,2017,37(6):1066-1072 犃犮狋犪犅狅狋. 犅狅狉犲犪犾. 犗犮犮犻犱犲狀狋. 犛犻狀. :1000 4025(2017)06 1066 07 犱狅犻 :10.7606/j.issn.1000 4025.2017.06.1066 "#$%& 6 ' ()*+ 1,2, 1, 1, 1, 1, 1 (1 <=,>$? 712100;2 @AB =,CD@A 663000), -:E +, 6 FGHIJKL( 犆犿犜犘犛 )M N1O,P QR N ST, UVDW 犜犘犛 MKL9+,KLXY O,)+,KLX Z[7\ 6 FGHIJK L,]^_`abc E 犆犿犜犘犛 1~7 3, 犆犿犜犘犛 4 犆犿犜犘犛 7E 1, 16\, _ 6 FGHIJ(TPS) ;_ 5\ 犆犿犜犘犛 KL E 2, 2~4\ ;` 7\+, TPS, CmTPS3 TPS,_ CmTPS TPS TPP1 UDP forming ; O 3,+, TPSM I (15.90% ~57.31%); Z,CmTPS1 CmTPS2 CmTPS6Z `,_ 4\ CmTPSZ ` 6 qrt PCR, +, 犆犿犜犘犛 4E ( TPS KL; 犆犿犜犘犛 KLMO E, ` ABA R E +, 犜犘犛 KLM Z K./0:+,; 6 FGHIJ;KLM; 1 :Q786;S652 2:A 犈狓狆狉犲狊犻狅狀犃狀犪犾狔狊犻狊犪狀犱犌犲狀狅犿犲 犠犻犱犲犐犱犲狀狋犻犳犻犮犪狋犻狅狀狅犳犜狉犲犺犪犾狅狊犲 6 犘犺狅狊狆犺犪狋犲犛狔狀狋犺犪狊犲犌犲狀犲犉犪犿犻犾狔犻狀犕犲犾狅狀 ( 犆狌犮狌犿犻狊犿犲犾狅犔.) YANG Yongchao 1,2,ZHANG Haifei 1,YANG Xiaozhen 1, WANGZhongyuan 1,WEIChunhua 1,ZHANG Xian 1 (1ColegeofHorticulture,NorthwestA &FUniversity,Yangling,Shaanxi712100,China;2WenshanAcademyofAgricultur alsciences,wenshan,yunnan663000,china) 犃犫狊狋狉犪犮狋 :Inordertoclarifytheinformationof 犆犿犜犘犛 genefamilyandthepaternsofresponsetoenviron mentcues,weidentifiedseven 犜犘犛 sin melongenomeinthiswork.these 犜犘犛狊 distributedonseven chromosomesanddesignated 犆犿犜犘犛 1-7.Phylogeneticanalysisindicatedthat 犆犿犜犘犛 geneswouldbe classifiedintoclassⅠandclassⅡ. 犆犿犜犘犛 4and 犆犿犜犘犛 7belongtoclassⅠwith16intronsandtheirpro teinsprobablypossesstpsactivity.theotherfive 犆犿犜犘犛 genes,containingtwotofourintrons,were classifiedintoclass Ⅱ.CmTPS3consistsofonly TPSdomain,whereastheremaining TPSscomprise TPS,TPPandUDP formingdomains.cmtpsproteinshavelowsimilarity (15.90% -57.31%)within familymembers.subcelularlocalizationpredictionindicatedthatcmtps1,cmtps2andcmtps6 were localizedtothenucleusandtheothersincytoplasm.theexpressionof 犆犿犜犘犛 genesundervariousabiotic stressesshowedthat 犆犿犜犘犛 4 mightbemaingeneencodingtpsinmelonleavesunderlowtemperature stress,whilemelon 犜犘犛 familygenesweremoresensitivetosaltstressandmightbeinvolvedinregula tionofabasignalpathway.ourresultslaythefoundationforfunctionalcharacterizationofmelon 犜犘犛 familygenesinthefuture. :2016 11 11; :2017 06 05 :!"#(CARS 26 18)!:$%&(1982-),',, ()*+,-./0 1 E mail:yangyongchaos@foxmail.com :2 3,,45, ()*+, 6789: ; E mail:zhangxian098@126.com
