ਏᔈೈқ - 效應蛋白RSp0213與PopP3在青枯病菌致病力之功能研究

࿶ಃΟࠠೈқϩݜس಍܌ϩݜǵᒡଌϐೈқᆀࣁਏᔈೈқ (effector)ǴځЬा

фૈӧܭυᘋ෌ނಒझғ౛ࢲ୏ǵቹៜ෌ނಒझխࣝϸᔈǴаׯᡂੰচ๵ठੰૈ

ΚǶਏᔈೈқӧ෌ނಒझޑբҔՏ࿼΢ϩࣁٿᜪǺ΋ᜪࢂ཮೏ଌΕ෌ނಒझϣޑ

ೈқǴٯӵ hop (hrp-dependent outer protein) ୷Ӣ܌ጓ᝿ޑೈқ (Alfano et al., 2000) ǹќ΋ᜪޑਏᔈೈқբҔܭ෌ނಒझ໔ሜǴό཮຾ΕಒझϣǴനԐว౜ޣ Ԗ P. syringae ޑ HrpZ Ϸ R. solanacearum ޑ PopA (Arlat et al., 1992)Ƕӧғނၗૻ

ϩ݋΢ޑࣴزᡉҢǴคፕ෌ނ܈୏ނੰচ๵ޑਏᔈೈқǴځ N ᆄ 25 ঁữ୷ለׇ

ӈࣣ൤֖Ԗ Ser ᆶ coiled ޑ୔ୱǵ٠લЮ Leu (Arnold et al., 2009; Samudrala et al., 2009)Ǵ௢ෳ೭ኬё៻Ԕޑೈқ่ᄬԖշܭਏᔈೈқ೯ၸ੟ઞޑ Hrp pilus (Büttner

and He, 2009)ǹԜѦǴਏᔈೈқӧϩݜၸำύǴӭኧޑਏᔈೈқѸ໪Ԗ੝ۓޑՔ ᒿೈқ (chaperone) ڐշωૈ຾ՉǴಃΟࠠϩݜೈқس಍ޑՔᒿೈқ೯தλܭ 170

ঁữ୷ለǵևለ܄ǵӧ C ᆄڀԖᚈཱུ܄ (amphipathic) ޑ α-helix ่ᄬ (Parsot et al., 2003) ǹ Ԗ ਔ Ք ᒿ ೈ қ ᗋ ת ᄽ ᆢ ࡭ ਏ ᔈ ೈ қ ᛙ ۓ ܄ ޑ ف Յ Ǵ ӵ Pectobacteria amylovora ޑՔᒿೈқ DspFǴ൩ڀԖᛙۓ၀ੰ๵ᜢᗖਏᔈೈқ DspA/E ޑфૈ

(Gaudriault et al., 2002)Ƕ

2. ߙߙ࢏ੰᆶߙ࢏ੰ๵

ߙ࢏ੰǴΞᆀಒ๵܄๺঒ੰ (bacterial wilt, BW)Ǵࢂҗߙ࢏ੰ๵ (Ralstonia solanacearum, Rs) ܌Їଆϐβᝆ໺ኞ܄ᆢᆅ״ੰ্ǶགࢉԜੰ্ޑ෌ਲ਼ಒਥፃϯ ᆭឳǵӦ΢೽ΠՏယ२ӃΠࠟǴԶࡕယТᅌԛ๺঒ǴӢ๺঒זೲࡺ෌ਲ਼ԝΫਔϝ և౜ߙᆘՅǴࡺᆀࣁȨߙ࢏ੰȩǶԜੰচ๵஌Ьጄൎ఼ᇂऊ 50 ࣽǴຬၸ 200 ᅿ෌

ނǴхࡴพनǵଭႍᖘϷ३ᑴ฻ख़ा࿶ᔮբނǴҞ߻ۘคԖਏϐϯᏢٛݯБݤǴ

ࢂШࣚ΢നᝄख़ޑ෌ނੰ্ϐ΋ (Poueymiro and Genin, 2009; Genin and Denny, 2012)Ƕ

ߙ ࢏ ੰ ๵ ࣁ ॠ ើ М ഍ ܄ (Gram-negative) ǵ ӳ ਼ (aerobic) ǵ ఎ ރ Ǵ ឦ β-proteobacteriaǴғߏޑཱུज़ྕࡋࣁ 4-40 °CǴคݤғߏܭ֖ 2%а΢ NaCl ᕉნǴӧ ଯᔼᎦ୻Ꭶ୷ΠܰౢғፃՅનǶߙ࢏ੰ๵ёߏԃӸࢲܭβᝆ܈НύǴ΋૓ރᄊΠ ёූӸܭβᝆऊ 14 ঁДǴനߏёၲ 25 ԃǴӧ 20-25 °C પНύ߾ёӸࢲߏၲ 40 ԃϐΦ (Denny, 2006)Ƕߙ࢏ੰ๵ޑ୷ӢಔӅऊ 5.8 MbǴҗ΋ঁᕉރࢉՅᡏ (3.7 Mb) аϷ΋ঁ୮ރѮ፦ᡏ (megaplasmid, 2.1 Mb) ܌ಔԋǴځύӸӧ೚ӭ mobile element Ϸ mosaic structureǴЪ୷Ӣಔύ֖ଯКٯ (66.5-68%) ޑ GC ᡵ୷ჹǶԾ 2002 ԃಃ΋ঁҗพनϩᚆԶٰϐߙ࢏ੰ๵๵ਲ਼ GMI1000 ୷Ӣಔ೏ֹ᏾ۓׇࡕ (Salanoubat et al., 2002)ǴҞ߻ӅԖ 5 ঁ๵ਲ਼ޑ୷Ӣಔֹ᏾ׇӈǴаϷ 17 ঁ๵ਲ਼ޑ

ׇӈ߃ዺ (genome draft) ೏ۓׇֹԋ (Boucher, NCBI website; Genin and Denny, 2012; Remenant et al., 2012; Cao et al., 2013; ߄΋ǵ߄Β) Ƕ

ߙ࢏ੰ๵ޑ๵س٩ྣځόӕ੝܄ёճҔΟᅿس಍຾ՉᅿϣϩဂǺਥᏵ஌Ьጄ ൎёϩԋϖᅿ racesǹ٩Ᏽჹόӕᚈᑗ/ᑗᎇϐճҔૈΚёϩԋϤঁ biovarsǹԶа DNA ׇӈϩ݋ᄽϯᜢ߯ёჄϩԋѤঁ phylotypes (Denny, 2006)Ƕphylotypes ᆶ๵

ਲ਼ޑӦ౛ٰྍεठ֍ӝǴϩձࣁ٥ࢪޑ phylotype IǴऍࢪޑ phylotype IIǴߚࢪޑ

phylotype IIIǴаϷٰԾӑѭǵᐞεճ٥کВҁޑ phylotype IV (Fegan and Prior, 2006)ǶԜѦǴᆶߙ࢏ੰ๵ᒃጔ࣬߈ޑ΍३ᐋੰচ๵ Ralstonia syzygii ᆶ३ᑴੰচ

๵ blood disease bacterium (BDB) Ψឦܭ phylotype IV (Remenant et al., 2011)Ƕ୷ ܭ΢ॊޑፄᚇ܄ǴҞ߻දၹа R. solanacearum species complex ٰᆀڥԜဂ๵ᅿ (Genin and Denny, 2012)Ƕ

3. ߙߙ࢏ੰ๵ޑϩηठੰᐒڋ

ߙ ࢏ ੰ ๵ ག ࢉ ஌ Ь ෌ ނ ၸ ำ Μ ϩ ፄ ᚇ Ǵ ӧ ᕉ ნ ڈ ᐟ ᆶ ဂ ᡏ ག ᔈ (quorum sensing) ޑૻ৲໺ሀၡ৩ΠǴठੰӢηڙډፄᚇЪᆒஏޑፓ௓Ǵ຾Զ௓ڋߙ࢏ੰ

๵᏾ᡏޑੰচ܄৖౜ (Valls et al., 2006; Vailleau et al., 2007; Genin and Denny, 2012)Ƕߙ࢏ੰ๵ޑགࢉၸำύǴ߃යූӸܭβᝆύޑߙ࢏ੰ๵གڙډ஌Ь෌ނਥ

೽ϩݜނϐࡕǴߡ཮ڙᖿϯ܄ (chemotaxis) ᇨᏤ౽୏Կ෌ނߕ߈Ǵ׎ԋғނጢ (biofilm) ຾Զߕ๱ឺᏵ (root attachment and colonization) (Schell, 2000; Yao and Allen, 2006)Ǵӕਔ߄౜ಃΟࠠೈқϩݜس಍ (type III secretion system, T3SS) ϩݜ ਏᔈೈқǴଌΕ෌ނಒझυᘋ෌ނ҅தғ౛фૈ (Poueymiro et al., 2009)Ƕ྽ᙖҗ

෌ނୁਥғߏೀ܈໾αߟΕࡕ (Yao and Allen, 2007)Ǵߡёس಍܄གࢉ᏾ਲ਼෌ނޑ ಳ ೽ Ǵ ݮ ๱ Е ፦ ೽ (xylem) ӛ ΢ ቚ ғ ᕷ ෗ ǵ ౢ ғ ε ໆ झ Ѧ ӭ ᗐ ᡏ (exopolysaccarides, EPS) ߔ༞Ր෌ނНϩၮᒡǴᕇڗ܌ሡᎦҽޑӕਔᏤठ෌ނԝ Ϋ (Denny, 2006)Ƕ

җܭߙ࢏ੰ๵ޑठੰၸำ౐ੋቶᗡǴ࣬ᜢޑठੰӢηཱུӭǴх֖ߙ࢏ੰ๵ޑ ᖿϯ܄ᆶݚ୏ૈΚ (motility)ǵझѦӭᗐᡏޑౢғǵಃΒࠠೈқϩݜس಍ (type II secretion system, T2SS) Ϸځ܌ϩݜϐಒझᏛНှሇન (cell wall degradation enzyme) (Liu et al., 2005)ǵᆶಃΟࠠೈқϩݜس಍฻ (Peeters et al., 2013)Ƕҁࣴزޑჹຝࣁ ߙ࢏ੰ๵ޑಃΟࠠϩݜਏᔈೈқ (Type III effectors, T3Es)Ƕ

4. ߙ࢏ੰ๵ಃΟࠠϩݜೈқ T3Es

ӧߙ࢏ੰ๵ύǴӧځ 1.9-Mb ޑε፦ᡏύಃΟࠠϩݜೈқፓ௓س಍ϐ hrp ୷Ӣ ဂεऊ՞Ԗ 23 kbǴϩԋԿϿϤঁᙯᒵൂՏǴጓ᝿рຬၸ 20 ঁᆫᴏ两᜘Ǵಔԋᐉ

ၠϣѦጢޑೈқϩݜس಍ (Arlat et al., 1992)Ƕੰচ๵२Ӄ೸ၸѦጢௗڙᡏ PrhA receptor གڙ෌ނૻဦϐࡕǴஒૻ৲໺ሀ๏ϣጢೈқ PrhRǴ٬চҁᆶ PrhR ่࣬ӝ

ޑ sigma factor PrhI ϩᚆញܫډಒझ፦྽ύǴ௴୏ prhJ ޑ߄౜ǴᙯᒵӢη PrhJ ཮

຾΋؁ࢲϯ hrpGǴHrpG ӆᇨᏤ hrpB ޑ߄౜Ǵനಖҗ HrpB ፓ௓ΠෞޑಃΟࠠϩ ݜೈқϐ߄౜ǴHrpB ཮ࢲϯ௴୏η஥Ԗ hrpII box ׇӈ TTCG-N16-TTCG ޑ୷ӢǴ

ࣴزว౜ऩׯᡂԜࢤߥӺ܄ׇӈǴ཮ቹៜډ௴୏ηڀԖ hrpII box ޑ popABCǵ hrpY ୷Ӣ߄౜ (Cunnac et al., 2004a)ǶЪ᏾ঁૻဦ໺ሀޑၡ৩೿཮ڙډठੰύኰፓ

௓Ӣη PhcA ܌ᆅ௓ (Peeters et al., 2013) (ߕკ΋)ǶԜѦǴHrpG ޑ਒سӕྍೈқ (paralogue) PrhG Ψ೏᛾ჴڀԖࢲϯ hrpB ޑфૈǴՠ prhG ѝ཮ӧ୷ҁ୻Ꭶ୷ (minimal medium) ύ೏ᇨᏤ߄౜Ǵࠅόڙډ෌ނಒझޑૻဦ܌ࢲϯ (Plener et al., 2010)ǶಃΟࠠϩݜೈқፓ௓س಍ჹܭߙ࢏ੰ๵ޑठੰߚதख़ाǴࣴزᡉҢஒߙ࢏

ੰ๵ޑҺ΋ hrp ୷ӢँᡂࡕǴ཮٬ߙ࢏ੰ๵ӧགੰ෌ނ΢ѨѐठੰΚǴӕਔΨ཮

Ѩѐӧלੰ෌ނ΢ᇨᏤၸ௵܄ϸᔈޑૈΚ (Van Gilsegem et al., 1995)Ƕ

ᒿ๱२ঁߙ࢏ੰ๵๵ਲ਼ GMI1000 ޑ୷ӢಔှׇǴ೸ၸғނၗૻϷᙯᒵᡏᏢޑ ϩ݋Ǵߙ࢏ੰ๵ޑਏᔈೈқ೏ว౜ڀԖаΠΟঁ੝܄ (Deslandes and Genin, 2014;

Kazan and Lyons, 2014)Ǻಃ΋Ǵൂ΋ߙ࢏ੰ๵๵ਲ਼ޑਏᔈೈқኧໆᇻӭܭځд෌

ނੰচ๵Ǵѳ֡΋ঁߙ࢏ੰ๵๵ਲ਼൩ڀԖ 60-80 ঁ T3Es ཮೏ଌΕ෌ނᡏ (Cunnac et al., 2004b)Ǵ࣬ၨܭ Xanthomonas spp.ᆶ P. syringae ޑ 30-45 ঁǴ೭ёૈΨࢂߙ

࢏ੰ๵஌ЬጄൎཱུቶޑচӢ (Poueymiro and Genin, 2009; Genin and Denny, 2012)Ǵ ՠ໻ԖϿኧೈқޑфૈ೏ϩ݋ǶಃΒǴߙ࢏ੰ๵Ӛ๵ਲ਼໔ᏱԖޑਏᔈೈқ۶Ԝৡ ౦܄ࡐଯǴ௢ෳࢂҗܭߙ࢏ੰ๵৒ܰ೸ၸНѳ୷Ӣᙯ౽ (horizontal gene transfer) ᕇளཥޑਏᔈೈқ (McCann and Guttman, 2008)ǴԖ٤ёૈٰԾ੿ਡಒझǴӵ஥Ԗ F-box domain ޑ GALA ೈқ (Angot et al., 2006)ǹԖ٤߾ёૈٰԾځд୏ǵ෌ނੰ

চ๵Ǵӵ TAL (transcriptional activator-like) ೈқǴځ೏ᇡࣁྍԾܭ Xanthomonas spp.ޑ TAL ೈқǴЪӕኬ೏᛾ჴ০ပӧ෌ނಒझਡ٠ڀԖӧ෌ނಒझύբࣁᙯᒵ ࢲϯӢη (transcriptional activator) ޑфૈ (Li et al., 2013)ǶಃΟǴߙ࢏ੰ๵ޑਏᔈ

ೈқ۳۳аৎ௼ (family) ޑ׎Ԅр౜Ǵӵ GALA ৎ௼ǵSKWP ৎ௼ (Poueymiro and Genin, 2009)Ǵ௢ෳߙ࢏ੰ๵ࣁΑ፾ᔈቶݱޑ஌Ь෌ނǴԶว৖р࣬՟ೈқࠅ фૈӭኬޑᄽϯ׎Ԅ (Remigi et al., 2011)Ƕ

