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A gene controlling the erect development of rice leaves and its application

A gene and rice technology, applied in the fields of application, genetic engineering, plant genetic improvement, etc., can solve the problems of unclear cellular and molecular mechanisms

Active Publication Date: 2017-09-15
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Brassinosterol synthesis and signaling pathways play an important and unique role in regulating the development of stem-leaf angle in monocots, but the underlying cellular and molecular mechanisms are still unclear

Method used

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  • A gene controlling the erect development of rice leaves and its application
  • A gene controlling the erect development of rice leaves and its application
  • A gene controlling the erect development of rice leaves and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Acquisition of a gene CYC U4;1 controlling the erect development of rice leaves:

[0032] The inventor constructed a gene-specific expression chip for rice leaves and leaf pillows, and selected the gene CYC U4 specifically expressed in the leaf pillows from the differentially expressed genes of the chip through the method of reverse genetics; 1. Using real-time qRT- PCR verified the specificity of CYC U4;1 expression in the occipital. details as follows:

[0033] A gene CYC U4; 1, which controls the erect development of rice leaves, is obtained by the following method:

[0034] The total volume of the reaction system is 50μl, the template is Nipponbare CDNA 1ul (about 50ng), 10×KOD enzyme reaction buffer 5μl, 25mM MgCL 2 2μl, 5mM dNTP 5μl, 5uM primer 5μl (by step-by-step PCR, use primers CYCU4-U and CYCU4-L-1 for the first time, use primers CYCU4-U and CYCU4-L-2 for the second time, each primer 2.5μl), 1μl KOD enzyme, add ddH 2 O (sterile deionized water) to 50 μl....

Embodiment 2

[0044] The application of CYC U4; 1 gene in controlling the uprightness of rice leaves is as follows:

[0045] 1) Construction of plant expression vector pCYCU4; 1-GUS

[0046] Amplify the CYC U4; 1 promoter region, and use the HindIII / BamH1 restriction site to clone the fragment into pCAMBIA1300GN GUS (Ren Z H, Gao J P, Li L G, et al. A rice quantitative traitlocus for salt tolerance encodes a sodium transporter[J ]. Nature genetics, 2005, 37(10): 1141-1146.) on the carrier.

[0047] The CYC U4; 1 promoter region is obtained by the following method:

[0048] The total volume of the reaction system is 50 μl, the template is Nipponbare Genomic DNA 1ul (about 50ng), 1×KOD enzyme reaction buffer 5μl, 25mM MgCL 2 2μl, 5mM dNTP 5μl, 5uM primer 5μl (primers pCYCU4-U and pCYCU4-L respectively 2.5μl), 1μl KOD enzyme, add ddH 2 O (sterile deionized water) to 50 μl. The reaction program was: denaturation at 94°C for 5min, 30s at 94°C, 1min at 55°C, 35cycles at 68°C for 2min, and ex...

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Abstract

The present invention provides a gene for controlling the establishment of grass crops and its application. The disclosed gene CYC U4;1 (Os10g41430) of the present invention is specifically expressed in the rice leaf pillow (it is believed that it is also expressed in other grass crops). Its CDS sequence is shown in SEQ ID NO.2, and the encoded amino acid sequence is shown in SEQ ID NO.3. The present invention also provides the promoter sequence of the gene, which is shown in SEQ ID NO.1. The promoter and full-length CDS of CYC U4;1 were cloned into pCAMBIA1301 and transformed into Nipponbare. The obtained transgenic lines showed that the stem-leaf angle was smaller than that of the wild type, so CYC U4 was increased or weakened by genetic engineering technology ; The expression level of the 1 gene can control the plant type development, thereby improving the plant type and increasing the germplasm density.

Description

technical field [0001] The invention belongs to the field of plant genetic engineering. It specifically relates to a gene that controls the upright development of rice leaves. Using the method of reverse genetics, the gene specifically expressed in the leaf pillow is screened from the chip analysis of rice leaf and leaf pillow specific gene expression, and the pCYCU4-GUS transgenic line is used to prove The promoter of CYC U4;1 is specifically expressed in the leaf pillow; the expression of CYC U4;1 was increased by transgene and the expression of CYC U4;1 was reduced by RNAi technology, which proves that the CYC U4;1 gene controls the upright development of rice leaves Gene. The present invention also relates to a vector containing the gene or its homologous gene and to the application of the gene or its functional analogue in agricultural production to regulate the uprightness of plant leaves. Background technique [0002] Plant architecture development is a key trait in...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/29C07K14/415C12N15/11A01H5/00
CPCC07K14/415C12N15/8225C12N15/8261Y02A40/146C12N15/8262
Inventor 王学路孙世勇
Owner HUAZHONG AGRI UNIV
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