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Clone and application of semi-dominant gene qGL3 capable of controlling grain length and grain weight of rice kernel

A rice grain and gene technology, applied in application, genetic engineering, plant genetic improvement and other directions, can solve the problem of not obtaining clone function verification and other problems

Active Publication Date: 2012-02-15
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Some other QTL loci controlling rice grain weight or grain shape have reached the level of fine mapping that can be selected by molecular markers, such as gw8.1 (Xie et al., 2006), gw9.1 (Xie et al., 2008) and GW6 (Guo et al., 2009), but have not yet been cloned and further functionally validated

Method used

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  • Clone and application of semi-dominant gene qGL3 capable of controlling grain length and grain weight of rice kernel
  • Clone and application of semi-dominant gene qGL3 capable of controlling grain length and grain weight of rice kernel
  • Clone and application of semi-dominant gene qGL3 capable of controlling grain length and grain weight of rice kernel

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Example 1 Isolation and acquisition of rice grain length gene qGL3

[0050] (1) Materials and methods:

[0051] 1. The construction of large-grained germplasm N411 and 05-643, 93-11 hybrid population (for specific procedures, see figure 1 ):

[0052] (1) Using rice large-grain germplasm N411 (preserved by the rice germplasm bank of Nanjing Agricultural University, whose dominant allele can be obtained from Nannongxian 5, variety application acceptance number 20110730.0) as a donor material for favorable genes, Crossed with 05-643 (bred by crossing rice varieties Yuefeng B and Longtepu B) and 93-11, planted hybrids, and obtained F by selfing 1 . Will F 1 The seeds obtained by selfing are planted into a single plant to form F 2 Population, used to map grain length, grain weight QTL.

[0053] (2)F 2 Select a single plant with longer grains in the population to continuously backcross to 05-643 or 93-11, and backcross to high-generation BCF 2 Populations were used t...

Embodiment 2

[0073] Example 2 Expression analysis of qGL3 and its homologous gene family

[0074] According to the analysis of the rice EST profiles database on NCBI, it was found that the three members of the rice qGL3 family were expressed in various tissues, especially in young tissues with vigorous division. In order to better understand the expression pattern of this gene family, we designed the specific expression primer pair qGL3Seq-F (SEQ ID NO: 14), qGL3Seq-R (SEQ ID NO: 15); qGL3-L1Seq-F (SEQ ID NO: 16), qGL3-L1Seq-R (SEQ ID NO: 17) and qGL3-L2Seq-F (SEQ ID NO: 18), qGL3-L2Seq-R (SEQ ID NO: 19). The expression of this gene family was analyzed by semi-quantitative analysis.

[0075] The roots (R), stems (C), fresh leaves (LB), leaf sheaths (LB), 2cm-3cm primary panicles (PP), 8cm-10cm young panicles were extracted from 93-11 and 93-11NIL-qGL3 respectively (YP), the RNA of the mature ear (MP) just about to be extracted (using the RNA extraction kit of Tianwei Company), and rever...

Embodiment 3

[0079] Example 3 Subcellular localization of qGL3 wild-type protein and dominant allelic protein qGL3-D

[0080] Using the primer pair qGL3GFP-F (SEQ ID NO: 32) and qGL3GFP-R (SEQ ID NO: 33), the qGL3 (see specific example 1) and its allelic variant gene qGL3-D cloned on the T vector were respectively The full-length coding region was amplified, connected to the pMD-18Tsimple vector (purchased from Takara) by T-A cloning, transformed Escherichia coli DH5a competent cells by heat shock method, coated with ampicillin-resistant LB plates, and single clones were picked by PCR For positive detection (using the primer pair qGL3Seq-F / qGL3Seq-R), the positive single clone was sent to the biological company for sequencing, and the strain with the correct sequence was shaken to extract the plasmid with the target fragment. The obtained plasmid was digested with Smal I and connected to the vector plasmid pBI121 which was also digested to form recombinant plasmids pBI121-qGL3 and pBI121-q...

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Abstract

The invention relates to the technical field of plant genetic engineering and discloses clone and application of semi-dominant gene qGL3 capable of controlling grain length and grain weight of rice kernel. In the invention, a main-effective QTL (quantitative trait loci) SEQ ID NO:1 which is capable of simultaneously controlling the grain length and grain weight of rice kernel and a semi-dominant allelic gene SEQ ID NO:3 having advantageous function are separated and cloned; the two genes have the capacities of modifying the yield and appearance quality of rice; and at the same time, the two homologous genes of the two genes in rice are cloned and a fact that the two homologous genes have regulation effect on the length of rice kernel as well is proved. Thus, the gene qGL3 and the advantageous allelic gene qGL3-D thereof as well as the two homologous genes of the gene qGL3 all can be applied to crop genetic modification.

Description

technical field [0001] The invention relates to the technical field of plant genetic engineering, and relates to the cloning and application of a semi-dominant gene qGL3 for controlling the grain length and grain weight of rice, and specifically relates to a control gene qGL3 located on the long arm of the third chromosome of rice. Gene cloning and crop improvement application of the heavy major semi-dominant gene qGL3. Background technique [0002] As the world's population grows, rice, as a major food crop, faces an urgent need to increase its yield. It is estimated that by 2030, rice production needs to increase by more than 40% to meet human needs. At the same time, with the improvement of human living standards, higher and higher requirements are placed on the appearance quality of rice. People in southern China, America and European countries tend to prefer slender grain type, while northern China, Japan and Korea tend to prefer short round grain type (Unnevehr et al...

Claims

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Application Information

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IPC IPC(8): C12N15/55C12N9/16C12N15/84A01H5/00A01H5/10
CPCA01H5/10C12Q1/6895C12N15/8261C12Q2600/13Y02A40/146A01H1/00A01H6/4636
Inventor 张红生张晓军蓝虹霞王建飞王才林唐海娟
Owner NANJING AGRICULTURAL UNIVERSITY
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