3)
4
Genetic Analysis for Bacterial Leaf Pustule Resistance for Improvement of Soybean Varieties Bacterial leaf pustule (BLP) caused by Xanthomonas axonopodis pv. glycines is one of the most prevalent bacterial diseases in plants. Early symptoms are characterized by small,yellow-to-brown lesions with a raised pustule in the center. The lesions progressively merge to form large necrotic areas, which cause premature defoliation and subsequently a loss of grain yield. The first goal of this research was performed to identify quantitative trait loci (QTL) for resistance to BLP as well as to understand the resistance of soybean genotypes to different BLP isolates. Also, candidate gene analysis and differential display between resistant and susceptible soybean genotypes was performed to develop BLP-specific DNA markers. 1. Genetic Analysis for Bacterial Leaf Pustule Resistance for Improvement of Soybean Varieties Quantitative trait loci (QTLs) for resistance to BLP in soybean (Glycine max [L.] Merr.) were identified using simple sequence repeat (SSR) markers, and the consistency of QTL detection across bacterial isolates as well as across greenhouse and field experiments were examined. Recombinant inbred lines (RILs) derived from a cross of BLP-susceptible 'Suwon157' and BLP-resistant 'Danbaekkong' were utilized to identify QTL conditioning the resistance to BLP with 76 SSR markers. For phenotypic analysis, six isolates of X. axonopodis were collected from different geographic origins (ocsf, ocsg and sd12178 from Korea, 8ra from USA, LMG7403 from Zambia, and LMG7404 from Zimbabwe). A combination of single-factor analysis of variance and multiple regression analysis revealed that three QTLs conditioned resistance to BLP in the field. Under greenhouse condition, one to four QTLs were revealed to be associated with resistance to each isolate. Satt372 on linkage group (LG) D2 was detected across six isolates under both greenhouse andfield conditions, an indication that resistance to BLP was mainly controlled by a single major gene tightly linked to Satt372. Several minor QTLson LG A1, C1, F, L, and O were also
found depending on the isolates. seventy-five different soybean cultivars were tested for the response to several different X.campestris pv. glycines races collected worldwide. Among tested cultivars, one cultivar, G. max cv. PI96188 showed very different response to all X. campestris pv. glycines races. It showed HR-like necrosis without halos but still carried pustules in the back of the necrosis symptom. Rxp gene conferring resistance to bacterial pustule was reported to be on linkage group (LG) D2. The gene controlling the novel response in PI96188 was mapped to whether the gene was the same as Rxp. An RIL population from a cross of PI96188 and Jinju 1 was constructed. Five markers on LG D2 were not associated with novel response in PI96188, indicating that the gene controlling this specific response was different from Rxp. Further mapping study revealed that Sat_108 on LG O was identified as being associated with novel response in PI96188. 2.
