Auxiliary screening method for indica rice and japonica rice

A technology for indica rice and japonica rice, applied in biochemical equipment and methods, microbe measurement/inspection, DNA/RNA fragments, etc., to achieve the effects of speeding up breeding, reducing breeding costs, and simple operation

Inactive Publication Date: 2010-06-02
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There is no report on the function of OsSRT2(LOC_Os12g07950)

Method used

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  • Auxiliary screening method for indica rice and japonica rice
  • Auxiliary screening method for indica rice and japonica rice
  • Auxiliary screening method for indica rice and japonica rice

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Example 1. Discovery and verification of different expression patterns of gene LOC_0s12g07950 in indica and japonica rice

[0021] First, by mining and analyzing the chip data of indica and japonica rice at different developmental stages and different tissue positions, it was found that the gene LOC_Os12g07950 has two sets of probes in the GengChip data, and the expression patterns of these two sets of probes in indica and japonica rice are opposite , The expression level of probe P1 in indica rice is very low or even not expressed, but the expression level in japonica rice is extremely high, while the expression level of the other probe P2 in indica rice is extremely high, but the expression level in japonica rice is extremely low . According to previous research results, the main reason for the difference in gene expression may be the difference in base sequence (InDel), or the regulation of other genes. Therefore, using the method of comparative genomics, firstly, the ...

Embodiment 2

[0022] Example 2. Design of primers for auxiliary screening of indica and japonica rice

[0023] In order to further verify whether there is a genomic difference between 93-11 and Nipponbare, based on the Nipponbare sequence, primers SIR2-2 were designed in areas where there may be genomic differences between the two. One primer sequence is shown in sequence 1 in the sequence table, and the other The sequence of a primer is shown in sequence 2 in the sequence table, and the genomic DNA of Nipponbare and 93-11 were used as templates for PCR amplification verification. The PCR amplification reaction system is: 20ng rice genomic DNA template, 0.5U Taq Plus DNA polymerase, 2.0μl 10×PCR buffer (100mM Tris·HClpH9.0, 500mM KCl, 15mM Mg 2+ , 1% Triton X-100), 100μM dNTPs, forward primer 4μM, reverse primer 4μM, supplement the reaction system with DEPC water to 20μl. The PCR reaction conditions are: first 94°C for 5 minutes; then 94°C for 30sec, 58°C for 45sec, 72°C for 1min30sec, a total...

Embodiment 3

[0025] Example 3 Verification of the feasibility of using primer SIR2-3 to assist in screening of subspecies of rice indica and japonica

[0026] Rice varieties and local species (all from the National Germplasm Bank): 93-11, Xianmazuo, Zinuo, Nepa (deep rice), Guichao No. 2, Aus6538 (Bamalaia), IR24, Teqing, Nipponbare, Dwarf Dazhong, Muli (light shell japonica rice), glutinous japonica rice, Yuefu, Qiuguang, Hongguang and Yunfeng 7.

[0027] First, 8 varieties and landraces of Indica and Japonica subspecies were randomly selected using morphological indicators and related literature search, as shown in Table 1:

[0028] Table 1. Identification results of indica and japonica subspecies of 16 rice varieties and landraces

[0029] Variety name

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PUM

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Abstract

The invention discloses a process of auxiliary screening hsien rice and japonica rice. The process for auxiliary screening hsien rice and japonica rice which is provided by the invention is used to detect whether rice losses ribonucleotide sequence of 67843-67878 from 5' end in GenBank accession AL731882, the detected rice is candidate hsien rice if lost, or the detected rice is candidate japonicarice if not lost. The process for auxiliary screening rice subspecies which is provided by the invention can be used to evaluate hsien rice and japonica rice breeds or local breed in early period, and plays important guiding roles for using heterosis of hsien rice and japonica rice and cultivating high-yield and high grade hybridization rice, thereby optimizing hybridized combination, accelerating breeding speed, reducing breeding cost, and having the advantages of simple operation, low cost and short cycle, which is suitable to be popularized and applied, and provides a fast choose method for the evaluation of rice subspecies germplasm.

Description

Technical field [0001] The invention relates to a method for auxiliary screening of indica and japonica rice. Background technique [0002] Cultivated rice is one of the most important food crops in the world. In its 11,500 years of domestication of cultivated rice, in order to adapt to different agro-ecological environments, cultivated rice has formed rich genetic diversity and obvious genetic differentiation, resulting in indica rice ( Indica and japonica (Morishima H, Oka HI. Phylogenetic differentiation of cultivated rice. XXII. Numerable evaluation of the Indica-Japonica differentiation. Japanese Journal of Breeding, 1981, 31: 402-413; Glaszmann J C. Isozymes and classification of Asian rice varieties. Theoretical and Applied Genetics, 1987, 74: 21-30; Wang ZY, Tanksley S D. Restriction fragment length polymorphism in Oryzasativa L. Genome, 1989, 32: 1113-1118; Blair MW, Panaud O, McCouch S R. Inter-simple sequence repeat (ISSR) amplification for analysis of microsatellite ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12N15/11
Inventor 苏震刘凤霞徐文英谭禄宾邸超张振海孙传清薛勇彪
Owner CHINA AGRI UNIV
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