KR101922420B1 - Composition comprising DNA marker derived from Nampyeongbyeo for selecting rice variety resistant to bakanae disease and method of selecting rice variety resistant to bakanae disease using the DNA marker - Google Patents

Abstract

The present invention relates to a composition for screening rice blast-resistant rice varieties comprising a DNA marker derived from Nambyeong Rice, and more particularly to a primer pair represented by SEQ ID NOs: 249 and 250 for 1625IND and SEQ ID NOs: 251 and 252 for 1675IND Wherein the primer pair comprises at least one primer pair selected from the group consisting of primer pairs to be displayed. Also, the present invention relates to a kit for screening a susceptible rice variety, which comprises the above composition, a method for screening a susceptible rice variety resistant to Kidari disease, and a method for producing a susceptible rice variety. The present invention relates to a primer set for cleaved amplified polymorphic sequences (CAPS) marker amplification for genetic mapping for determining quantitative trait loci (QTLs) associated with resistance to a keypad disease.
The DNA marker of the present invention can be efficiently distinguished from the susceptibility of susceptible individuals by using the composition for screening the susceptible rice varieties comprising the DNA markers derived from the Nambyeong plant according to the present invention. It can be used for rice breeding with improved disease resistance. Therefore, the promotion of the development of Rice Kidney Disease Resistant Varieties can reduce the damage to the Kidari disease and reduce the amount of pesticide use.

Description

[0001] The present invention relates to a composition for screening a variety of rice-resistant rice varieties including DNA markers derived from Nambyeong Rice, and a method for screening the rice-resistant rice varieties using the DNA markers. resistant to bakanae disease using the DNA marker}

The present invention relates to a composition for screening rice blast-resistant rice varieties comprising a DNA marker derived from Nambyeong Rice, and more particularly to a primer pair represented by SEQ ID NOs: 249 and 250 for 1625IND and SEQ ID NOs: 251 and 252 for 1675IND Wherein the primer pair comprises at least one primer pair selected from the group consisting of primer pairs to be displayed. Also, the present invention relates to a kit for screening a susceptible rice variety, which comprises the above composition, a method for screening a susceptible rice variety resistant to Kidari disease, and a method for producing a susceptible rice variety. The present invention relates to a primer set for cleaved amplified polymorphic sequences (CAPS) marker amplification for genetic mapping for determining quantitative trait loci (QTLs) associated with resistance to a keypad disease.

Rice Kidari disease is a common infectious disease caused by Fusarium fujikuroi , a fungal pathogen. It is named Bakanae disease due to abnormal growth of the seedlings. It was first reported in Japan in 1898. In Korea, it was a serious problem in some farms in the 1960s. In recent years, severe infections in rice seeds and the emergence of resistant bacteria to medicines have led to a rapid increase in the incidence, It is a tendency to develop. In addition, since rice varieties with high resistance to rice seedling disease have not been developed, enormous pesticides and labor force are consumed for spraying pesticides for control, which is a major obstacle to production cost reduction and production of environmentally friendly agricultural products.

In order to sterilize rice seeds, i) a hot water bath at an appropriate temperature is made to drench sowing seeds for a predetermined period of time, thereby conducting a hot water immersion method, which is a method of sterilizing and preventing pathogenic spores and harmful microorganisms, or ii) New seed disinfectants such as hexaconazole and tebuconazole have been developed and used. However, there is a limitation in that it is difficult to treat a large amount of seeds at a uniform temperature, and the use of the seed disinfectant , There is a concern that the appearance of a drug resistance-resistant microorganism may occur. Therefore, it is necessary to identify resistant cultivars against rice pathogens in order to solve the root disease of rice.

The conventional method for assaying resistance to rice hind pandemic disease includes: i) a method of seeding seedlings in a spore culture of a pathogenic bacterium and then seeding the seedlings, A method for investigating the onset of Kidari disease in a seedling by sowing seeds on a soil mixed with a pathogenic bacterium cultivated in a medium, iii) an indoor test method using a test tube culture, iv) a method in which rice seeds are immersed in a spore culture medium, And the like. However, these methods have a limitation in that accuracy is decreased according to the method, a large area is required, and a long time is required for the verification.

Generally, it takes 8 ~ 12 years to develop and produce new varieties, and a wide cultivation area is needed for system development and selection. Most of the rice cultivars cultivated in Korea are weak against rice pandemic, so the development of resistant cultivars is very demanding. Therefore, there is a need for a molecular labeling method, which is the most effective method for obtaining precise and objective test results, in order to reduce the time and cost required for selection of resistance resources and lines for rice genetic resources and mating groups. Molecular labeling methods are widely used in molecular breeding systems for the search of useful traits, identification of species of organisms, identification of breed classification, and analysis of the relationship of group populations. Since the rice seedlings using the molecular markers can be traced in a young period without being affected by the environmental variation, it is possible to analyze a large amount of resources accurately and quickly (Korean Patent No. 10-1509076).

Recently, the development of Next Generation Sequencing has resulted in a decrease in the cost of large-scale sequencing and acceleration of speed. In addition, when the rice genome is decoded, whole genome re-sequencing is performed based on a standard genome (reference sequence), and various markers such as SNP, insertion and deletion are mass- It can be developed in a short time. In China, a large number of SNPs were searched for by genome sequence analysis of native rice plants to produce a high-density molecular genetic map ( Nature Genet. , 42: 961-967, 2010).

Agronomically important traits in crop breeding are not independent single genes but quantitative traits that are controlled by numerous genes on the entire chromosome. Quantitative traits are generally affected not only by a large number of genes but also by various environmental factors. Thus, in quantitative traits, it is extremely difficult to qualify traits such as individual gene actions or traits that affect them. Therefore, statistical methods have been used as a powerful tool for studying quantitative traits, and studies have been conducted by several scholars to effectively isolate genetic factors affecting quantitative traits from various environmental factors. The use of molecular genetic techniques for breed improvement will enable the study of the genetic predisposition of individuals at the DNA level, and it will be possible to identify genes related to the target traits to be improved, that is, major genes or loci of quantitative traits (QTL), or selection of genetic markers associated with quantitative trait loci (QTLs) can provide a tool for realizing a genetic locus. By using molecular genetic information rather than relying solely on phenotypic information, genetic improvement can be accelerated. Therefore, in order to study such a quantitative trait locus today, many approaches such as finding a gene through high density molecular genetic mapping and revealing its function are being performed. To date, various DNA markers have been developed, including restriction fragment-length polymorphism (RFLP), randomly amplified polymorphism (RAPD) and amplified fragment length polymorphism (AFLP) to generate high density molecular genetic maps ( Genetics , 148: 479-494, 1998 ).

Conventional DNA markers are not easy to identify, costly and take a long time. Moreover, the frequency of appearance is low. Recently developed SNPs (single nucleotide polymorphism) show a much higher polymorphism among varieties or individuals, and are widely used as markers because of their high utilization value because they are distributed evenly.

Under these circumstances, the present inventors have made intensive efforts to develop DNA markers that can be efficiently used to distinguish rice cultivars resistant to Kidney Disease. As a result, they have developed a new CAPS marker for determining QTL associated with Kidney Disease Resistance, In addition, it is used for the mapping, and the succeeding group derived from the crossing between the Kindari disease resistant varieties such as Nambyeongbyeon and Kidari disease and the mutant strain Dongjin AD, which is sensitive to the Kindari disease, is cultivated and PCR and agarose gel And developed an Indel (insertion / deletion) marker which can be easily analyzed by using the strain and accurately select the strain resistant to the genus.

Korean Patent No. 10-1509076

Nature Genet., 42: 961-967, 2010 Genetics, 148: 479-494, 1998

It is an object of the present invention to provide a composition for screening a rice blast resistance resistant rice variety, a kit for screening a susceptible rice variety with the above-mentioned composition, a method for screening a susceptible rice variety for a susceptible disease, and a method for producing a susceptible rice varieties .

Another object of the present invention is to provide cleaved amplified polymorphic sequences (CAPS) markers for genetic mapping to determine the quantitative trait loci associated with the resistance of the tympanic resistance of the succeeding population derived from crosses between Nampwa and Dongjin AD And a method for producing a genetic map for determining a quantitative trait locus related to a susceptibility of a succeeding population of the Nam Pyeong and Dongjin AD.

In one aspect to accomplish the above object, the present invention provides a primer pair comprising a pair of primers represented by SEQ ID NOS: 249 and 250 for 1625 IND, and a pair of primers represented by SEQ ID NOS: 251 and 252 for 1675 IND The present invention provides a composition for screening a rice blast resistance resistant rice variety comprising the above primer pair.

In the present invention, the term " bakanae disease " is a representative seed infectious disease caused by Fusarium fujikuroi , and the leaves become pale yellow immediately after the seedling of the rice or the transplant The length of the pool is close to twice that of healthy. In the pathogenesis of seed transmission, the fungus of the pathogen formed on the fungus after the flowering of the rice is scattered and becomes infected. In addition, new infections occur when the seeds are mixed when immersed in water to pre-sow. If pathogens are invading, they do not germinate or die soon. The untreated is painted as described above and exhibits typical symptoms, but is terminated at the end.

In the present invention, the term " rice paddy rice resistance " refers to a rice plant resistant to pandemic disease, which has a trait that prevents the occurrence of pandemic disease in rice. The term resistance refers to the property that when a pathogen is infected, it does not easily catch disease, and the term disease resistance is also used.

In the present invention, the term " rice ( Oryza stiva "is an annual plant of the monocotyledonous liana, and it is planted in rice field and field with the edible crops of the East Indies. It is about 1m high and the leaves are thin and long. Rice is the main food of 40% of the world's population and more than 20 kinds of plants belong to the rice family. (O. sativa) are mostly cultivated. Kidari disease, blast disease, blight of blight, blight of stripe leaf blight, mildew disease, and mite (moth, moth) are known.

The term " primer " in the present invention is a short nucleic acid sequence having a free 3 'hydroxyl group at the free 3' end and can form a base pair with a complementary template. Means a nucleic acid sequence functioning as a starting point. Primers can initiate DNA synthesis in the presence of reagents for polymerization (i. E., DNA polymerase or reverse transcriptase) and four different nucleotide triphosphates at the appropriate buffer solution and temperature.

The primers used in the present invention include a hybridization nucleotide sequence complementary to the target nucleic acid. The term " complementary " means that under certain annealing or hybridization conditions the primer is sufficiently complementary to selectively hybridize to the target nucleic acid sequence, and is substantially complementary and perfectly complementary, , And preferably means completely complementary.

In another aspect, the present invention provides a method of producing a rice plant, comprising: (a) separating genomic DNA from a rice sample; (b) a pair of primers represented by SEQ ID NOS: 249 and 250 for the 1625 IND using the separated genomic DNA as a template; And amplifying the target sequence by performing amplification reaction using at least one primer pair selected from the group consisting of primers represented by SEQ ID NOS: 251 and 252 for 1675 IND; And (c) detecting the amplification product.

