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Gene for controlling rice stem-leaf angle and application of gene
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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: 2015-09-02
HUAZHONG AGRI UNIV
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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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Embodiment 1
[0031] Obtaining of a gene CYC U4;1 that controls the erect development of rice leaves:
[0032] The inventor constructed a gene-specific expression chip for rice leaves and leaf occiputs, and selected the gene CYC U4 specifically expressed at the leaf occipital from the differentially expressed genes of the chip by means of reverse genetics; 1, using real-time qRT- PCR verified the specificity of CYC U4;1 expression in the occipital region. details as follows:
[0033] A gene CYC U4;1, which controls the erect development of rice leaves, was 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 (using stepwise PCR, the first use of primers CYCU4-U and CYCU4-L-1, the second use of primers CYCU4-U and CYCU4-L-2, each primer is 2.5μl), 1μl KOD enzyme, add ddH 2 O (sterile deionized water) to 50μl.
[0035] The reactio...
Embodiment 2
[0044] The application of CYC U4;1 gene in the control of rice leaf erection, the application process is as follows:
[0046] Amplify the CYC U4; 1 promoter region, and clone the fragment into pCAMBIA1300GN GUS(Ren ZH,Gao JP,Li LG,et al.A rice quantitative trait locus for salt tolerance encodes a sodiumtransporter[] by using the HindIII / BamH1 restriction site J]. Nature genetics, 2005, 37(10): 1141-1146.).
[0047] The CYC U4; 1 promoter region is obtained in the following way:
[0048] The total volume of the reaction system is 50μl, 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 2.5μl respectively), 1μl KOD enzyme, add ddH 2 O (sterile deionized water) to 50μl. The reaction procedure is: denaturation at 94°C for 5 min, 94°C for 30s, 55°C for 1 min, 68°C for 2 min, 35 cycles, and 68°C for extensio...
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Abstract
The invention provides a gene for controlling a rice stem-leaf angle and application of the gene. The gene CYC U4;1(Os10g41430) disclosed by the invention specifically expresses in a rice pulvinus (believe that the gene also specifically expresses in other graminaceous crops), the CDS sequence of the gene is shown in the SEQ ID No.2, and a coded amino acid sequence is shown in the SEQ ID No.3. The invention further provides a promoter sequence, shown in the SEQ ID No.1, for the gene. A promoter of CYC U4;1 and overall length CDS are cloned to pCAMBIA1301, and then converted into Nipponbare, leaf-stem angles of the obtained transgenic lines are less than those of wild rice, therefore, plant type growth of plants can be controlled by increasing or decreasing expression quantity of CYC U4;1 gene through a genetic engineering technology, plant types are improved and germplasm density is increased.
Description
Technical field [0001] The invention belongs to the field of plantgenetic engineering. Specifically, it relates to a gene that controls the erect development of rice leaves. Using reverse genetics methods, the genes specifically expressed in the leaf occipital region are selected from the rice leaf and leaf occipital-specific gene expressionchip analysis, and the pCYCU4-GUS transgenic line is used to prove The promoter of CYC U4;1 is specifically expressed in the leaf occiput; the expression of CYC U4;1 is increased by transgene and RNAi technology is used to reduce the expression of CYC U4;1. Both prove that CYC U4;1 gene controls the erect development of rice leaves gene. The invention also relates to a vector containing the gene or its homologous gene and to the application of the gene or its functional analogue to regulate the erection of plant leaves in agricultural production. Background technique [0002] Plant type development of crops is a key trait for high-yield br...
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