3. Search for Molecular Markers for Bacterial Leaf Pustule Resistance through the Analysis of Disease resistance and Phathogenesis-Related Genes
Genetic Analysis for Bacterial Leaf Pustule Resistance for Improvement of Soybean Varieties
Search for Molecular Markers for Bacterial Leaf Pustule Resistance through the Analysis of Disease resistance and Phathogenesis-Related Genes
1. 기술적측면
1. 국내연구현황 8ra F G 980 2178 2179 + ++ ++ + ++ ++ ± ± ± ± ± ± - ++ ± ± ± ± + +++ +++ ++ + + +++ +++ +++ + ++ ++ + ++ ± +++ + ++ + ++ ++ + + + + + + + + + + +++ +++ +++ +++ +++ +++ +++ ++ ± +++ +++ ± +++ +++ +++ ++ + - +++ + ++ ++ +++ - + + + + + ± ++ ++ ± ± ++ +++ +++ +++ +++ +++ ++ - ++ + + ± ± - ± ± - ± ± (+++; very susceptible, ++; susceptible, +; mid, ±; resistant, -; very resistant)
제 1 세부과제 내병성계통유전형질분석 RIL 집단양성 ( 태국 ) Marker-Assisted Selection 내병성계통육성 협제 2 동세과부과제제 병균침입시발현되는특이적저항성콩유전자 식물병저항성유전자유사체 PCR 마커개발 식물유전자개발 콩의병저항성병저항성유전자와비병원성유전자의상호이해에의한내병성계통선발체계선발체계확립 불마름병균유전자연구 병원균 계열 지표작성 제 1 세부과제 불마름병저항성저항성콩계통개발
제 1 세부과제 ( 서울대이석하 ) 제 2 세부과제 ( 서울대백남천 ) 협동과제 ( 전북대윤성중 ) 제 1 세부과제 ( 농과원허성기 ) 콩불마름병저항성유전분석및내병성콩계통육성 콩불마름병특이적저항성콩유전자분리및마커개발 콩불마름병저항성관련후보식물유전자분석에의한마커개발 콩불마름병 생리적 균계분화 및 발병기작 연구 제 1 차년도 저항성과감수성품종간 F1 양성및 F2 종자를태국으로송부, 세대진전 콩유전자원 60 여점불마름병균계별저항성평가및모부본간 polymorphism 조사, 150 여 SSR 마커선발 불마름병특이적 저항성콩유전자의분리 ( DDRT-PCR, SSH) 저항성콩유전자발현특성조사 Full-length cdna 분리 타식물에서이미분리및보고된 병저항성유전자들의염기서열비교분석 염기서열의비교분석에의한 Primer 제작및 PCR cloning 으로유전자유사체분리 우리나라에서콩불마름병수집, 분리, 동정 및분화형조사 균계별품종간의 친화정도를조사 제 2 차년도 콩유전자원 60 여점불마름병균계별저항성평가및모부본간 polymorphism 조사, 150 여 SSR 마커선발 후기세대 집단양성 RIL(F 6) 분리된 유전자 염기서열 확인및 타식물의 병저항성 유전자들과 비교분석 분리된콩유전자의저항성기능검정 ( 세균감수성인담배와아기장대모델식물로형질전환 ) 타식물병저항성 유전자유사체 콩 cdna 의 발현특성 조사 cdna library 에서 full-length cdna 분리하여 염기서열확인 분화형을구분하여균계분화지표를작성 제 3 차년도 콩유전자원 60 여점 저항성 평가, 150 여 SSR 마커선발 SSR 마커에 의한 저항성과 감수성 품종간의 450 여 마커 polymorphism 조사및연관조사 내병성콩계통개발 불마름병 특이적 저항성 콩유전자들과 저항성 분리집단을 이용한 RFLP 연관도 조사 및 98% 이상정확도의 제한효소 확립 콩불마름병 저항성 육종을 위한 유전자 마커개발 분리집단을 이용하여 저항성 유전자유사체 clone 들의 RFLP map 과 연관도조사및 내병성육종의 마커개발 콩후기세대에서콩불마름병저항성계통육성 ( 5 여계통확립 ) 콩불마름병특이적저항성콩유전자다수확보및유전자마커개발콩의타식물병저항성유전자유사체를이용한마커 2 종이상개발우리나라콩불마름병균계분화지표작성