The term " sample " in the present invention refers to a tissue of rice that can be infected with a pox, and includes all organs and cells derived from the tissue of a plant including seed, root, leaf, stem or flower, A callus, a plant tissue selection, a culture, and a transformed tissue for regeneration of the plant.

The term " amplification reaction " in the present invention means a reaction for amplifying a nucleic acid molecule. A variety of amplification reactions have been reported in the art, including polymerase chain reaction (PCR) (US Pat. Nos. 4,683,195, 4,683,202 and 4,800,159), reverse-transcription polymerase chain reaction (RT-PCR) (Sambrook et al., Molecular Cloning. (LCR) method, the method of Lambert, HI (WO 89/06700) and Davey, C. et al (EP 329,822) Gap-LCR (WO 90/01069), repair chain reaction (EP 439,182), transcription-mediated amplification (TMA) (WO 88/10315), self sustained sequence (WO 90/06995), selective amplification of target polynucleotide sequences (US Patent No. 6,410, 276), consensus sequence primed polymerase chain reaction (CPPCR) (U.S. Patent No. 4,437,975), arbitrary priming (U.S. Patent Nos. 5,413,909 and 5,861,245), nucleic acid sequence based amplification (NASBA) (U.S. Patent Nos. 5,130,238, 5,409,818 5,554,517, and 6,063,603), strand displacement amplification and loop-mediated isothermal amplification. LAMP), but is not limited thereto. Other amplification methods that may be used are described in U.S. Patent Nos. 5,242,794, 5,494,810, 4,988,617 and U.S. Patent No. 09 / 854,317.

As used herein, the term " target sequence " may be Indel markers specifically detected in resistant cultivars. Preferably, the target sequence comprises a pair of primers represented by SEQ ID NOS: 249 and 250; And the primer pairs represented by SEQ ID NOS: 251 and 252, respectively.

The term " Indel (insertion / deletion) marker " in the present invention collectively refers to a mutation in which some bases are inserted or deleted in the nucleotide sequence of DNA. The Indel marker searches for insertion or deletion regions of the standard dielectric by comparing and analyzing the genomic information of the genotypes used in the experiment and the primers based on the information. Therefore, the amplification result can show two types of insertion and deletion in comparison with the standard genome. In the present invention, the term " marker " means a base sequence used as a reference point when identifying genetically unrelated loci, and the term " locus " means a position on a genetic map of a molecular marker.

In the present invention, the term " standard dielectric " refers to a genome of a variety of crops which is a standard for recognizing the breeds of the present invention. But it is not limited thereto.

In the present invention, the term " detection " refers to confirming the presence or absence of the Indel marker 1625IND or 1675IND used for confirming the presence of the keypad disease resistance gene in the sample.

In one embodiment of the present invention, 1625IND and 1675IND Indel markers, which are DNA markers derived from Nambyeong Rice, were developed and the selection efficiency of the rice cultivars resistant to Kinderian disease was tested using the developed markers. As a result, on the agarose gel, (Fig. 9). It was confirmed that the strain of the present invention can be efficiently screened for resistance to the insect resistance.

Specifically, the step (a) is a step of isolating genomic DNA from a rice sample. Examples of samples that can be used are as described above, but are not limited thereto. A DNA extraction kit can be used as a method of isolating genomic DNA from a rice sample, and a method known in the art can be easily modified by those skilled in the art.

The step (b) comprises amplifying the target sequence by using the genomic DNA isolated in step (a) as a template and performing amplification reaction using a primer pair capable of specifically binding to a gene having resistance to a tympanic disease or a fragment thereof . In one embodiment of the present invention, a pair of primers represented by SEQ ID NOS: 249 and 250; And a pair of primers represented by SEQ ID NOS: 251 and 252 were used to amplify the gene in a sample having a keypad disease resistance gene, and when the primer pair was used, the target sequence Was amplified. The method for amplifying the nucleic acid sequence may be selected from the group consisting of a polymerase chain reaction (PCR), a ligase chain reaction, nucleic acid sequence-based amplification, a transcription-based amplification system, Any other suitable method for amplifying nucleic acid molecules known in the art can be used, such as by strand displacement amplification or amplification through Q-replicase, and methods known in the art can be used by the skilled person And can be used. In one embodiment of the present invention, the target sequence was amplified using a polymerase chain reaction.

In addition, the amplified nucleic acid sequence may further be labeled with a detectable labeling substance. The labeling substance may be selected from the group consisting of an enzyme, a ligand, a coloring material, a microparticle, a radioactive isotope, a fluorescence, and a phosphorescent material, but is not limited thereto.

Examples of the enzyme used as a detection marker include acetylcholinesterase, alkaline phosphatase,? -D-galactosidase, horseradish peroxidase,? -Lactamase and the like, and ligands include biotin derivatives and the like And colloid, gold, colored latex and the like, and the radioactive isotope includes 37Co, 3H, 125I, and 125I-Bolton ( Bonton) Hunter reagent and the like, and the fluorescent material includes Cy3, Cy5, fluororesin isothiocyanate (FITC), tetramethylrhodamine isothiocyanate (RITC), Alexa, 4,4 -Difluoro-4-boro-3a, 4a-diaza-s-indacene (BODIPY), Texas Red or biotin.

Wherein said step (c) comprises the step of detecting the amplification product of step (b). The method for detecting the amplification product can be detected by electrophoresis of the amplification product. Specific examples of electrophoresis include, but are not limited to, automated electrophoresis (including chip and capillary methods), acrylamide gel, high performance liquid chromatography (HPLC), or agarose gel electrophoresis.

In yet another embodiment, the present invention provides a primer pair comprising at least one primer pair selected from the group consisting of a pair of primers represented by SEQ ID NOS: 249 and 250 for 1625 IND and a pair of primers represented by SEQ ID NOS: 251 and 252 for 1675 IND; And a reagent for carrying out an amplification reaction.

The kit of the present invention can identify the resistance to rice plants by confirming the presence or absence of Indel which can confirm the presence of the resistance gene of the rice blast resistance. The kit of the present invention may include not only a primer for measuring the presence of the Indel but also one or more other component compositions, solutions or devices suitable for the assay method. Specifically, in the present invention, the kit for measuring the presence or absence of the Indel comprises a sequence selected from the group consisting of a primer pair represented by SEQ ID NO: 249 and 250, and a primer pair represented by SEQ ID NO: 251 and 252 against 1675 IND And may be a kit that may include one or more primer pairs.

The tympanic resistance-resistant rice selection kit according to the present invention may further include DNA polymerase, dNTPs and / or a reaction buffer, but is not limited thereto. Wherein the buffer may be required for the PCR reaction, which has a conventionally known composition in the art and may include, for example, Tris-HCl, MgCl2, KCl, and the like. The kit of the present invention may further comprise components necessary for electrophoresis to confirm amplification of the PCR product.

In yet another aspect, the present invention provides a method of producing a medicament for the treatment of (a) obtaining a progeny population derived from breeding between a Kidney Disease resistant strain and a Kidney Disease susceptible variety; (b) separating the genomic DNA from the rice samples obtained in the later group; (c) one or more primers selected from the group consisting of a pair of primers represented by SEQ ID NOS: 249 and 250 for 1625 IND and a pair of primers represented by SEQ ID NOS: 251 and 252 for 1675 IND using the separated genomic DNA as a template Amplifying the target sequence by performing amplification reaction using the pair; And (d) detecting the amplification product to determine resistance.

In the present invention, the term " later group " is a hybrid group produced by cross breeding, including, but not limited to, crossbreeding of the susceptible strain of the present invention, (F2) or third generation (F3) line (pedigree) produced by the manufacturer.

The resistant strain of the present invention which can be used in the production of the rice blast resistance resistant rice varieties of the present invention may be a rice blast resistant strain among the second generation (F2) or third generation (F3) strain produced by crossing between the Nam Pyong rice and the Nam Pyung rice.

On the other hand, the susceptible varieties of Kidari disease include all rice varieties which are not proved to be resistant to Kidari disease. Or the rice varieties determined to have susceptibility to Kidari disease according to the method for screening the susceptible rice varieties using the DNA marker according to the present invention.

In the present invention, the terms "sample", "resistance to keypad disease" and "detection" are as described above.

In addition, the method of amplifying and detecting the target sequence is as described above.

Preferably, if the size of the amplified product is confirmed to be the same as the size of the amplified product of Nampegwa, a resistant strain of Kidari disease, by amplification and detection of the target sequence, the corresponding strain can be judged as a resistant strain of Kidari disease. That is, it is possible to judge whether or not the resistance to the keypad disease is caused by the difference in size of the amplified product.

In one embodiment of the present invention, the susceptible varieties of Kidari disease on agarose gels were selected using 1625IND and 1675IND Indel markers, which are DNA markers derived from Nambyeong Rice. As a result, susceptible varieties of Kidari disease, Dongjinbyeon, Anambi, Ilhami, Haami, Samgwangbyeon, Ungwangbyeol, Sogyeongjangbyeol, Chungbokpyeon, Ohdabyeong, Chowun, Hyeonpyeong, (Fig. 9). It can be seen from Fig. 9 that the rice bran of the present invention exhibits electrophoretic bands different in size from those of the susceptible rice cultivars such as Kidari disease and Shanori rice.

