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Application of OsNRT2.3b to increase of yield and improvement of quality of oryza sativa

A yield and rice technology, applied in the field of genetic engineering, can solve the problems of high food production cost, poor yield increase effect and slow fertilizer efficiency.

Active Publication Date: 2020-10-20
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

my country's traditional agriculture attaches great importance to the use of organic fertilizers, which can promote crop yields, improve quality, and improve soil quality. However, there are disadvantages such as slow fertilizer efficiency, low nutrient content, labor-intensive application, and poor yield-increasing effects (Zhou Jiangming, 2012)
Although inorganic fertilizers have the advantages of fast increase in production, high nutrient content, and low dosage, the current excessive use by farmers has caused problems such as high food production costs, soil quality degradation, and serious agricultural non-point source pollution.

Method used

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  • Application of OsNRT2.3b to increase of yield and improvement of quality of oryza sativa
  • Application of OsNRT2.3b to increase of yield and improvement of quality of oryza sativa
  • Application of OsNRT2.3b to increase of yield and improvement of quality of oryza sativa

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Example 1 Construction of OsNRT2.3b Gene Overexpression Vector

[0017] According to the full-length cDNA sequence of the OsNRT2.3b gene (searched on the NCBI website), the primers for amplifying the full-length ORF were designed with the software Primer 5.0, and two different restriction sites, SpeI and BglII, were designed at the 5' end of the primers respectively. For different expression vectors pCAMBIA1302, the sense strand is overNRT2.3b-F: 5'-caACTAGTGCTACCACGTGTTGGAGATG-3', the antisense strand overNRT2.3b-R: 5'-gaAGATCTGAGCAAACCACCAACAAG-3', purchased from National Institute of Agrobiological Sciences ( NIAS) OsNRT2.3b gene cDNA full-length cloning plasmid was amplified as a template to obtain the open reading frame (ORF) sequence. After recovery and purification, the PCR product containing the target gene was connected into pMD19-Tcloning Vector, verified by enzyme digestion and sequenced. The correctly sequenced plasmids were digested with speI and BglII res...

Embodiment 2

[0018] Example 2 Obtaining and Molecular Biological Identification of OsNRT2.3b Overexpression Plants

[0019] transgenic T 0 After the seedlings were screened with hygromycin or GUS staining, 50 lines positive for the OsNRT2.3b-Ox (35S promoter) transgene were obtained, and the obtained transgenic seedlings were transplanted to the field for propagation in order to obtain T 0 generation seeds. Among the positive lines, we obtained 18 OsNRT2.3b-Ox transgenic lines whose phenotypes were significantly different from those of the wild type. In order to further determine whether the target gene in these lines was enhanced, T 0 Total RNA was extracted from the leaves of transgenic seedlings for preliminary RT-PCR identification, and several strains with enhanced expression of the target gene were obtained. According to the detection results, the enhanced expression strain was selected as the material for the next research, named O8 (35S promoter).

[0020] In order to verify the ...

Embodiment 3

[0021] Embodiment 3 experimental design

[0022] The test set three fertilizer application levels (M: organic fertilizer treatment, NPK: inorganic fertilizer treatment, NPKM: organic fertilizer), two fertilizer gradients (1: low, 2: high), that is, organic low (M1), organic high (M2 ), inorganic low (NPK1), inorganic high (NPK2), inorganic fertilizer + organic fertilizer low (NPKM1), inorganic fertilizer + organic fertilizer high (NPKM2), a total of 6 treatments, each treatment set 3 replicates, A total of 18 plots, arranged in random blocks, each with an area of ​​3.75m 2 (3m×1.25m), the depth of the plots is 1.2 m, and the plots are separated by brick concrete to prevent the exchange of water and fertilizer between plots. Nitrogen fertilizer is applied to urea (containing N 46%) according to base fertilizer: tiller fertilizer: panicle fertilizer = 5: 2: 3, organic fertilizer is applied once at the base fertilizer stage, and no fertilization is applied in the later stage. A...

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Abstract

The invention discloses application of OsNRT2.3b to increase of yield and improvement of quality of oryza sativa. Under different fertilizer treatment conditions, effective tillering, weight per ear,yield per plant, brown rice rate and head rice yield of a total OsNRT2.3b overexpression material are higher than those of Nipponbare wild oryza sativa; and moisture content and length-width ratio arehigher than those of the Nipponbare wild oryza sativa, and chalky grain percentage and chalkiness degree are both lower than those of the Nipponbare wild oryza sativa.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering and relates to the application of OsNRT2.3b in improving yield and rice quality. Background technique [0002] Rice (Oryza sativa L.) is an important staple food for billions of people (Zeng, et al., 2017). As a large agricultural country, my country is one of the earliest countries in the world to grow rice, and it is also one of the most important food crops in my country (Chen Ligang, 2019, Situsong, et al., 2000). For a long time, in order to increase rice yield, farmers have always regarded fertilization as the most important physical and chemical technical measure for rice production. Traditional agriculture in my country attaches great importance to the use of organic fertilizers, which can increase crop yields, improve quality, and improve soil quality, but have disadvantages such as slow fertilizer efficiency, low nutrient content, labor-intensive application, and poor yield-...

Claims

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Application Information

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IPC IPC(8): C12N15/29C12N15/82A01H5/10A01H6/46
CPCC07K14/415C12N15/8261
Inventor 范晓荣赵利梅钱开芸
Owner NANJING AGRICULTURAL UNIVERSITY
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