ᜢܭߙ࢏ੰ๵ਏᔈೈқޑࣴزǴ߈ԃٰ໣ύӧағނၗૻБݤǵ܈೸ၸфૈ

܄ᑔᒧٰ൨פ҂ޕޑਏᔈೈқ (Deslandes and Genin, 2014; Kazan and Lyons, 2014)Ǵ

୤໻ԖϿኧਏᔈೈқԖ೏ుΕࣴزǴᖐٯ௶ॊӵΠǺ

4.1 Harpin

೚ӭॠើМ഍܄෌ނੰচ๵ޑ hrp ୷Ӣဂ཮ᙯ᝿р΋ᅿᆀࣁ harpin ޑೈқǴ ၀ೈқޑӅӕ੝܄ࣁǺ዗ᛙۓ (heat-stable)ǵữ୷ለׇӈύ൤֖ glycine Ъ཮ᇨᏤ לੰ෌ނޑၸ௵܄ϸᔈ (Kanda et al., 2003)ǶPopA (Pseudomonas outer protein A) ࣁߙ࢏ੰ๵ύಃ΋ঁ೏ว౜ޑ harpin (Arlat et al., 1992)ǴԜਏᔈೈқբҔܭ෌ނ ಒझ໔ሜǴёᇨᏤ๨૛ޑၸ௵܄ϸᔈǴ٠ᆶ popB (Pseudomonas outer protein B)ǵ popC (Pseudomonas outer protein C) Ӆࣁፓ௓ಔ (operon)Ǵڙ HrpB ܌ፓ௓Ƕࣴز ว౜ݙ৔஥Ԗ popA ϐߙ࢏ੰ๵ܭ๨૛ယύǴpopA ܭъλਔϣ൩཮ଯࡋ߄౜Ǵՠ ࠅ཮ѨѐᇨᏤၸ௵܄ϸᔈޑૈΚǹԶ࡭ុ߄౜ popA ϐߙ࢏ੰ๵ӧ๨૛዆ឲ၂ᡍ ύठੰΚΠफ़Ǵՠޔௗಳ೽ݙ৔၂ᡍࠅόቹៜځठੰૈΚǶᡉҢ PopA ӧߙ࢏ੰ

๵Ԑයགࢉਔ཮ڙډ๨૛܌ᒣᇡǴ٬๨૛ӧᒣᇡډੰচ๵ΕߟਔջЇวٛᑇϸᔈ (Kanda et al., 2003)Ƕ

4.2 GALA ৎ௼

GALA ৎ௼ϐਏᔈೈқӧߙ࢏ੰ๵ύ֖ԖΎঁԋ঩Ǵϩձࣁ GALA1-7Ǵځ Ӆӕ੝܄ӧܭữ୷ለׇӈύ֖Ԗ෌ނ੝Ԗϐ leucine-rich repeats (LRR) ᆶ੿ਡғނ (eukaryote) ϐ F-box domainǹځύӧ LRR ׇӈ္Ǵ׳ڀԖߥӺ܄ GAxALA ޑׇ

ӈǴӢԶᆀࣁ GALA ৎ௼ (Angot et al., 2006)ǶΔgala1-7 ϐँᡂਲ਼ӧߓ܎դ޺ᆶพ नύ཮फ़եठੰΚ (Angot et al., 2006)ǴځύΞа GALA2ǵ3ǵ6ǵ7 ჹགࢉߓ܎դ

޺ၨࣁख़ाǴGALA3ǵ7 ߾ࣁགࢉพन܌Ѹ໪ (Remigi et al., 2011)ǹԜѦǴ GALA7 ࣁ೷ԋरጴ (Medicago truncatula) ठੰ܌όё܈લޑೈқǴЪځ F-box domain ჹܭ GALA7 ޑठੰΨΜϩᜢᗖǴ F-box ೈқ೏ᇡࣁࢂ੿ਡೈқύᆶ SKP1-like ೈқҬϕբҔϐ୔ୱǴԶ SKP1-like ೈқΞ཮ᆶ Cullin1 ೈқҬϕբҔǴ Ӆӕ׎ԋ SCF-type E3 ubiquitin ligaseǴ೷ԋೈқ፦ޑݱનϯ (ubiquitination) ᆶफ़

ှ (Angot et al., 2006)ǶӢԜǴGALA ৎ௼೏ᇡࣁ཮೸ၸυᘋ஌Ь෌ނޑݱનϯफ़

ှ೼৩Ǵѐቹៜ෌ނޑғ౛фૈǴԶόӕޑ GALA ৎ௼ԋ঩ӧόӕ෌ނ΢܌೷ԋ ޑठੰૈΚΨόӕǴ௢ෳ GALA ৎ௼ԋ঩໔ϝԖфૈৡ౦Ǵࣁޑࢂ፾ᔈቶݱޑ஌

Ь෌ނǶ

4.3 AvrA

ߙ࢏ੰ๵ AvrA ਏᔈೈқǴ೏ว౜ࢂ๨૛ N. tabacum ᆶ N. benthamiana ύന Ьाޑᒣ᛽ჹຝǴavrA ँᡂਲ਼཮Ѩѐӧ N. tabacum ᆶ N. benthamiana ΢ᇨᏤၸ௵

܄ϸᔈޑૈΚǴځύữ୷ለಃ 32 ဦډಃ 50 ဦׇӈǴԜ 18 ঁữ୷ለࣁౢғၸ௵ϸ ᔈޑख़ा୔ୱǶҗܭ AvrA ׇӈύӸӧԖεໆޑख़ፄׇӈ (tandem repeats)Ǵ؂ঁ

ׇӈх֖ 12 ঁᡵ୷ჹ (base pairs)ǴЪ೽ҽׇӈᙯ᝿ࡕখӳՏܭಃ 32 ډ 50 ဦữ୷ ለϐ໔ǴᡉҢ೭٤ख़ፄׇӈӧᄽϯ΢നಖڙډ๨૛܌ᒣᇡ (Poueymiro et al., 2009)Ƕ

4.4 YopJ superfamily

ӧߙ࢏ੰ๵ख़ाЪ೏ుΕࣴزϐਏᔈೈқύǴхࡴ PopP ৎ௼Ǵᗧឦܭ YopJ (Yersinia outer protein J) superfamilyǴࣁಃΟࠠϩݜೈқύޑ΋ঁѮεৎ௼Ǵ നԐӧ୏ނޑဉၰࣽੰচ๵ Yersinia pseudotuberculosis ύ೏ว౜ (Monack et al., 1997)ǴԶځӕسނ (homologue) ቶݱӸӧܭӚᅿόӕޑ୏ǵ෌ނੰচ๵྽ύǴх

ࡴ୏ނੰচ๵ Salmonella spp.ޑ AvrAǹ෌ނੰচ๵ Pseudomonas spp.ޑ HopZ ৎ

௼ǴXanthomonas spp.ޑ AvrRxvǵAvrXv4ǵAvrBsTǵXopJǴPectobacterium spp.

ޑ ORFBǴR. solanacearum ޑ PopP ৎ௼ǹаϷ෌ނӅғ๵ Rhizobium spp.ޑ Y4LO (Lewis et al., 2011)Ƕ

ӧ ߙ ࢏ ੰ ๵ ύ Ԗ Ο ঁ ᗧ ឦ YopJ superfamily ޑ ਏ ᔈ ೈ қ ಍ ᆀ ࣁ PopP (Pseudomonas outer protein P)Ǵϩձࣁ PopP1ǵPopP2 Ϸ PopP3Ƕࣴزว౜ߙ࢏ੰ

๵ GMI1000 popP1::Tn ँᡂਲ਼ӧགࢉלੰ෌ނ࿖౐Ф (Petunia) ࡕ཮೷ԋ෌ނԝΫǵ གੰ෌ނߓ܎դ޺ᆶพनޑ๺঒௃׎߾཮ౣ༾ගϲǴԶচҁؒԖ PopP1 ޑߙ࢏ੰ

๵ Rd15 ๵ਲ਼Ǵӧ߄౜ PopP1 ࡕ཮Ꮴठ၀๵ਲ਼ӧচҁགੰޑ࿖౐Ф΢ࢥΚΠफ़ (Lavie et al., 2002)ǴЪаၭఎ๵อኩ߄౜ PopP1 ܭ๨૛ယύ཮ౢғၸ௵܄ϸᔈǴ ߄Ң PopP1 ڙ๨૛܌ᒣ᛽ǴᡉҢ PopP1 ӧߙ࢏ੰ๵ύࣁ avirulence protein ޑفՅ (Poueymiro et al., 2009)Ƕ

PopP2 ߾ӧߓ܎դ޺ύ೏ుΕ௖૸ǺGMI1000 ΔpopP2 ӧགࢉלੰޑߓ܎դ޺

ࠔس Nd-1 ࡕǴ཮Ꮴठ Nd-1 ޑԝΫǴЪࣴز᛾ჴ Nd-1 ύڀԖёޔௗᆶ PopP2 Ҭ ϕբҔޑ R ೈқ RRS1-RǴ၀ೈқ C ᆄڀԖӧ R ೈқύவ҂ว౜ޑ WRKY motif (Deslandes et al., 2003)ǹӕਔǴҗ RRS1-R ܌ᇨᏤޑ෌ނל܄ሡा΋ঁ cysteine ೈ қ䁙 Rd19 ٰӅӕ௴୏ǴRd19 ᆶ RRS1-R ΋ኬ೏᛾ჴᆶ PopP2 ёޔௗҬϕբҔ

(Bernoux et al., 2008)ǶPopP2 ӧ N ᆄ 34-53 ঁữ୷ለׇӈ೏ႣෳڀԖΕਡׇӈ (nuclear localization signal, NLS)Ǵჴᡍ่݀Ψᡍ᛾ PopP2 ዴჴ཮຾Ε෌ނಒझਡ ύǴᆶ RRS1-R ڀԖ࣬ӕޑ০ပՏ࿼ (Deslandes et al., 2003)ǹќѦǴPopP2 ೏ว౜

ёԾךΌㄽϯ (auto-acetylation) ޑૈΚǴΌㄽϯՏᗺӧ K383Ǵ၀ữ୷ለՏᗺӧ YopJ superfamily ԋ঩ύߥӺӸӧǴЪ PopP2 ԾךΌㄽϯޑࢲ܄ჹܭ RRS1-R ܌ᇨ Ꮴޑ෌ނל܄ࢂߚதख़ाޑ (Tasset et al., 2010)Ƕ

5. ෌෌ނੰ্ٛᑇϸᔈ

෌ނޑխࣝϸᔈ߯җ΋঺ፄᚇޑགޕၸำᆶૻဦ໺ሀୖᆶځύǴ΋૓ԶقǴ Ҟ߻෌ނޑٛᑇϸᔈё୔ϩࣁٿεᜪǺҗੰচ๵ي΢ޑߥӺ܄ϩη pathogen-associated molecular patterns (PAMP) ܌ᇨᏤޑ PTI (PAMP-triggered immunity) ٛᑇ ϸᔈǴаϷ೸ၸלੰ R ೈқѐᒣ᛽ੰচ๵ ਏᔈೈқԶ௴୏ޑ ETI (effector-triggered immunity) ٛᑇϸᔈ (Pieterse et al., 2009)ǶԶੰচ๵ࣁΑၲԋགࢉ෌ނҞ ޑǴᄽϯр׭ڋ෌ނٛᑇϸᔈޑਏᔈೈқǴҔаυᘋ෌ނޑғ౛фૈǵ׭ڋ஌Ь ϐխࣝᆀࣁ ETS (effector-triggered susceptibility)Ƕനಖੰচ๵ᏱԖӭኬৡ౦ϯޑ ਏᔈೈқǴԶ෌ނᡏύࢂցڀԖ஑΋܄ёᒣᇡੰ๵ਏᔈೈқϐלੰ R ೈқǴ߾،

ۓΑ෌ނૈց௴୏஑΋܄ၨଯޑխࣝس಍ǵౢғၸ௵܄ϸᔈϷ PCD ฻аज़ڋੰ๵

ᘉණԶ،ۓஎЬޑלੰૈΚ (Deslandes and Genin, 2014; Kazan and Lyons, 2014)Ƕ

5.1 ෌ނ PTI ٛᑇϸᔈ

ੰচ๵ي΢ёᇨᏤ PTI ٛᑇϸᔈޑ PAMPǴ೯தࣁੰ๵ߥӺ (conserved) Ъ ό ё ܈ લ ޑ ่ ᄬ ೈ қ Ǵ ӵ ಒ ๵ ᚎ Л ೈ қ flagellin ǵ ಒ ๵ ಒ झ Ꮫ ԋ ҽ peptidoglycan (PGN) ǵ চਡғނ两᜘ۯߏӢ η (elongation factor Tu, EFTu) ฻ (Boller and Felix, 2009) Ƕ ੰ চ ๵ ௗ ᝻ ډ ෌ ނ ਔ Ǵ ෌ ނ ཮ ೸ ၸ ጢ ΢ ޑ pattern recognition receptors (PRRs) གڙډੰ๵ي΢ޑ PAMPǴ଺ࣁᑈᐟ䁙 (kinase) ޑ PRR ೈқߡ཮௴୏ᕗለϯϸᔈ (phosphorylation)Ǵ֎Їځдೈқ፦่ӝ܈ׯᡂԾ ي่ᄬញрҬϕբҔೈқǴࢲϯΠෞޑխࣝᐒڋǶ྽෌ނௗԏډੰচ๵ૻဦǴ२ Ӄಒझጢޑᚆη೯೸܄཮ӧࡐอޑਔ໔ϣวғׯᡂǴගϲಒझ፦ϣޑ້ᚆη֖ໆǴ ᇨว෌ނ਻Ͼ (stomata) ᜢഈǴᗉխ෌ނᝩុௗ᝻ੰচ๵ǹ້ᚆηӕਔΨёᇨᏤಒ झϣ ROS ޑғԋǴROS ନޔௗ೷ԋಒ๵ғӸӒ্ѦǴ׳ёࢲϯΠෞޑ MAPK

cascades ޑૻ৲໺ሀၡ৩Ǵ຾Զ௴୏ٛᑇ୷Ӣ߄౜Ǵ܈ᘰ໾ဟᆫᑗ (callose) ғӝ ԋ୷ӢǴӧ෌ނಒझ໔ሜϣಕᑈᘰ໾ဟᆫᑗǴа߳ज़ੰচ๵ޑΕߟǶΨӢԜǴ PTI ٛᑇϸᔈࣁ෌ނჹбੰচ๵ޑ२ၰٛᑇس಍ǴځЬा੝ቻࣁᜢഈ਻Ͼǵ້ᚆ ηޑቚуǵזೲᇨᏤ ROS аϷ୴ᑈᘰ໾ဟᆫᑗ (Muthamilarasan and Prasad, 2013)Ƕ

5.2 ෌ނ ETI ٛᑇϸᔈ

Ꮓᆅ෌ނ PTI ϸᔈӧᄽϯ΢ԋфܢᏲΑε೽ϩੰচ๵ǴࠅΨԖੰচ๵ᄽϯр

࣬ᔈޑჹ฼ٰँઇ PTI ٛጕǴҭջᄽϯрբࣁ Virulence factor ޑӭᅿόӕਏᔈೈ

қٰυᘋ෌ނஎЬޑ҅தғ౛ᐒڋǵ׭ڋ஌ЬϐխࣝǶฅԶ΋ѿਏᔈೈқ೏෌ނ ޑ R ೈқ܌ᒣᇡǴٗ൩཮ᇨᏤ෌ނಃΒၰٛᑇϸᔈǴΨ൩ࢂ ETI ϸᔈǶETI ϸᔈ ନёᇨᏤಒझϣ ROS ޑғԋǴΨ཮ࢲϯΠෞޑ MAPK ૻ৲໺ሀၡ৩Ǵ຾Զ௴୏