In yet another embodiment, the present invention provides a primer pair represented by SEQ ID NOs: 1 and 2 for JNC010055, a primer pair represented by SEQ ID NOs: 3 and 4 for JNC010084, a primer pair represented by SEQ ID NOs: 5 and 6 for JNC010086 Pair, a pair of primers represented by SEQ ID NOs: 7 and 8 for JNS01057, a pair of primers represented by SEQ ID NOs: 9 and 10 for JNC010139, a pair of primers represented by SEQ ID NOs: 11 and 12 for JNS01030, And 14, a pair of primers represented by SEQ ID NOs: 15 and 16 for JNC010809, a pair of primers represented by SEQ ID NOs: 17 and 18 for JNC011008, a pair of primers represented by SEQ ID NOs: 19 and 20 for JNC011512, The primer pair shown in SEQ ID NOs: 21 and 22 for JNC011753, the primer pair shown in SEQ ID NOs: 23 and 24 for JNC011767, the sequence for JNC011804 A pair of primers represented by SEQ ID NOS: 27 and 28 for JNC011850, a pair of primers represented by SEQ ID NOS: 29 and 30 for JNC011887, a pair of primers represented by SEQ ID NOS: 31 and 32 for JNC020021, Pair, a pair of primers represented by SEQ ID NOS: 33 and 34 for JNC020047, a pair of primers represented by SEQ ID NOS: 35 and 36 for JNC020052, a pair of primers represented by SEQ ID NOS: 37 and 38 for JNC020058, a pair of primers represented by SEQ ID NO: 39 And 40, a pair of primers represented by SEQ ID NOs: 41 and 42 for JNC020149, a pair of primers represented by SEQ ID NOs: 43 and 44 for JNC020176, a pair of primers represented by SEQ ID NOs: 45 and 46 for JNC020186, The primer pair shown in SEQ ID NO: 47 and 48 for JNC030012, the primer pair shown in SEQ ID NO: 49 and 50 for JNC030028, the JNC0301 51 and 52 for JNC030134, a pair of primers represented by SEQ ID NOs: 53 and 54 for JNC030134, a pair of primers represented by SEQ ID NOs: 55 and 56 for JNC030270, a pair of primers represented by SEQ ID NOs: 57 and 58 for JNC030275 59 and 60 for JNC030408, a pair of primers represented by SEQ ID NOs: 61 and 62 for JNC030411, a pair of primers represented by SEQ ID NOs: 63 and 64 for JNC030606, a pair of primers represented by SEQ ID NO: The primer pair shown in SEQ ID NO: 65 and 66, the primer pair shown in SEQ ID NO: 67 and 68 for JNC040055, the primer pair shown in SEQ ID NO: 69 and 70 for JNC040202, and the SEQ ID NO: 71 and 72 for JNC040270 Primer pair shown in SEQ ID NO: 73 and 74 for JNC040277, primer pair shown in SEQ ID NO: 75 and 76 for JNC040334 79 and 80 for JNC050127, a pair of primers represented by SEQ ID NOs: 81 and 82 for JNC050148, a pair of primers represented by SEQ ID NO: 77 and 78 for JNC050091, a pair of primers represented by SEQ ID NO: 83, and 84, a pair of primers represented by SEQ ID NOs: 85 and 86 for JNC050166, a pair of primers represented by SEQ ID NOs: 87 and 88 for JNC050174, a pair of primers represented by SEQ ID NOs: 89 and 90 for JNC050178 The primer pair shown in SEQ ID NO: 91 and 92 for JNC050194, the primer pair shown in SEQ ID NO: 93 and 94 for JNC050197, the primer pair shown in SEQ ID NO: 95 and 96 for JNC050199, the SEQ ID NO: 97 for JNC060015, 98, a pair of primers represented by SEQ ID NOs: 99 and 100 for JNC060019, a pair of primers represented by SEQ ID NOs: 101 and 102 for JNC060286 The primer pair shown in SEQ ID NO: 103 and 104 for JNC060295, the primer pair shown in SEQ ID NO: 105 and 106 for JNC060434, the primer pair shown in SEQ ID NO: 107 and 108 for JNC060623, the primer pair for JNC062136 A pair of primers represented by SEQ ID NOs: 109 and 110, a pair of primers represented by SEQ ID NOs: 111 and 112 for JNC064053, a pair of primers represented by SEQ ID NOs: 113 and 114 for JNC064080, and a pair of SEQ ID NOs: 115 and 116 for JNC064090 Primer pair shown in SEQ ID NOs: 117 and 118 for JNC070004, primer pair shown in SEQ ID NOs: 119 and 120 for JNC070005, primer pair shown in SEQ ID NOs: 121 and 122 for JNC070013, SEQ ID NO: 123 and 124, a pair of primers represented by SEQ ID NOs: 125 and 126 for JNC070134, a pair of primers represented by JNC070337 129 and 130 for JNC070424, a pair of primers represented by SEQ ID Nos. 131 and 132 for JNC070431, and a pair of primers represented by SEQ ID NOS: 133 and 134 for JNC070437, respectively Primer pair shown in SEQ ID NO: 135 and 136 for JNC070474, primer pair shown in SEQ ID NO: 137 and 138 for JNC080171, primer pair shown in SEQ ID NO: 139 and 140 for JNC080409, primer pair shown in JNC080486 for JNC080486 A pair of primers represented by SEQ ID NOs: 141 and 142, a pair of primers represented by SEQ ID NOs: 143 and 144 for JNC080498, a pair of primers represented by SEQ ID NOs: 145 and 146 for JNC080501, and a pair of SEQ ID NOs: 147 and 148 for JNC080505 Primer pair, the primer pair shown in SEQ ID NO: 149 and 150 for JNC080506, SEQ ID NO: 151 and 152 for JNC090001 The primer pair shown in SEQ ID NOs: 153 and 154 for JNC090015, the primer pair shown in SEQ ID NOs: 155 and 156 for JNC090019, the primer pair shown in SEQ ID NOs: 157 and 158 for JNC090024, the primer pair for JNC090029 The primer pair shown in SEQ ID NOs: 159 and 160, the primer pair shown in SEQ ID NOs: 161 and 162 for JNC090043, the primer pair shown in SEQ ID NOs: 163 and 164 for JNC090046, the SEQ ID NOs: 165 and 166 for JNC090053 Primer pair shown in SEQ ID NOs: 167 and 168 for JNC090060, primer pair shown in SEQ ID NOs: 169 and 170 for JNC100027, primer pair shown in SEQ ID NOs: 171 and 172 for JNC100057, and primer pair shown in SEQ ID NO: 173 and 174, a pair of primers represented by SEQ ID NOs: 175 and 176 for JNC100164, a pair of primers represented by JNC1 179 and 180 for JNC100202, a pair of primers represented by SEQ ID NOs: 181 and 182 for JNC100206, and a pair of primers represented by SEQ ID NOS: 183 and 184 for JNC110013 Primer pair shown in SEQ ID NO: 185 and 186 for JNC110050, primer pair shown in SEQ ID NO: 187 and 188 for JNC110432, primer pair shown in SEQ ID NO: 189 and 190 for JNC111059, primer pair shown in JNC111269 191 and 192 for JNC111346, a pair of primers represented by SEQ ID NOS: 193 and 194 for JNC111346, a pair of primers represented by SEQ ID NOS: 195 and 196 for JNC111531, and SEQ ID NOS: 197 and 198 for JNC111652 Primer pair shown in SEQ ID NO: 199 and 200 for JNC111851, SEQ ID NO: 201 for JNC112037 and 202, a pair of primers represented by SEQ ID NOs: 203 and 204 for JNC112157, a pair of primers represented by SEQ ID NOs: 205 and 206 for JNC112511, a pair of primers represented by SEQ ID NOs: 207 and 208 for JNC112653, a pair of primers represented by JNC112736 The primer pair shown in SEQ ID NO: 209 and 210 for JNC112853, the primer pair shown in SEQ ID NO: 211 and 212 for JNC112853, the primer pair shown in SEQ ID NO: 213 and 214 for JNC112884, and the SEQ ID NOs: 215 and 216 for JNC113082 Primer pair shown in SEQ ID NOs: 217 and 218 for JNC113160, primer pair shown in SEQ ID NOs: 219 and 220 for JNC113212, primer pair shown in SEQ ID NO: 221 and 222 for JNC120005, primer pair shown in JNC120091 for JNC120091 A pair of primers represented by SEQ ID NOS: 223 and 224, a pair of primers represented by SEQ ID NOS: 225 and 226 for JNC120123, The primer pair shown in SEQ ID NOs: 227 and 228 for JNC120149, the primer pair shown in SEQ ID NOs: 229 and 230 for JNC120161, the primer pair shown in SEQ ID NOs: 231 and 232 for JNC120176, SEQ ID NOs: 233 and 234 for JNC120219 , Primer pairs represented by SEQ ID NOS: 235 and 236 for JNC120491, primer pairs represented by SEQ ID NOS: 237 and 238 for JNC120546, primer pairs represented by SEQ ID NOS: 239 and 240 for JNC120785, primer pairs represented by JNC120803 243 and 244 for JNC120907, a pair of primers represented by SEQ ID Nos. 245 and 246 for JNC120937, and a pair of primers represented by SEQ ID NOS: 247 and 248 for JNC12053 Derived from the mating between Nampyeong and Dongjin AD, including all of the displayed primer pairs. It provides a quantitative trait loci (QTLs, quantitative trait loci) genetic map for creating CAPS (cleaved amplified polymorphic sequences) markers for amplification primers for determining the associated performance.

In the present invention, the term "a group derived from mating between Nampyeong and Dongjin AD" is used in the present specification in combination with "F2 group of mating hybrid of Nampeung and Dongjin AD" It is a group of child lines that were breed by crossing Dongjin AD.

In the present invention, the term " quantitative trait loci (QTL) " refers to a trait that is expressed by a mutation of alleles present in a locus associated with resistance to a keypad disease.

The term " cleaved amplified polymorphic sequence (CAPS) marker " in the present invention refers to a SNP located in a restriction enzyme site in a single nucleotide polymorphism (SNP). In one embodiment of the present invention, the CAPS marker is selected from the restriction enzymes (ApaL I, BamHI, Bgl II, BspT104I, Cla I, EcoR I, EcoR V, EcoT22 I, Fok I , Hind III, Kpn I, Mlu I, Nde I, Not I, PshB I, Pvu II, Sac I, Sac II, Sca I, SnaB I, Stu I, Xba I and Xho I) It can be developed as a candidate CAPS marker.

Meanwhile, in the present invention, a specific SNP between CAPS (cleaved amplified polymorphic sequences) markers was developed through Nest Generation Sequencing (NGS) to carry out a QTL study on the resistance to Kidney Disease 5 to determine the locus associated with the resistance to Kidney disease of the succeeding group derived from the cross between the Nongpyeong and the Dongjin AD. The genetic map can be generated by identifying the physical location of the DNA marker on the rice rice genome.

The DNA marker of the present invention can be efficiently distinguished from the susceptibility of susceptible individuals by using the composition for screening the susceptible rice varieties comprising the DNA markers derived from the Nambyeong plant according to the present invention. It can be used for rice breeding with improved disease resistance. Therefore, the promotion of the development of Rice Kidney Disease Resistant Varieties can reduce the damage to the Kidari disease and reduce the amount of pesticide use.

FIG. 1 is a photograph showing the results of rice typhus resistance test. FIG. 4 (A) shows the in-vitro culture after inoculation with Kidari disease and (B) 4th day after inoculation of Kidneybirds, which are susceptible rice varieties, This is a photograph showing the degree of death of a person.
Fig. 2 is a photograph showing the cultivation of the F2 group of the mating hybrid between the Nambu paddy, a susceptible rice variety, and the susceptible strain, Dongjin AD.
FIG. 3 is a graph showing the results of single nucleotide polymorphism (SNP) detection for Nam-Pyeong and Dongjin ADs by resequencing and analysis of resequencing data of (A) Nampyeong and Dongjin AD And (B) chromosomal distribution of single nucleotide sequence variants (SNPs) between Nampyeong and Dongjin AD.
4 is an electrophoresis image showing a result of a polymorphism test of (A) using the novel CAPS (Cleaved Amplified Polymorphic Sequences) marker of the present invention and (B) a genotype analysis result of the F2 population of Nampyeong and Dongjin AD.
FIG. 5 is a diagram showing the genetic map of the F2 group of the succeeding mating of Nampyeong and Dongjin AD according to an embodiment of the present invention. Here, chromosome numbers are shown on each map, with the name of each marker on the right side of the chromosome, and the genetic distance from the top of each chromosome to the first marker is shown on the left. The genetic distance unit is cM (centimorgan), which is calculated by the Kosambi function.
FIG. 6 is a graph showing a mortality distribution of the F2 lines of the hybridization of Nambu paddy rice and Dongjin AD of the present invention, according to the in-flight test of rice typhus.
FIG. 7 is a graph showing the results of quantitative trait loci (QTL) analysis of rice hind resistance against rice, which was measured in the F2 group of the mating seedlings of Nampyeong and Dongjin AD according to an embodiment of the present invention.
FIG. 8 is a graph showing the relationship between the positions of two genes (Os01g0601625, Os01g0601675) belonging to the LRR (leucine rich repeat) family of chromosome 1 QTL region related to the resistance to rice tiller disease according to the embodiment of the present invention, (Insertion / Deletion) region of the LRR gene derived from the region (1625IND, 1675IND). Here, the InDel region is shown in red.
FIG. 9 is an electrophoresis image showing genotyping results of domestic Japonica rice cultivars using 1625IND and 1675IND, which are the selection markers of the rice blight-resistant rice varieties of the present invention.

