Paddy rice NRT 1.1A gene and application of encoded protein of paddy rice NRT 1.1A gene to improve plant yield in breeding

A technology for transgenic plants and yield, applied in plant peptides, genetic engineering, recombinant DNA technology, etc., can solve problems such as environmental degradation, excessive use of nitrogen fertilizer, and low nitrogen use efficiency

Active Publication Date: 2018-03-09
INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are a series of problems in my country, such as excessive use of nitrogen fertilizer, low nitrogen use efficiency and environmental degradation caused by nitrogen fertilizer loss.

Method used

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  • Paddy rice NRT 1.1A gene and application of encoded protein of paddy rice NRT 1.1A gene to improve plant yield in breeding
  • Paddy rice NRT 1.1A gene and application of encoded protein of paddy rice NRT 1.1A gene to improve plant yield in breeding
  • Paddy rice NRT 1.1A gene and application of encoded protein of paddy rice NRT 1.1A gene to improve plant yield in breeding

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0077] Example 1, Analysis of the expression of NRT1.1A in rice tissues

[0078] The NRT1.1A gene involved in this example is derived from rice (Oryza.sativa L.), its genome sequence is shown in sequence 2 in the sequence listing, sequence 2 consists of 8143 nucleotides, and its cDNA sequence is the sequence listing Middle sequence 3, sequence 3 consists of 1812 nucleotides. Sequence 2 and Sequence 3 encode the protein (NRT1.1A protein) shown in Sequence 1 in the Sequence Listing, and Sequence 1 consists of 603 amino acid residues.

[0079] Total RNA was extracted from the root, stem, leaf sheath, leaf and ear of rice variety Dongjin and cDNA was obtained by reverse transcription. Furthermore, using the obtained cDNA as a template, real-time quantitative fluorescent PCR was carried out for the NRT1.1A gene to detect the expression level of the NRT1.1A gene at the transcription level in various tissues of rice. The experiment was repeated 3 times, and the results were averaged....

Embodiment 2

[0088] Example 2. Verification of NRT1.1A's in vitro nitrate transport activity

[0089] The NRT1.1A gene involved in this example is derived from rice (Oryza.sativa L.), its genome sequence is shown in sequence 2 in the sequence listing, sequence 2 consists of 8143 nucleotides, and its cDNA sequence is the sequence listing Middle sequence 3, sequence 3 consists of 1812 nucleotides. Sequence 2 and Sequence 3 encode the protein (NRT1.1A protein) shown in Sequence 1 in the Sequence Listing, and Sequence 1 consists of 603 amino acid residues.

[0090] 1. Xenopus laevis recombinant expression vector pCS2 + / Build of NRT1.1A

[0091] The total RNA of japonica rice variety Dongjin was extracted and reverse transcribed into cDNA. The obtained cDNA was used as a template, and the NRT1.1A cDNA was amplified by PCR using the following primer sequences. The two ends of the primers used for amplification were respectively introduced into the recognition sites of the restriction endonu...

Embodiment 3

[0101] Example 3. Obtainment and functional verification of rice nrt1.1a mutant

[0102] The NRT1.1A gene involved in this example is derived from rice (Oryza.sativa L.), its genome sequence is shown in sequence 2 in the sequence listing, sequence 2 consists of 8143 nucleotides, and its cDNA sequence is the sequence listing Middle sequence 3, sequence 3 consists of 1812 nucleotides. Sequence 2 and Sequence 3 encode the protein (NRT1.1A protein) shown in Sequence 1 in the Sequence Listing, and Sequence 1 consists of 603 amino acid residues.

[0103] 1. Obtaining and identification of rice nrt1.1a mutant

[0104] Rice nrt1.1a mutant: from Korean Rice Mutant Bank (Crop Biotech Institute, KyungHee University, Republic of Korea, http: / / www.postech.ac.kr / life / pfg / risd), the order URL is http: / / signal.salk.edu / cgi-bin / RiceGE5, the corresponding number of the rice nrt1.1a mutant is PFG_1E-00433.L. The specific construction method is described in the article "Jakyung Yi, Gynheung A...

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Abstract

The invention discloses a paddy rice NRT 1.1A gene and an application of encoded protein of the paddy rice NRT 1.1A gene to improve the plant yield in breeding. Concretely, protein or an encoded genethereof is applied to regulate the growth and development of a plant. The growth and development is reflected as the single plant yield and/or the strain height and/or grain number and/or flowering time and/or bolting time and/or size of a rosette leaf. The protein is any one of (1) protein having the amino acid sequence shown as in the sequence 1 in a sequence list; and (2) protein which is derived from the sequence 1 by replacing and/or deleting and/or adding one or more amino acid residues in and/or from and/or to the amino acid sequence in the sequence 1 in a sequence list, and is relatedto control of the growth and development of a plant. The paddy rice NRT 1.1A gene and encoded protein are of great importance in breeding of new high-yield paddy rice variety.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to the application of a rice NRT1.1A gene and its coded protein in breeding for improving plant yield. Background technique [0002] Plants need a variety of mineral nutrients to maintain their normal growth, and nitrogen is the mineral nutrient element that plants need the most. Generally speaking, nitrogen can account for 1.5-2% of the dry weight of the plant body. Nitrogen is a constituent of proteins, nucleic acids, phospholipids, and organic nitrogen compounds necessary for plant growth and development, and these substances are structural or functional components for living cells to survive, so nitrogen is also called a life element. In order to pursue the high yield of crops, a large amount of nitrogen fertilizer is often applied in agricultural production. Among them, the amount of nitrogen fertilizer applied to rice far exceeds that of any other crops, and the loss of nitrogen f...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82A01H6/20A01H6/46
CPCC07K14/415C12N15/8243C12N15/8261C12N15/82Y02A40/146
Inventor 储成才王威胡斌李华张志华刘永强
Owner INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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