೚ӭس಍܄ٛᑇລᅟᆾ࣬ᜢ୷Ӣޑ߄౜ǴᇨᏤхࡴНླྀለ (salicylic acid, SA) GluA Ϸ PR1a ฻࣬ᜢ኱ᇞ୷Ӣ߄౜ (Milling et al., 2011)ǵΌ౎ (ethylene, ET) PR1b1 ک Osmotin ฻ғӝԋ࣬ᜢ኱ᇞ୷Ӣޑ߄౜ (Milling et al., 2011) (ߕ߄Ύ)Ǵ٬

෌ނౢғس಍܄խࣝϸᔈ (systemic acquired resistance, SAR)Ǵ٠ᇨᏤ୔ୱ܄ޑಒ झԝΫ (programmed cell death, PCD) ฻ૻ৲໺ᏤǴаߔЗੰচ๵ᝩុΕߟ෌ނ (Deslandes and Genin, 2014; Kazan and Lyons, 2014)Ƕ

5.3 ෌ނಒझጢᄬ೷ೈқୖᆶٛᑇϸᔈ

೚ӭࣴزࣣࡰр෌ނಒझጢᆶځ΢ protein complexes ჹܭੰ্ٛᑇϸᔈתᄽ ᜢᗖفՅ (Poueymiro and Genin, 2009; Bohm et al., 2014)ǴٯӵǺᒣᇡੰ๵ PAMPs ϐ PRRs ᆶ೚ӭᒣᇡੰ๵ਏᔈೈқϐ R proteins ࣣՏܭ෌ނಒझጢ (Bohm et al., 2014) Ǵ ٠ Ъ ӧ ೚ ӭ ࣴ ز ύ ೿ ࡰ р Α PRR ᆶ PAMP ٿ ޣ ϐ ໔ ޑ ჹ ᔈ ᜢ ߯ (Muthamilarasan and Prasad, 2013)Ǵՠჹܭ PRRs ޑ࣬ᜢڐӕբҔೈқ (association proteins)Ǵ߾ࢂӧ߈൳ԃഌុ೏ుΕࣴزǴٯӵ PRR FLS2Ǵࣁ΋ᅿ leucine-rich receptor kinaseǴӧௗڙډಒ๵ᚎЛࡕǴջሡाӧ෌ނಒझጢᆶќ΋ঁ receptor-like kinase BIK1 ӅӕբҔǴۈૈᇨᏤΠෞ࣬ᜢ PTI ໺ሀၡ৩ (Lu et al., 2010)ǶԶ ჹܭԖᜢ෌ނಒझጢୖᆶ෌ނ࣬ᜢٛᑇϸᔈޑࣴزǴନΑᚈቫᕗિޑᄬ೷ගٮΑ ނ౛܄ࡀም (physical barrier) аѦǴಒझጢ΢ޑᕗિжᖴፓ௓ҭ཮ჹಒझٛᑇϸ ᔈౢғख़ाቹៜǴӵፓ௓ಒझᕗિޑሇન phospholipaseǴջ೏᛾ჴჹܭ෌ނಒझ

ޑ PCD ϸᔈϷٛᑇૻ৲໺Ꮴתᄽख़ाفՅ (Kim et al., 2014)ǴΞӵಒझጢ΢ޑ੝

ۓ่ᄬિใ (lipid raft)ǴӢځୖᆶፓ௓ಒझϣޑિ፦жᖴ (Bhat and Panstruga, 2005;

Lingwood and Simons, 2010)ǴЪς᛾ჴᆶᇨᏤ PCD ౢғϐ ROS ಕᑈஏϪ࣬ᜢ (Mersmann et al., 2010)Ǵࡺӧ෌ނٛᑇϸᔈޑፓ௓΢ڀԖᜢᗖفՅ (Fallahi-Sichani and Linderman, 2009)Ƕ

િใ߯җᕗિǵڰᎇᜪ (sterols) аϷ࣬ᜢ੝ਸфૈೈқ܌ᄬԋǴቶݱӸӧܭ

੿ਡғނύхࡴ෌ނಒझǶ࣬ᜢࣴزࡰрિใӧಒझጢޑᄬԋᛙۓ΢ǴаϷಒझ ϣ࣬ᜢિ፦жᖴޑፓ௓΢ڀԖԿᜢख़ाޑفՅ (Lingwood and Simons, 2010)ǴԶ ಒझޑ࣬ᜢિ፦жᖴፓ௓߾΋ޔ೏ຎࣁᆶ෌ނޑٛᑇϸᔈஏϪ࣬ᜢ (Munnik, 2001)ǶᖐٯԶقǴӧ෌ނ༾ғނҬϕբҔύǴԖ٤ੰচ๵཮ӧ෌ނಒझύញрિ

፦Нှ䁙 (lipid hydrolases) Ǵаઇᚯ஌Ьಒझޑٛᑇ่ᄬǴ٠Ъ٬எЬಒझሡा

ᙖҗಒझጢೈқ܌Їว܈ቚமޑ࣬ᜢٛᑇᐒڋфૈ഼ѨǴٰၲډߔ๊෌ނಒझౢ

ғس಍܄ٛᑇϸᔈϐҞޑ (Shah, 2005)ǶԶੰচ๵ନΑаሇનНှޑБԄઇᚯ෌

ނ ಒ झ ጢ Ѧ Ǵ ҭ ཮ ᙖ җ υ ᘋ ෌ ނ ಒ झ ޑ ਼ ϯ ᄊ ޑ િ ፦ (oxidized fatty acid) Oxylipins ޑբҔǴٰቹៜ෌ނಒझғߏᆶٛᑇϸᔈ (Vellosillo et al., 2007)Ƕ

6. ੰੰচಒ๵ਏᔈೈқӧ෌ނᡏϐբҔ

٩ᏵҞ߻ςޕϐਏᔈೈқӧ෌ނᡏբҔޑՏ࿼Ǵёεౣஒੰ๵ϐਏᔈೈқϩ ᜪǺϩձࣁբҔܭಒझጢǵಈጕᡏ (mitochondria)ǵယᆘᡏ (chloroplast)ǵ៶ݰ (vesicle) ၮᒡǵMAPK ૻ৲໺ሀǵᆶಒझਡϐਏᔈೈқ (Deslandes and Rivas, 2012;

Bohm et al., 2014)Ƕ

6.1 բҔܭಒझጢϐਏᔈೈқǺ

෌ނಒझጢ΢ڀԖ PRRs ฻ڙᡏೈқǴаϷ೚ӭᇨᏤΠෞٛᑇϸᔈϷፓ௓ᕗ

િжᖴޑᄬ೷ǴӢԜஎЬಒझጢதԋࣁੰচ๵ਏᔈೈқޑբҔҞ኱Ǵӵߓ܎դ޺

ಒझጢ΢ޑ RIN4 (RPM1-interacting protein 4)ǴRIN4 ࣁ෌ނխࣝϸᔈύॄፓ௓ೈ

қ (negative regulator)ǴPseudomonas ਏᔈೈқ AvrB ᆶ AvrRpm1 ൩೸ၸᇨᏤ RIPK (RIN4-interacting receptor-like protein kinase) ޑᕗለϯ߄౜Ǵ຾Զᕗለϯځ ҬϕբҔϐೈқ RIN4Ǵ׭ڋ PTI ϸᔈǹځ࣬ჹᔈϐ R ೈқ RPM1 Ψς೏פډ (Liu et al., 2011)ǶPseudomonas ਏᔈೈқ AvrRpt2 ߾ࣁ cysteine ೈқ䁙 (protease)Ǵ

཮೷ԋ RIN4 ޑϩှǴа׭ڋ෌ނ PTI ϸᔈ (Afzal et al., 2011)Ǵՠځϩှޑ࿗Т ࠅΨ཮೏ R ೈқ RPS2 ܌ᒣᇡǵ௴୏ ETI ٛᑇǴࡺќ΋ P. syringae HopF2 ਏᔈೈ

қǴ཮ᆶ RIN4 ҬϕբҔǴᗉխ AvrRpt2 ϩှ RIN4 ೷ԋ ETI ޑࢲϯǶ

ќѦǴP. syringae ޑ AvrPto ᆶ AvrPtoB ΨࢂբҔӧ෌ނಒझጢ΢ޑਏᔈೈқǶ AvrPto ཮׭ڋߓ܎դ޺Ϸพन FLS2 ޑᕗለϯфૈǴߔЗΠෞޑ PTI ૻဦ໺ሀ (Xiang et al., 2008)ǹAvrPtoB ߾ࢂ཮ᆶ FLS2ǵFLS2 ϐӅௗڙᡏ (co-receptor) BAK1 (BRI1-associated kinase 1) ҬϕբҔǴ׭ڋჹಒ๵ᚎЛೈқޑགޕௗڙ (Gohre et al., 2008)ǶԶӧพनύǴAvrPto ᆶ AvrPtoB ߾཮ᆶพनಒझύޑ Pto ᐟ 䁙ҬϕբҔǴࣴز௢ෳ Pto ёૈתᄽᝡݾ่ӝ (competitive binding) ޑفՅǴ٬

AvrPto ᇻᚆ PRR ೈқǴ຾ԶᇨᏤ෌ނޑ ETI ٛᑇϸᔈ (Xiang et al., 2008)Ƕ

6.2 բҔܭಈጕᡏϐਏᔈೈқǺ

੿ਡғނಈጕᡏࢂᆅ௓ PCD ϸᔈޑύኰ (Lam et al., 2001)ǴPCD ϸᔈՔᒿޑ

౜ຝхࡴǺಒझ፦๺ᕭǵਡࢉՅ፦ (chromatin) ᆫ໣ǵಈጕᡏᑩεǵనݰ (vacuole) ᆶယᆘᡏ (chloroplast) ڙཞ฻ (Muthamilarasan and Prasad, 2013)Ƕੰচ๵གࢉၸำ ύதᇨว੿ਡғނ PCD ϸᔈаٛᑇੰচ๵ߟࢉǴ෌ނޑ HR ϸᔈߡࢂന٫ٯηϐ

΋ǶӢԜಈጕᡏࣁੰচ๵ਏᔈೈқբҔޑख़ाՏ࿼ϐ΋ǴӃ߻ࣴز൩ว౜୏ނੰ

চ๵ޑਏᔈೈқ཮೸ၸࢲϯಒझғӸૻဦ (cell survival signals) υᘋஎЬಈጕᡏբ Ҕǵϩှ௴୏ಒझԾఠ (apoptosis) ޑૻဦೈқ (pro-apoptotic proteins)Ǵ׭ڋಒझ Ծఠаڐշੰচ๵Εߟ (Faherty and Maurelli, 2008)Ƕ

6.3 բҔܭယᆘᡏϐਏᔈೈқǺ

෌ނੰচ๵ύ೏ว౜০ပܭယᆘᡏύޑ ਏᔈೈқǺP. syringae HopI1 ᆶ HopN1ǶHopI1 ຾Εߓ܎դ޺෌ނಒझࡕǴ཮׎ԋ΋ঁεޑፄӝᡏ (complex)Ǵځ

ೈқ፦ C ᆄᆶ Hsp70 (heat shock protein 70) ่࣬ӝǴஒಒझ፦ύණပޑ Hsp70 ໣ ύԿယᆘᡏύǴယᆘᡏࣁ SA ғӝԋ൑܌ǴHsp70 ೏ᇡࣁ཮ቹៜ SA ޑғӝԋ܈ၮ ᒡ࣬ᜢфૈ (Jelenska et al., 2010)ǶԶӧ๨૛ᆶพनύǴHopN1 ཮෧եੰচ๵܌ᇨ Ꮴޑ PCDǵROS ౢғǵᆶᘰ໾ဟᆫᑗಕᑈǹࣴزว౜ HopN1 ཮ᆶ෌ނӀس಍

(photosystem II, PSII) ϐख़ाԋ঩ PsbQ ่ӝǴ೷ԋ PsbQ ޑफ़ှ຾Զ׭ڋӀس಍ޑ ࢲ܄ǵ׭ڋ ROS ғԋǴυᘋ෌ނ҅தғ౛ࢲ୏ (Rodriguez-Herva et al., 2012)Ƕ

6.4 բҔܭ៶ݰၮᒡϐਏᔈೈқǺ

ӧߓ܎դ޺ύǴP. syringae ϐਏᔈೈқ HopM1 ཮ಕᑈܭ෌ނಒझޑϸԄଯ୷ Мᡏᆛ๎ (trans-Golgi network ,TGN)Ǵᆶ AtMIN7 (Arabidopsis thaliana HopM1 interactor7) ่ӝǴӕਔ೷ԋځफ़ှ (Nomura et al., 2011)ǶAtMIN7 ࣁ៶ݰၮᒡၸำ ϐᜢᗖೈқǴჹܭ෌ނᘰ໾ဟᆫᑗϐಕᑈתᄽख़ाفՅǴࡺ HopM1 ཮׭ڋ෌ނ ಒझᏛ࣬ᜢϐٛᑇᐒڋ (Nomura et al., 2006)Ƕ

6.5 բҔܭ MAPK ૻ৲໺ሀϐਏᔈೈқǺ

MAPK ૻ৲໺ሀࣁ௴୏ PTI Ϸ ETI ٛᑇόё܈લޑ΋ᕉǴၸำፄᚇǵ౐ੋ೚

ӭᕗለϯೈқ䁙ୖᆶځύǴԋࣁਏᔈೈқբҔޑख़ᗺҞ኱ϐ΋ǺP. syringae ਏᔈ

ೈқ HopAI1 ڀԖ phosphothreonine lyase ޑࢲ܄Ǵ٬ளߓ܎դ޺ MPK3ǵMPK4 ᆶ MPK6 ѐᕗለϯ (dephosphorylation)Ǵՠ׭ڋ MPK4 ࢲ܄ޑ่݀཮Ꮴठ HopAI1

೏ R ೈқ SUMM2 ܌ᒣ᛽ǵ௴୏ ETI ٛᑇ (Zhang et al., 2012)ǶP. syringae ќ΋ਏ ᔈ ೈ қ HopF2 ߾ڀԖ mono-ADP-ribosyltransferase ޑࢲ܄Ǵ཮೸ၸਡᑗ୷ϯ (ribosylate) MKK5Ǵ׭ڋځᕗለϯфૈǴυᘋߓ܎դ޺ޑٛᑇϸᔈ (Wang et al., 2010)Ƕ

6.6 բҔܭಒझਡϐਏᔈೈқǺ

೚ӭੰচ๵཮೸ၸޔௗυႣ෌ނٛᑇ୷Ӣޑ߄౜ٰቹៜ෌ނϐխࣝϸᔈǴࡺ

ಒझਡதԋࣁਏᔈೈқբҔޑჹຝǴӵ TAL (transcription activator-like) ਏᔈೈқǺ TAL ਏᔈೈқࣁ DNA ่ӝೈқ (DNA binding protein)Ǵӧ Xanthomonas spp.ᆶ R.

solanacearum ύࣣӸӧǴTAL ೈқ೸ၸኳү੿ਡғނϐᙯᒵӢηǴޔௗᆶ஌Ь DNA ϐ௴୏η (promoter)่࣬ӝǴࢲϯ෌ނ୷Ӣǵᏹ௓խࣝϸᔈǶX. oryzae pv.

oryzae (Xoo)ϐ PthXo1 ཮ᇨวፓ௓ዿԯᗐᜪၮᒡޑ OsSWEET11 ߄౜Ǵஒᗐᜪᙯ ଌ๏ੰ๵٬Ҕ (Yang et al., 2006; Chen et al., 2010)ǹӧच෍ (pepper) ύǴX.

campestris pv. vesicatoria ϐ AvrBs3 ่ӝӧ UPA20 ϐ௴୏ηޑ UPA box ΢Ǵ UPA20 ࣁᙯᒵӢηǴљᙍ෌ނಒझᘉ஭ (enlargement) ፓ௓ǴלੰࠔسӸӧԖ R