Hereinafter, the constitution and effects of the present invention will be described in more detail through examples. These examples are only for illustrating the present invention, and the scope of the present invention is not limited by these examples.

Example  1: rice Kidari bottle  Resistance test

In order to test the degree of death of the cultivars according to the infecting strain of Kidari, the inoculated cultivar of Kidari was inoculated at 4th week. Survey method was as follows: The rate of mortality was evaluated as the percentage of dryness, wilt, and blight of more than 3/4 of the leaves.

As shown in Table 1, the results of in-flight tests on the degree of resistance to Kidari disease indicate that Kidari disease is relatively resistant among the rice cultivars cultivated in Korea (Jeonnam Agricultural Research and Extension Services (2002), National Agricultural Research Service (2005) , 2011)), the mortality rate was 30.8%. On the other hand, other varieties (Chowun, Unbong 40, Haiami, Inwol Rice, Paddal Rice, Hwangyoung Rice, Ungwang Rice, Hwasung Rice, Jinheung Rice, The rice cultivars of Namwolbyeon were lower than those of other cultivars by the ratio of 42.9% to 100.0%, which was higher than that of other cultivars. Respectively. As a result, it was found that the Nampega rice was resistant to Kidari disease as previously reported (Fig. 1).

On the other hand, Dongjin AD developed by the Ministry of Agriculture and Life Resources of the National Academy of Agricultural Science using the original varieties of Dongjinbyeong is a line of the rice insertion mutation group ( Plant J. , 39 (2): 252-263, 2004) And 96.7%, respectively, as a result of the in vitro test for the resistance to the disease.

(%) Of rice varieties after 4th week of inoculation with Kidari disease kind Mortality rate % ) Nam Pyung 30.8 Joan 42.9 Unbong 40 43.3 High ami 48.3 Inwol Rice 63.6 Pallad Rice 64.3 Hwadong 65.5 Ugwanghwa 71.4 Hwaseong 71.4 Jinheung Rice 93.3 Il-myeong 96.6 Sign rice 96.6 Rice paddy 100.0 Osaka 100.0 Jindo No. 43 100.0 Dong Jin-Pyo 100.0 Kumo rice 100.0 Clover rice 100.0 During the rice 100.0 Suh Hyang-chul 100.0 An Mibyeong 100.0 Consumption rice 100.0 Young An Rice 100.0 Naktong 100.0 Juana 100.0 Dongjin AD 96.7

Example  2: rice Kidari bottle  Resistance gene Mapping mapping and Kidari bottle  Development of selective markers for resistant varieties

The results of the in vitro tests on the rice resistance of rice hind legs in Example 1 confirmed that the rice hind pods were resistant to Kidari disease.

In order to investigate the location of rice resistance genes, we have developed F2 group of mating hybrid of Nipyeongbyeo, which is a rice - resistant strain of rice, and Dongjin AD, which is a susceptible strain, and resequencing of the cultivated Nongpyeong and Dongjin AD And the CAPS markers for genetic mapping were derived to determine the QTL related to the resistance to Kidari disease. In addition, we tested the resistance of the F2 group of the mating seedling of Nippei and Dongjin AD to the rice seedlings and performed the QTL mapping of rice seedling resistance. On the other hand, the QDAR resistance resistant breed marker was identified based on the QTL mapping related to the resistance to Kidari disease.

Example  2-1: Mutation area search and CAPS Marker  Development, based on this Nam Pyung  And Dongjin AD  Mating Kidari bottle  Resistance-related Quantitative trait Locus ( QTLs , quantitative trait loci)

To detect the mutation region, a single nucleotide polymorphism (SNP) was detected using a computer program or the like, comparing the nucleotide sequence information of the variety to be analyzed with the standard genome.

Specifically, resequencing was performed based on the Nipponbare sequence of Japonica (Nipponbare), which is a member of the recombinant heritable group, Nambuki and Dongjin AD. Genomic DNA sequence data of about 30x each were produced, and about 170,000 single nucleotide sequence variants (SNPs) were found between Nampyeong and Dongjin AD through comparative analysis between them (Fig. 3).

Among the above identified SNPs, restriction enzymes (ApaL Ⅰ, BamH Ⅰ, Bgl Ⅱ, BspT104 Ⅰ, Cla Ⅰ, EcoR Ⅰ, EcoR Ⅴ, EcoT22 Ⅰ, Fok Ⅰ, Hind Ⅲ, Kpn Ⅰ, Mlu Ⅰ, Nde Ⅰ, We have developed 124 new Cleaved Amplified Polymorphic Sequence (CAPS) markers for SNPs located at recognition sites of PshB Ⅰ, Pvu Ⅱ, Sac Ⅰ, Sac Ⅱ, Sca Ⅰ, SnaB Ⅰ, Stu Ⅰ, Xba Ⅰ and Xho Ⅰ Table 2 and Table 3). Genotypes were analyzed by genotyping the F2 group of the mating hybrid of Nampyeong and Dongjin AD (FIG. 4 and FIG. 5). Genetic maps were generated using MapDisto (v. 1.7) program (Lorieux M (2012) MapDisto: fast and efficient computation of genetic linkage maps. Molecular Breeding 30: 1231-1235 (DOI 10.1007 / s11032-012-9706 -y)).

Dongjin AD and Nampyeong specific CAPS marker information Marker name
(Marker name) chromosome
(chromosome) Location (bp)
(position) SNP Restriction enzyme
(Restriction enzyme) Dongjin AD Nam Pyung JNC010055 One 6,637,160 A G BamHI JNC010084 One 10,922,110 T C EcoR I JNC010086 One 11,737,804 C T PvuII JNS01057 One 20,118,452 G A BspT104I JNC010139 One 21,781,634 G T BglII JNS01030 One 23,362,397 T C HindIII JNC010565 One 23,727,134 A T EcoR I JNC010809 One 24,100,191 A C EcoRV JNC011008 One 24,466,866 C T HindIII JNC011512 One 31,955,367 T C BglII JNC011753 One 33,311,109 G A EcoRV JNC011767 One 34,349,772 A C BglII JNC011804 One 35,080,918 C A HindIII JNC011850 One 37,620,858 T C PvuII JNC011887 One 38, 493, 513 A G EcoR I JNC020021 2 3,231,863 A T EcoRV JNC020047 2 3,506,507 G A StuI JNC020052 2 4,372,712 T G EcoR I JNC020058 2 4,961,282 G A Nde I JNC020069 2 8,042,374 T C Mlu I JNC020149 2 24,683,419 A G EcoRV JNC020176 2 26,281,925 G T BamHI JNC020186 2 28,085,081 A G EcoR I JNC030012 3 4,262,859 G A EcoR I JNC030028 3 4,321,251 G A PvuII JNC030131 3 4,803,086 G A BamHI JNC030134 3 4,871,579 G A HindIII JNC030270 3 5,798,131 T C Kpn I JNC030275 3 5,818,943 G T EcoR I JNC030408 3 6,218,556 G C BamHI JNC030411 3 6,226,091 G A BamHI JNC030606 3 32,346,717 C A EcoR I JNC040003 4 981,521 C T EcoRV JNC040055 4 1,283,486 T C HindIII JNC040202 4 5,112,849 C T PvuII JNC040270 4 17,321,777 A G EcoRV JNC040277 4 18,706,206 C T Kpn I JNC040334 5 30,754,439 C T Sac I JNC050091 5 2,315,301 G A HindIII JNC050127 5 4,663,967 A G Sac I JNC050148 5 7,480,049 A G EcoRV JNC050159 5 9,442,985 T G Mlu I JNC050166 5 12,092,388 T A BspT104I JNC050174 5 17,340,757 G C FokI JNC050178 5 17,593,585 G A SnaBI JNC050194 5 20,982,835 A T Nde I JNC050197 5 23,451,217 T C SacII JNC050199 5 23,847,833 C T PvuII JNC060015 6 6,635,225 A G FokI JNC060019 6 7,752,980 C T PshBI JNC060286 6 11,251,904 G A HindIII JNC060295 6 12,644,942 T G FokI JNC060434 6 13,182,981 G A EcoR I JNC060623 6 13,479,469 A C EcoRV JNC062136 6 16,033,712 A G EcoRV JNC064053 6 20,452,558 T G PvuII JNC064080 6 20,976,276 C T StuI JNC064090 6 23,842,828 T C EcoT22I JNC070004 7 3,610,370 T G Cla I JNC070005 7 7,862,742 G T Sca I JNC070013 7 17,277,936 A C Xba I JNC070112 7 18,992,451 C T Cla I JNC070134 7 19,369,066 G A HindIII JNC070337 7 20,600,790 T C HindIII JNC070424 7 21,098,445 T A EcoR I JNC070431 7 21,356,842 G A Sac I JNC070437 7 23,104,842 A G Kpn I JNC070474 7 25,614,751 A G BamHI JNC080171 8 5,273,811 A G HindIII JNC080409 8 7,639,612 G A EcoR I JNC080486 8 8,374,383 C T Not I JNC080498 8 13,005,851 A T PshBI JNC080501 8 15,673,343 C T EcoR I JNC080505 8 18,626,100 A T StuI JNC080506 8 19,203,455 T A StuI JNC090001 9 1,742,371 G A Xho I JNC090015 9 9,306,379 A T PvuII JNC090019 9 12,001,828 A T StuI JNC090024 9 12,691,110 G A EcoT22I JNC090029 9 14,626,583 A G FokI JNC090043 9 17,218,983 T C EcoRV JNC090046 9 17,760,398 A G FokI JNC090053 9 19,579,315 G T HindIII JNC090060 9 21,208,251 A G EcoR I JNC100027 10 3,852,518 A G FokI JNC100057 10 15,431,656 G T Sac I JNC100137 10 15,632,832 - T HindIII JNC100164 10 16,682,908 G T EcoT22I JNC100175 10 17,351,413 T A ApaL I JNC100202 10 21,256,797 T C Mlu I JNC100206 10 22,389,675 T G EcoR I JNC110013 11 1,113,959 T C Xba I JNC110050 11 2,931,977 A G EcoR I JNC110432 11 3,903,612 A C EcoRV JNC111059 11 5,386,025 A G HindIII JNC111269 11 7,505,190 C T Xba I JNC111346 11 19,263,788 A T EcoRV JNC111531 11 19,930,112 T C EcoR I JNC111652 11 20,451,561 T A EcoR I JNC111851 11 20,924,926 G T HindIII JNC112037 11 21,467,425 A T EcoR I JNC112157 11 21,929,856 T C EcoRV JNC112511 11 23,042,145 A T EcoR I JNC112653 11 23,615,956 A G HindIII JNC112736 11 24,002,739 A G EcoR I JNC112853 11 25,834,302 G C BglII JNC112884 11 26,261,267 G A EcoRV JNC113082 11 27,391,298 T C HindIII JNC113160 11 27,815,552 G A Sac I JNC113212 11 28,485,932 G T PvuII JNC120005 12 3,984,418 G A HindIII JNC120091 12 4,465,298 G C EcoRV JNC120123 12 6,808,447 A T Xba I JNC120149 12 14,170,963 T C EcoRV JNC120161 12 14,586,078 T G EcoR I JNC120176 12 16,995,171 G C EcoRV JNC120219 12 18,063,117 G A HindIII JNC120491 12 19,130,580 C T EcoRV JNC120546 12 20,039,780 T C EcoR I JNC120785 12 21,113,697 G A EcoR I JNC120803 12 23,727,022 C T EcoRV JNC120907 12 25,296,067 A C EcoRV JNC120937 12 26,998,100 A G BamHI JNC121053 12 27,204,864 A G EcoR I