6 $%&,G:+, 6 FGHIJKL Z1 1067 犓犲狔狑狅狉犱狊 :melon;tps;genefamily;expressionanalysis Uα,α 1,1 I `, HI J : 6 FGHI J(TPS) ) UDP 6 FG I 6 FG(T6P), 6 FGFGJ (TPP) F G K I [1] O 6 R,` ( R [2 3],_ T6P 9 H R I 1!"HI,# $ %9 & (, TPS T6P H I U ' \ ( H I J, TPS [4] U 犗狊犜犘犛 1 KL ()O *.1 +0,-, -./01 3; [5] ` KL U VDW 犜犘犛 KL, 3,2 K L,. 3 4 5 [6] 6,78 9: 9 犜犘犛 KL ;, KL - [7 9] 犜犘犛 KLO - ( R1` ; 5 TPS TPS TPP,< TPS./ TPP [10 11] 犜犘犛 \M K L,= _ 犜犘犛 I Y > =?,@VD W K L X 11 \ 犜犘犛 K L [10], E 9 \ [11],$ A E 12 \ [4],< +, 犜犘犛 K L M BC D +, (,, E, E F 3GHI,J_`KL Mc E NO =, [12] N 1+,KLX PI [13],E+, 1 Q R ST U TR N S T,R V D W 犜犘犛 M K L ` +, KLXY V O, Z[7\+, 犜犘犛 KL,. 7 \ 犜犘犛 K L K L W M 1`= XY, E+,- / 1- Q0 K 1 9ST 1.1 3 4 +,Z[ NantE \ ],2 2016 ^ 3 ~7 _ 2 `, a XYE b28 /20,16h/8h( c/ ) ` () 7 ) K 6. Q R Hoag land a V ` +, ) V (4 ) (250mmol L -1 NaCl)1 ABA (100μmol L -1 )+ 0, ^ S T [14] + 0 0(CK) 1 3 6 12124h 3 8, ;.`-80 +03 1.2 5 6 1.2.1 "# 犜犘犛 78 ()9: ; V DW TPS (AtTPS1 11)E,`+,K L X Y (htps://melonomics.net/) V Blastp O(E=0.0001), TR Pfam (htp://pfam.xfam.org/)1 NCBI CDD(htp:// www.ncbi.nlm.nih.gov/cdd/)` W J R ExPASy(htp://web.expasy.org/ protparam/)o+, TPS V K G Y 0 G I V JRSubcelularlo calizationpredictor (htp://celo.life.nctu.edu. tw/)` V Z +, 犜犘犛 KLI `abc V,T R MapInspect V a b Z J R MEME (htp://meme suite.org/) V. 2\XY 8KL *Y [15] :(1) O &b & 75% c;(2) O 270% J R Clustal W V O, J R MEGA6.0 YQ R T A, ] Y BootstrapE1000 VDW TPS (AtTPS) ) TAIR(htps://www.arabidopsis.org/), TPS (OsTPS)) K L X Y (htp://rice.plantbiology.msu.edu/) 1.2.2 "# 犜犘犛 *+ ; RNA J R RNAPlantPlusReagent (c ), Q R FastQuantRT Kit (with gdaase),. ] J R Primer Premier6.0! \ Z, HI C 1 QR ABIStepOnePlus\ Z V+, TPSKL, JR '( ) SYBRPremix Ex 犜犪狆 Ⅱ E SYBRmix10μL,cDNA2μL,c 0.8μL,