ೈқ Bs3Ǵૈ୼٬෌ނౢғၸ௵܄ϸᔈ (Kay et al., 2007)Ƕ

6.7 װᔐᙯ᝿ϐਏᔈೈқǺ

Զќ΋ᜪਏᔈೈқǴӵ P. syringe ޑ HopU1Ǵ཮ቹៜ෌ނ RNA ޑжᖴ (metabolism) ၸำǴ ࣴ زว౜ mono-ADP-ribosyltransferse ਏᔈೈқ HopU1 ཮ ribosylate ෌ނύΟঁယᆘᡏ RNA ่ӝೈқ (chloroplast RNA-binding protein, CP-RBPs)ǵٿঁ൤֖ glycine ϐ RNA ่ӝೈқ (GR-RBPs)ǴЪ཮೷ԋځჹܭ RNA ޑ ࣁǺଯࢥΚϐ Pss190 ᆶ Pss1308ǵύࢥΚϐ Pss4 аϷեࢥΚϐ Pss216ǶଯࢥΚ๵

ਲ਼ᆶύǵեࢥΚ๵ਲ਼ޑϩᜪ٩Ᏽࣁځӧόӕพनࠔسύ܌೷ԋϐ๺঒౗ (ߕ߄΋)Ƕ

زი໗Ϸځд୯Ѧࣴزი໗ӝբǴа comparative genomic analyses ฼ౣКၨόӕ ࢥΚϐߙ࢏ੰ๵๵ਲ਼ޑ୷Ӣ౦ӕǴЪᆫขܭӧੰ๵-஌Ь෌ނҬϕբҔύڀᜢᗖف ՅޑಃΟࠠೈқϩݜس಍ᒡрϐਏᔈೈқ (Type III effectors, T3Es)Ƕ

ӧᆶݤ୯ Christian Boucher ࣴز঩ (INRA-CNRS, Toulouse) ϐࣴزი໗ޑӝ բࣴزύǴճҔύࢥΚϐ type strain GMI1000 Ⴃෳϐ୷Ӣׇӈ೛ीޑ microarray

຾ՉޑКၨϩ݋่݀ว౜΋٤ύ/եࢥΚ੝Ԗϐ୷Ӣ (ߕკΒ)Ǵځύхࡴ೏Ⴃෳࣁ T3Es ޑ RSp0213 ᆶ RSc3174Ƕҁࣴز࠻ޑӃ߻ࣴز᛾ჴԜΒ T3Es ڀଯࡋߥӺ܄

Ӹӧܭύեࢥٯ๵ਲ਼ǵЪڙ T3SS ፓ௓ (ߕკΟǵѤǵϖ)Ǵ٠௦Ҕ୷Ӣᙯ౽߄౜

฼ౣǴஒեࢥΚ๵ਲ਼ᐱԖޑ T3Es ᙯ౽߄౜ԿଯࢥΚ Pss190 Ϸ Pss1308 ޑᒪ໺ङ

ඳΠ߄౜Ǵ่݀ᡉҢᙯ౽߄౜٠҂ቹៜځӧ୻Ꭶ୷΢ғߏǵ෌ނᡏϣቚғǵӧ๨

૛΢ޑၸ௵܄ϸᔈǵғނጢ׎ԋаϷݚ୏ૈΚǴՠӧ Pss190 ङඳΠޑ୷Ӣᙯ౽๵

ਲ਼ӧਥ೽ߕ๱ૈΚԖΠफ़ޑ௃׎ǴPss1308 ङඳΠ߾คቹៜ (ߕ߄Β)Ƕҁࣴزԑ ӧ຾΋؁ᔠෳӧ T3Es ୷Ӣᙯ౽߄౜ΠǴߙ࢏ੰ๵ӧพन΢ޑࢥΚ߄౜Ǵ٠Ъғ ނၗૻϩ݋ᇶշΠ (ߕკϤ)ǴᕕှԜΒ T3Es ࢂց཮ڙ෌ނᒣᇡکӵՖᇨᏤ෌ނ ޑ࣬ᜢٛᑇϸᔈᐒڋǶ

ӧᆶम୯ Exeter εᏢ Murray Grant ௲௤ࣴزი໗ޑӝբύǴଞჹόӕࢥΚޑ ߙ࢏ੰ๵๵ਲ਼ Pss190ǵPss1308ǵPss4 Ϸ Pss216 (ߕ߄Ο)Ǵךॺςֹԋځӄ୷Ӣ ಔ (х֖ࢉՅᡏ DNA ᆶ megaplasmid) ޑۓׇǴ٠Ъว౜ځύ΋ঁଯࢥΚ๵ਲ਼ Pss190 ੝Ԗޑ୷Ӣࢂ೏Ⴃෳࣁ T3E ϐ΋ޑ PopP3 (ߕკΎǵΖǵΐ)Ƕҁࣴز࠻ޑ Ӄ߻ࣴز᛾ჴԜ T3E ཮ڙ T3SS ፓ௓ (ߕკΜǵΜ΋)Ǵࣴز่݀ҭᡉҢ୷Ӣᙯ౽ ߄౜ԿύࢥΚ๵ਲ਼ Pss4 ёቚமځӧพन΢ޑࢥΚ (ߕკΜΒ)ǴԶஒ PopP3 ߄౜ܭ

๨૛ယТคੰቻౢғՠԖճܭߙ࢏ੰ๵ϐғߏ (ߕკΜΟǵΜѤǵΜϖ)Ǵ٠௢ෳ

ځӧ෌ނಈጕᡏޑ০ပՏ࿼ᆶځфૈԖᜢǴёૈᙖԜቹៜพन PTI ࡰ኱ϐ΋ޑᘰ

໾ဟᆫᑗಕᑈ (callose deposition)Ǵа׭ڋ෌ނ PTI ϸᔈ (ߕკΜϤǵΜΎǵΜΖ)Ƕ ҁࣴزԑӧ຾΋؁᛾ჴ PopP3 ಈጕᡏ০ပՏ࿼ჹځфૈϐख़ा܄Ǵ٠ᔠෳԜ T3E ӧ୷Ӣᙯ౽߄౜ΠǴߙ࢏ੰ๵ӧพन΢ޑࢥΚ߄౜Ǵکځфૈᆶ HR ϐ࣬ᜢ܄Ǵ аឍܴ PopP3 ӵՖ׭ڋ෌ނޑ࣬ᜢٛᑇϸᔈᐒڋǶ

ಃ

ಃΒക ׷਑ᆶБݤ

1. ٮ၂๵ਲ਼ǵ፦ᡏϷ๵ਲ਼୻Ꭶచҹ

ҁࣴز܌٬Ҕϐ๵ਲ਼ᆶ፦ᡏޑ੝܄کٰྍӈܭߕ߄ѤǴ୻Ꭶ୷ଛБӈܭߕ߄ ϖǶ܌Ԗ๵ਲ਼ࣣҗ-80°C ߥӸڗрǴჄ๵ܭӝ፾ϐڰᄊ୻Ꭶ୷܈֖፾྽ᐚࡋלғ ન (ߕ߄Ϥ) ϐ୻Ꭶ୷Ǵࡷڗ๵ပࡕ຾ՉჴᡍǶεဉఎ๵ (Escherichia coli) а Luria Bertani (LB) ୻Ꭶ୷ܭ 37°C ୻Ꭶ΋Ϻǹߙ࢏ੰ๵ (R. solanacearum, Rs) а CPG ୻ Ꭶ୷ܭ 28°C ୻ᎦٿϺǹၭఎ๵а YEP ୻Ꭶ୷ܭ 28°C ୻ᎦٿϺǶ

2. ٮ၂෌ނ୻ػచҹ

ҁࣴز٬Ҕϐ෌ނࣁߙ࢏ੰלੰࠔسพन Hawaii 7996 (BW resistant, BW^R) ᆶ གੰࠔسพन L390 (BW susceptible, BW^S)Ǵҗ٥Ԁጫ๼ύЈ (Asian Vegetable Research and Development Center) Цᆬ޹Դৣ܌ගٮǶஒᅿηݰН᎜ᕏΟϺࡕǴ ኞܭ 3 मәࣧǴа 2Ǻ1 షکϐ಻ើβᆶਥ୷ܮ୻ᎦβύǴܫ࿼ܭࡡྕ 25°CǴ12 λਔӀྣǴ12 λਔ໵སޑΓπྕ࠻୻ػǴ٩ჴᡍሡ؃Կ፾྽ελࡕ຾ՉჴᡍǶ๨

૛ࠔسࣁ Nicotiana benthamiana ک Nicotiana tabacum W38Ǵҗ٥ࢪጫ๼ύЈЦᆬ

޹ԴৣගٮǴҁჴᡍ࠻ԾՉᕷ෗ǴᅿηӃаኜኞБԄኞܭ୻ᎦβǴܭΓπғߏጃ

୻ػว޵Ǵ΋ຼࡕ౽भԿ 3 मәࣧǴ୻ػܭࡡྕ 25°CǴ12 λਔӀྣǴ12 λਔ໵

སޑΓπྕ࠻Ǵය໔аᐚࡋऊ 1 g/L ϐ޸ᝊ 2 ဦࡼޥǴNicotiana benthamiana Ѥຼ

ࡕ຾ՉჴᡍǴNicotiana tabacum Ύຼࡕ຾ՉჴᡍǶ

3. ғނ಍ीϩ݋

ҁࣴزа SAS ಍ी೬ᡏ຾Չ಍ीϩ݋Ƕ܌ԖჴᡍϐኧᏵ࿶಍ी Student’s t-test ϩ݋Ǻ^ɀ߄Ңڀᡉ๱ৡ౦ (p<0.05) Ǵ^ɀɀ߄Ңڀཱུᡉ๱ৡ౦ (p<0.01)Ƕ

4. ख़ಔ፦ᡏᄬᑐᆶ୷Ӣၸໆ߄౜๵ਲ਼ࡌᄬ

4.1. DNA ᛏિᑗᏉጤႝݚ (Agarose Gel Electrophoresis) 4.1.1. ѦࢉᛏિᑗᏉጤႝݚϩ݋Ǻ

٩Ᏽటϩ݋ϐ DNA Тࢤελᆀڗ 0.8-1.2%ό฻ϐᛏિᑗᏉጤǴуΕ 0.5X TBE ጗ፂన (Tris-borate-EDTA buffer)Ǵ༾ݢу዗Կֹӄྋှࡕ᠗ኳǴࡑᏉڰϐࡕ ܭ 0.5X TBE ጗ፂనύа 100V/30 ϩដ຾Չႝݚϩ݋Ǵϩ݋ֹ౥ޑᛏિᑗᏉጤǴ

੆ݰܭऊ0.5 μg/mL Ethidium Bromide (EtBr) ྋనύ຾ՉѦࢉևՅ 15 ϩដǴܭ UV ᐩΠᔠຎ DNA ТࢤՏ࿼ᆶߝࡋǶ

4.1.2. ϣࢉᛏિᑗᏉጤႝݚપϯǺ

ӵటӣԏጤᡏϣ DNAǴ߾٬ҔϣࢉᛏિᑗᏉጤǶ٩Ᏽటપϯϐ DNA Тࢤε λᆀڗ 0.8-1.2%ό฻ϐᛏિᑗᏉጤǴуΕ 0.5X TAE ጗ፂన (Tris-acetate-EDTA buffer)Ǵ༾ݢу዗ԿֹӄྋှǴัհࠅࡕ؂ 10 mL уΕ 0.1 μL ޑ 50 mg/mL EtBrǴ འϬ᠗ኳǴࡑᏉڰࡕܭ 0.5X TAE ጗ፂనύа 100V/30 ϩដ຾Չႝݚϩ݋Ǵܭ UV ᐩΠᔠຎ DNA ТࢤՏ࿼ᆶߝࡋǶӵటϩ݋ъۓໆ RT-PCRǴ߾௦΢ॊБԄᇙբ TBE ϣࢉᛏિᑗᏉጤǶ

4.2. ᆫᆫӝ䁙ೱᙹϸᔈ (Polymerase Chain Reaction, PCR) 4.2.1. Taq PCR (Violet)Ǻ

٬Ҕ੝ۓЇηଛჹ (ߕ߄Ύ) аᆫӝ䁙ೱᙹϸᔈቚ൯Ҟ኱ DNA ТࢤǴϸᔈన ύа 1 ng ࢉՅᡏ DNA բࣁኳ݈ǴуΕ 2 μL ޑ 10X Taq PCR bufferǴ1.6 μL ޑ 2.5mM dNTPǴ1 μL ޑ 10 μM forward ЇηǴ1 μL ޑ 10 μM reverse ЇηǴ2 μL DMSOǴ0.2 μL ޑ Taq DNA ᆫӝ䁙 (5 U/μL)ǴуΕค๵ѐᚆηНԿ 20μLǶషӝ֡

Ϭࡕаᆫӝ䁙ೱᙹϸᔈᏔ຾ՉϸᔈǶϸᔈྕࡋᆶਔ໔ӵΠ܌ҢǺ

Cycle# Denaturation Annealing Polymerization

1 94°C /5 ϩដ

30 94°C /30 ࣾ T°C /30 ࣾ 72°C /t ϩដ

1 72°C /5 ϩដ

TǺྕࡋ٩ᏵЇη Tm ॶǹtǺਔ໔٩Ᏽቚ൯ߏࡋǴ1kb/minǶ

4.2.2. Q5® Taq PCR (Invitrogen)Ǻ

17 Cycle# Denaturation Annealing Polymerization

1 95°C /2 ϩដ

30 95°C /12 ࣾ T°C /30 ࣾ 72°C /t ϩដ

1 72°C /3 ϩដ

TǺྕࡋ٩ᏵЇη Tm ॶǴtǺਔ໔٩Ᏽቚ൯ߏࡋǴ2kb/minǶ

࿶ PCR ᘉቚϐౢނǴڗ 2 μL ຾Չ 1% ᛏિᑗᏉጤႝݚϩ݋Ǵаዴۓ DNA ߏࡋǶ

4.3. ፦፦ᡏ๧ڗ (Plasmid Purification) 4.3.1. εဉఎ๵፦ᡏ๧ڗǺ

ճҔ Gene-SpinTM Miniprep Purification Kit (Protech technology) ຾Չεဉఎ

๵፦ᡏ๧ڗǴӆа 1 % ᛏિᑗᏉጤႝݚϩ݋ዴۓ DNA ᐚࡋǴߥӸܭ-20°C ഢҔǶ уΕ฻ᡏᑈ phenol: chloroform: isoamyl alcohol (25: 24: 1)Ǵ΢ΠϸᙯషϬǴа 13000 rpm ࠻ྕᚆЈ 10 ϩដ٬ྋనϩቫǶλЈڗр΢ቫనԿཥޑ༾ໆᚆЈᆅǴу Ε฻ᡏᑈޑ isopropanol Ϸ 1/10 ᡏᑈޑ 3M NaOAcǴܫ࿼ܭ-20°CǴ10 ϩដࡕа 13000 rpm ࠻ྕᚆЈ 10 ϩដǴѐନ΢మనǴуΕ 500 μL 75%ଚᆒؑࢱ؈ᐘނǴӆ ᚆЈ 10 ϩដࡕѐନ΢మనǴѺ໒༾ໆᚆЈᆅ΢ᇂ٬ଚᆒචว 5 ϩដǴуΕ 40 μL ค๵ѐᚆηНӣྋ፦ᡏ DNAǴ٠уΕ 2 μL RNaseA (20 μg/mL)Ǵ࿼ܭ 37ɗ 30 ϩ ដǴஒ๧ڗޑ፦ᡏ DNA а 1 % ᛏિᑗᏉጤႝݚ຾Չϩ݋ (Sambrook et al., 1989)Ƕ

