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Application of rice PHI1 gene in regulation and control of plant photosynthesis

A photosynthesis, plant technology, applied in the field of plant genetic engineering, can solve problems such as little known

Active Publication Date: 2021-06-04
THE INST OF BIOTECHNOLOGY OF THE CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, little is known about the molecular basis controlling rice leaf traits

Method used

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  • Application of rice PHI1 gene in regulation and control of plant photosynthesis
  • Application of rice PHI1 gene in regulation and control of plant photosynthesis
  • Application of rice PHI1 gene in regulation and control of plant photosynthesis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0082] Example 1, the acquisition of PHI1 protein and its coding gene

[0083] 1. Phenotypic analysis of mutant scd1

[0084] The plant height and leaf curl degree of mutant scd1 and wild-type rice Nipponbare were measured, and the chlorophyll A content, chlorophyll B content and photosynthetic rate were detected at the same time.

[0085] Determination of plant height: measured at maturity, the height from the ground surface to the top of the highest ear (excluding awn) of a single plant. The data of each section is also deduced in the same way (sections are counted from top to bottom).

[0086] Curling degree (Leaf rolling index LRI) = Determination of maturity period, [Lw (leaf edge spacing after deployment)-Ln (measured at the same point the natural distance from the leaf to the leaf edge)] / Lw (leaf edge spacing after deployment) × 100 %.

[0087] Determination of photosynthetic rate: One week after the initial ear, select a sunny day with air temperature (daily average...

Embodiment 2

[0106] Example 2, construction of transgene complementary expression plants and detection of their traits

[0107] 1. Construction of pCambia23A-PHI1 recombinant expression vector

[0108] Utilize TWI1-F and TWI1-R to amplify SEQ ID No.3 (derived from rice cDNA), the obtained product is digested with SmaI / XbaI, and then used with pCambia23A vector (Beijing Dingguochangsheng Biotechnology Co., Ltd.) The recombinant expression vector pCambia23A-PHI1 was obtained by digestion with XbaI and SmaI enzymes, and was verified to be correct by sequencing.

[0109] TWI1-F-5'SmaI:5'-TT CCCGGG GTGTAAATAGTAGGCTTGTTGGAG-3';

[0110] TWI1-R-3'XbaI:5'-CC TCTAGA AAGCTTCTCTCCACTAGTCAAGTC-3'.

[0111] 2. The pCambia23A-PHI1 recombinant expression vector obtained in step 1 was introduced into Agrobacterium AGL1 (Beijing Dingguo Changsheng Biotechnology Co., Ltd.) to obtain recombinant Agrobacterium.

[0112] 3. Introduce the recombinant Agrobacterium obtained in step 2 into the transformati...

Embodiment 3

[0122] Example 3, Establishment and Phenotype Observation of PHI1 Functional Loss Transgenic Plants

[0123] 1. Interfering with plant construction

[0124] 1. Using the cDNA of wild-type rice Nipponbare as a template, PCR amplification is performed with primers P5utrF and primers P5utrR to obtain PCR amplification products.

[0125] P5utrF: 5'-GG GGTACCACTAGT GCTTTACCTAAGCGAATTCT-3';

[0126] P5utrR: 5'-CG GGATCCGAGCTC ATACATTATTACCATTTCC-3'.

[0127] In the primer P5utrF, the underline marks KpnI and SpeI restriction sites;

[0128] In the primer P5utrR, the underline marks are the restriction sites of BamH I and Sac I.

[0129] 2. Ligate the PCR amplification product obtained in step 1 with the pEasy vector (Beijing Dingguo Changsheng Biotechnology Co., Ltd.) to obtain the ligation product pEasy-PHI1.

[0130] 3. The ligation product pEasy-PHI1 in step 2 was double-digested with SpeI and SacI restriction endonucleases, and the digested product of about 0.3 Kbp was r...

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Abstract

The invention discloses application of a rice PHI1 gene in regulation and control of plant photosynthesis. The invention provides application of a PHI1 protein or a related biological material thereof in regulation and control of plant photosynthesis. The related biological material is a nucleic acid molecule expressing the PHI1 protein or an expression cassette, a recombinant vector, a recombinant bacterium or a transgenic cell line containing the nucleic acid molecule; and the PHI1 protein is shown as SEQ ID No. 1 or is substituted and / or deleted and / or added by one or more amino acid residues, or is a protein with 80% or more homology of the sequence and the same function, or is a fusion protein obtained by connecting a tag to the N end and / or C end of the PHI1 protein. In a normal growth state and under a drought condition, the expression quantity of the PHI1 gene in a target plant is reduced, and the photosynthetic efficiency of the plant can be obviously improved. The application has important theoretical and practical significance for cultivating a high-photosynthetic-efficiency rice material under an adverse condition.

Description

technical field [0001] The invention relates to the field of plant genetic engineering, in particular to an application of rice PHI1 gene in regulating plant photosynthesis. Background technique [0002] In our country, food security has always been one of the important issues. Due to the increase in population and the continuous reduction of arable land area, for important food crops such as rice, breeding higher-yield varieties has always been the primary breeding goal. Every breakthrough in rice production is inseparable from the discovery and utilization of new favorable resources. In the 1950s, the discovery of rice dwarf resources and the success of dwarf breeding achieved the first leap in rice yield; in the 1960s and 1970s, the discovery of rice sterile cytoplasm and the successful matching of three lines made my country's rice Yield achieved a second leap; at the end of the 20th century, the successful breeding of super hybrid rice increased the yield of hybrid ric...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/82A01H5/00A01H5/04A01H5/12A01H6/46
CPCC07K14/415C12N15/8218C12N15/8269C12N15/8273C12N15/8261C12N15/825
Inventor 张治国崔学安邓晨孙晶吴金霞路铁刚威廉 保罗·奎克
Owner THE INST OF BIOTECHNOLOGY OF THE CHINESE ACAD OF AGRI SCI
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