Gene capable of increasing nitrogen fertilizer utilization efficiency and yield of rice and application thereof

A high-nitrogen fertilizer, high-yield technology, applied in application, genetic engineering, plant genetic improvement, etc., can solve problems such as limited understanding

Active Publication Date: 2018-04-20
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

Although several candidate genes related to efficient nitrogen use have been obtained in rice and maize (Gallais and Hirel, 2004; Martin et al., 2006; Obara et al. 2001; Tabuchi et al., 2005), the control The understanding of the genetic regulatory network of plant nitrogen uptake and use efficiency is still very limited

Method used

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  • Gene capable of increasing nitrogen fertilizer utilization efficiency and yield of rice and application thereof
  • Gene capable of increasing nitrogen fertilizer utilization efficiency and yield of rice and application thereof
  • Gene capable of increasing nitrogen fertilizer utilization efficiency and yield of rice and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0132] Example 1: Identification of the main QTL-qNGR2 controlling the nitrogen absorption efficiency of rice

[0133] The inventors carried out hydroponic cultivation on 36 different rice resource materials carrying the "green revolution" gene sd1 for 4 weeks; after that, the pair of different varieties of materials was measured 15 The absorption rate of N-labeled ammonium nitrogen was screened and obtained a rice strain NM73 ( figure 1 a). NM73 is an intermediate breeding material selected by Anhui Quanyin Hi-Tech Seed Industry Co., Ltd.

[0134] Studies have shown that the plant height of NM73 is not sensitive to external application of nitrogen fertilizer: after two different fertilization treatments of low nitrogen and high nitrogen, its plant height does not change significantly. On the contrary, the plant height and yield of the indica rice variety Nanjing 6 (NJ6) are sensitive to exogenous nitrogen fertilizer application: under low-nitrogen treatments, Nanjing 6 exhibits a...

Embodiment 2

[0144] Example 2: Fine mapping and map-based cloning of qNGR2

[0145] The inventors used NM73 and NJ6 hybrid F 1 Materials, and NJ6 as the reincarnation parent, backcrossed continuously for multiple generations to construct BC 4 F 2 group. On this basis, we carried out fine mapping and map-based cloning, and successfully isolated and cloned the candidate gene NGR2. The sequences of the polymorphic marker primers used for fine mapping and map-based cloning are shown in Table 3, and the method for detecting polymorphic markers is as described in Example 1.

[0146] Table 3. Primers and sequences used for fine mapping and map-based cloning

[0147]

[0148]

[0149] In the following examples, it is confirmed that the NGR2 candidate gene is OsGRF4 through the comparative analysis of the candidate gene sequencing in the card position segment and the genetic complementation verification experiment.

[0150] In addition, the inventors found through sequence comparison and analysis that the...

Embodiment 3

[0151] Example 3: Excellent allele OsGRF4 ngr2 SNPs analysis of the promoter region

[0152] Compared with the 1kb promoter (SEQ ID NO: 1) of the OsGRF4 gene in the NJ6 variety, the allele OsGRF4 ngr2 There are 8 SNPs differences in its 1kb promoter (SEQ ID NO: 4). The inventors sequenced and analyzed the promoter region within 1 kb of the OsGRF4 gene in the different rice resource varieties described in Example 1, and the results showed that the allele OsGRF4 ngr2 There are 3 specific SNPs (c.-884T>A, c.-847C>T, c.-801C>T) in the 1kb promoter region. Further studies have shown that these SNP changes are related to increasing the expression level of OsGRF4 gene and the rate of ammonia nitrogen uptake by rice roots.

[0153] On this basis, the inventors performed haplotype analysis on the SNPs in the promoter region of the OsGRF4 gene within 1 kb in 225 cultivated rice varieties. The results of the study showed that there are 3 different haplotypes, namely Hap.A (such as high-yield...

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Abstract

The invention discloses a gene OsGRF4 capable of controlling nitrogen fertilizer utilization efficiency and yield trait as well as an allele gene and application thereof. OsGRF4 is capable of accelerating absorption and transfer of nitrogen fertilizer in rice and expression and enzyme activity of an assimilation gene, so that the nitrogen fertilizer utilization efficiency of the rice is improved;meanwhile, OsGRF4 is capable of accelerating cell division and improving the photosynthetic efficiency so as to increase the yield of the rice. The allele gene OsGRF4<ngr2> is capable of synergistically increasing the nitrogen fertilizer utilization efficiency and the yield of the rice under the condition that the plant height character is not influenced. The invention also discloses expression ofOsGRF4<ngr2> in high-yield wheat varieties for further increasing the nitrogen fertilizer utilization efficiency and the yield increase of the wheat. A promoter of the OsGRF4 gene and SNP polymorphism of a coding area can be used as molecular markers for identifying efficient utilization of nitrogen fertilizer and high-yield varieties of the rice; and OsGRF4 has broad application prospects in breeding on the basis of the increase of nitrogen fertilizer utilization efficiency and yield of the rice and the wheat.

Description

Technical field [0001] The invention belongs to the field of biotechnology. Specifically, the present invention relates to a gene and its alleles for controlling crop nitrogen utilization efficiency and yield, and applications thereof. The present invention also relates to the promoter sequence of the gene and the polypeptide sequence encoded by the gene, as well as a method for cultivating high-yield rice varieties with high-efficiency nitrogen fertilizer utilization by using molecular marker-assisted selection, genome editing and transgene technology. Background technique [0002] Rice (Oryza sativa L.) is a very important food crop in the world. Its total output accounts for 1 / 4 of the world's total food output. More than half of the world's population feeds on rice as the staple food. On the one hand, as the world’s population has increased year by year, the rigid demand for food has also been increasing; on the other hand, the arable land area has been decreasing year by ye...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C12N1/21C12N15/113C12Q1/6895C07K14/415A01H5/00A01H1/02A01H6/46
CPCC07K14/415C12N15/8243C12N15/8261C12Q1/6895C12Q2600/13C12Q2600/156Y02A40/10
Inventor 傅向东李姍吴昆田水航叶亚峰刘倩
Owner INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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