4.4. DNA પપϯ (DNA Purification)

ճҔ Micro-Elute DNA clean/Extraction Kit (GeneMark) ຾ՉጤᡏપϯǴӆа 1

% ᛏિᑗᏉጤႝݚϩ݋аዴۓ DNA ᐚࡋǶ

4.5. DNA ज़ڋ䁙੃ϯНှ (DNA Digestion)

ట຾Չख़ಔ፦ᡏϐ DNAǴၩᡏᆶҞ኱ DNA Тࢤ຾Չज़ڋ䁙 (New England Biolabs) ੃ϯНှǴϸᔈనᕴᡏᑈࣁ 50 μLǹటϩ݋፦ᡏ DNA ТࢤελǴϸᔈన ᕴᡏᑈ߾ࣁ20 μLǶϸᔈనϣ৒ӵΠǺ

Component 20 μL reaction 50 μL reaction

DNA 0.5 μg 4 μg

10X NEB Buffer 2 μL 5 μL

100X BSA 0.2 μL 0.5 μL

Restriction enzymes 2-4 units 4-8 units

ddH2O ံԿᕴᡏᑈ20 μL ံԿᕴᡏᑈ50 μL

ϸᔈనషӝ֡ϬࡕǴܭሇનϸᔈྕࡋ 37°C Πϸᔈ 2-4 λਔ (੝ਸሇનϸᔈྕࡋǺ SmaI ࣁ 25°C/ SfiI ࣁ 50°C)Ƕӧᄬᑐख़ಔ፦ᡏਔǴ٩ჴᡍሡ؃ஒҞ኱ DNA у዗

65°C ܈ 80°C ࡭ ុ 20 ϩ ដ ѐ ନ ज़ ڋ 䁙 ሇ ન ࢲ ܄ Ǵ ӆ у Ε ᡵ ܄ ᕗ ለ 䁙 (calf instestinal alkaline phosphatase, CIP) (New England Biolabs) 2 units ѐନ DNA 5 ᆄ΢

ޑᕗለਥǴܭ 37°C Πϸᔈ 1 λਔǶനࡕа 1 % ᛏિᑗᏉጤႝݚϩ݋Ƕ

4.6. DNA Тࢤௗӝ (DNA Ligation)

ஒज़ڋ䁙ሇનϪപપϯࡕޑҞ኱ DNA ᆶၩᡏ DNA ϐವԸКٯፓ᏾ࣁ 5: 1 ܈ 20: 1ǴDNA ᕴᐚࡋεܭ 100 ngǴуΕ 1 μL ޑ 10X T4 ligation buffer Ϸ 0.2 μL ޑ T4 DNA ligase (400 U/μL) (New England Biolabs)Ǵаค๵ѐᚆηНံԿᕴᡏᑈ 10 μLǴ֡Ϭషӝࡕܭ࠻ྕΠ຾Չௗӝϸᔈ 16 λਔǶ

4.7. TOPO® ፦ᡏᄬᑐ (TOPO® Cloning)

ஒ PCR ᘉቚϐౢނճҔ pCR®8/GW/TOPO® TA Cloning® Kit (Invitrogen)Ǵ ஒҞ኱ DNA ᄬᑐܭ pCR®8/GW/TOPO®ၩᡏ΢Ƕஒཥᗲޑ PCR ౢނ 4.5 μL уΕ 1 μL salt solution ᆶ 0.5 μL pCR®8/GW/TOPO®ၩᡏǴ࠻ྕΠϸᔈԿϿ 1 λਔǶ

4.8. LR ख़ख़ಔϕඤϸᔈ (LR Recombination)

ճҔGateway® LR Clonase™ II Enzyme Mix (Invitrogen) ஒҞ኱ DNA аӕྍ

ख़ಔϕඤޑБԄඤԿట٬Ҕޑၩᡏ΢Ƕҁࣴز٬Ҕ pCR8®/GW/TOPO® ၩᡏ຾Չ ӕྍख़ಔϕඤǴϸᔈనಔԋӵΠǺ

LR recombination components Amount (μL) Entry clone (50-150 ng)

Destination vector (150 ng/μL) LR Clonase^TMII enzyme mix TE buffer, pH 8.0

1-8 μL 1 μL 1 μL

ံԿᕴᡏᑈ10 μL ஒϸᔈనషӝ֡ϬࡕǴ࿼ܭ࠻ྕԿϿ 1 λਔ຾ՉϸᔈǶ

4.9. εဉఎ๵യҺಒझ዗ҶլᙯࠠբҔ (Heat Shock Transformation)

ҁࣴزаεဉఎ๵ DH5α ๵ਲ਼຾Չ፦ᡏख़ಔჴᡍǶڗᕴᡏᑈ΋ъϐௗӝౢނ

4.11. ႝႝऀϾᙯࠠբҔ (Electroporation Transformation)

ஒയҺಒझᆶऊ 50 ng ፦ᡏ DNA షӝࡕǴܫΕ٣ӃܭӇ΢Ⴃհޑ cuvette ύǴ

YEP ୻Ꭶ୷ /εဉఎ๵ǺLB ୻Ꭶ୷)Ǵߙ࢏ੰ๵ᆶၭఎ๵ܭ 28°C Π୻Ꭶ 2-3 ϺǴ εဉఎ๵߾ܭ 37°C Π୻Ꭶ 16 λਔǴࡷڗൂ΋๵ပǶ

4.12. ߙߙ࢏ੰ๵ࢉՅᡏ DNA ๧ڗ (Bacterial Genomic DNA Purification)

ࡷᒧൂ΋๵ပаబу੝ۓלғનϐ 523 నᡏ୻Ꭶ୷ܭ 28°C Π 200 rpm ႖ڹ᎜ Purification and Protein Localization Assay)

ڗ 4 ຼε N. benthamiana ϐԃᇸယТǴаΘТჄрኧၰ໾αǴ࿼ܭς֖ 25 mL enzyme solution ϐ୻ᎦҝύǴenzyme solution ሡӃа 50°C Ⴃ዗ 10 ϩដࡕհࠅ Կ࠻ྕǴ๨૛ယТӧ࠻ྕΠа 50 rpm ໵ས᎜ᕏ 3 ঁλਔࡕǴа༟ጤᅀᆅ֎ڗ֖চ Plasmid Midiprep Purification Kit (GeneMark) ๧ڗϐଯᐚࡋ፦ᡏ DNA (p2GWF7.0-RSp0213-GFP ᆶಒझጢ኱૶ pm-rkǵp2GWF7.0-PopP3-GFP ᆶಈဏᡏ኱૶ pBIN-mt-rk) (Nelson et al., 2007) 30 mg Ъᡏᑈόεܭ 20 uLǴуԿཥ༟ጤᚆЈᆅǴ٠у

22 microscope ᢀჸ (modified from Treuter et al., 1993)Ƕ

5.2. ਏਏᔈೈқܭ๨૛ယТಒझ০ပՏ࿼ᔠෳ (N. benthamiana Leaves Protein Localization Assay) λਔ໵སޑ෌ނғߏጃΒԿΟϺǴаᑻӀᡉ༾᜔ Confocal microscope ᢀჸ๨૛ယ ङǶ

5.3. ߙ࢏ੰ๵ࢥΚෳ၂ (Virulence Test)

ஒߙ࢏ੰ๵ Pss4 ܈ Pss190 ഁғࠠ๵ਲ਼܈୷Ӣকନϐ๵ਲ਼ǴճҔ߄౜ၩᡏ

๺঒ǹ2 ࣁΒԿΟТယТ๺঒ǹ3 ࣁନΑഗယѦځᎩယТࣣ๺঒ǹ4 ࣁӄ೽ယТࣣ

๺঒ǹ5 ࣁ෌ਲ਼ॹҷԝΫǶ

6. ෌෌ނᡏอኩ߄౜ਏᔈೈқ୷Ӣᆶځфૈ܄ࣴز

6.1. ๨૛ယ೽୷Ӣอኩ߄౜ (Transient Overexpression in Tobacco)

ճҔ஥ԖΒϡ߄౜ၩᡏ (binary vector) pCAMBIA1300 ϐၭఎ๵ GV3101 ຾Չ

୷Ӣอኩ߄౜Ƕ-80°C ཥᗲჄ๵Ǵܭ 28°C ғߏጃ୻ᎦٿϺǴаబу kanamycin ᆶ

6.2. พनอኩ܄ੰࢥᇨᏤ୷Ӣၸໆ߄౜ (Virus-mediated Gene Overexpression) а߄౜ potato virus X (PVX) ϐΒϡၩᡏ (pSfinX) းၩਏᔈೈқ୷ӢǴӆճҔ

஥ԖՔᒿೈқ (chaperon) ߄౜፦ᡏ (pSoup) ϐၭఎ๵ MOG101 ๵ਲ਼߄౜ (Takken et al., 2000)Ƕ-80°C ཥᗲჄ๵Ǵܭ 28°C ғߏጃ୻ᎦٿϺǴаబу kanamycinǵ tetracyclin ᆶ rifampin ϐ 3 mL YEP నᡏ୻Ꭶ୷Ǵ28°C Π 200 rpm ႖ڹ᎜ᕏ୻Ꭶǹ

6.3. ෌෌ނᡏอኩ߄౜୷Ӣᔠෳ

6.3.1 ෌ނ RNA ๧ڗ (RNA Extraction from Plants)Ǻ

ԏڗаၭఎ๵อኩ୷Ӣ߄౜ٿϺϐ๨૛ယТǴ܈อኩ܄ੰࢥᇨᏤ୷Ӣၸໆ߄

౜ࡕ 14 ϺϐพनယТǴऊ 0.1 g ෌ނಔᙃǴа Total RNA Mini Kit (LabPrep^TM) ܜ ڗ෌ނ RNAǶ٠а RNase-free DNase I kit (Promega) ຾Չ DNA ੃ϯНှբҔǺڗ 5 μg ޑ RNAǴуΕ 5 μL (1U/μL) ϐ DNase ሇનǵ5 μL RNase-free DNase 10X bufferǴа DEPC-H2O ံԿനಖᡏᑈ 50 μLǴషӝ֡ϬࡕуΕ༾ໆᚆЈᆅϣǴ࿼ܭ 37 ɗ ϸ ᔈ 1.5 λ ਔ Ƕ у Ε 150 μL ϐ DEPC-H2O ᆶ 200 μL RNA Ҕ phenol:chloroform (3:1) (pH= 4)Ǵϸᙯ֡Ϭషӝ 5 ϩដǴа 13000 rpm 4°C ᚆЈ 15 ϩដǴڗ΢మనԿཥޑ 1.5 mL ༾ໆᚆЈᆅǴуΕ 20 μL ޑ 3 M ᎉለ໊ (pH 5.3) Ϸ 1 mL 100%ଚᆒ٠షӝ֡ϬǴ࿼ܭ-80°C ႖ڹᓉ࿼؈ᐘǶа 4°C 13000 rpm ᚆЈ 20 ϩដǴѐନ΢మనǴуΕ500 μL 75% RNA Ҕଚᆒమࢱ؇ᐘނǴа 13000 rpm ᚆ Ј 2 ϩដࡕѐନ΢మనǴख़ፄԜ؁ᡯ΋ԛǴᄇۭ౽ନ΢మనࡕǴܭค๵ᏹբѠϣ චวଚᆒ 2 ϩដǴа 15 μL DEPC-H2O ӣྋ RNAǴߥӸԿ-80°C ഢҔǶ

6.3.2 ϸᙯᒵᆫӝ䁙ೱᙹϸᔈ (Reverse Transcription PCR, RT-PCR)Ǻ

а Reverse Transcription System kit (Promega) ຾Չϸᙯᒵᆫӝ䁙ೱᙹϸᔈǶڗ 1 μg RNAǴа DEPC-H2O ံԿ 10.2 μLǴа 70°C у዗ 10 ϩដǴҥջ࿼ܭӇ΢ 3 ϩដǶуΕ4 μL 25 mM MgCl2ǵ2 μL 10X reverse transcription bufferǵ2 μL 10 mM dNTP mixǵ1 μL Oligo (dT)15 (0.5 μg/μL)ǵ0.4 μL recombinant RNasin ribonuclease inhibitorǵ0.4 μL Avian myeloblastosis virus (AMV) reverse transcriptase (15U/μL)Ǵ షӝ֡ϬࡕܫΕᆫӝ䁙ೱᙹϸᔈᏔύǴа 42°Cǵ1.5 λਔ຾ՉϸᔈǴϐࡕа 95°Cǵ 5 ϩដಖЗሇનϸᔈǶӝԋϐ cDNA аค๵ѐᚆηНံԿᕴᡏᑈ 80 μLǴߥӸܭ-20°C ഢҔǶ

6.3.3 ъۓໆ RT-PCR (Semi-quantitative RT-PCR, sqRT-PCR)Ǻ

ڗ 100-200 ng cDNA բࣁኳ݈ǴуΕ 2 μL ޑ 10X Taq PCR bufferǴ1.6 μL ޑ 2.5mM dNTPǴ1 μL ޑ 10 μM forward ЇηǴ1 μL ޑ 10 μM reverse ЇηǴ0.2 μL ޑ

Taq DNA ᆫӝ䁙 (5 U/μL)ǴуΕค๵ѐᚆηНԿ 50μLǶషӝ֡Ϭࡕаᆫӝ䁙ೱᙹ ϸᔈᏔ຾ՉϸᔈǶϸᔈྕࡋᆶਔ໔ӵΠ܌ҢǺ

Cycle# Denaturation Annealing Polymerization

1 94°C /5 ϩដ

6.3.4 ջਔۓໆ Real-time PCRǺ

! ҁჴᡍࣁ٬Ҕྕࡋఊࡋਡለջਔۓໆୀෳس಍ (Bio-Rad Real-Time PCR Detection SystemsǴࠠဦ BIO-RAD MyiQTM)ǴReal-time PCR ϸᔈ၂Ꮚ߾ࢂᖼວ KAPA SYBR® qPCR Kit (Universal, ABI Prism®, Bio-Rad iCycler™, or Roche LightCycler™ 480)Ƕჴᡍ߻Ѹ໪Ӄ຾Չ No Template Control (NTC) ෳ၂Ǵዴᇡࢂ

ցԖ primer dimmerǶҁࣴزϐჴᡍϸᔈᕴᡏᑈࣁ 18 μLǴჴᡍБݤӵΠǴ२Ӄ cDNA ีញࣁ 30 ng/μL ࡕڗ 8 μLǴуΕ٣Ӄଛ࿼ӳޑ cocktailǴϣ֖Ԗ 9 μL 2x KAPA SYBR^® FAST qPCR Master MixǴ0.5 μL ޑ forward primer (10 μM)Ǵ0.5 μL ޑreverse primer (10 μM)Ǵషӝ֡ϬࡕܫΕ Real-Time PCR ϸᔈᏔύ຾Չ PCR ϸ ᔈǴӆа Bio-Rad ࣴวϐ iQ™5 Optical system version 2.0 ހ೬ᡏǴ຾Չϩ݋Ƕ

Real-time PCR (qPCR)

Reaction Component Amount (μL) Template cDNA (100 ~200 ng) 8 μL

Forward primer (10 μM) 0.5 μL Reverse primer (10 μM) 0.5 μL 2x KAPA SYBR® FAST qPCR Master Mix 9 μL

Total volume 18 μL

Real-time PCR program Cycling step Temperature & time Hot start Initial denaturation 3 min at 95°C PCR Denaturation 10 sec at 95°C

40 cycles Annealing 30 sec at 55°C (ຎ primer Զۓ)

