure 11 , purchased from Beijing Quanshijin Biotechnology Co., Ltd.) and sequenced correctly, digested with KpnI-XbaI (purchased from Treasure Bioengineering Dalian Co., Ltd.), and connected to the 1301S-GAL4BD carrier treated with the same enzyme. After correct digestion The vectors were named GAL4BD-GFP and GAL4BD-LAX1, respectively. Desig...

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Abstract

The invention belongs to the technical field of plant genetic engineering, and particularly relates to a function and application of a transcription inhibitor LIP1 for regulating and controlling rice yield. A cloned LIP1 protein gene interacts with a LAX1 gene, and is expressed in the later developing stage of the meristem of a lateral spikelet and inhibits the transcriptional activity of the LAX1 protein by interaction with the LAX1. The deletion or space-time ectopic expression of the LIP1 gene functions can change the space-time activity of the LAX1 protein and influence formation of rice spikelet. The spatio-temporal expression of the LIP1 defines the activation range of LAX1 in spikelet lateral meristem, and ensures formation of rice lateral spikelet so as to control the yield. By utilizing the cloned LIP1 gene and the encoding protein thereof, the space-time activity of LAX1 can be changed by regulating and controlling the expression activity of the LIP1 gene through a gene manipulation means, so that the formation of spikelet is regulated and controlled, and important significance is achieved on genetic improvement of rice yield traits.

Description

technical field [0001] The invention belongs to the technical field of plant genetic engineering, and specifically relates to the function and application of a transcription inhibitor LIP1 regulating rice yield. LAX1 is a protein repressor of rice yield trait gene interaction. The cloned LIP1 gene of the present invention is expressed in the later stage of the development of the lateral spikelet meristem, and inhibits the protein activity of LAX1 by interacting with the LAX1 gene. The spatiotemporal expression of LIP1 defines the role of LAX1 in the lateral spikelet meristem. range, ensuring the formation of lateral spikelets in rice and controlling yield. Utilizing the LIP1 gene and its encoded protein cloned in the present invention, regulating its spatiotemporal expression through genetic manipulation means can change the spatiotemporal activity of LAX1, thereby regulating the formation of lateral spikelets, which is of great significance to the genetic improvement of rice...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/84A01H5/00A01H6/46
CPCC07K14/415C12N15/8261
Inventor 吴昌银符德保
Owner HUAZHONG AGRI UNIV
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