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Plant leaf curl related protein OsRoc8 and coding gene and application of OsRoc8

A curl-related protein and plant leaf technology, which is applied to the plant leaf curl-related protein OsRoc8 and its encoding gene and application fields, and can solve problems such as complex mechanism and the like

Pending Publication Date: 2019-11-12
JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the morphological structure of the leaf curl seems simple, the mechanism of its formation is very complicated

Method used

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  • Plant leaf curl related protein OsRoc8 and coding gene and application of OsRoc8
  • Plant leaf curl related protein OsRoc8 and coding gene and application of OsRoc8
  • Plant leaf curl related protein OsRoc8 and coding gene and application of OsRoc8

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1 Homologous cloning of plant leaf curl control-related proteins and genes encoding them

[0037] 1. Phylogenetic tree analysis of rice HD-Zip IV transcription factors

[0038] Previous reports have confirmed that HD-Zip transcription factors play an important role in the growth and development of Arabidopsis and maize. The rice HD-Zip IV family gene OsRoc5 has been reported to regulate leaf curling and further determine the Leaf type and plant type ultimately affect the photosynthetic efficiency of rice. BLAST was performed using the reported OsRoc5 protein sequence and the Bioinformatics website (https: / / blast.ncbi.nlm.nih.gov / Blast.cgi). In rice, multiple proteins that are relatively homologous to the target protein were obtained. On the basis of multiple comparison analysis, the corresponding phylogenetic tree was established using MEGA 3.0 software. The result is as figure 1As shown, the obtained HD-Zip subfamily members are distributed on different bra...

Embodiment 2

[0049] Embodiment 2: Obtaining of transgenic plants

[0050] 1. Construction of binary plant overexpression vector pCAMBIA1305-2×35s-OsRoc8

[0051] The schematic diagram of pCAMBIA1305-2×35s-OsRoc8 vector is as follows image 3 shown. Firstly, the pCAMBIA1305.1 vector (http: / / www.cambia.org / daisy / cambia / 585) was digested with SacI, and the restriction endonucleases used were purchased from ThermoFisher. The enzyme digestion system was as follows:

[0052]

[0053] After digestion at 37°C for 1 h, agarose gel electrophoresis and gel slicing were carried out, and the SacI-digested vector was recovered using the OMEGA DNA Recovery Kit and set aside for later use.

[0054] Secondly, entrust Nanjing KingScript Biotechnology Co., Ltd. to synthesize double-copy cauliflower mosaic virus (CAMV) 35S promoter DNA sequence, and add a linker sequence 5'-TTACGAATTC at the 5' end GAGCTC -3' (SEQ ID NO: 9), add linker sequence 5'-CCCGGGTACC at the 3' end GAGCTC -3' (SEQ ID NO: 10), ...

Embodiment 3

[0080] Example 3: Identification of transgenic plants

[0081] 1. Expression identification

[0082] T 1 Transplant pCAMBIA1305-2×35s-OsRoc8 plants, T 1 The empty vector control plants and wild-type Kitaake were planted in the greenhouse of the Food Crops Research Institute of the Jiangsu Academy of Agricultural Sciences, and the leaves were observed from the seedling stage to the heading stage. The result is as Figure 5 As shown, OE-1, OE-2, OE-3 three T 1 The pCAMBIA1305-2×35s-OsRoc8 plants showed a stable leaf curling phenotype throughout the growth period ( Figure 5 A). The three pCAMBIA1305-2×35s-OsRoc8 plants and wild-type Kitaake were further identified by Real-time PCR. For the method used, please refer to the above-mentioned "OsRoc8 gene expression analysis" section, and the rice ubiquitin gene (LOC_Os03g13170) was still selected as the primer. Internal reference, RT-OsRoc8-F (SEQ ID NO: 3) and RT-OsRoc8-R (SEQ ID NO: 4) are OsRoc8 quantitative primers. The r...

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Abstract

The invention discloses an OsRoc8 gene of a plant leaf curl related protein and the protein, and application of an expression kit, an recombinant vector or cell in obtaining a plant with curled leaves. According to the present invention, the OsRoc8 gene is homologously cloned from rice to a transcription, the complete cDNA of the OsRoc8 gene is homologously recombined into a plant vector with overexpression, the plant is transformed by an Agrobacterium infection method, the corresponding transgenic plant is relative upright with leaves curled to the front, the photosynthetic efficiency increases, and then transgenic plants with high photosythetic efficiency and curled leaves can be cultivated.

Description

technical field [0001] The invention belongs to the field of genetic engineering. Specifically, the present invention relates to plant leaf curl-related protein OsRoc8 and its coding gene and application. Background technique [0002] As the main organ of photosynthesis in green plants, leaves play an important role in their life activities. Rice is one of the important food crops in my country, and more than 50% of the population takes it as the main ration. Ensuring rice production and production safety is the top priority of my country's agricultural production. Leaf curling is an important agronomic trait closely related to yield in rice plant type indicators. In-depth study of leaf curling is of great significance for rice leaf type, plant type improvement and yield potential breakthrough. [0003] Leaf is an important indicator of plant morphogenesis. Moderate curling can cause the upper leaves to stand upright, increase the light transmittance of the group, and thus...

Claims

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Application Information

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IPC IPC(8): C12N15/29C12N15/82A01H5/12A01H6/46
CPCC07K14/415C12N15/8261
Inventor 许扬杨杰王芳权陈智慧王军范方军李文奇陶亚军
Owner JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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