LG χ 1 satt155 A1 38 29 67 1.209 2 satt236 A1 31 38 69 0.710 3 satt276 A1 35 37 72 0.056 4 satt545 A1 36 32 68 0.235 5 satt177 A2 36 30 66 0.545 6 satt187 A2 37 34 71 0.127 7 satt409 A2 31 19 50 2.880 8 satt197 B1/S 38 28 66 1.515 9 satt415 B1/S 36 30 66 0.545 10 satt426 B1/S 28 30 58 0.069 11 satt509 B1/S 39 28 67 1.806 12 satt168 B2/P 40 27 67 2.522 13 satt556 B2/P 37 31 68 0.529 14 satt577 B2/P 32 37 69 0.362 15 sat-085 C1 17 33 50 5.120
LG 16 satt180 C1 30 35 65 0.385 17 satt294 C1 34 35 69 0.014 18 satt396 C1 38 30 68 0.941 19 satt134 C2 40 34 74 0.486 20 satt202 C2 37 27 64 1.563 21 satt277 C2 38 34 72 0.222 22 satt286 C2 33 34 67 0.015 23 satt291 C2 41 30 71 1.704 24 satt322 C2 41 27 68 2.882 25 satt147 D1a/Q 33 33 66 0.000 26 satt179 D1a/Q 32 34 66 0.061 27 satt408 D1a/Q 30 34 64 0.250 28 satt532 D1a/Q 35 34 69 0.014 29 satt189 D1b/W 33 30 63 0.143 30 satt290 D1b/W 32 31 63 0.016 31 satt186 D2/R 36 28 64 1.000 32 satt226 D2/R 36 30 66 0.545 33 satt372 D2/R 36 30 66 0.545 34 satt458 D2/R 38 33 71 0.352 35 satt389 D2/R 31 24 55 0.891 36 satt397 D2/R 36 33 69 0.130 37 satt154 D2/R 33 19 52 3.769 38 sat-112 E 40 23 63 4.587 39 satt185 E 30 39 69 1.174 40 satt411 E 36 26 62 1.613 41 sat-074 F 26 36 62 1.613 42 satt114 F 30 39 69 1.174 43 satt160 F 40 28 68 2.118 44 satt269 F 32 33 65 0.015 45 satt224 G 36 24 60 2.400 46 satt288 G 38 35 73 0.123 47 satt324 G 29 40 69 1.754 48 sat_122 H 24 39 63 3.571 49 satt192 H 25 42 67 4.313 50 satt279 H 33 37 70 0.229 χ
LG 51 satt317 H 28 34 62 0.581 52 satt353 H 30 37 67 0.731 53 satt434 H 18 30 48 3.000 54 satt442 H 24 37 61 2.770 55 satt541 H 29 33 62 0.258 56 satt239 I 40 28 68 2.118 57 satt292 I 38 34 72 0.222 58 satt571 I 37 23 60 3.267 59 satt183 J 37 29 66 0.970 60 satt215 J 32 30 62 0.065 61 satt244 J 29 40 69 1.754 62 satt285 J 35 30 65 0.385 63 satt414 J 34 33 67 0.015 64 satt431 J 27 42 69 3.261 65 satt167 K 32 36 68 0.235 66 satt196 K 35 30 65 0.385 67 satt441 K 34 36 70 0.057 68 satt143 L 26 35 61 1.328 69 satt156 L 35 36 71 0.014 70 satt373 L 36 33 69 0.130 71 satt462 L 29 39 68 1.471 72 satt336 M 29 39 68 1.471 73 satt590 M 33 37 70 0.229 74 GMABAB N 33 23 56 1.786 75 satt584 N 34 38 72 0.222 76 satt188 O 37 35 72 0.056 77 satt243 O 25 38 63 2.683 78 satt259 O 31 32 63 0.016 79 satt445 O 44 28 72 3.556 χ
표 1-2. 수원157 호 단백콩의유전자지도특성 그림 1-1. 유전자지도( 수원157호x 단백콩) LGA1 LGA2 LGB1 Satt276 Satt155 16.8 35.2 Satt177 Satt187 27.6 Satt197 Satt509 18.0 29.8 Satt545 57.6 Satt415 22.7 Satt409 LGC1 Satt236 LGB2 Satt396 Satt556 40.7 Satt168 20.2 Satt294 Sat_085 7.2
LGD1a+Q Satt147 Satt408 1.6 3.3 Satt179 Satt532 LGD1a+Q Satt147 Satt408 1.6 3.3 Satt179 Satt532 LGD1b+W 0.9 Satt189 Satt290 LGD1b+W 0.9 Satt189 Satt290 LGC2 16.8 24.5 Satt291 Satt322 Satt286 9.0 Satt134 1.4 31.8 Satt277 Satt202 LGC2 16.8 24.5 Satt291 Satt322 Satt286 9.0 Satt134 1.4 31.8 Satt277 Satt202 LGE Sat_112 Satt411 1.9 LGE Sat_112 Satt411 1.9 LGF 47.1 11.2 Satt269 Satt160 Satt114 Sat_074 LGF 47.1 11.2 Satt269 Satt160 Satt114 Sat_074 LGG 39.4 23.1 Satt288 Satt224 Satt324 LGG 39.4 23.1 Satt288 Satt224 Satt324 LGD2 8.7 Satt458 Satt154 Satt226 7.9 Satt186 Satt372 Satt397 Satt389 13.1 12.7 30.6 12.7 LGD2 8.7 Satt458 Satt154 Satt226 7.9 Satt186 Satt372 Satt397 Satt389 13.1 12.7 30.6 12.7
