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Molecular marker for determining rice cross affinity

A molecular marker, rice hybridization technology, applied in hybridization, biochemical equipment and methods, microbial determination/inspection, etc., can solve the problem of low fertility of indica-japonica hybrid F1, and achieve the effect of rapid and stable family

Pending Publication Date: 2022-07-08
CAS CENT FOR EXCELLENCE IN MOLECULAR PLANT SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] Indica-japonica hybrids have strong heterosis, but the fertility of F1 offspring of indica-japonica hybrids is very low (Qiu et al.2005)

Method used

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  • Molecular marker for determining rice cross affinity
  • Molecular marker for determining rice cross affinity
  • Molecular marker for determining rice cross affinity

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Main instruments: PCR machine, ABI 3730 sequencer.

[0031] Rice gene donor varieties: 32 rice varieties were donated by Shanghai Institute of Plant Physiology and Ecology, Chinese Academy of Sciences.

[0032] Molecular marker screening includes the following steps:

[0033] 1) extracting the genomic DNA of the rice plant to be tested;

[0034] 2) PCR amplification of rice genomic DNA with primers for each gene S5-ORF3, S5-ORF4 and S5-ORF5 respectively;

[0035] PCR reaction system is calculated as 20μl: 10X PCR reaction buffer 2μ1, 25mM MgSO 4 0.8μl, 2mM dNTP 2μ1, 5μM forward primer (-F) and reverse primer (-R) each 1.2μ1, genomic DNA 20ng, KOD-Plus polymerase 0.4μl, add ddH 2 O is made up to 20 μl.

[0036] PCR reaction conditions were: 94°C pre-denaturation for 2 minutes, 94°C denaturation for 15 seconds, 58°C annealing for 30 seconds, 68°C extension for 1 minute, a total of 35 cycles; 68°C incubation for 5 minutes.

[0037]3) purify the PCR products of the SN...

Embodiment 2

[0071] Example 2: Molecular markers for determining rice hybrid affinity

[0072] The genotypes of the three genes S5-ORF3, S5-ORF4 and S5-ORF5 of 32 rice materials were sequenced, which was in line with the fact that the hybrid affinity identification had been determined through traditional breeding experiments. The results are shown in Table 1.

[0073] Table 1. Genotypes and broad-compatibility relationships of rice genes S5-ORF3, S5-ORF4 and S5-ORF5

[0074] Rice varieties ORF3 ORF4 ORF5 broad affinity HP486 ORF3+ ORF4- ORF5-i no broad affinity HP274 ORF3+ ORF4- ORF5-i no broad affinity HP119 ORF3+ ORF4- ORF5-i no broad affinity HP341 ORF3+ ORF4- ORF5-i no broad affinity HP362 ORF3+ ORF4- ORF5-i no broad affinity HP327 ORF3+ ORF4- ORF5-i no broad affinity HP492 ORF3+ ORF4- ORF5-i no broad affinity HP577 ORF3+ ORF4- ORF5-i no broad affinity HP396 ORF3...

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Abstract

The invention discloses a molecular marker for determining the cross affinity of rice, the molecular marker is the genotype of a rice gene S5-ORF5, and when the genotype of the S5-ORF5 is ORF5-n as shown in a nucleotide sequence SEQ ID NO: 1, the rice is prompted to have wide affinity. The molecular marker can be used for guiding rice crossbreeding.

Description

technical field [0001] The invention belongs to the field of gene detection, and in particular relates to a molecular marker for determining hybridization affinity of rice and its application. Background technique [0002] Indica-japonica hybrids have strong heterosis, but indica-japonica hybrids have low F1 fertility (Qiu et al. 2005). This phenomenon is mainly controlled by three genes S5-ORF3, S5-ORF4 and S5-ORF5 on the sixth chromosome of rice. [0003] ORF4 and ORF5 are only 832bp apart, they are arranged head-to-head, the promoters share the same sequence, and the directions are opposite, so the transcription directions of the two genes are also opposite. ORF5 acts as a killer, it can transmit signals to ORF4, causing endoplasmic reticulum stress, leading to programmed cell death and embryo sac abortion (Chen et al. 2008). ORF3+ plays a protective role. In indica-japonica hybrids, female gametes carrying the guard gene ORF3+ can prevent the stress response of the end...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12N15/11G16B5/00G16B20/20G16B20/40G16B25/20
CPCC12Q1/6895G16B5/00G16B25/20G16B20/20G16B20/40C12Q2600/13C12Q2600/156
Inventor 韩斌司丽珍吕威赵强上官颖颖朱静洁
Owner CAS CENT FOR EXCELLENCE IN MOLECULAR PLANT SCI