The primer sequence for CAPS marker detection of the present invention Marker name
(Marker name) Forward primer
(Forward primer) SEQ ID NO: Reverse primer
(Reverse primer) SEQ ID NO: JNC010055 AGGAAATCCATCTGGACCAA One TATGCAACCTCGATGAGCAA 2 JNC010084 GCGTCTCTAACGATGCCTTC 3 GTGGTCATGGATGACGAGTG 4 JNC010086 CTGCAACACGGGTATTCAGA 5 TCAGCGATGTTCATCAGGAG 6 JNS01057 GTAGGGCTTGGGATCGGTTC 7 TGTGGCGGATTTCTAGAAGGA 8 JNC010139 TTCTTTCTTCCCCACACACA 9 CTGATGACGCTACAGCCAAA 10 JNS01030 AATGGCGAGCTCAACTCCAA 11 TGCACCACCTGTACCAGGAA 12 JNC010565 TGCATTTCCAGCCCTTTAAC 13 TGGAAGACTTGGGAATCCAT 14 JNC010809 CCACATCATCACCCCTCTTT 15 TTCCTAACAACGTCGCTCCT 16 JNC011008 TCTTGGCCTTCTTTGAAACG 17 CGGCGACTAGTACACCAACG 18 JNC011512 CAGCTGTTAGGTGCGTTGTG 19 TGGAATGTCGCAAAACTACG 20 JNC011753 GCATATGGTCATTGGTCGAT 21 GCCATTTCGATAGCCATGTT 22 JNC011767 ACCGTCTGTCTGTTGCGTTA 23 CTGCATGGTTTCACATGGAC 24 JNC011804 GAGCAAAAGGGTAGGTGCTG 25 CCGTTGACCCTGTGGAATAG 26 JNC011850 ATGCGCATTTGTTGTGTCAT 27 GAATCGAGTTGCCCTAGCTG 28 JNC011887 TCGATTTGGTCATTTGGTGA 29 CTTCCGGTTTGTGCGTACTT 30 JNC020021 CCCATTAAGCATTTGCCTTT 31 TGCGGAATATGTTGCCTAGA 32 JNC020047 GTTGCCGTCAAGGTGCTATC 33 TTGCCTTAGCTTTTCCACTGA 34 JNC020052 CTCCTTTCCGTTGACAGCTC 35 TGCCTTGTCAGACGATTGAA 36 JNC020058 ATCCGGCACTTATGTGTGGT 37 GGAGAAAAGTCCGGTTTGGT 38 JNC020069 TGTGTTGAAATGGGGATTCA 39 GCGAGGTTTCTCGTAAGTGC 40 JNC020149 GGGGATGTTCCCTCGTTTAC 41 CCGAGAAGAGCAGGTACGTC 42 JNC020176 GGTGATATGCCTCAACGACA 43 AGGCTCACCTTCTGCACAGT 44 JNC020186 CGCGTTGGTCCCACTATTAT 45 GGGCTACACAAGCTGCCTTA 46 JNC030012 TCTTACTCCCTCGCTCATGG 47 AACAGCAGCCACAAAGAACA 48 JNC030028 CATCAAAGCTGCTCGATTCA 49 ATAGTATGCTCCCCGGGTTT 50 JNC030131 AAGGCAAAGAGTGCCACAAC 51 CACCCTAGCAGAGGATCTCG 52 JNC030134 CATGAGCCACCTCCTTTGAT 53 TACATTGTATGCCGCATGGT 54 JNC030270 AGACCGTTTGCTGTTGCTCT 55 TCGTCGAGAGGAAGAAGACC 56 JNC030275 CTCAGCCAGTCCAACAAGGT 57 CCCAACAACGACCAGACCTA 58 JNC030408 CCCGGACATTGAACTTGACT 59 AGTGGTTTTCCCCAACAGTG 60 JNC030411 CCCATCACCGGTACACTTCT 61 GGTTGAACGCTCCTTCAGTT 62 JNC030606 CGCACACTAGGGAGAGAGGA 63 GAATGAGTTTTGTCGCCTACG 64 JNC040003 TAGATCGCGTCAAGGGCTAC 65 ATTTAAAGGCGCTTTGGATG 66 JNC040055 CAGGATTCGCCAGTACATCA 67 ATGAGCATTGTTGGTGCAAA 68 JNC040202 GTCTGGGACATACCGCTGTT 69 GAGCTCAGGAAGATCCACGA 70 JNC040270 TGTTGTCCCCCGAGTAACTT 71 CACAAGCATTGGTTGATGGT 72 JNC040277 CATCCATGTTGCATGGCTAA 73 CCTTTCCCAGTCACCTTTCA 74 JNC040334 TGAGACGGCGACTGATAGTG 75 AAATGCGAGACGCATCTTTT 76 JNC050091 AAGGGATATGTGCCTCTTGG 77 ACCAATCCTTTTCCCTGCTT 78 JNC050127 CGTGAAACCCACATGTCAAC 79 GGAGGGAGGAGGAGAACAAG 80 JNC050148 GCGGTGGGGTAGTTTCTTCT 81 CAACACGACGCACTAAGCAT 82 JNC050159 GTGCGTTTGGTAGGAGCATT 83 ATTCTCCAGCATCCCACATT 84 JNC050166 TGGGCTTCTTTGGGACTAGA 85 GGATAAACATGCCGGGTTTT 86 JNC050174 GTAGGGCATGAGCGATGTTT 87 TGCGTGGAACATTAAATATGGA 88 JNC050178 TCACGCGAGAGACTTCCATA 89 GGGGATGTTCACTTGTGAGG 90 JNC050194 CCGAATAAGCCTGAAGTTGG 91 CCTCCCAAAAGTTGGAATCA 92 JNC050197 GCAGTGGAGAGAAGGGTGAC 93 GGGAGGAGGCAAGAAGTAGG 94 JNC050199 TGCGCAAGATTATTCCAGTG 95 ATGTCACACCCCAATCTGGT 96 JNC060015 TCGATTTGGTGGAAACTTGA 97 CATGCTTGGGTGATGAAAAAA 98 JNC060019 TGAAGCTAGGGGGAAGAACA 99 AGGAGGGCCACTGGAAGTAT 100 JNC060286 TAGGATCGCTTGATCCGAAC 101 AAGACATGCAAGAGGCAACC 102 JNC060295 AATGGAGCGCGCTAAAGTAA 103 TTGGAAGTCTACCGGTTTCC 104 JNC060434 TTGGCTTCTGCTATCCCAGT 105 GCAAGTGAATAAACCCCTGCT 106 JNC060623 GGCTTTGGTCAAACCTCATT 107 CCAAGCCAAAGATTCCACAT 108 JNC062136 GATCGAAGCTGAACCACCAT 109 TAGCGGGTGATTGAGAGTCC 110 JNC064053 CAAGATTCAAAACAACACGTTCA 111 GAAAGTTTTCGCACGGACTG 112 JNC064080 GTGGAGTTGCATTGCTCAGA 113 AACTTCCCGTGTGTTCTGTGT 114 JNC064090 AGGAGTATTGGCCCATGTGA 115 GGTGGTGGTGGGATTAAATG 116 JNC070004 TGGTGTAGAGCATTGCCTTG 117 CGTCCACACACAGGGATTTT 118 JNC070005 TTCAACGGGTGGGATGTATT 119 ATGGGTGTGTCACCCTGTCT 120 JNC070013 CCTAAATGAGCCGTGCCTAA 121 TCATGTCTTGCCATTTTTGC 122 JNC070112 ATCGTCGAGTTTTCCGTCAA 123 TCCACCAGTACATGCGAAGA 124 JNC070134 CGATGGTTTCATGCTCACAA 125 GCAGCAGTGGCTCCTGTAG 126 JNC070337 CACGTCGATCCGGTTAGTCT 127 TCGTGCGACAGAGGAATCTA 128 JNC070424 ATCGCTGCATTTACCGAGTC 129 GCAGCAGTGATGAAGTCCAA 130 JNC070431 GAACGTGCTGTTCTCCGAGT 131 CTGGTTGTTTTGACGACGTT 132 JNC070437 AGACTGGGCACCTTGTATCG 133 TGGGACAAACTATGCAGTCG 134 JNC070474 GCTGCTGCTTCTCTTTTCCA 135 GCGAAATTTGGATGGGATTA 136 JNC080171 TGACGCTTACGTGGCAGTTT 137 GTACACGGGGTGCGGTTATC 138 JNC080409 TGCACACGACCACTGGAGTA 139 CCCTAACACGGTTCATGTGC 140 JNC080486 CGAAGACGAAGACGGTGACTT 141 GCCTCCTCCGGCTAATCTTT 142 JNC080498 GCGACTGACGAAGTGACGAG 143 TGTGGAGGGAACCGGTAACT 144 JNC080501 CCGTGGGTACAAATCATGGA 145 AGTGCACGGAACCCACTACA 146 JNC080505 ATCCGAGCCATGGAAAAATG 147 ACCTGCCGAGATGTTTGAGG 148 JNC080506 AATCCGCATGGAATTCGGTA 149 GGCGATAGTGAGGTCGGTTC 150 JNC090001 CACAGTGGGCAAGGTCAGAT 151 ATAGCCTTTTGGGCGTGTTG 152 JNC090015 TGGGTCGTCGACGTATCGTA 153 AAGATGAGGACGTGGCATCC 154 JNC090019 TGGAGATTGCCTTCTGACAAGA 155 CAACGGTGTAATCGGATTGT 156 JNC090024 GGGTGGGTCGATGATAAGGA 157 TTGCGAGCTGCTTGTTTAGC 158 JNC090029 CCTCGGCAAAAAGAAACGAC 159 AGCCTGTTCTGCAGGACCTC 160 JNC090043 GAGGAAGCTTCGGTTTGCTG 161 TCGTCCATATCCTGCCTGTG 162 JNC090046 AGGACCCATTACGCTGATGC 163 ACGGCCATTTCAATTCCCTA 164 JNC090053 CGACACCCTCACCTTCACAA 165 TGGCTGAGTGCGTCATGTAA 166 JNC090060 CGGAGCTAAGGCCTGATTTG 167 CGAGTTTTTGCGGCTTTTTA 168 JNC100027 TGTGGAAGAATGGAGAGTCACG 169 TGTGTGTTTGGCCTTTGGTC 170 JNC100057 CGAGCTCCTCGACCCATCTA 171 AGTTCCGGTCCGCTTTGATT 172 JNC100137 CCATCCCTGCCAATTCTGAG 173 ATCCCCAACGTGATCTTCCA 174 JNC100164 TGGCAATTTTCCCCTATTTCG 175 TGAGACGGAGGGAGTAACATGC 176 JNC100175 CAGCGAGAAATGCCCAGAAG 177 TGCCACGTCAGCAAAACTAGG 178 JNC100202 CGCCCATTGATCCAGTGAAC 179 TTGATTCCACATGGCTGCTC 180 JNC100206 CCAGAAGCTGGTGCAGCAAT 181 TGGTTTTCAAGGGCCACAAT 182 JNC110013 CACGGCGGTGAACAGACAT 183 TACGGGCTTGGTTCGGTATG 184 JNC110050 TGGCAATGGCAATCAATGTG 185 TGGGTTGTTGGTGTGTCCAT 186 JNC110432 CTCCTGCCATTCTGCCACTG 187 TGGCAGCAGCAATGAGTTCT 188 JNC111059 CGATGAGCCAATAGTCGGTTTC 189 CCTTCTCTGACCGTGCTCCT 190 JNC111269 TCGCCTCTCACCGTATCGTT 191 AGGGGCAGTAAAGCCAGGAG 192 JNC111346 GGCCATCCACATGGAGTACG 193 ACAGCACCCTGGTGGTTGAT 194 JNC111531 AGCCGTACGTCCAATGGAGA 195 TCGATTCGTATTCGCTCCGTA 196 JNC111652 AAGGCGGTCAAAAGCTCGTT 197 CCCTTCCTCCTCCCCTTCTT 198 JNC111851 CCCGAGTGAGAAGCACTCCA 199 GCTCAAGGTCCCCCTCAGTT 200 JNC112037 TGTTTCCTCACTTTGGCTCTCC 201 CCAGGTTGGAAAGGTTGGTTT 202 JNC112157 CACATGCATGCAACGAGACA 203 TGCTTTTTGAAAGGGGGAGA 204 JNC112511 TGCACGACCAATGTCGGATA 205 CGCAAGGCGTAGCTTCAGAC 206 JNC112653 TCGAAGGAAGTCGACAGCAA 207 ACAGAAGCAGTGGCCCATGT 208 JNC112736 ACCAGGAAACTGCGAGACGA 209 GACCTCGCAGCTGATGTGTG 210 JNC112853 TCATATGGAGGCCGTTCGTT 211 GCGTGTTTGATCGAGGCTTT 212 JNC112884 CGGCTTGGAGAGCCCCTTATT 213 CGCAGGGGAACACCACTATC 214 JNC113082 CAGCCGGATCAGAACCAGAC 215 TCGGCTACCGAGCTTGAATC 216 JNC113160 GCCTGATGAACCGTTGGTGT 217 TGGGGAATTCATGGAAGGAA 218 JNC113212 TCTCCTGCCCAAGAACCAGA 219 GGGACGAACACGCTCTTGAA 220 JNC120005 ACCTGGAAGCCGGTGATCTT 221 GTGAGGCTTTGGGACACCAC 222 JNC120091 TGCAGGAGGAGGAAGCTAAGG 223 CGCAAATATTGGCCGCTTTA 224 JNC120123 GCGCGTACTATTTCGGTTGG 225 AGCACATTTGGTGGGTGTGA 226 JNC120149 CCTGGCATCTAGTTCCACCTG 227 TGATGTTGACAAAGGGGTTGG 228 JNC120161 TACGGTCAGCCACCGAGTTT 229 GCCTGCACCGTCAACGTAT 230 JNC120176 GGTCATTTGCCCTTCCACAAA 231 TTGTGCCGCCTAATCAAAGC 232 JNC120219 CCCACCGCTCCATATTGGT 233 GGAGAACATGGCATCGATCA 234 JNC120491 GTGTTGCGCTGTTGTTGCTC 235 GCACACCGCCAACCAACTA 236 JNC120546 CTTCCATGGCTGACCAGCTT 237 TTCAAACCCAGGTCGGAGAA 238 JNC120785 AGATGGCACTTTGGGGTTGA 239 CGGTCAAATGGCAAATCCTG 240 JNC120803 CGTCATACCACCCTGCACAA 241 TTGGTTGCATGGTTTCTTGG 242 JNC120907 TCAGCGAATACCACCGTCAA 243 CGTGCTCTTGTCCAAAACGA 244 JNC120937 TGATGCCTGAAGTAGCTTGTGC 245 TGCAACAATTGAAGCCTTGCT 246 JNC121053 GATGGACATTTGGGCATGTG 247 AGCAGTGGTGCGCTCAATTT 248