LGJ 35.8 1.6 Satt285 Satt414 Satt183 Satt215 Satt244 Satt431 4.6 16.0 13.7 LGJ 35.8 1.6 Satt285 Satt414 Satt183 Satt215 Satt244 Satt431 4.6 16.0 13.7 LGI 9.8 Satt571 Satt239 Satt292 LGI 9.8 Satt571 Satt239 Satt292 LGH Satt353 Satt192 38.2 Satt442 Satt541 Sat_122 Satt279 Satt317 Satt434 48.1 11.7 0.8 2.7 24.7 8.2 LGH Satt353 Satt192 38.2 Satt442 Satt541 Sat_122 Satt279 Satt317 Satt434 48.1 11.7 0.8 2.7 24.7 8.2 LGO 26.9 17.9 Satt445 Satt259 Satt188 LGO 26.9 17.9 Satt445 Satt259 Satt188 LGL 12.7 12.5 Satt143 Satt462 Satt373 39.1 Satt156 LGL 12.7 12.5 Satt143 Satt462 Satt373 39.1 Satt156 LGK Satt441 Satt167 0.8 LGK Satt441 Satt167 0.8
표 1. 수원157 단백콩 F2개체에대한 Satt372 마커의분리비와병원균의표현형. Satt372 N Mean S/S 80 3.32 S/D 150 3.16 D/D 60 2.07 개체수 40 35 30 25 20 15 10 5 0 수원 157 호 단백콩 1 1.5 2 2.5 3 3.5 4 4.5 5 병저항성 그림 1. 수원157 단백콩 F2개체에대한 ocsf 병원균의표현형분리.
표2. Means and ranges of parents and RIL progenies for the resistance to bacterial leaf pustule in soybean Isolate a 8ra OCS-F OCS-G SDL2178 LMG7403 LMG7404 Field b Female parent Male Parent 1.3 2.7 3.7 2.4 4.2 3.3 4.1 0.8 1.2 1.3 1.5 2.3 1.3 1.0 Progeny 1.0~ 3.3 1.2~3.9 1.0~4.9 0.8~3.5 1.0~3.8 0.3~3.5 1.0~ 4.8 LSD 0.05 1.2 1.2 1.3 1.2 1.3 1.1 1.3 Progeny Mean 2.0 2.3 2.5 2.2 2.2 2.0 2.9 H2(%) 47 66 60 41 38 59 82 a Disease severity was scored on the basis of the number of small, yellow-to-brown lesions with a raised pustule per leaflet on a scale of 0 to 5 (0 = no legion, 1 = 1 to 25 lesions, 2 = 26 to 50 lesions, 3 = 51 to 75 lesions, 4 = 76 to 100 lesions, and 5 = more than 100 lesions). b Disease severity was scored on a scale of 0 to 5(0=no visible symptoms to 5=heavily infected)
표 3. Markers linked to QTLs associated with the resistance to BLP disease under greenhouse experiment using an RIL population derived from a cross of the susceptible Suwon157 and the resistant Danbaekkong in soybean SF-ANOVA a SLG-Regr a MLG-Regr a Isaolate Locus LG Allelic means b P R2 P R2 P R2 S/S D/D % ---score-- % % 8ra Satt509 B1/S 0.027 7 2.1 1.8 NA c - - - Satt372 D2/R <0.001 17 2.2 1.7 <0.001 22 0.002 20 Satt458 D2/R <0.001 19 2.2 1.7 0.004 5 - - Satt156 L 0.042 6 2.1 1.8 NA - - - Satt243 O 0.022 9 2.2 1.9 NA - 0.010 11 Total 31 OCS-F Satt134 C2 0.035 6 2.5 2.1 NA - - - Satt372 D2/R <0.001 44 2.8 1.8 <0.001 48 <0.001 43 Satt458 D2/R <0.001 28 2.6 1.9 - - - - Satt353 H 0.047 6 2.5 2.1 NA - - - Satt156 L 0.030 7 2.5 2.1 NA - - - Total 43 OCS-G Satt177 A2 0.011 10 2.7 2.3 NA - - - Satt372 D2/R <0.001 40 2.8 2.0 <0.001 44 <0.001 39 Satt458 D2/R <0.001 19 2.7 2.1 - - - - Satt167 K 0.017 8 2.3 2.7 - - - - Satt441 K 0.026 7 2.3 2.7 0.018 8 - - Satt143 L 0.011 10 2.7 2.3 NA - 0.036 6 Total 45 SDL2178 Satt294 C1 0.019 8 2.4 2.1 0.019 9 0.027 7 Satt396 C1 0.033 7 2.3 2.0 - - - - Satt291 C2 0.016 8 2.4 2.0 NA - - - Satt372 D2/R <0.001 28 2.5 1.9 <0.001 29 <0.001 25 Satt458 D2/R 0.008 10 2.4 2.0 - - - - Satt336 M 0.025 7 2.4 2.1 NA - - - Total 32 LMG7403 Satt155 A1 0.019 9 2.1 2.5 NA - 0.007 13 Satt134 C2 0.043 6 2.4 2.0 NA - - - Satt372 D2/R <0.001 24 2.5 1.9 <0.001 28 0.004 1.8 Satt458 D2/R <0.001 19 2.5 1.9 0.043 5 - - Satt269 F 0.025 8 2.4 2.1 NA - 0.015 8 Satt156 L 0.021 8 2.4 2.1 NA - 0.008 11 Satt259 O 0.046 7 2.4 2.1 NA - - - Total 50 LMG7404 Satt372 D2/R <0.001 37 2.4 1.6 <0.001 39 <0.001 39 Satt458 D2/R <0.001 21 2.3 1.7 - - - - Total 39
a SF-ANOVA: single factor analysis of variance SLG-Regr: Multiple regression with markers on each linkage group MLG-Regr: multiple regression with all significant markers from the SLG-Regr model b S/S: homozygous Suwon 157, D/D : homozygous Danbaekkong c Not applicable. Not linked to other markers Satt276 16.8 Satt155 35.2 Satt545 22.7 Satt236 Satt396 40.7 Satt294 7.2 Sat_085 Satt226 7.6 Satt186 Satt372 11.5 Satt458 LG A1 LG C1 LGD2 Satt269 11.2 Satt160 Satt143 12.5 Field Satt462 Satt445 12.7 26.9 75.3 Satt156 Satt259 17.9 39.1 Satt188 Satt114 Satt373 Satt243 Field 8ra ocsf ocsg 2178 7403 7404 LG F LG L LG O 그림2. Soybean linkage groups (LG) showing marker positions and estimated map distance with the independent QTL conditioning the resistance to bacterial leaf pustule disease. Field resistance was evaluated through inoculation (a filled bar) with the mixture of three isolates(ocs-f, SDL2178, LMG7404), while greenhouse resistance was evaluated through individual inoculation with six strains(an open, a dotted, and four hatched bars). Length of horizontal bars indicates R 2 value for the
loci.
BamHI 17.5x34 amino acid repeats BamHI
장엽대원장원소원잎깍지잎깍지잎깍지줄기잎깍지
avrbs3::eztn WT MT WT MT Williams 82 CNS
Clone No. 1 1D02 2 1A05 검색결과 Glycine max glycine-rich RNA-binding protein (GRP) mrna Glycine max ribulose-1,5-bisphosphate carboxylase small subunit SS2-2 특이적발현 (by northern blot) N Y 3 1B08 unknown N 4 1D15 Glycine max 28 kda protein mrna Y 5 2A19 Glycine max ribulose-1,5-bisphosphate carboxylase small subunit Y 6 3K02 Glycine max ribulose-1,5-bisphosphate carboxylase small subunit Y 7 3N02 G.hirsutum light regulated unknown reading frame DNA Y 8 3I05 9 3O14 10 3A15 Glycine max vegetative storage protein mrna (VSP25 