Melting curve 1 min at 95°C 1 min at 55°C

10 sec at 95°C 81 cycles

ಃ

ಃΟക ่݀

I. ե/ύࢥΚ๵ਲ਼੝Ԗϐ T3Es RSp0213 ᆶ RSc3174 ӧߙ࢏ੰ๵ठੰΚϐфૈࣴز

1. RSp0213 ᆶ RSc3174 ӧςֹӄှׇϐߙ࢏ੰ๵๵ਲ਼ύϐϩթ

ࣁ߃؁ᕕှ RSp0213 ᆶ RSc3174 ϐӸӧᆶցࢂցᆶ๵ਲ਼ࢥΚ߄౜Ԗ࣬ᜢ܄Ǵ

ࡺаҞ߻ςှׇֹ᏾܈ςԖ୷Ӣಔ૛ዺ (genome draft) ޑ 22 ਲ਼ߙ࢏ੰ๵๵ਲ਼ࣁჹ ຝǴϩ݋ RSp0213 ᆶ RSc3174 ϐӸӧ௃׎ (߄΋)Ƕ่݀ᡉҢӧϩ݋ޑ๵ਲ਼ύǴύ ࢥΚ๵ਲ਼ GMI1000 ᆶ Pss4ǵեࢥΚ๵ਲ਼ Pss216 Ϸ҂ޕࢥΚ๵ਲ਼ PSI07 ڀԖ RSp0213ǴԶ GMI1000ǵPss216 Ϸ PSI07 ๵ਲ਼ΨڀԖ RSc3174 (߄΋)Ƕҁࣴز࠻

Ӄ߻ճҔ Southern blotting ᆶ genomic DNA PCR ᔠෳ܌ள่݀Ψᕇ΋ठ่݀ (ߕ კΟ)ǴЪӧեࢥΚ๵ਲ਼ᆶ GMI1000 ύ RSp0213 ᆶ RSc3174 ϐữ୷ለׇӈ൳Яֹ

ӄ࣬ӕ (ߕკѤ)Ǵࡺ೭ٿঁ T3Es ӧύࢥΚϷեࢥΚ๵ਲ਼ύࣣڀԖଯࡋߥӺ܄Ƕ

2. RSp0213 ک RSc3174 ჹߙ࢏ੰ๵ठੰΚϐቹៜ

ࣁΑ຾΋؁ᡍ᛾ RSp0213 ᆶ RSc3174 ϐӸӧᆶ๵ਲ਼ठੰΚϐ߄౜ዴჴԖᜢೱǴ

ࡺаၸໆ߄౜ᆶ୷Ӣকନ฻ࣴز฼ౣ຾Չځфૈϩ݋Ƕ२ӃǴ೭ٿঁ T3E ୷Ӣঁ

ձᒧ෗ډ low-copy-number ϐ pUFR047 ၩᡏǴ٠ϩձᙯ౽ډচҁόڀԖ၀ T3Es ޑଯࢥΚ๵ਲ਼ Pss190 ύ߄౜ǴҔޜၩᡏ (empty vector) ᙯࠠϐ๵ਲ਼଺ࣁჹྣಔǴ

ௗ๱ෳ၂ᙯࠠ๵ਲ਼ӧלੰพनࠔس H7996 ΢ϐठੰΚ߄౜ (კ΋)Ƕ่݀ᡉҢ Pss190 ๵ਲ਼߄౜ RSp0213 ࡕځठੰΚዴჴ཮ᡉ๱Ӧफ़ե (p<0.01)Ǵՠ Pss190 ๵ਲ਼ ߄౜ RSc3174 ࡕ߾ځठੰΚᗨԖΠफ़ޑᖿ༈Ǵՠᆶޜၩᡏᙯࠠ๵ਲ਼໔٠όڀᡉ๱

ৡ౦ (p>0.05)Ƕࡺࡕុჴᡍஒа RSp0213 ࣁЬाࣴزჹຝǶ

࿶ճҔ NCBI Genome ၗ਑৤຾ՉׇӈКჹ (blastp) ว౜ӧߙ࢏ੰ๵ Pss4 ύ ٠ ค RSp0213 ϐ paralogs Ǵ ࡺ ߃ ؁ ௨ ନ Ԗ ᆶ RSp0213 ф ૈ ख़ ፄ (functional redundancy) ϐ paralogs ޑёૈ܄ǶࣁΑ຾΋؁ዴᇡ RSp0213 ჹߙ࢏ੰ๵ठੰΚޑ ख़ा܄ǴӧύࢥΚ๵ਲ਼ Pss4 ύஒ RSp0213 কନǴ٠Кၨ܌ள๵ਲ਼ᆶഁғࠠ๵ਲ਼ӧ གੰพनࠔس L390 ΢ϐठੰΚ߄౜Ƕ่݀ᡉҢஒ RSp0213 কନࡕځठੰΚܴᡉ ଯܭഁғ๵ਲ਼ (კΒ)Ǵᆕ΢่݀ǴRSp0213 ϐӸӧόճܭߙ࢏ੰ๵ϐठੰΚ߄౜Ƕ

3. RSp0213 ೈೈқӧ෌ނಒझύϐ০ပՏ࿼

ࣁΑ߃؁ᕕှ RSp0213 ຾Ε෌ނಒझࡕёૈЇଆޑբҔǴӃճҔ Softberry ᆛ ઠ (http://linux1.softberry.com/berry.phtml) ϩ݋ RSp0213 ёૈڀԖϐ functional domains ᆶӧ෌ނಒझύёૈޑ০ပՏ࿼Ƕ่݀ᡉҢ RSp0213 ޑ N ᆄ 47 ঁữ୷ለ ёૈࢂᏤӛᴏ两 (targeting peptide) Ϸૻ৲ᴏ两 (signal peptide, SP)ǴЪႣෳӧԜࢤ

ૻ৲ᴏ两ύڀԖၠጢТࢤ (transmembrane segments)Ǵ௢ෳ RSp0213 ёૈՏܭಒझ ጢ΢ (ߕკϤ)Ƕ

ࣁΑᡍ᛾ RSp0213 ӧ෌ނಒझϣϐ০ပՏ࿼Ǵࡺаᆶพनӕࣁनࣽ෌ނޑ༝

ယ๨૛ (Nicotiana benthamiana) ଺ࣁჴᡍ׷਑຾Չ RSp0213 ϐۓՏϩ݋Ƕ่݀ᡉ Ң RSp0213::GFP ӧ๨૛চғ፦ᡏ (protoplasts) ύϐ০ပՏ࿼ᆶಒझጢ኱૶ೈқ (membrane marker) PIP2A::mcherry ϐϩթ΋ठ (კΟ)ǴԶЪǴӧ຾ՉቹႽ᠄კೀ

౛ࡕ (fold over) ёᢀჸډ RSp0213::GFP և౜ಈރ่ᄬϩթǴ௢ෳ RSp0213 ᔈࢂ

০ပӧ෌ނಒझጢ΢ϐೈқ፦໣ဂ (protein cluster) ԶևᗺރϩթǶќѦǴаၭఎ

๵ ᇶ շ ݙ ৔ ݤ ஒ RSp0213::GFP ߄ ౜ ܭ N. benthamiana ယ ೽ ࡕ Ψ ว ౜ RSp0213::GFP ϐ০ပՏ࿼ᆶ PIP2A::mcherry ϩթ΋ठ (კѤ)ǴᡉҢ RSp0213 ᔈ

ࢂ০ပӧ෌ނಒझጢ΢Ƕ

4. RSp0213 ၸໆ߄౜ჹ෌ނϐቹៜ

ࣁΑᕕှ RSp0213 ჹ෌ނϐёૈբҔǴࡺอኩ߄౜ஒჹྣಔᆶ 35S::RSp0213 ᏤΕ๨૛ᆶพनǴ٠ᢀჸ෌ނϐϸᔈǶӧ N. benthamiana ᆶ N. tabacum W38 ယТ

΢ճҔၭఎ๵ᇶշݙ৔ݤၸໆ߄౜ RSp0213Ǵ่݀ᡉҢǴӧ N. benthamiana ယТ

΢߄౜ RSp0213 ࡕ 48 λਔϣջ཮ЇଆܴᡉޑಒझԝΫ (programmed cell death, PCD) (კϖ A)ǴԶޜၩᡏჹྣಔೀ౛߾҂೷ԋቹៜǹӧ N. tabacum W38 ယ೽߄

౜ RSp0213 ёᢀჸډಒझԝΫ౜ຝ (კϖ BǴѰΒკ)Ǵՠӧ߄౜ RSp0213 ޑచҹ Πӕਔೀ౛ߙ࢏ੰ๵ँᡂਲ਼ Pss4 hrpG^-߾཮ౢғቃਗ਼ޑ PCD ϸᔈ (კϖ BǴѓΒ კ)Ƕ

аੰࢥᇨᏤ୷Ӣ߄౜ (Virus-mediated gene overexpression, VMGO) ฼ౣǴ٬Ҕ PVX (Potato virus X) ୷ Ӣ ߄ ౜ ၩ ᡏ Ǵ ӧ ל ੰ พ न ࠔ س H7996 س ಍ ܄ ߄ ౜ 35S::RSp0213 ࡕ (კϤ)ǴΨӕኬ཮ӧพनယ೽ЇଆܴᡉಒझԝΫ (კΎ A)Ǵ٠ᢀ

ჸ ډ ೷ ԋ พ न ෌ ਲ਼ ғߏ ڙ ׭ Զ ᡂ ࿖ λ (კΎ B)ǴᡉҢพनёૈڀԖёᒣᇡ RSp0213 ϐᐒڋǶ

5. ӧӧพन߄౜ RSp0213 ჹੰ্ϸᔈϐቹៜ

ࣁΑᕕှ߄౜ RSp0213 ࢂց཮ቹៜพनϐߙ࢏ੰϸᔈǴճҔ PVX ୷Ӣ߄౜

ၩᡏǴӧלੰࠔسพन H7996 س಍܄߄౜ 35S::RSp0213 ࡕǴаβᝆ዆ឲБԄௗᅿ ଯࢥΚߙ࢏ੰ๵ਲ਼ Pss190Ǵ٠ᢀჸੰำว৖Ƕ่݀ᡉҢس಍܄߄౜ RSp0213 ཮ཱུ

ᡉ๱Ӧफ़ե Pss190 ೷ԋϐพन๺঒௃׎ (p<0.01) (კΖ)Ƕ

6. ӧพन߄౜ RSp0213 ჹٛᑇϸᔈ࣬ᜢ኱ᇞ୷Ӣ (marker genes) ߄౜ϐቹៜ җ΢ॊჴᡍёޕǴ߄౜ RSp0213 ཮ቹៜพनϐߙ࢏ੰϸᔈǴࣁΑ຾΋؁ᙶమ

ੰ্ٛᑇૻ৲໺ᏤӵՖڙቹៜǴࡺᔠෳس಍܄ၸໆ߄౜ RSp0213 ϐלੰพनࠔس H7996 ύس಍܄ٛᑇ಻ᅟᆾᆶಒझԝΫ࣬ᜢࡰ኱܄኱ᇞ୷Ӣϐ߄౜ (ߕ߄Ύ)Ƕ่

݀ᡉҢǴᆶ߄౜ gfp ϐ௓ڋಔ࣬КǴ߄౜ RSp0213 ཮ε൯ࡋӦᇨᏤ SAǵET Ϸಒ झ ԝ Ϋ (PCD) ฻ૻ৲ ໺ Ꮴ ϐ ኱ᇞ ୷ Ӣޑ߄ ౜ ( კ ΐ Aǵ C ǵD) Ǵ Զ फ ಹ ለ (jasmonic acid, JA) ӝԋϐ࣬ᜢ኱ᇞ୷Ӣޑ߄౜߾ڙډ׭ڋ (კΐ B)ǶᡉҢӧพन ߄౜ RSp0213 ཮ᇨᏤ෌ނޑٛᑇϸᔈ୷Ӣϐ߄౜Ƕ

7. ،ۓ RSp0213 ೈқӧ෌ނಒझύϐ০ပՏ࿼ޑख़ा୔ࢤϩ݋

ࣁΑᔠᡍ RSp0213 ޑ N ᆄ 47 ঁữ୷ለᄬԋϐૻ৲ᴏ两 (SP) ჹܭځ০ပܭ෌

ނಒझጢϐѸा܄Ǵࡺ຾ՉТࢤմନ (კΜ A)Ǵ٠ᢀჸ RSp0213::GFP ӧ N.

benthamiana protoplast ᆶယ೽ϐϩթǶ่݀ᡉҢӄߏ RSp0213::GFP ϐϩթᆶಒझ ጢ኱૶ೈқ PIP2A::mcherry ΋ठǴԶմନ N ᆄૻ৲ᴏ两 21 ܈ 47 ঁữ୷ለϐ RSp0213::GFPǴځϩթ߾ᆶ free GFP ΋ठ (კΜ BǵკΜ΋)Ƕࡺ RSp0213 ޑ N ᆄ 47 ঁữ୷ለᄬԋϐૻ৲ᴏ两ӧनࣽ෌ނಒझϐಒझጢ০ပՏ࿼ዴჴ࣬྽ख़ाǴЪ Тࢤմନ N ᆄനڀߥӺ܄ޑ 21 ঁữ୷ለϐ RSp0213 ջѨѐځ০ပӧ෌ނಒझϐ ಒझጢՏ࿼Ƕ

8. RSp0213 ০ပܭ෌ނಒझጢჹځфૈϐख़ा܄

ࣁΑឍܴ RSp0213 Տܭ෌ނಒझጢჹځфૈࢂցڀख़ा܄Ǵࡺа VMGO ஒ ӄߏᆶТࢤմନϐ 35S::RSp0213 ӧלੰพनࠔس H7996 س಍܄Ӧ߄౜Ǵ٠ᔠෳพ नϐғߏᆶߙ࢏ੰϸᔈࢂցׯᡂǶ่݀ᡉҢӧ߄౜ӄߏϐ RSp0213 ӧพनယ೽೷ PopP3ǴЪឦܭӧੰচ๵ቶݱӸӧޑ YopJ superfamily ԋ঩ (Lewis et al., 2011)Ƕߙ

࢏ੰ๵ GMI1000 ڀԖΟঁឦܭ YopJ superfamily ޑԋ঩Ǵϩձࣁ PopP1ǵPopP2 ᆶ PopP3Ǵځύ PopP1 ᆶ PopP2 ڀ Avr protein ੝܄ (Lavie et al., 2002; Deslandes et al., 2003)Ǵՠ PopP3 ߾фૈ҂ޕǶҁࣴزӃ߃؁ϩ݋ YopJ superfamily ԋ঩ϐӸ ӧᆶցࢂցёૈᆶߙ࢏ੰ๵ठੰΚ߄౜ԖᜢᖄǴЪᕕှ၀ৎ௼ԋ঩໔ϐׇӈ࣬՟

܄ᆶёૈϐфૈߥӺ܄Ƕ

२ӃǴаҞ߻ςֹӄှׇޑ 22 ਲ਼ߙ࢏ੰ๵๵ਲ਼ࣁჹຝǴϩ݋ PopP ৎ௼ԋ঩

ϐӸӧ௃׎ (߄Β)Ƕ่݀ว౜ӧ Pss190ǵPss1308ǵGMI1000ǵPss216ǵFJAT_91ǵ FJAT_1458 ǵ Po82 Ϸ SD54 ๵ ਲ਼ ڀ Ԗ PopP1 ǹ Pss1308 ǵ GMI1000 ǵ Pss216 ǵ FQY_40ǵFJAT_91ǵFJAT_1458ǵPo82ǵK60ǵP673ǵCMR15 Ϸ SD54 ๵ਲ਼ڀԖ