Example  2-2: Nam Pyung  And Dongjin AD  Mating Kidari bottle  The results of the resistance test and QTL mapping related to the resistance to the secondary resistance based on this result

For the QTL mapping related to the resistance to Kidney disease, the mortality rate of the F2 group of the mating hybrid group of Nampyeong and Dongjin AD was determined by preference to Kidari disease. Specifically, 20 F3 seeds harvested from each individual were seeded on MS medium for 235 individuals in the F2 group of the succeeding matings of Nambwa and Dongjin AD, and then infected with the Kidari germs and subjected to in vitro tests to determine the fatal rate of the F2 strain Respectively.

As shown in FIG. 6, the number of F2 lines showing a high rate of 0-10% was high, and the majority of F2 lineages were 50 %. ≪ / RTI > As a result, it was found that the mating group of Myeongpyeong and Dongjin AD had a high resistance to Kidari disease.

The QTL mapping of rice hind pod resistance was performed using the genetic maps generated based on the Kindergarten reaction data of the F2 group of the next generation of the Mt. Pyeong and the Dongjin AD and the CAPS marker derived from the Example 2-1. As a result, as shown in FIG. 7, the major QTL (major quantitative trait loci) was detected in the 70.7 cM region of chromosome 1.

Example  2-3: Kidari bottle  Resistant varieties Select marker  Development

Two genes (Os01g0601625, Os01g0601675) belonging to the family of LRR (leucine rich repeat) reported to be related to disease resistance were isolated from the QTL region discovered in Example 2-2, (Fig. 8). Based on this, two kinds of InDel markers (1625IND, 1675IND) were developed in the InDel (Insertion / Deletion) region between Dongjin AD and Nampyeong rice, which were found in the two LRR genes (Table 4).

In order to confirm that the new InDel marker, 1625IND and 1675IND, which were excavated from the InDel (Insertion / Deletion) site between the above-mentioned Dongjin AD and Nampyeong Rice, could be useful as a selection marker for the resistance to rice seedlings, 26 varieties of domestic rice Japonica PCR was carried out using the genomic DNA extracted from the leaves of rice leaves as a template and using the two new InDel markers. At this time, the PCR was repeated for 35 cycles in total under the conditions of PCR amplification at 94 ° C for 3 minutes, followed by heating at 94 ° C for 40 seconds, 62 ° C for 40 seconds, and 72 ° C for 100 seconds After that, it was further heated at 72 캜 for 5 minutes. After the PCR reaction, the amplified PCR products were developed on a 1% agarose gel using an electrophoresis apparatus and the band of the reaction product developed under UV was confirmed.

This study was carried out to investigate the effect of rice cultivar on the growth of rice varieties of Japonica japonica (27 varieties) (Dongjinbyeon, Dongjin AD, Nambyeongbyeon, Hwajeongbyeon, Juanbyeon, Hapyeongbyeon, Anmyeongbyeon, The genotypes of Kumari rice, Hwasungbyeon, Nakdongbyeon, Sinjungbyeon, Paddalbyeon, Chinheungbyeong, Chojungbyeon, Tamagumi, All of the remaining 25 species of susceptible rice varieties showed electrophoretic bands of different size from those of Nambung and Sawnori rice (Fig. 9).

Therefore, it was found from the above results that the InDel markers newly discovered from the InDel (Insertion / Deletion) region between the Dongjin AD and the Nampyeong Rice, that is, the two 1625IND and 1675IND markers, .

InDel (Insertion / Deletion) marker primer sequence for selection of rice blight-resistant rice varieties of the present invention Marker name
(Marker name) Forward primer
(Forward primer) SEQ ID NO: Reverse primer
(Reverse primer) SEQ ID NO: 1625IND AAACAAGTTGGTTGGCGAGCTAC 249 AGATTACGCCTTGGAACCTGTTA 250 1675IND TTTCTACTAAGTCACGTAGCATGCTCC 251 ATGTTCGTCGTATGCATAGCCAAAC 252