1068 37 *1 ( 犘犆犚 <=> Table1 PrimersforqRT PCR KL Gene c Forwardprimer(5 3 ) Reverseprimer(5 3 ) 犆犿犜犘犛 1 GCACATTGCCGAGCCTGTTATG TTGCCTGGGATGATCCGAAACC 犆犿犜犘犛 2 GTGGTGCGGTCAGGGTAAATCC TGCGAGCCCAGTAAGCAACATC 犆犿犜犘犛 3 CTGTGGCTGGTGTCTTGTCTGAG CAGTCTCCATTCTCGTTCCGATTCG 犆犿犜犘犛 4 GCCAATAGGTTGCCCGTCTCTG ACACCAGCCCAACCAATCCATC 犆犿犜犘犛 5 CAAGTGACGCTGGCTGGTGATT AGAAATCCAAGACGCTGACGGTTC 犆犿犜犘犛 6 AAGACACCGAGCCCTGAAGTTATCT TCACAAGAGACGAACCACTGACCTA 犆犿犜犘犛 7 GGTTGCCTGTGTCTGCTGTTAGAA CCAATCCATCTTGCCTCAAACTCCT β 犃犮狋犻狀 TGCCCAGAAGTTCTATTCCAGC CATAGTTGAACCACCACTGAGGAC ddh 2O6.4μL, +, 犃犮狋犻狀 E KL X Y:95 3min,94 30s,60 30 s,72 10s, 40\, ]^ ' QR2 -ΔΔCt T VY 2 9 2.1 "# 犜犘犛 78?)@AB C U O1W,`+,KLXY Z[7\ 犜犘犛 KL,]^`abc E 犆犿犜犘犛 1~7,_ ` 1 2 6 7 8 10 12 abc( 1) CmTPS1~7 442~935\ KG,0 G I5.64~6.46, 49.94~105.54kD Z 3 \Z `,4\Z ` 6 ( 2) 2.2 "# 犜犘犛 78?DE ) O7\+, TPS V M O. A( 2), A3 +, 犜犘犛 KLM I E 2, 1 犆犿犜犘犛 4 犆犿犜犘犛 7, 16\ _ I E 2, 2~4 \ CmTPS3 TPS,_ 6\ CmTPS TPS TPP UDP forming,_ UDP forming TPS TPP ( 3) O +, TPSKLMI V O,_ 3 I (15.90%~57.31%), 8KL *Y Z,+, TPSKLM ` U' *Y E +, 犜犘犛 KLM 犜犘犛 K LM 8,JR+, VDW TPS M A( 3) _ 3 CmTPS3 9 AtTPS5,CmTPS1 9 AtTPS7,CmTPS4 9 AtTPS1 ` VDW,OsTPS1 1 AtTPS1 TPS, CmTPS4 CmTPS79 OsTPS1 AtTPS1` \ c, CmTPS4 CmTPS7 TPS 1 +, 犜犘犛 KL`abc Fig.1 Distributionof 犆犿犜犘犛狊 onchromosomes *2 "# 犜犘犛 F Table2 BasiccharacteristicsofCmTPSproteins KL Genename Accessionnumber KGY Amino acids(aa) 0 G I Theoretical isoelectricpoint Molecular weight/kd Z Predictedsubcelularlocalization 犆犿犜犘犛 1 MELO3C018715 843 5.64 95.80 Nuclear 犆犿犜犘犛 2 MELO3C017104 855 6.36 97.09 Nuclear 犆犿犜犘犛 3 MELO3C013837 442 6.29 49.94 6 Cytoplasmic 犆犿犜犘犛 4 MELO3C025673 926 6.33 104.81 6 Cytoplasmic 犆犿犜犘犛 5 MELO3C007197 850 5.50 96.23 6 Cytoplasmic 犆犿犜犘犛 6 MELO3C012010 860 6.21 97.20 Nuclear 犆犿犜犘犛 7 MELO3C002694 935 6.46 105.54 6 Cytoplasmic