PopP2ǹPss190ǵGMI1000 Ϸ Y45 ๵ਲ਼ڀԖ PopP3Ǵՠ GMI1000 ޑ popP3 Ԗ΋ၢ

៌ηකΕځύǶҗܭନ PopP3 ϐѦޑځд PopPs ৎ௼ԋ঩ϩණܭӚߙ࢏ੰ๵๵ਲ਼ ύǴคݤᘜયѬॺޑӸӧᆶߙ࢏ੰ๵๵ਲ਼੝܄ϐᜢᖄ܄Ƕ

аߙ࢏ੰ๵ GMI1000 ๵ਲ਼ޑ PopP1ǵPopP2 ᆶ Pss190 ๵ਲ਼ޑ PopP3 ຾Չữ

୷ለׇӈٿٿКჹǴ่݀ᡉҢӄߏׇӈ࣬ӕࡋ (identity)ǺPopP1 ᆶ PopP2 ࣁ 11.6%ǴPopP1 ᆶ PopP3 ࣁ 15%ǴPopP2 ᆶ PopP3 ࣁ 11.7%ǹԶሇન໽ϯ୔ޑׇӈ

࣬ӕࡋ߾ࢂǺPopP1 ᆶ PopP2 ࣁ 30%ǴPopP1 ᆶ PopP3 ࣁ 33%ǴPopP2 ᆶ PopP3 ࣁ 26% (ߕკΎ)ǶΟޣϐ໔คፕӄߏ܈ሇન໽ϯ୔ޑữ୷ለׇӈ೿ৡ౦ࣗεǴ௢

ෳ YopJ superfamily ޑԋ঩ӧߙ࢏ੰ๵ύΨ೚ӚԖځфૈǶ

຾΋؁Ǵӧ NCBI Genome ၗ਑৤ڀԖ PopP3 ޑ܌Ԗߙ࢏ੰ๵๵ਲ਼ (х֖୷Ӣ ಔ҂ૈֹӄှׇޑߙ࢏ੰ๵๵ਲ਼) ύǴ12 ਲ਼ߙ࢏ੰ๵๵ਲ਼ (х֖ Pss190) ڀԖ PopP3ǴԶჹ PopP3 ೈқ፦ϐׇӈ຾ՉׇӈКჹ߾ว౜ PopP3 ׇӈӧӚ๵ਲ਼ύڀ ԖଯࡋߥӺ܄ (ߕკΖ)Ƕ

2. ӧӧଯࢥΚ๵ਲ਼ PopP3 ϐϩթ

ࣁΑᕕှଯࢥΚ Pss190 ๵ਲ਼܌੝Ԗޑ PopP3 ӧځдଯࢥΚ๵ਲ਼ύޑϩթ௃׎Ǵ

߻Γࣴزᒧ᏷ 3 ਲ਼ phylotype I ޑଯࢥΚ๵ਲ਼ Pss158ǵPss365ǵPss749ǴаϷ 14 ਲ਼ phylotype II ޑଯࢥΚ๵ਲ਼ Pss525ǵPss526ǵPss1327ǵPss1351ǵPss1361ǵPss1370ǵ Pss1475ǵPss1482ǵPss1569ǵPss1586ǵPss1696ǵPss1697ǵPss1703ǵPss1710Ǵ

຾ՉࢉՅᡏ PCR ϩ݋Ƕ่݀ว౜ phylotype I ύନচҁςޕڀԖ PopP3 ϐ Pss190 ᆶ GMI1000 (GMI1000 ࣁύࢥΚ๵ਲ਼ǴӧԜբࣁჴᡍ҅௓ڋಔ) ѦǴۘԖ Pss365 ڀԖ PopP3 (ߕკΐ)ǴԶ phylotype II ߾҂ว౜ԖҺՖ๵ਲ਼ڀԖ PopP3ǶᡉҢѝԖ ϿኧଯࢥΚ๵ਲ਼ڀԖ PopP3ǴځόӸӧܭ܌ԖଯࢥΚ๵ਲ਼྽ύǶ

3. PopP2 ჹߙ࢏ੰ๵ӧพन΢ठੰΚϐቹៜ

ߙ࢏ੰ๵ଯࢥΚ๵ਲ਼ Pss190 ٠όڀԖӧߓ܎դ޺΢ࣁ Avr protein ੝܄ϐ PopP2 (Deslandes et al., 2003)Ǵՠ PopP2 ჹܭߙ࢏ੰ๵ӧพन΢ϐठੰΚ߄౜ޑቹ ៜۘ҂ޕǶࡺஒ popP2 ୷Ӣᒧ෗ډ pUFR047 ၩᡏ٠ᙯΕډ Pss190 ߄౜Ǵ࿶ෳ၂ ᙯࠠ๵ਲ਼ӧלੰพनࠔس H7996 ΢ϐठੰΚ߄౜ࡕว౜Ǵεໆ߄౜ popP2 ཮फ़ե ଯࢥΚ๵ਲ਼ Pss190 ӧלੰࠔسพनϐठੰΚ߄౜ (კΜϖ)Ƕ

ࣁΑ௖૸ PopP3 ຾Ε෌ނಒझࡕǴځфૈ܄୔ࢤ (functional domain) ჹ PopP3 ӧ෌ނಒझύϐ০ပՏ࿼ޑख़ा܄ǴךॺӃа Softberry ᆛઠϩ݋ PopP3 ӧ෌ނಒ झύёૈޑ০ပՏ࿼Ǵ่݀ว౜ PopP3 നԖёૈՏܭಈጕᡏ (mitochondria) ΢ǹ

຾΋؁а iPSORT ᆛઠ (http://ipsort.hgc.jp/) ϩ݋ PopP3 ࢂց஥Ԗૻ৲ᴏ两 (signal

peptide)Ǵว౜ PopP3 ޑ N ᆄ 30 ঁữ୷ለᄬԋΑಈጕᡏᏤӛᴏ两 (mitochondria targeting peptide, MT) Ϸૻ৲ᴏ两 (SP)ǴӕਔႣෳ PopP3 ຾ΕಈጕᡏࡕϪନᴏ两 Տ࿼ (mitochondria cleavage site) ࣁ N ᆄ߻ 10 ঁữ୷ለ (კΜΐ A)Ƕ

ԶࣁΑᡍ᛾ PopP3 ޑ N ᆄ 30 ঁữ୷ለᄬԋϐಈጕᡏᏤӛᴏ两ჹܭځ০ပܭ

෌ނಈጕᡏϐѸा܄ǴךॺஒಈጕᡏᏤӛᴏ两Тࢤմନ (sequence-truncated) ᆶӄ ߏϐ PopP3::GFP ϩձճҔ߄౜ၩᡏ p2GWF7.0ǴᙯΕ N. benthamiana চғ፦ᡏ຾

Չอኩ߄౜Ǵ่݀ᡉҢ୤Ԗ߄౜ӄߏϐ PopP3::GFP ӧ๨૛চғ፦ᡏύև౜Տ࿼

ᆶಈጕᡏ኱૶ (mitochondria marker) ScCOX4::RFP ϩթ΋ठǴԶ߄౜մନಈጕᡏ Ꮴӛᴏ两ϐ PopP3::GFP ӧ๨૛চғ፦ᡏύև౜Տ࿼߾ᆶ߄౜ free GFP ޑޜၩᡏ

௓ڋಔ࣬ӕ (კΜΐ B)Ƕ

7. ТТࢤմନࡕϐ PopP3 ჹߙ࢏ੰ๵ठੰΚϷჹพनੰ্ϸᔈϐቹៜ

຾΋؁ǴࣁΑᡍ᛾ PopP3 ϐಈጕᡏ০ပՏ࿼ჹځӧพनϐфૈࢂցڀख़ा܄Ǵ а VMGO ஒӄߏᆶТࢤմନ MT ϐ 35S::popP3 ӧགੰพनࠔس L390 س಍܄Ӧ߄

౜Ǵ٠ᔠෳพनϐߙ࢏ੰϸᔈࢂցׯᡂǶ่݀ᡉҢ߄౜ӄߏϐ PopP3 ቚமพनӧ Pss4 ೀ౛Πޑ๺঒௃׎ (კΒΜ)ǴԶ߄౜ಈጕᡏᏤӛᴏ两մନࡕϐ PopP3 ߾Ѩѐ ೭ᅿቹៜǴᆶ߄౜ GFP ϐ௓ڋಔคᡉ๱ৡ౦ (p>0.05)Ƕ

ќѦǴךॺΨஒӄߏᆶТࢤմନ MT ϐ popP3 ୷Ӣᒧ෗ډ pUFR047 ၩᡏǴ٠ ϩձᙯ౽߄౜ԿচҁόڀԖ၀ T3E ޑύࢥΚ Pss4 ᒪ໺ङඳ๵ਲ਼ύǴෳ၂ӧགੰ

ࠔسพन L390 ΢ϐठੰΚ߄౜Ǵ٠аޜၩᡏ (empty vector) ᙯࠠ๵ਲ਼଺ࣁ௓ڋಔǶ

่݀ҭว౜Ǵ୤Ԗӧ Pss4 ߄౜ӄߏϐ PopP3 ཮ቚம၀๵ਲ਼ϐठੰΚ߄౜ǴԶ߄౜

ಈጕᡏᏤӛᴏ两 (MT) մନࡕϐ PopP3 ߾Ѩѐ೭ᅿቹៜ (კΒΜ΋)ǶᡉҢ PopP3 N ᆄϐಈጕᡏૻ৲ᴏ两ᆶ෌ނಈጕᡏϐ০ပ੝܄ǴჹԜ T3E ӧ෌ނύϐфૈཱུࣁ ख़ाǶ

ಃ

ಃѤക ૸ፕ

ੰচ๵གࢉ෌ނਔଌΕϐਏᔈೈқǴ٩Ᏽ஌Ь෌ނޑᒣ᛽ૈΚёᏤठόӕޑ

่݀ (Alfano and Collmer, 2004)Ǻऩ T3E ೏෌ނޑ R protein ᒣᇡǴ෌ނ཮௴୏ז ೲԶᐟਗ਼ޑխࣝϸᔈǴ೭ᜪޑਏᔈೈқߡёᆀࣁ avirulence (Avr) proteinǹ࣬ϸӦǴ ऩਏᔈೈқ҂೏஌Ь෌ނᒣᇡǴ߾ёૈԋф׭ڋ஌Ьٛᑇϸᔈ܈υᘋஎЬғ౛ф

ૈǴаճੰচ๵ԋфགࢉ෌ނԶ೷ԋੰ্Ǵ೭ᜪޑਏᔈೈқߡёᆀࣁ virulence protein (Deslandes and Genin, 2014; Kazan and Lyons, 2014)Ƕ

΋ǵߙ࢏ੰ๵ RSp0213 ϐфૈࣴز

1. RSp0213 फ़եߙ࢏ੰ๵ჹพनϐठੰΚ

ߙ࢏ੰ๵གࢉ஌Ь෌ނၸำፄᚇǴӧᕉნڈᐟᆶဂᡏགᔈ (quorum sensing) ޑૻ৲໺ሀၡ৩ΠǴठੰӢηڙډፄᚇЪᆒஏޑፓ௓ (Valls et al., 2006; Vailleau et al., 2007; Genin and Denny, 2012)Ǵ٠߄౜ಃΟࠠೈқϩݜس಍ (T3SS) ϩݜਏᔈೈ

қǴଌΕ෌ނಒझׯᡂஎЬੰ্ϸᔈ (Poueymiro et al., 2009)ǴՠҞ߻ᜢܭߙ࢏ੰ

๵ޑࢥΚࣴزߚதϿǴ࣬ᜢၗૻલЮǶߙ࢏ੰ๵ RSp0213 ϐ߄౜ڙ T3SS ܌ፓ௓

(Cunnac et al., 2004a) (ߕკϖ)Ǵࡺ௢ෳ RSp0213 ᔈࢂಃΟࠠϩݜೈқ (T3E)Ǵՠ

҂ٰۘሡ຾΋؁ዴᇡځ೏ϩݜϐ੝܄Ƕҁࣴز຾΋؁ว౜ RSp0213 ޑӸӧё٬ଯ ࢥΚϐߙ࢏ੰ๵ӧพन΢ޑठੰΚΠफ़ (კ΋)ǴԶஒ RSp0213 ԾύࢥΚ๵ਲ਼ Pss4 ύকନ߾ёගଯੰ๵ӧพनϐठੰΚ (კΒ)Ǵ᛾ჴ RSp0213 ࣁቹៜߙ࢏ੰ๵ठੰ

ΚϐᜢᗖӢηǶ߻ΓࣴزΨว౜Ǵӧচҁόڀ PopP1 ޑߙ࢏ੰ๵๵ਲ਼ Rd15 ύ߄

౜ PopP1 ཮Ꮴठ၀๵ਲ਼ӧགੰ஌Ь࿖౐Ф΢ठੰΚΠफ़ǴЪᇡࣁ PopP1 ӧߙ࢏ੰ

๵ύࣁ avirulence protein ޑفՅ (Lavie et al., 2002)Ƕࡺךॺ௢ෳǺύ/եࢥΚ๵ਲ਼

੝Ԗϐ RSp0213 ᙖҗ T3SS ೏ଌΕ෌ނಒझࡕǴёૈ཮೏෌ނٛᑇس಍ᒣᇡԶᇨ Ꮴ෌ނౢғלੰϸᔈǴ٬ੰচ๵ޑठੰૈΚڙډ׭ڋԶև౜ठੰΚၨեޑ߄౜ࠠǴ Զ phylotype I ߙ࢏ੰ๵ଯࢥΚ๵ਲ਼ӢόڀԖ RSp0213Ǵளаխܭ೏෌ނᒣᇡ຾Զ ԖଯठੰΚǶќѦǴҗܭ߈ԃϩᚆளϐ phylotype I ߙ࢏ੰ๵ଯࢥΚ๵ਲ਼٠όڀԖ RSp0213Ǵࡺךॺ௢ෳӧߙ࢏ੰ๵ޑ๵ᅿᄽϯၸำύǴRSp0213 ᔈឦܭӅ೯Ъၨ

ࣁԐයޑ effectorǴԶ஌Ь෌ނΨς࿶ᄽϯр࣬ᜢޑٛᑇᐒڋୀෳϐǴ٠ளаԋф

ჹלϐ (ፎـаΠ૸ፕ)ǶԜѦǴ߈යࣴزࡰрߙ࢏ੰ๵ϐ T3SS ӧੰ๵གࢉ߃ය ᆶࡕය೿཮࡭ុ߄౜ǴᡉҢ T3SS ӧԜ๵ϐس಍܄ठੰၸำύᔈࣣԖཱུᜢᗖфૈ

(Poueymiro and Genin, 2009; Monteiro et al., 2012; Stotz et al., 2014)ǹ RSp0213 ࣁߙ

࢏ੰ๵ठੰΚ࣬ᜢӢηǴՠӧߙ࢏ੰ๵ठੰΚ΢ڀԖᜢᗖفՅޑ RSp0213 زഖࢂ

benthamiana ᆶ N. tabacum W38 ယТݙΕ஥Ԗ RSp0213 ߄౜ၩᡏϐၭఎ๵ 72 λਔ ࡕࣣ཮ӧݙΕಔᙃЇଆֽ೽ޑಒझԝΫϸᔈ (კϖ)ǴЪӧ N. tabacum W38 ယ೽ೀ พनٛᑇس಍ᒣᇡԶЇว෌ނלੰϸᔈ (Deslandes and Genin, 2014; Kazan and Lyons, 2014)Ǵࡺӧ N. benthamianaǵN. tabacum Ϸพनύᔈ၀೿Ԗёаᒣᇡ RSp0213 ϐӅӕჹᔈלੰೈқ፦ǶԜѦǴ߈ԃࣴزᡉҢพनߙ࢏ੰϐלੰᐒڋᆶ ࢲϯ SA ᆶ ET ૻ৲໺ᏤϷ፾྽ፓ௓ࢲϯ਼ϩη (reactive oxygen speciesǴᙁᆀ ROS) ಕᑈ֖ໆ৲৲࣬ᜢ (Mersmann et al., 2010; Coll et al., 2011)ǴΨᆶҁࣴز܌ள