gene) N unknown(arabidopsis thaliana photosystem I PSI-N mrna) Y Glycine max vegetative storage protein mrna (VSP25 gene) N 11 3A19 Glycine max vspa gene Y 12 4P04 M.sativa 26S rrna Choristylis rhamnoides 26S ribosomal RNA gene N 13 4J06 unknown (Pseudomonas aeruginosa) N 14 4N14 G.max vegetative storage protien mrna (VSP B) Y 15 4O15 Glycine max repressor protein (Dr1) mrna, complete cds Y 16 4I16 Arabidopsis thaliana unknown protein (F19P19.2) mrna N 17 4C23 Glycine max 28 kda protein mrna Y 18 4G23 unknown N 19 5F2 Arabidopsis thaliana chromosome 2 N 20 5M6 Glycine max 28 kda protein mrna Y
21 5N08 Glycine max 28 kda protein mrna Y 22 5M11 G.max vegetative storage protein mrna (VSP25 gene) Y 23 5A15 Glycine max 28 kda protein mrna Y 24 Glycine max gene encoding 5B17 ribulose-1,5-bisphosphate carboxylase small subunit Y 25 5B22 Glycine max 28 kda protein mrna Y 26 6L12 Vigna radiata chlorophyll a/b-binding protein CP24 precursor (CipCp24) mrna Y 27 6H23 Atlantic salmon mrna for myostatin, isoform II Y 28 7C05 Pseudomonas aeruginosa Y 29 7D06 hypothetical protein XP_104599 [Homo sapiens] N 30 7O07 unknown N 31 7F20 unknown N 32 7L22 Arabidopsis thaliana mrna for putative beta-galactosidas Y 33 7L23 putative senescence-associated protein [Pisum sativum] N 34 7M24 Glycine max Homeodomain Leucine zipper protein Y 35 8A02 Arabidopsis thaliana clone 37689 mrna Y 36 8I07 unknown N 37 8I13 (NM_115671) putative protein [Arabidopsis thaliana] N 38 8D14 Arabidopsis thaliana unknown protein mrna Y 39 8J14 cysteine proteinase tpp (EC 3.4.22.-) - garden pea N (AC022472) Contains similarity to CaLB protein from 40 8C16 A. thaliana gb X96598 and contains multiple C2 N PF 00168 domains 41 8K16 N 42 8L16 imidazoleglycerol-phosphate synthase subunit H - like [Arabidopsis thaliana] N 43 8G21 Arabidopsis thaliana unknown protein (MSJ11.24/AT3g15840) mrna, N 44 8A22 (NM_101113) ATP-dependent Clp protease proteolytic subunit (ClpR2)[Arabidopsis thaliana] N 45 8L24 (AC005169) hypothetical protein [Arabidopsis thaliana] N
46 9I01 (AF136636) PR1a precursor [Glycine max] reverse 49 9C23 50 10E13 51 10O17 52 10G19 (NM_115343) vacuolar protein sorting protein 33a-like protein [Arabidopsis thaliana] Arabidopsis thaliana putative RING zinc finger protein (At1g63900) (AP002460) gene_id:f1d9.26~unknown protein [Arabidopsis thaliana] gene_id:f1d9.26~unknown protein [Arabidopsis thaliana] N Y N Y 53 1B05 (X85130) catalase [Campylobacter jejuni] N
β β
