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Method of imparting or controlling fertility with the use of fertility restoring gene for rice BT-male sterility cytoplasm and method of judging the existence of fertility restoring gene

A technology for restoring genes and rice, applied in the direction of genetic engineering, plant genetic improvement, chemical instruments and methods, etc., can solve the problems that cannot be excluded, cannot be completely guaranteed, and cannot correctly identify individual plants.

Inactive Publication Date: 2005-03-02
JAPAN TOBACCO INC
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

[0011] The second problem is that since the gene map distance between the RFLP marker G291 site and the G127 site is about 30cM (equivalent to about 9000kbp in rice DNA), the probability of double recombination in this region is considered to be 100%. Therefore, it cannot be fully guaranteed to correctly determine the genotype of the Rf-1 gene locus through these markers
[0012] The third issue is that if the presence of the Rf-1 gene is inferred by detecting the RFLP markers G291 and G127, not only the Rf-1 gene but also the gene region between these loci is introduced into the screened fertility restorer
Therefore, even if the combination of co-dominant markers and dominant markers is used to determine the genotype of the Rf-1 gene locus, it is not possible to correctly identify whether the individual plants are homozygous or heterozygous for the Rf-1 gene
In addition, the PCR reaction with a dominant marker did not get a PCR product, and it cannot be ruled out that it may be caused by a problem with the experimental procedure.

Method used

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  • Method of imparting or controlling fertility with the use of fertility restoring gene for rice BT-male sterility cytoplasm and method of judging the existence of fertility restoring gene
  • Method of imparting or controlling fertility with the use of fertility restoring gene for rice BT-male sterility cytoplasm and method of judging the existence of fertility restoring gene
  • Method of imparting or controlling fertility with the use of fertility restoring gene for rice BT-male sterility cytoplasm and method of judging the existence of fertility restoring gene

Examples

Experimental program
Comparison scheme
Effect test

Embodiment approach

[0209] A preferred mode of the method for identifying whether the tested rice has the Rf-1 gene is illustrated by the following example description. In the example here, it was found that the nucleotide sequence of the Rf-1 gene (SEQ ID NO: 27) carried by the indica rice variety IR24 has at least the following 1)-8) polymorphisms compared with the corresponding region of the japonica rice variety:

[0210] 1) The base corresponding to the 1239th base of SEQ ID NO: 27 is A;

[0211] 2) The base corresponding to the 6227th base of SEQ ID NO: 27 is A;

[0212] 3) The base corresponding to the 20680th base of SEQ ID NO: 27 is G;

[0213] 4) The base corresponding to base 45461 of SEQ ID NO: 27 is A;

[0214] 5) The base corresponding to the 49609th base of SEQ ID NO: 27 is A;

[0215] 6) The base corresponding to base 56368 of SEQ ID NO: 27 is T;

[0216] 7) The base corresponding to base 57629 of SEQ ID NO: 27 is C; and

[0217] 8) The base corresponding to base 66267 of SEQ...

Embodiment 1

[0392] Example 1: Obtaining recombinants close to the Rf-1 site

[0393] (Materials and methods)

[0394] Pollination of MS Koshihikari (BC10F1 generation) with pollen from MS-FR Koshihikari (BC9F1 generation, heterozygous at the Rf-1 gene locus) produced a BC10F1 population consisting of 4103 individuals, at the S12564 Tsp509I and C1361 MwoI locus The genotypes of the dots were the same as those described in Reference Example 2. Among them, the Koshihikari individual homozygous at the S12564 Tsp509I site is considered to be due to the recombination between Rf-1 and S12564 Tsp509I, while the Koshihikari individual homozygous at the C1361 MwoI site gene is considered to be due to the recombination between Rf-1 and S12564 Tsp509I. -1 and C1361 MwoI due to recombination. DNA was extracted from each of these 4103 individuals.

[0395] (Results and discussion)

Embodiment 2

[0398] Example 2 Chromosome Walking

[0399] (1) The first chromosome walk

[0400] (Materials and methods)

[0401] According to the method described in Reference Example 1, a genomic library of genomic DNA of Asominori japonica rice (without Rf-1) was constructed using the Lambda DASH II vector, and detected by chromosome walking.

[0402] According to conventional methods, the total DNA of Asominori was used as a template, and the following primer pairs were used for PCR reaction:

[0403] 5'-atcaggagccttcaaattgggaac-3' (SEQ ID NO: 29) and

[0404] 5'-ctcgcaaattgcttaattttgacc-3' (SEQ ID NO: 30)

[0405] The two primers are designed according to the partial base sequence of RFLP probe S12564 (Accession No. D47284). Agarose gel electrophoresis showed that the amplified product was about 1200bp, and it was purified with QIAEXII (QIAGEN). The purified DNA was labeled with the Rediprime DNA labeling system (Amersham Pharmacia) to form library screening probes (probe A, f...

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Abstract

The purpose of the present invention is to provide a method for providing and inhibiting the rice fertility to the rice BT type cytoplasmic male sterility, and discerning the presence of the rice restorer gene. The present invention employs a nucleic acid having the base sequence of SEQ ID NO.27, or a nucleic acid having a base sequence which is identical to at least 70% of the base sequence of SEQ ID NO.27, and which functions to restore fertility. Alternatively, a nucleic acid having the base sequence of bases 38538-54123 of SEQ ID NO.27, or having a base sequence which is identical to at least 70% of the base sequence of bases 38538-54123 of SEQ ID NO.27, and which functions to restore fertility, is used.

Description

technical field [0001] The invention relates to a method for obtaining or inhibiting rice fertility by utilizing a restorer gene of rice BT-type cytoplasmic male sterility and identifying the restorer gene of rice. [0002] This application claims Japanese Patent Application No.2001-285247 filed on September 19, 2001, Japanese Patent Application No.2001-309135 filed on October 4, 2001 and Japanese Patent Application No. 2002 filed on June 26, 2002 Priority of .2002-185709. The full disclosures of these three patent applications are hereby incorporated herein Background technique [0003] Rice is a self-pollinated plant. When crossing between rice varieties, first all the stamens in the spikelets must be removed before rice flowering to avoid self-pollination, and then the pollen from the parent variety is used to obtain the hybrid variety. Fertility. However, this method of artificial hybridization is totally unsuitable for commercial purposes to produce large quantities ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/82
CPCC12N15/8289C07K14/415
Inventor 小森俊之太田象三村井宣彦樋江井佑弘
Owner JAPAN TOBACCO INC
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