Application of CYP78A gene in increasing of plant height of corns and growth vigor of plants

A corn and gene technology, applied in the application field of CYP78A gene, can solve the problems of increasing plant height, increasing plant height, etc.

Inactive Publication Date: 2016-12-07
HANGZHOU RUIFENG BIOTECH LIMITED
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For example, previous related studies on rice did not find that the gene significantly increases plant height and enhances the function of growth vigor, but the present invention discloses for the first time that increasing the expression of CYP78A gene in maize can increase plant height and enhance plant growth vigor

Method used

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  • Application of CYP78A gene in increasing of plant height of corns and growth vigor of plants
  • Application of CYP78A gene in increasing of plant height of corns and growth vigor of plants
  • Application of CYP78A gene in increasing of plant height of corns and growth vigor of plants

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1. Construction of corn CYP78A gene--CYP78A1 overexpression vector

[0042] Maize CYP78A1 gene coding region (nucleotide sequence as shown in SEQ ID No.4) and terminator DNA fragment obtained by artificial synthesis, its nucleotide sequence as SEQ ID No.7, this fragment can be digested by BamHI and KpnI Get it later.

[0043] The promoter fragment pCYP78A1 of CYP78A1 gene was obtained by PCR. The two primers for PCR were: pCYP78A1-F (5'-AAGCTTCTCAAGACTCCTAATTCTAGC-3') and pCYP78A1-R (5'-AGATCTTTCTGGCTAGCTGCTTCTAGTAGC-3'). HindIII and BglII restriction sites were designed in the primers respectively. Using the corn (B73) genome as a template, the DNA fragment of about 1.9kb obtained by PCR was cloned into the pMD-18-T-Vector vector (TaKaRa), and the sequence was determined. Finally, the nucleotide sequence obtained as SEQ ID NO.9 For the promoter sequence shown, this fragment can be obtained by digestion with HindIII and BglII.

[0044] The reaction system of...

Embodiment 2

[0048] Example 2. Construction of Rice CYP78A Gene CYP78A11 Overexpression Vector

[0049] The rice CYP78A11 gene coding region (the nucleotide sequence is shown in SEQ ID NO.5) and the terminator DNA fragment are obtained by PCR. The two primers for PCR were: CYP78A11-F1 (5'-GGATCCAACAATGGCAATGGCCACCGCCAC-3') and CYP78A11-R1 (5'-GGTACCCATCTCACAAAGCTCACACGGC-3'). Using the genome of Xiushui 134, the main rice variety in Zhejiang, as a template, the DNA fragment of about 2.2 kb obtained by PCR was cloned into the pMD-18-T-Vector vector (TaKaRa), and the sequence was determined to obtain the DNA shown in SEQ ID NO.8 Fragment, this DNA fragment contains an intron and a terminator in addition to the DNA sequence encoding the rice CYP78A11 amino acid. The reaction system of PCR is: 5x PrimeSTAR TM Buffer (Mg 2+ plus) (purchased from TaKaRa Company), 10 μl; dNTP Mixture (each 2.5 mM), 4 μl; primer CYP78A11-F1 (10 μM), 1 μl; primer CYP78A11-R1 (10 μM), 1 μl; template DNA 100 ng; ...

Embodiment 3

[0054] (1) Corn transformation

[0055] Maize transformation methods have been relatively mature, for example, Frame et al. described the method of using Agrobacterium to transform maize (Frame et al., (2002) Plant Physiol, 129:13-22). Take the Agrobacterium containing the vector pCambia1300-G10-pCYP78A1-CYP78A1 and pCambia1300-G10-pCYP78A11-CYP78A11 respectively (that is, the Agrobacterium containing the T-DNA vector) to draw a plate, pick a single colony and inoculate it, and prepare the Agrobacterium for transformation. Take Hi-II corn ears 8-10 days after pollination. All immature embryos (1.0-1.5 mm in size) were collected. Co-cultivate the Agrobacterium containing T-DNA vector and immature embryos for 2-3 days (22°C). Transfer the immature embryos to the callus induction medium (the medium contains 200mg / L of Timentin for killing Agrobacterium, referring to (Frame et al., (2002) PlantPhysiol, 129:13-22)), 28 Cultivate in the dark for 10-14 days. All calli were transf...

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Abstract

The invention discloses application of a CYP78A gene in the increasing of the plant height of corns and the growth vigor of plants. An amino acid sequence coded by the CYP78A gene is one of the following sequences or an amino acid sequence with 75% of homology with the following sequences: SEQ ID NO:1, SEQ ID NO:2 or SEQ ID NO:3. According to the novel application of the CYP78A gene, the CYP78A gene is over-expressed in the corns, so that the plant height and the growth vigor of the corns are increased, the biomass of the corns is further increased, and finally, the yield of the corns is increased; and meanwhile, as an improvement, the CYP78A gene and a TEL gene are combined to be introduced into the corn plants, so that the plant height of the corns is increased, leaves of the corns are widened, the growth vigor of the corns is increased, the biomass of the corns is increased, and furthermore, the phenotypic changes including the largening and the increasing of corn cobs and seeds are realized.

Description

(1) Technical field [0001] The invention relates to the application of a CYP78A gene, in particular to the application in increasing the height of maize plants and enhancing the growth of plants. (2) Background technology [0002] As the human demand for food continues to increase, the cultivation of high-yield crops is becoming more and more important. Transgenic technology has been widely used to improve the traits of crops, such as acquiring insect resistance, herbicide resistance, enhancing drought resistance, and disease resistance. High yield is also an important content of transgenic crop improvement. For example, the yield of crops can be improved by transgenic expression of a plant's transcription factor (U.S. Pat. No. 7,598,529,4). Crop yield is a complex quantitative trait regulated by many genes (Xing and Zhang, (2010) Annual Review of Plant Biology, 61:421-442). Therefore, it is still very necessary to study and discover new genes with great application value...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/53C12N15/82A01H5/00
Inventor 沈志成张先文王东芳
Owner HANGZHOU RUIFENG BIOTECH LIMITED
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