Flax L6 ( 241) DDHITAVLEKLSLDSENVTMVGLYGMGGIGKTTTAKAVYNKI-SSCFDCCCFIDNIRETQ Tobacco N ( 192) DTHLEKIESLLEIGINGVRIMGIWGMGGVGKTTIARAIFDTLLGRMDSSYQFDGACFLKD Arab RPS2 ( 159) TTMMEQVLEFLSEEEERGIIGVYGPGGVGKTTLMQSINNELITKGHQYDVLIWVQMSREF Arab RP1 ( 180) DAAKGKLIGWLLSPEPQRIVVSVVGMGGSGKTTLSANIFKSQTVRKHFASYAWVTISKSY Flax L6 ( 300) EKDGVVVLQKKLVSEILRIDSGSVGFNNDSGGRKTIKER-VSRFKILVVLDDVDEKFKFE Tobacco N ( 252) IKENKRGMHSLQNALLSELLREKANYNNEEDGKHQMASR-LRSKKVLIVLDDIDNKDHYL Arab RPS2 ( 219) GECTIQQAVGARLGLSWDEKETGENRALKIYRALRQKRFLLLLDDVWEEIDLEKTGVPRP Arab RP1 ( 240) VIEDVFRTMIKEFYKEAETQIPGELYSLTYRELVEKLVEYLHSKRYFVMLDDVWNTGLWR Flax L6 ( 359) DMLGSPKDFISQSRFIITSRSMRVLGTLNENQCKLYEVGSMSKPRSLELFSKHAFKKNTP Tobacco N ( 311) EYLAGDLDWFGNGSRIIITTRDKHL---IEKNDIIYEVTALPDHESIQLFKQHAFGKEVP Arab RPS2 ( 279) DRENKCKVMF-TTRSIALCNNMGAEYKLRVEFLEKKHAWELFCSKVWRKDLLE------- Arab RP1 ( 300) EISIALPDGI-SGSRVMVTTRSNNMASFSYGSGSRKHEIELLKEDEAWALFCNKAFSGSL Flax L6 ( 419) PSYYETLANDVVDTTAGLPLTLKVIGSLLFKQEIAVWEDTLEQLRRTLNLDEVYDRLKIS Tobacco N ( 368) NENFEKLSLEVVNYAKGLPLALKVWGSLLHNLRLTEWKSAIEHMK-NNSYSGIIDKLKIS Arab RPS2 ( 331) ----SSSIRRLAEIIVSKCGGLPLALITLGGAMAHRETEEEWIHASEVLTRFPAEMKGMN Arab RP1 ( 359) EECRRRNLEVVARKLVERCQGLPLAIASLGSMMSTKRLESEWKQVYNSLNWELNNNLELK
(1) 병저항성유전자유사체의불마름병균에대한반응 Glycinemax cv. Suwon157 RGA3 X. campestrispv. glycines8ra 10 mm MgCl 2 Hours after inoculation Hours after inoculation 0 2 4 6 12 24 48 72 120168216288 0 2 4 6 12 24 48 72 120168216288 RGA4 RGA5 RGA6 RGA7 RGA8
Glycinemax cv. Danbaek RGA3 X. campestrispv. glycines8ra 10 mm MgCl 2 Hours after inoculation Hours after inoculation 0 2 4 6 12 24 48 72 120168216288 0 2 4 6 12 24 48 72 120168216288 RGA4 RGA5 RGA6 RGA7 RGA8
RGA3,4 5,6,7,8 DB SW CTL YE PQ H 2 0 2 WD UV JA ABA ET SA rrna DB SW RGA3,4 SS 5,6,7,8 JY rrna SS JY
Glycinemax cv. Suwon157 PR-1a Per PAL CS IFS rrna X. campestrispv. glycines8ra 10 mm MgCl 2 Hours after inoculation Hours after inoculation 0 2 4 6 12 24 48 72 120168216288 0 2 4 6 12 24 48 72 120168216288 Glycinemaxcv. Danbaek PR-1a Per PAL CS IFS rrna X. campestrispv. glycines8ra 10 mm MgCl 2 Hours after inoculation Hours after inoculation 0 2 4 6 12 24 48 72 120168216288 0 2 4 6 12 24 48 72 120168216288
PAL DB SW DB IFS SW CTL YE PQ H 2 0 2 WD UV JA ABA ET DB CS SW DB rrna SW IFS CS SS JY SS JY CTL YE PQ H 2 0 2 WD UV JA ABA ET SA rrna SS JY
Jinju 1 PI96188 CTL YE H 2 0 2 JA SA CTL YE H 2 0 2 JA SA PAL IFS rrna
JY IFS mrna rrna 12 10 8 6 4 2 0 CTL YE PQ H 2 0 2 WD UV JA ABA ET SA rrna IFS mrna SS2-2 10 8 0 2 4 6
CTR YE PQ H 2 O 2 WD UV ABA ETH SA DB SW JY SS PI JJ rrna
연구 1차년도에는병저항성유전자유사체 5종이상을개발이었으나유전자유사체 6종 을개발하여연구목표를달성하였다. 유전자유사체 6종의불마름병및각종산화스트레스 호르몬에대한반응여부를불마름병저항성과감수성품종에서조사한결과, 유전자유사 체는처리및품종에불구하고발현이유도되지않았다. 불마름병저항성콩으로부터 full-length cdna 2종을분리하여그발
현특성을조사하고자하였으며, PAL, IFS, CS, SAM 등의유전자를분리하였다. 이들병 저항성유전자의불마름병및각종산화스트레스호르몬에대한반응여부를조사하였으 나불마름병저항성과감수성품종에서의반응이유사하여이들유전자는불마름병저항 성과직접적인관련이없는것으로나타났다.
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