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A Molecular Marker of Rice Panicle Length Gene qpnl‑12

A molecular marker, qpnl-12 technology, applied in the field of rice genetic improvement and agricultural biotechnology applications, to achieve strong specificity, high stability, and improve the efficiency of breeding and selection

Active Publication Date: 2018-03-02
JIANGSU ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Based on the above statement, there is no report about the location of rice panicle length-related genes on the 12th chromosome

Method used

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  • A Molecular Marker of Rice Panicle Length Gene qpnl‑12
  • A Molecular Marker of Rice Panicle Length Gene qpnl‑12
  • A Molecular Marker of Rice Panicle Length Gene qpnl‑12

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1 Development of Molecular Marker Tightly Linked with Rice Neck Length New Gene qPNL-12

[0024] 1) Phenotype of substitution line C115 and construction of segregation population

[0025] Using a set of high-generation backcross replacement lines with 119 lines constructed with the indica rice variety 9311 as the recipient parent and the japonica rice variety Nipponbare as the donor parent, a very short panicle neck was detected through the investigation of the panicle neck length traits The phenotype of the replacement line C115 has a very significant difference in the neck length of 9311 ( figure 1a, b), and there are also significant differences in plant height between C115 and 9311 ( figure 1 c), other agronomic traits such as panicle length, panicle number, grains per panicle and seed setting rate were not significantly different ( figure 1 d-g), the relative length of the inverted I stem node and the plant height of C115 has a very significant difference...

Embodiment 2

[0033] Example 2 Polymorphism Detection and Linkage Analysis of a Molecular Marker Tightly Linked to the New Rice Panicle Length Gene qPNL-12

[0034] 1) Genomic DNA extraction

[0035] Leaf samples were taken from rice materials to be extracted for DNA at the peak tillering stage of rice. Genomic DNA of parents 9311, C115 and 30 F2 individual plants were extracted for polymorphism screening of 22 InDel markers; 306 individual plants with short neck phenotype were extracted from F2 population for linkage analysis of polymorphic markers and qPNL-12 gene fine mapping; extract 100 F3 individual plants for subsequent effect verification of molecular markers.

[0036] Genomic DNA was extracted from rice leaves using the SDS method, and the operation steps were referred to Della porta S L, et al., Plant Mol Biol Rep, 1983, 1(1):19221.

[0037] The specific steps are: take the rice seedling leaves, grind them with liquid nitrogen in a mortar pre-cooled at -20°C and put them into a ...

Embodiment 3

[0045] Example 3 Cloning and sequencing of molecular markers Ind1529 and Ind1561

[0046] Using the parental 9311 and C115 genomic DNA as templates, the forward and reverse primers of molecular markers Ind1529 and Ind1561 were used as Ind1529-F / Ind1529-R and Ind1561-F / Ind1561-R primer pairs, respectively, and high-fidelity enzymes were used for PCR amplification . Take 1 μL of PCR product and connect with pMD19-T vector at room temperature for 10 minutes, transform Escherichia coli competent cells DH5α by heat shock, and culture the transformed bacteria in 100ml LB liquid medium for 1 hour with simple shaking, and then spread it on the medium containing 100μg / ml ampicillin LB solid plates were incubated at 37°C for 14 hours. After colony PCR detection, positive clones were selected and sent to Shanghai Handsome Biotech Co., Ltd. for DNA sequence determination.

[0047] The sequencing results showed that the length of the amplified band of the primers Ind1529-F / Ind1529-R in C...

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Abstract

The invention discloses a molecular marker of rice spike neck length gene qPNL-12 which is divided into an upstream molecular marker Ind1529 and a downstream molecular marker Ind1561 respectively aiming to specific insertion / deletion locus polymorphism represented as the SEQ ID No.1 and the SEQ ID No.2. The invention also discloses specific primers comprising SEQ ID No.3, SEQ ID No.4, SEQ ID No.5 and SEQ ID No.6, which are used for amplification on the molecular marker. The molecular marker or the specific primers can be applied in marker assisted selective breeding of the character of spike neck length of a progeny and derivative materials thereof containing the qPNL-12 gene, can save cost and can increase efficiency of breeding and selection.

Description

technical field [0001] The invention belongs to the fields of rice genetic improvement and agricultural biotechnology application, and relates to a molecular marker of the rice panicle neck length gene qPNL-12, in particular to a nucleotide sequence closely linked with the qPNL-12 gene molecular marker and a molecule used for amplification Labeled forward and reverse specific primers. Background technique [0002] Ear neck length, also known as ear extraction, refers to the distance from the flag leaf sheath to the ear base. It is an important agronomic trait connecting the stalk and ear. During the transportation of nutrients and water from the stalk and leaves to the ear play an extremely crucial role. Most of the rice sterile lines currently used in production have different degrees of ear wrapping characteristics (caused by too short ear extraction), which greatly affects the improvement of hybrid rice seed production. In the 1980s, the discovery of the rice long panic...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12N15/11
Inventor 赵春芳梁彦丽王才林周丽慧陈涛张亚东赵庆勇
Owner JIANGSU ACAD OF AGRI SCI
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