ϐ่݀΋ठǶ҂ٰᔈ຾΋؁ཛྷ൨ᒣᇡ RSp0213 ϐբҔೈқǴаճ຾΋؁ᕕှनࣽ

෌ނჹלߙ࢏ੰ๵ϐϩηᐒڋǶ

3. RSp0213 ০০ပܭ෌ނಒझጢჹځфૈԿᜢख़ा

ੰচ๵ϐਏᔈೈқ຾Ε஌Ьಒझࡕ཮౽Տډ੝ۓϐझᏔа୺ՉځբҔǴ٠٩ ځфૈٰቹៜΠෞϐૻ৲໺ሀၡ৩ (Poueymiro and Genin, 2009; Deslandes and Rivas, 2012)Ƕҁࣴزว౜ RSp0213 ޑ N ᆄڀԖ targeting peptide ᆶ signal peptide (SP)Ǵ࿶ᆶ NCBI Genome ၗ਑৤ύ຾Չ blastp ׇӈКჹǴว౜хࡴ PSI07 Ϸ R229

฻ΐਲ਼ߙ࢏ੰ๵ਲ਼ޑ putative T3E N ᆄࣣڀԖԜ SP Тࢤ (E value < 10^-5)ǴЪႣෳ

ӧԜࢤૻ৲ᴏ两ύёૈڀԖၠጢТࢤ (transmembrane segments) (ߕკϤ)Ƕჴᡍ่

݀ᡉҢ RSp0213 ০ပܭ෌ނಒझጢ΢ǴЪև౜ᗺރϩѲ (კΟǵѤ)ǴԶځ N ᆄന ڀߥӺ܄ޑ 21 ঁữ୷ለࣁԜ০ပ੝܄ϐѸሡТࢤ (კΜǵკΜ΋)ǴЪჹځӧพन

΢ᇨᏤלੰϸᔈϐ੝܄Ԗ،ۓ܄ޑቹៜ (კΜΒǵΜΟǵΜѤ)Ǵ׳ᡉҢԜ SP Т ࢤёૈჹځѬ T3Es ০ပ੝܄ࣣڀख़ा܄ǴӢԜ҂ٰёаԜ SP Тࢤௗӝ GFP ೈ қᢀჸ০ပՏ࿼Ǵᡍ᛾ځዴϪфૈǶᆕӝ΢ॊჴᡍ่݀Ǵךॺ௢ෳ RSp0213 ᔈࢂ

ሡा০ပӧ෌ނಒझጢ΢٠׎ԋೈқ፦໣ဂ (protein cluster) Бૈ৖౜ځਏᔈǴԶ ӧᄽϯޑၸำύǴ෌ނҭςว৖р੝ۓϐᒣᇡೈқᆶಒझጢ࣬ᜢٛᑇس಍Ǵᒣ᛽

RSp0213 ٠ᇨวٛᑇϸᔈǶ

೚ӭࣴزࣣࡰр෌ނಒझጢᆶځ΢ protein complexes ჹܭੰ্ٛᑇϸᔈתᄽ ᜢᗖفՅ (Poueymiro and Genin, 2009; Bohm et al., 2014)ǴٯӵǺᒣᇡੰ๵ PAMPs ϐ PRRs ᆶ೚ӭᒣᇡੰ๵ਏᔈೈқϐ R proteins ࣣՏܭ෌ނಒझጢ (Bohm et al., 2014)ǹќѦǴಒझጢ΢ޑᕗિжᖴፓ௓ҭ཮ჹಒझٛᑇϸᔈౢғख़ाቹៜǴӵፓ

௓ಒझᕗિޑሇન phospholipaseǴջ೏᛾ჴჹܭ෌ނಒझޑ PCD ϸᔈϷٛᑇૻ৲

໺Ꮴתᄽख़ाفՅ (Kim et al., 2014)ǴΞӵಒझጢ΢ޑ੝ۓ่ᄬિใ (lipid raft)Ǵ Ӣځୖᆶፓ௓ಒझϣޑિ፦жᖴ (Lingwood and Simons, 2010)ǴЪς᛾ჴᆶᇨᏤ PCD ౢғϐ ROS ಕᑈஏϪ࣬ᜢ (Mersmann et al., 2010)Ǵࡺӧ෌ނٛᑇϸᔈޑፓ

௓΢ڀԖᜢᗖفՅ (Fallahi-Sichani and Linderman, 2009)ǶԜѦǴᜢܭਏᔈೈқբ Ҕܭ෌ނಒझጢ΢ςԖ೚ӭਢٯǴٯӵ Pseudomonas syringae pv. tomato DC3000 ޑ T3E AvrPto ᆶ AvrPtoB ջࣁբҔӧ෌ނಒझጢ΢ޑਏᔈೈқǹAvrPto ཮׭ڋߓ

܎դ޺ FLS2 ޑᕗለϯфૈ (Xiang et al., 2008)ǴߔЗΠෞޑ PTI ૻဦ໺ሀǹ

AvrPtoB ߾ࢂ཮ᆶ FLS2 Ϸ FLS2 ϐӅௗڙᡏ (co-receptor) BAK1 (BRI1-associated kinase 1) ҬϕբҔǴ׭ڋჹಒ๵ᚎЛೈқޑགޕௗڙ (Gohre et al., 2008)ǹฅԶӧ พनύǴAvrPto ᆶ AvrPtoB ߾཮ᆶพनϐ Pto ᐟ䁙ҬϕբҔǴЪ Pto ёૈ཮֎Ї AvrPto ٬ځᇻᚆ PRR ೈқǴЪёૈ೸ၸᕗለϯ AvrPtoB Զ׭ڋځݱનϯࢲ܄

(Gohre et al., 2008)Ǵ຾ԶᇨᏤ෌ނޑ ETI ٛᑇϸᔈǶ҂ٰёа຾΋؁ഄڗᆶ RSp0213 ԖҬϕբҔϐ෌ނೈқဂǴаයёཛྷ൨ᒣᇡ RSp0213 ϐբҔೈқᆶځд

ૻ৲໺Ꮴ࣬ᜢೈқ፦Ǵ٠຾΋؁௖૸ځҬϕբҔᆶ࣬ᜢϩηᐒڋǶ Զ N. benthamianaǵN. tabacum Ϸพन฻नࣽ෌ނ߾ςᄽϯрёᒣᇡ RSp0213 ϐ

࣬ჹᔈբҔೈқǴܭགࢉ୔Їว PCD ϸᔈǴ٠ࢲϯٛᑇລᅟᆾ SA ᆶ ET ϐس಍

܄ૻ৲໺Ꮴၡ৩Ǵᇨวس಍܄խࣝϸᔈǴаफ़եੰচ๵ӧ෌ނᡏޑठੰૈΚǶ҂

ٰᔈ຾΋؁ཛྷ൨ᒣᇡ RSp0213 ϐբҔೈқᆶځд interacting proteinsǴаճ຾΋؁

ᕕှनࣽ෌ނჹלߙ࢏ੰ๵ϐϩηᐒڋǴڀᡏឍܴځዴϪϩηᐒڋǴБૈᙶమ RSp0213 ϐڀᡏբҔᆶ෌ނٛᑇϸᔈޑૻ৲໺Ꮴၡ৩ (Deslandes and Genin, 2014;

Kazan and Lyons, 2014)ǶќѦǴճҔ RSp0213 ᆶځբҔೈқϐᒣᇡس಍Ǵ҂ٰҭ ёஒҗቶݱੰ๵ᇨᏤ܄୷Ӣ௴୏η (Lin et al., 2014) ௓ڋ߄౜ϐ RSp0213 ᏤΕ஥

ԖԜբҔೈқϐ፾྽नࣽբނύǴว৖ڀቶݱל๵܄ϐੰ্ٛᑇ฼ౣǶ

Βǵߙ࢏ੰ๵ PopP3 ϐфૈࣴز

1. ଯ ࢥ Κ ๵ ਲ਼ Pss190 ੝ Ԗ ϐ T3E PopP3 ࣁ ߙ ࢏ ੰ ๵ ख़ ा ޑ ठ ੰ Κ Ӣ η (Virulence factor)

Нѳ୷Ӣᙯ౽ (horizontal gene transfer, HGT) ೏ຎࣁߙ࢏ੰ๵זೲᄽϯǵ፾ᔈ όӕғᄊᕉნϷளаӧӭᅿ஌Ьس಍ԋфठੰޑᜢᗖӢન (Peeter et al., 2013)Ƕ T3E ࢂߙ࢏ੰ๵ठੰޑᜢᗖӢηǴӚ๵ਲ਼໔ڀଯݔ౦ࡋǴԶ T3E ϐНѳ୷Ӣᙯ౽

೏ᇡࣁёૈࢂᏤठߙ࢏ੰ๵ठੰΚৡ౦ޑচӢǶӧߙ࢏ੰ๵ GMI1000 ๵ਲ਼ύ popP1 ൩೏ᇡࣁࢂ೸ၸ HGT ᕇளޑ୷Ӣ (Lavie et al., 2002)Ǵӧ GMI1000ǵPo82 ᆶ CMR15 Οঁ๵ਲ਼ϐ୷Ӣಔύ popP2 ೿ᆶচᏘ๵ᡏ (prophage) ϐТࢤ࣬ᎃ (Genin and Denny, 2012)ǴԶҁࣴز࠻ᔠຎςှׇߙ࢏ੰ๵๵ਲ਼Ψว౜ӧ Pss190ǵ GMI1000 Ϸ Y45 ๵ਲ਼ϐ୷Ӣಔύ popP3 ᎃ߈୷Ӣҭᆶ prophage Тࢤ࣬ᎃǴࡺך (Pss1308ǵPss749) ᛙۓǴࡺ௢ෳ PopP3 ёૈࢂߙ࢏ੰ๵߈ය࿶ HGT Զᕇளޑཥ ᑉਏᔈೈқǴΨӢԜ෌ނۘ҂ᄽϯрىаᒣ᛽ PopP3 ϐלੰ R ೈқ (ߕკΜΟǵ ΜѤ)ǴԶ٬ PopP3 ளаԋࣁߙ࢏ੰ๵ޑख़ा virulence factorǴගଯߙ࢏ੰ๵ϐठ

ੰΚ߄౜Ƕ

ߙ࢏ੰ๵ଯࢥΚ๵ਲ਼ Pss190 ٠คӧߓ܎դ޺ύ཮೏ R protein ᒣᇡޑ Avr protein PopP2 (Deslandes et al., 2003)ǴԶҁࣴزΨ᛾ჴ PopP2 ཮फ़եଯࢥΚ๵ਲ਼ Pss190 ӧלੰࠔسพनϐठੰΚ (კΜϖ)Ƕ೭٤ၗ਑ᡉҢǴӧߙ࢏ੰ๵ύǴPopP ৎ௼ԋ঩ჹܭ Pss190 ӧพन΢ϐଯठੰΚ߄౜ڀख़ाቹៜфૈǴPss190 ନΑڀԖ virulence protein PopP3 ϐѦǴΨӢࣁόڀԖӧพन΢ёૈڀ Avr protein ੝܄ϐ PopP2ǴԶ٬ Pss190 ёೕᗉพनלੰᐒڋޑᒣᇡǴ຾Զ৖౜ Pss190 ӧלੰࠔسพ न΢ϐଯठੰΚǶ ϐᘰ໾ဟᆫᗐಕᑈ (callose deposition)Ǵ٠Ъ٬พनჹ೬ᆭੰಒ๵ (Pcc) ׳ࣁགੰǴ

ՠόቹៜ flg22 ᇨᏤϐ ROS ಕᑈ (ߕკΜϤǵΜΎǵΜΖ)Ƕҗܭਏᔈೈқ׭ڋ HR ٠ߚ׭ڋ෌ނ PTI ϸᔈޑѸाచҹ (Kazan and Lyons, 2014)ǴӢԜ௢ෳ PopP3

׭ڋ PTI ϸᔈϐၡ৩ёૈբҔܭ HR ᇨᏤၡ৩ϐѦ (independently)Ǵ܈ёૈځբҔ ᐒڋՏܭ ROS ૻ৲໺ᏤϐΠෞа׭ڋᘰ໾ဟᆫᗐϐಕᑈ (Muthamilarasan and Prasad, 2013)Ǵࡺคੋܭፓ௓ ROS ಕᑈᆶ HR ϸᔈǶฅԶǴӢӃ߻ޑ ROS ಕᑈჴ

ࢂߙ࢏ੰ๵ޑख़ा virulence factorǶՠԖ፪ޑࢂǴӧלੰพनࠔس H7996 ߄౜

popP3 ࠅό཮ቹៜ H7996 ჹߙ࢏ੰ๵ Pss190 ϐੰ্ϸᔈ (კΜΖ)ǹҗܭ PopP3 ϐ

΋ǶӢԜಈጕᡏࣁੰচ๵ਏᔈೈқբҔޑख़ाՏ࿼ϐ΋ǴӃ߻ࣴز൩ว౜୏ނੰ

ӧ੿ਡғނಒझύǴڀԖಈጕᡏᏤӛᴏ两 (mitochondria targeting peptide, MT) ϐೈқ፦཮೏ಈጕᡏ߄य़ڙᡏ (receptor) ᒣ᛽٠ᆶځ่ӝǴೈқ፦ω཮೏ଌΕಈ ጕᡏϣǹۘ҂೏໺ᒡԿಈጕᡏϣޑೈқև౜҂ᄙ᠄ޑރᄊ (unfolded form)ǴѝԖ

྽຾ΕಈጕᡏࡕǴಈጕᡏૻ৲ᴏ两೏ϪନǴೈқω཮ӧՔᒿೈқ (chaperon) ޑᔅ շΠᄙ᠄ǵౢғфૈ (Hicks and Galán, 2013)ǶҁࣴزճҔғނၗૻϩ݋ว౜

PopP3 ޑ N ᆄ 30 ঁữ୷ለᄬԋΑ MT ᆶૻ৲ᴏ两 (SP) (კΜΐ A)ǴԶ຾΋؁ჴᡍ ΨዴᇡԜ MT Тࢤࣁ PopP3 ০ပܭ෌ނಈጕᡏϐѸाׇӈ (კΜΐ B)ǴЪჹܭ PopP3 ෧եพनჹߙ࢏ੰ๵ Pss4 ל܄ᆶቚம Pss4 ϐࢥΚڀᜢᗖфૈ (კΒΜǵΒ Μ΋)ǶҞ߻೏ว౜০ပܭ੿ਡғނಈጕᡏϐੰচ๵ਏᔈೈқཱུϿǴ୏ނੰচ๵໻

Ԗ E. coli ϐ MapǵEspF Ϸ SopA (Hicks and Galán, 2013)Ǵ෌ނੰচ๵߾໻Ԗ P.

syringe ϐ HopG1 (Hicks and Galán, 2013)ǴԶځύѝԖ E. coli Map ೏᛾ჴ཮຾Ε ಈጕᡏύǴЪ཮Ѻ໶ಈጕᡏϣѦጢޑᚆη೯೸ࡋ٠೷ԋಈጕᡏᘐ຋ґှ (Hicks and Galán, 2013)ǴԶ P. syringe ޑ HopG1 ऩ҂০ပܭ෌ނಈጕᡏߡคݤ׭ڋ෌ނ ғߏวػ (Block et al., 2010)Ƕךॺ௢ෳ PopP3 ёૈᙖҗυᘋ෌ނಈጕᡏϐၮբǴ PopP3 ϐ interacting proteinsǴаճ຾΋؁ᕕှ PopP3 υᘋพनלੰϸᔈϐڀᡏᐒ ڋǴឍܴځዴϪϩηᐒڋᆶ࣬ᜢૻ৲໺Ꮴၡ৩ (Gohre et al., 2008; Gimenez-Ibanes

et al., 2009)Ǵ٠යఈёૈᙖԜࣴᔕԖշܭբނჹלଯࢥΚߙ࢏ੰ๵๵ਲ਼Ӓ্ϐੰ

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在文檔中效應蛋白RSp0213與PopP3在青枯病菌致病力之功能研究 (頁 13-0)