<110> Republic of Korea <120> Composition of DNA marker derived from Nampyeongbyeo for          selecting rice variety resistant to bakanae disease and method of          selecting rice variety resistant to bakanae disease using the DNA          marker <130> P16R12D1610 <160> 252 <170> KoPatentin 3.0 <210> 1 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC010055_F <400> 1 aggaaatcca tctggaccaa 20 <210> 2 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC010055_R <400> 2 tatgcaacct cgatgagcaa 20 <210> 3 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC010084_F <400> 3 gcgtctctaa cgatgccttc 20 <210> 4 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC010084_R <400> 4 gtggtcatgg atgacgagtg 20 <210> 5 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC010086_F <400> 5 ctgcaacacg ggtattcaga 20 <210> 6 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC010086_R <400> 6 tcagcgatgt tcatcaggag 20 <210> 7 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNS01057_F <400> 7 gtagggcttg ggatcggttc 20 <210> 8 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> JNS01057_R <400> 8 tgtggcggat ttctagaagg a 21 <210> 9 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC010139_F <400> 9 ttctttcttc cccacacaca 20 <210> 10 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC010139_R <400> 10 ctgatgacgc tacagccaaa 20 <210> 11 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNS01030_F <400> 11 aatggcgagc tcaactccaa 20 <210> 12 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNS01030_R <400> 12 tgcaccacct gtaccaggaa 20 <210> 13 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC010565_F <400> 13 tgcatttcca gccctttaac 20 <210> 14 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC010565_R <400> 14 tggaagactt gggaatccat 20 <210> 15 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC010809_F <400> 15 ccacatcatc acccctcttt 20 <210> 16 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC010809_R <400> 16 ttcctaacaa cgtcgctcct 20 <210> 17 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC011008_F <400> 17 tcttggcctt ctttgaaacg 20 <210> 18 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC011008_R <400> 18 cggcgactag tacaccaacg 20 <210> 19 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC011512_F <400> 19 cagctgttag gtgcgttgtg 20 <210> 20 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC011512_R <400> 20 tggaatgtcg caaaactacg 20 <210> 21 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC011753_F <400> 21 gcatatggtc attggtcgat 20 <210> 22 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC011753_R <400> 22 gccatttcga tagccatgtt 20 <210> 23 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC011767_F <400> 23 accgtctgtc tgttgcgtta 20 <210> 24 <211> 33 <212> DNA <213> Artificial Sequence <220> <223> JNC011767_R <400> 24 ctgcatggtt tcacatggac suncdscrpt ion 33 <210> 25 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC011804_F <400> 25 gagcaaaagg gtaggtgctg 20 <210> 26 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC011804_R <400> 26 ccgttgaccc tgtggaatag 20 <210> 27 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC011850_F <400> 27 atgcgcattt gttgtgtcat 20 <210> 28 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC011850_R <400> 28 gaatcgagtt gccctagctg 20 <210> 29 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC011887_F <400> 29 tcgatttggt catttggtga 20 <210> 30 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC011887_R <400> 30 cttccggttt gtgcgtactt 20 <210> 31 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC020021_F <400> 31 cccattaagc atttgccttt 20 <210> 32 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC020021_R <400> 32 tgcggaatat gttgcctaga 20 <210> 33 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC020047_F <400> 33 gttgccgtca aggtgctatc 20 <210> 34 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> JNC020047_R <400> 34 ttgccttagc ttttccactg a 21 <210> 35 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC020052_F <400> 35 ctcctttccg ttgacagctc 20 <210> 36 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC020052_R <400> 36 tgccttgtca gacgattgaa 20 <210> 37 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC020058_F <400> 37 atccggcact tatgtgtggt 20 <210> 38 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC020058_R <400> 38 ggagaaaagt ccggtttggt 20 <210> 39 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC020069_F <400> 39 tgtgttgaaa tggggattca 20 <210> 40 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC020069_R <400> 40 gcgaggtttc tcgtaagtgc 20 <210> 41 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC020149_F <400> 41 ggggatgttc cctcgtttac 20 <210> 42 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC020149_R <400> 42 ccgagaagag caggtacgtc 20 <210> 43 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC020176_F <400> 43 ggtgatatgc ctcaacgaca 20 <210> 44 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC020176_R <400> 44 aggctcacct tctgcacagt 20 <210> 45 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC020186_F <400> 45 cgcgttggtc ccactattat 20 <210> 46 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC020186_R <400> 46 gggctacaca agctgcctta 20 <210> 47 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC030012_F <400> 47 tcttactccc tcgctcatgg 20 <210> 48 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC030012_R <400> 48 aacagcagcc acaaagaaca 20 <210> 49 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC030028_F <400> 49 catcaaagct gctcgattca 20 <210> 50 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC030028_R <400> 50 atagtatgct ccccgggttt 20 <210> 51 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC030131_F <400> 51 aaggcaaaga gtgccacaac 20 <210> 52 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC030131_R <400> 52 caccctagca gaggatctcg 20 <210> 53 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC030134_F <400> 53 catgagccac ctcctttgat 20 <210> 54 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC030134_R <400> 54 tacattgtat gccgcatggt 20 <210> 55 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC030270_F <400> 55 agaccgtttg ctgttgctct 20 <210> 56 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC030270_R <400> 56 tcgtcgagag gaagaagacc 20 <210> 57 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC030275_F <400> 57 ctcagccagt ccaacaaggt 20 <210> 58 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC030275_R <400> 58 cccaacaacg accagaccta 20 <210> 59 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC030408_F <400> 59 cccggacatt gaacttgact 20 <210> 60 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC030408_R <400> 60 agtggttttc cccaacagtg 20 <210> 61 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC030411_F <400> 61 cccatcaccg gtacacttct 20 <210> 62 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC030411_R <400> 62 ggttgaacgc tccttcagtt 20 <210> 63 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC030606_F <400> 63 cgcacactag ggagagagga 20 <210> 64 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> JNC030606_R <400> 64 gaatgagttt tgtcgcctac g 21 <210> 65 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC040003_F <400> 65 tagatcgcgt caagggctac 20 <210> 66 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC040003_R <400> 66 atttaaaggc gctttggatg 20 <210> 67 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC040055_F <400> 67 caggattcgc cagtacatca 20 <210> 68 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC040055_R <400> 68 atgagcattg ttggtgcaaa 20 <210> 69 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC040202_F <400> 69 gtctgggaca taccgctgtt 20 <210> 70 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC040202_R <400> 70 gagctcagga agatccacga 20 <210> 71 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC040270_F <400> 71 tgttgtcccc cgagtaactt 20 <210> 72 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC040270_R <400> 72 cacaagcatt ggttgatggt 20 <210> 73 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC040277_F <400> 73 catccatgtt gcatggctaa 20 <210> 74 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC040277_R <400> 74 cctttcccag tcacctttca 20 <210> 75 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC040334_F <400> 75 tgagacggcg actgatagtg 20 <210> 76 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC040334_R <400> 76 aaatgcgaga cgcatctttt 20 <210> 77 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC050091_F <400> 77 aagggatatg tgcctcttgg 20 <210> 78 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC050091_R <400> 78 accaatcctt ttccctgctt 20 <210> 79 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC050127_F <400> 79 cgtgaaaccc acatgtcaac 20 <210> 80 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC050127_R <400> 80 ggagggagga ggagaacaag 20 <210> 81 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC050148_F <400> 81 gcggtggggt agtttcttct 20 <210> 82 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC050148_R <400> 82 caacacgacg cactaagcat 20 <210> 83 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC050159_F <400> 83 gtgcgtttgg taggagcatt 20 <210> 84 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC050159_R <400> 84 attctccagc atcccacatt 20 <210> 85 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC050166_F <400> 85 tgggcttctt tgggactaga 20 <210> 86 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC050166_R <400> 86 ggataaacat gccgggtttt 20 <210> 87 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC050174_F <400> 87 gtagggcatg agcgatgttt 20 <210> 88 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> JNC050174_R <400> 88 tgcgtggaac attaaatatg ga 22 <210> 89 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC050178_F <400> 89 tcacgcgaga gacttccata 20 <210> 90 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC050178_R <400> 90 ggggatgttc acttgtgagg 20 <210> 91 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC050194_F <400> 91 ccgaataagc ctgaagttgg 20 <210> 92 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC050194_R <400> 92 cctcccaaaa gttggaatca 20 <210> 93 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC050197_F <400> 93 gcagtggaga gaagggtgac 20 <210> 94 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC050197_R <400> 94 gggaggaggc aagaagtagg 20 <210> 95 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC050199_F <400> 95 tgcgcaagat tattccagtg 20 <210> 96 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC050199_R <400> 96 atgtcacacc ccaatctggt 20 <210> 97 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC060015_F <400> 97 tcgatttggt ggaaacttga 20 <210> 98 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC060015_R <400> 98 catgcttggg tgatgaaaaa 20 <210> 99 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC060019_F <400> 99 tgaagctagg gggaagaaca 20 <210> 100 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC060019_R <400> 100 aggagggcca ctggaagtat 20 <210> 101 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC060286_F <400> 101 taggatcgct tgatccgaac 20 <210> 102 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC060286_R <400> 102 aagacatgca agaggcaacc 20 <210> 103 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC060295_F <400> 103 aatggagcgc gctaaagtaa 20 <210> 104 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC060295_R <400> 104 ttggaagtct accggtttcc 20 <210> 105 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC060434_F <400> 105 ttggcttctg ctatcccagt 20 <210> 106 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> JNC060434_R <400> 106 gcaagtgaat aaacccctgc t 21 <210> 107 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC060623_F <400> 107 ggctttggtc aaacctcatt 20 <210> 108 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC060623_R <400> 108 ccaagccaaa gattccacat 20 <210> 109 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC062136_F <400> 109 gatcgaagct gaaccaccat 20 <210> 110 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC062136_R <400> 110 tagcgggtga ttgagagtcc 20 <210> 111 <211> 23 <212> DNA <213> Artificial Sequence <220> <223> JNC064053_F <400> 111 caagattcaa aacaacacgt tca 23 <210> 112 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC064053_R <400> 112 gaaagttttc gcacggactg 20 <210> 113 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC064080_F <400> 113 gtggagttgc attgctcaga 20 <210> 114 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> JNC064080_R <400> 114 aacttcccgt gtgttctgtg t 21 <210> 115 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC064090_F <400> 115 aggagtattg gcccatgtga 20 <210> 116 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC064090_R <400> 116 ggtggtggtg ggattaaatg 20 <210> 117 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC070004_F <400> 117 tggtgtagag cattgccttg 20 <210> 118 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC070004_R <400> 118 cgtccacaca cagggatttt 20 <210> 119 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC070005_F <400> 119 ttcaacgggt gggatgtatt 20 <210> 120 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC070005_R <400> 120 atgggtgtgt caccctgtct 20 <210> 121 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC070013_F <400> 121 cctaaatgag ccgtgcctaa 20 <210> 122 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC070013_R <400> 122 tcatgtcttg ccatttttgc 20 <210> 123 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC070112_F <400> 123 atcgtcgagt tttccgtcaa 20 <210> 124 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC070112_R <400> 124 tccaccagta catgcgaaga 20 <210> 125 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC070134_F <400> 125 cgatggtttc atgctcacaa 20 <210> 126 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> JNC070134_R <400> 126 gcagcagtgg ctcctgtag 19 <210> 127 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC070337_F <400> 127 cacgtcgatc cggttagtct 20 <210> 128 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC070337_R <400> 128 tcgtgcgaca gaggaatcta 20 <210> 129 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC070424_F <400> 129 atcgctgcat ttaccgagtc 20 <210> 130 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC070424_R <400> 130 gcagcagtga tgaagtccaa 20 <210> 131 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC070431_F <400> 131 gaacgtgctg ttctccgagt 20 <210> 132 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC070431_R <400> 132 ctggttgttt tgacgacgtt 20 <210> 133 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC070437_F <400> 133 agactgggca ccttgtatcg 20 <210> 134 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC070437_R <400> 134 tgggacaaac tatgcagtcg 20 <210> 135 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC070474_F <400> 135 gctgctgctt ctcttttcca 20 <210> 136 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC070474_R <400> 136 gcgaaatttg gatgggatta 20 <210> 137 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC080171_F <400> 137 tgacgcttac gtggcagttt 20 <210> 138 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC080171_R <400> 138 gtacacgggg tgcggttatc 20 <210> 139 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC080409_F <400> 139 tgcacacgac cactggagta 20 <210> 140 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC080409_R <400> 140 ccctaacacg gttcatgtgc 20 <210> 141 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> JNC080486_F <400> 141 cgaagacgaa gacggtgact t 21 <210> 142 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC080486_R <400> 142 gcctcctccg gctaatcttt 20 <210> 143 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC080498_F <400> 143 gcgactgacg aagtgacgag 20 <210> 144 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC080498_R <400> 144 tgtggaggga accggtaact 20 <210> 145 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC080501_F <400> 145 ccgtgggtac aaatcatgga 20 <210> 146 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC080501_R <400> 146 agtgcacgga acccactaca 20 <210> 147 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC080505_F <400> 147 atccgagcca tggaaaaatg 20 <210> 148 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC080505_R <400> 148 acctgccgag atgtttgagg 20 <210> 149 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC080506_F <400> 149 aatccgcatg gaattcggta 20 <210> 150 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC080506_R <400> 150 ggcgatagtg aggtcggttc 20 <210> 151 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC090001_F <400> 151 cacagtgggc aaggtcagat 20 <210> 152 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC090001_R <400> 152 atagcctttt gggcgtgttg 20 <210> 153 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC090015_F <400> 153 tgggtcgtcg acgtatcgta 20 <210> 154 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC090015_R <400> 154 aagatgagga cgtggcatcc 20 <210> 155 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> JNC090019_F <400> 155 tggagattgc cttctgacaa ga 22 <210> 156 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC090019_R <400> 156 caacggtgta atcggattgt 20 <210> 157 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC090024_F <400> 157 gggtgggtcg atgataagga 20 <210> 158 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC090024_R <400> 158 ttgcgagctg cttgtttagc 20 <210> 159 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC090029_F <400> 159 cctcggcaaa aagaaacgac 20 <210> 160 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC090029_R <400> 160 agcctgttct gcaggacctc 20 <210> 161 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC090043_F <400> 161 gaggaagctt cggtttgctg 20 <210> 162 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC090043_R <400> 162 tcgtccatat cctgcctgtg 20 <210> 163 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC090046_F <400> 163 aggacccatt acgctgatgc 20 <210> 164 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC090046_R <400> 164 acggccattt caattcccta 20 <210> 165 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC090053_F <400> 165 cgacaccctc accttcacaa 20 <210> 166 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC090053_R <400> 166 tggctgagtg cgtcatgtaa 20 <210> 167 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC090060_F <400> 167 cggagctaag gcctgatttg 20 <210> 168 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC090060_R <400> 168 cgagtttttg cggcttttta 20 <210> 169 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> JNC100027_F <400> 169 tgtggaagaa tggagagtca cg 22 <210> 170 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC100027_R <400> 170 tgtgtgtttg gcctttggtc 20 <210> 171 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC100057_F <400> 171 cgagctcctc gacccatcta 20 <210> 172 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC100057_R <400> 172 agttccggtc cgctttgatt 20 <210> 173 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC100137_F <400> 173 ccatccctgc caattctgag 20 <210> 174 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC100137_R <400> 174 atccccaacg tgatcttcca 20 <210> 175 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> JNC100164_F <400> 175 tggcaatttt cccctatttc g 21 <210> 176 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> JNC100164_R <400> 176 tgagacggag ggagtaacat gc 22 <210> 177 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC100175_F <400> 177 cagcgagaaa tgcccagaag 20 <210> 178 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> JNC100175_R <400> 178 tgccacgtca gcaaaactag g 21 <210> 179 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC100202_F <400> 179 cgcccattga tccagtgaac 20 <210> 180 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC100202_R <400> 180 ttgattccac atggctgctc 20 <210> 181 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC100206_F <400> 181 ccagaagctg gtgcagcaat 20 <210> 182 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC100206_R <400> 182 tggttttcaa gggccacaat 20 <210> 183 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> JNC110013_F <400> 183 cacggcggtg aacagacat 19 <210> 184 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC110013_R <400> 184 tacgggcttg gttcggtatg 20 <210> 185 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC110050_F <400> 185 tggcaatggc aatcaatgtg 20 <210> 186 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC110050_R <400> 186 tgggttgttg gtgtgtccat 20 <210> 187 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC110432_F <400> 187 ctcctgccat tctgccactg 20 <210> 188 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC110432_R <400> 188 tggcagcagc aatgagttct 20 <210> 189 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> JNC111059_F <400> 189 cgatgagcca atagtcggtt tc 22 <210> 190 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC111059_R <400> 190 ccttctctga ccgtgctcct 20 <210> 191 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC111269_F <400> 191 tcgcctctca ccgtatcgtt 20 <210> 192 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC111269_R <400> 192 aggggcagta aagccaggag 20 <210> 193 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC111346_F <400> 193 ggccatccac atggagtacg 20 <210> 194 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC111346_R <400> 194 acagcaccct ggtggttgat 20 <210> 195 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC111531_F <400> 195 agccgtacgt ccaatggaga 20 <210> 196 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> JNC111531_R <400> 196 tcgattcgta ttcgctccgt a 21 <210> 197 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC111652_F <400> 197 aaggcggtca aaagctcgtt 20 <210> 198 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC111652_R <400> 198 cccttcctcc tccccttctt 20 <210> 199 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC111851_F <400> 199 cccgagtgag aagcactcca 20 <210> 200 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC111851_R <400> 200 gctcaaggtc cccctcagtt 20 <210> 201 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> JNC112037_F <400> 201 tgtttcctca ctttggctct cc 22 <210> 202 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> JNC112037_R <400> 202 ccaggttgga aaggttggtt t 21 <210> 203 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC112157_F <400> 203 cacatgcatg caacgagaca 20 <210> 204 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC112157_R <400> 204 tgctttttga aagggggaga 20 <210> 205 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC112511_F <400> 205 tgcacgacca atgtcggata 20 <210> 206 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC112511_R <400> 206 cgcaaggcgt agcttcagac 20 <210> 207 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC112653_F <400> 207 tcgaaggaag tcgacagcaa 20 <210> 208 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC112653_R <400> 208 acagaagcag tggcccatgt 20 <210> 209 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC112736_F <400> 209 accaggaaac tgcgagacga 20 <210> 210 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC112736_R <400> 210 gacctcgcag ctgatgtgtg 20 <210> 211 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC112853_F <400> 211 tcatatggag gccgttcgtt 20 <210> 212 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC112853_R <400> 212 gcgtgtttga tcgaggcttt 20 <210> 213 <211> 33 <212> DNA <213> Artificial Sequence <220> <223> JNC112884_F <400> 213 cggcttggag agcccttatt suncdscrpt ion 33 <210> 214 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC112884_R <400> 214 cgcaggggaa caccactatc 20 <210> 215 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC113082_F <400> 215 cagccggatc agaaccagac 20 <210> 216 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC113082_R <400> 216 tcggctaccg agcttgaatc 20 <210> 217 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC113160_F <400> 217 gcctgatgaa ccgttggtgt 20 <210> 218 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC113160_R <400> 218 tggggaattc atggaaggaa 20 <210> 219 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC113212_F <400> 219 tctcctgccc aagaaccaga 20 <210> 220 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC113212_R <400> 220 gggacgaaca cgctcttgaa 20 <210> 221 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC120005_F <400> 221 acctggaagc cggtgatctt 20 <210> 222 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC120005_R <400> 222 gtgaggcttt gggacaccac 20 <210> 223 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> JNC120091_F <400> 223 tgcaggagga ggaagctaag g 21 <210> 224 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC120091_R <400> 224 cgcaaatatt ggccgcttta 20 <210> 225 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC120123_F <400> 225 gcgcgtacta tttcggttgg 20 <210> 226 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC120123_R <400> 226 agcacatttg gtgggtgtga 20 <210> 227 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> JNC120149_F <400> 227 cctggcatct agttccacct g 21 <210> 228 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> JNC120149_R <400> 228 tgatgttgac aaaggggttg g 21 <210> 229 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC120161_F <400> 229 tacggtcagc caccgagttt 20 <210> 230 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> JNC120161_R <400> 230 gcctgcaccg tcaacgtat 19 <210> 231 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC120176_F <400> 231 ggtcatttgc ccttccacaa 20 <210> 232 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC120176_R <400> 232 ttgtgccgcc taatcaaagc 20 <210> 233 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> JNC120219_F <400> 233 cccaccgctc catattggt 19 <210> 234 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC120219_R <400> 234 ggagaacatg gcatcgatca 20 <210> 235 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC120491_F <400> 235 gtgttgcgct gttgttgctc 20 <210> 236 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> JNC120491_R <400> 236 gcacaccgcc aaccaacta 19 <210> 237 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC120546_F <400> 237 cttccatggc tgaccagctt 20 <210> 238 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC120546_R <400> 238 ttcaaaccca ggtcggagaa 20 <210> 239 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC120785_F <400> 239 agatggcact ttggggttga 20 <210> 240 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC120785_R <400> 240 cggtcaaatg gcaaatcctg 20 <210> 241 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> JNC120803_F <400> 241 cgtcatacca ccctgcaca 19 <210> 242 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC120803_R <400> 242 ttggttgcat ggtttcttgg 20 <210> 243 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC120907_F <400> 243 tcagcgaata ccaccgtcaa 20 <210> 244 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC120907_R <400> 244 cgtgctcttg tccaaaacga 20 <210> 245 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> JNC120937_F <400> 245 tgatgcctga agtagcttgt gc 22 <210> 246 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> JNC120937_R <400> 246 tgcaacaatt gaagccttgc t 21 <210> 247 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC121053_F <400> 247 gatggacatt tgggcatgtg 20 <210> 248 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> JNC121053_R <400> 248 agcagtggtg cgctcaattt 20 <210> 249 <211> 23 <212> DNA <213> Artificial Sequence <220> <223> 1625IND_F <400> 249 aaacaagttg gttggcgagc tac 23 <210> 250 <211> 23 <212> DNA <213> Artificial Sequence <220> <223> 1625IND_R <400> 250 agattacgcc ttggaacctg tta 23 <210> 251 <211> 27 <212> DNA <213> Artificial Sequence <220> <223> 1675IND_F <400> 251 tttctactaa gtcacgtagc atgctcc 27 <210> 252 <211> 25 <212> DNA <213> Artificial Sequence <220> <223> 1675IND_R <400> 252 atgttcgtcg tatgcatagc caaac 25