6 $%&,G:+, 6 FGHIJKL Z1 1069 2 +, TPSM (A)1 KL (B) Fig.2 PhylogeneticanalysisofCmTPSproteins(A)andgenestructure(B)of 犆犿犜犘犛 s *3 "# 犜犘犛 GH I ; Table3 DomainanalysisofCmTPSproteins +, TPS CmTPSprotein CmTPS1 Domain Locationofdomain (aa) e e value Accessionofdomain TPS 60~530 0 cd03788 TPP 580~815 3.51E 79 pfam02358 UDP forming 1~832 0 PLN02205 TPS 56~539 0 cd03788 CmTPS2 TPP 589~823 5.46E 86 pfam02358 UDP forming 25~841 0 PLN02205 CmTPS3 TPS 63~432 3.50E 148 cd03788 TPS 46~442 2.08E 94 COG0380 TPS 95~560 0 cd03788 CmTPS4 TPP 595~840 2.97E 64 pfam02358 UDP forming 1~922 0 PLN03064 TPS 60~543 0 cd03788 CmTPS5 TPP 593~827 1.85E 85 pfam02358 UDP forming 1~846 0 PLN02205 TPS 61~544 0 cd03788 CmTPS6 TPP 594~829 1.90E 91 pfam02358 UDP forming 1~842 0 PLN02205 TPS 95~560 0 cd03788 CmTPS7 TPP 619~827 1.02E 61 pfam02358 UDP forming 1~932 0 PLN03064 2.3 "# 犜犘犛 78*+JK 4 3,` 250 mmol L -1 NaCl + 0,+, 犜犘犛 MKL 0 犆犿犜犘犛 1 犆犿犜犘犛 2 犆犿犜犘犛 6, 0~12h,, 犆犿犜犘犛 1 犆犿犜犘犛 2 犆犿犜犘犛 6O E,`+012h O^c 34.2 96.9 23.4 TPS J K L 犆犿犜犘犛 4 犆犿犜犘犛 7 _, `1~6hc, 5,` 4 + 0, 犆犿犜犘犛 1 犆犿犜犘犛 4 犆犿犜犘犛 5 0 c,. c c,.`24h _ O 犆犿犜犘犛 7`+01hc, ; 犆犿犜犘犛 6`0~6h, c CmTPS4 CmTPS7 TPS, ` 犆犿犜犘犛 7E ( TPS K L, 犆犿犜犘犛 4E ( TPS KL 6 3,` ABA + 0 1h, 犆犿犜犘犛 3,+, 犜犘犛 MKL = 'b c, _ 犆犿犜犘犛 1 犆犿犜犘犛 2 犆犿犜犘犛 5 犆犿犜犘犛 6 犆犿犜犘犛 7 `+01h, 犆犿犜犘犛 1 犆犿犜犘犛 2 c 2.3 3 O TPS 犆犿犜犘犛 4 犆犿犜犘犛 7, 2 \ K L = 犆犿犜犘犛 4`+03 6h,`+01 12
1070 37 24h c 犆犿犜犘犛 7`1 12h c,_ 3\ I 3 A +Y P b,y,, AtTPS1.VDW(At1g78580);AtTPS2.VDW(At1g16980); AtTPS3.VDW(At1g17000);AtTPS4.VDW(At4g27550); AtTPS5.VDW(At4g17770);AtTPS6.VDW(At1g68020); AtTPS7.VDW(At1g06410);AtTPS8.VDW(At1g70290); AtTPS9.VDW(At1g23870);AtTPS10.VDW(At1g60140); AtTPS11.VDW(At2g18700);OsTPS1. (HM050424); OsTPS2. (HM050425);OsTPS3. (HM050426); OsTPS4. (HM050427);OsTPS5. (HM050428); OsTPS6. (HM050434);OsTPS7. (HM050429); OsTPS8. (HM050430);OsTPS9. (HM050431); OsTPS10. (HM050432);OsTPS11. (HM050433) CmTPS1~CmTPS7,U Z[ 7\ TPS 3 +, VDW TPSKLM Thenumbersinthephylogenetictreeindicatetheconfidencevalues ofthebreach.attps1. 犃狉犪犫犻犱狅狆狊犻狊狋犺犪犾犻犪狀犪 (At1g78580); AtTPS2. 犃狉犪犫犻犱狅狆狊犻狊狋犺犪犾犻犪狀犪 (At1g16980);AtTPS3. 犃狉犪犫犻犱狅狆狊犻狊狋犺犪犾犻犪狀犪 (At1g17000);AtTPS4. 犃狉犪犫犻犱狅狆狊犻狊狋犺犪犾犻犪狀犪 (At4g27550); AtTPS5. 