Claims (9)

A composition for screening a rice blast resistance-resistant rice variety comprising a pair of primers represented by SEQ ID NOs: 249 and 250.
(a) separating genomic DNA from a rice sample;
(b) amplifying the target sequence by performing amplification reaction using the separated genomic DNA as a template and using primer pairs represented by SEQ ID NOs: 249 and 250; And
(c) detecting the amplification product.
3. The method according to claim 2, wherein the sample is derived from rice seeds, roots, leaves, stems, flowers or ear tissue.
A pair of primers represented by SEQ ID NOS: 249 and 250, and a reagent for carrying out an amplification reaction.
5. The kit of claim 4, wherein the reagent for carrying out the amplification reaction comprises a DNA polymerase, dNTPs, and a reaction buffer.
(a) obtaining a progeny population derived from crossbreeding between a susceptible and resistant strain;
(b) separating the genomic DNA from the rice samples obtained in the later group;
(c) amplifying the target sequence by performing amplification reaction using the separated genomic DNA as a template and using primer pairs represented by SEQ ID NOS: 249 and 250; And
(d) detecting the amplified product to determine resistance.
[Claim 7] The method according to claim 6, Among the breeding species produced by crossing with Nampyeong, breeds showing resistance; And the second step of the method according to claim 2, wherein the strain is selected from the group consisting of rice bran-resistant rice varieties.
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