犃狉犪犫犻犱狅狆狊犻狊狋犺犪犾犻犪狀犪 (At4g17770);AtTPS6. 犃狉犪犫犻犱狅狆狊犻狊狋犺犪犾犻犪狀犪 (At1g68020);AtTPS7. 犃狉犪犫犻犱狅狆狊犻狊狋犺犪犾犻犪狀犪 (At1g06410); AtTPS8. 犃狉犪犫犻犱狅狆狊犻狊狋犺犪犾犻犪狀犪 (At1g70290);AtTPS9. 犃狉犪犫犻犱狅狆狊犻狊狋犺犪犾犻犪狀犪 (At1g23870);AtTPS10. 犃狉犪犫犻犱狅狆狊犻狊狋犺犪犾犻犪狀犪 (At1g60140); AtTPS11. 犃狉犪犫犻犱狅狆狊犻狊狋犺犪犾犻犪狀犪 (At2g18700).OsTPS1. 犗狉狔狕犪狊犪狋犻狏犪 (HM050424);OsTPS2. 犗狉狔狕犪狊犪狋犻狏犪 (HM050425);OsTPS3. 犗狉狔狕犪狊犪狋犻狏犪 (HM050426);OsTPS4. 犗狉狔狕犪狊犪狋犻狏犪 (HM050427);OsTPS5. 犗狉狔狕犪狊犪狋犻狏犪 (HM050428);OsTPS6. 犗狉狔狕犪狊犪狋犻狏犪 (HM050434); OsTPS7. 犗狉狔狕犪狊犪狋犻狏犪 (HM050429);OsTPS8. 犗狉狔狕犪狊犪狋犻狏犪 (HM050430);OsTPS9. 犗狉狔狕犪狊犪狋犻狏犪 (HM050431);OsTPS10. 犗狉狔狕犪狊犪狋犻狏犪 (HM050432);OsTPS11. 犗狉狔狕犪狊犪狋犻狏犪 (HM050433);CmTPS1 -CmTPS7,7CmTPSproteinswereindentifiedinthiswork. Fig.3 PhylogeneticanalysisoftheTPSgenesfrom melon, 犃狉犪犫犻犱狅狆狊犻狊 andrice TPS 1 T6PHI (KL,` % ( R, 犜犘犛 K L ` ; ( R ` VDW $A, 犜犘犛 KLM Z KL 1 [14,16] < `+,, 犜犘犛 K L M Z 1 B C D U TR NST,`+,KLXY Z[ 7 \ 犜犘犛 KL ` VDW 9\ 11 \ 犜犘犛 KLI, U, 3\ 犜犘犛 KL U S, `VDW 1\ 犜犘犛 KL 2KKLX, 犃狋犜犘犛 2 犃狋犜犘犛 3 K L U 犃狋犜犘犛 1 [16] ` +, K L X U'./ KKLX *Y [13], U, 犆犿犜犘犛 K L M / $ : *Y, 7\ 犆犿犜犘犛 7\ KL, ` `7\ 犜犘犛 KL [16] W,7 \ CmTPS TPS, CmTPS3,_ TPP 1 UDP forming ` U ' CmTPS3 TPP, ` TPS M [14] ` VDW,R [ S OsTPS1 AtTPS1 TPS [11,17] ) A!,CmTPS4 CmTPS7 E 1 TPS M,9 OsTPS1 AtTPS1` \ c, CmTPS4 CmTPS7 TPS 9 " # 犜犘犛 K L M 2 \ K L ( 犣犿犜犘犛 1, 犣犿犜犘犛 3) TPS [18] U,+, 犜犘犛 K L M O = +0 = ` +0, TPS KL 犆犿犜犘犛 4 犆犿犜犘犛 7`1 ~6h c, 01 T6P c `,T6P E ( $ % J 1(SnRK1), SnRK1 &H 9' % (%J [19] 2 01,( H R,< \ U' 01 ) H O * [20], T6P 1 +b ` U', % R O V,_ TPS
６l 678 q CDmno ６ pqrstuv IÎÐ １０７１ aýâeø1î ０ ０５ >FQGµHI ÔÝ ４ CD ＴＰＳ (wuv ÚÒÓÔ}ÎÐ Ｄ ｎ ｎｏ ｍ ｕｄ ｃ ｄ ｃ ｎ ｄ ｎｃ ０ ０５ ｖ ｈ ｍ ｂ ｏｗ ｇｎ Ｆ ４ Ｅｘｐ ｏｎｐ ｏ ｏ 犆犿犜犘犛 ｎ ｐｏｎ ｏ ｇ ５ CD 犜犘犛 uv ÉÑÒÓÔ}ÎÐ Ｆ ５ Ｅｘｐ ｏｎｐ ｏ ｏ 犆犿犜犘犛 ｕｎｄ ｏｗ ｍｐ ｕ ｇ ６ CD 犜犘犛 uv ＡＢＡ ÒÓÔ}ÎÐ Ｆ ６ Ｅｘｐ ｏｎｐ ｏ ｏ 犆犿犜犘犛 ｎ ｐｏｎ ｏａｂａ ｍｎ ｇ µ}犞狏犜犘犛１ Fa;ÉÑéH ２１ ñcd ÉÑ $K } a Ý < ＡＢＡ C H " 犆犿犜犘犛７ ３ ６ ÒÓ õ l 犆犿犜犘犛７ â Î Ð 犆犿犜犘犛４ " l Î [ ２４ｈ Þ ã ñ １ [ １２ｈ û ü â 犆犿犜犘犛４ Ð â ２４ｈ Ð ÿ ５ ５ î K Î Ð H I Ø J ÎÐK a k Q Î Ð 犃狋犜犘犛１ ý u v h
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