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Characterization of a CBL interacting Protein Kinase Gene TaCIPK16 in Wheat

小麦CBL结合蛋白激酶基因TaCIPK16的克隆及特征分析



全 文 :书!"#:10004025(2016)06107307               犱狅犻:10.7606/j.issn.10004025.2016.06.1073
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犆犺犪狉犪犮狋犲狉犻狕犪狋犻狅狀狅犳犪犆犅犔犻狀狋犲狉犪犮狋犻狀犵犘狉狅狋犲犻狀犓犻狀犪狊犲犌犲狀犲
犜犪犆犐犘犓16犻狀犠犺犲犪狋
MYOTHWIN,LIUPeng,MAWei,XUEQinghe,GUOJun,KANGZhensheng
(StateKeyLaboratoryofCropStressBiologyforAridAreas,ColegeofPlantProtection,NorthwestA&FUniversity,Yan
gling,Shaanxi712100,China)
犃犫狊狋狉犪犮狋:ThecalcineurinBlike(CBL)CBLinteractingproteinkinase(CIPK)networkplaysapivotalrole
inregulatingthephysiologicalaswelasdevelopmentalprocessesinplants.Inthisstudy,weobtaineda
CIPKgene,犜犪犆犐犘犓16,fromthewheatleavesinfectedby犘狌犮犮犻狀犻犪狊狋狉犻犻犳狅狉犿犻狊f.sp.狋狉犻狋犻犮犻(犘狊狋)using
犻狀狊犻犾犻犮狅cloningandRTPCR.ThefullengthcDNAof犜犪犆犐犘犓16was1606bp,whichencoding433a
minoacids.MultisequencealignmentshowedthatTaCIPK16sharehighsimilaritywithOsCIPK16and
AtCIPK16inriceand犃狉犪犫犻犱狅狆狊犻狊,respectively.Analysisoftheproteindomainfeaturesindicatedthat
TaCIPK16containedconservedregulatorydomaininCterminalandproteinkinasesdomaininNterminal.
SubcelularlocalizationassaysrevealedthatTaCIPK16displayedalocalizationpatternsimilartothatofthe
GFPcontrol,indicatingacytoplasmicandnuclearlocalization.Yeasttwohybridassaysshowedthat
TaCIPK16stronglyinteractswithTaCBL4andTaCBL9,respectively.qRTPCRassaysindicatedthat
犜犪犆犐犘犓16wasinducedby犘狊狋infectionanddifferentialyexpressedduringincompatibleandcompatible
interactionsbetweenwheatand犘狊狋.Ourresultssuggestthat犜犪犆犐犘犓16hasapositiveroleinregulating
wheatresistanceagainst犘狊狋.
犓犲狔狑狅狉犱狊:wheat;CBL;proteinkinase;TaCIPK16;犘狌犮犮犻狀犻犪狊狋狉犻犻犳狅狉犿犻狊f.sp.狋狉犻狋犻犮犻
  †æåç£èé+,êj]닍,#ìí ¬îï]N‡Mð 。Ca2+ñN‡òVkF]ó ôNõ , †#]SŠÌSö‚Œ÷økF]O


#†ùåúû]N‡Ìüýþ , ¦§] Ca2+¿ÿ!ŽSÇå]"é[1]。#Í†3#t
~€z{

ªñtu%CaM (Calmodulin)¥ ­Ç&z{ 、 tˆ‰±z{„x (Ca2+dependent proteinkinases,CDPK)、tuvwx B sz{ (calcineurinBlikeprotein,CBL)[2]。­CBL ()ńx*± , +,Ì-z{CIPK€ìíç .9¯ŽO [3]。CIPKñ}›#°Å]/
¸w0¸wz{„x(š,12ó3s]SnRK3
„x
(SNF1relatedproteinkinase3,SnRK3)[4]。
á34#ÁÂÃCIPKz{]€½ª«´µ,5 Å]CIPKz{ºÅ N6]„xz{¾ÌC6] u7¾ , 8P|CBL°Ø9]NAF€½¾[5]Ì ¯ƒPP2CÇÐO]PPI€½¾[6]。 :;KL´µCIPK(š.œàƒùå<<、 =>?P@güý , Aüý , B¥CK+、Dw<、C E=­Füý [7]。 †ÁÂÍ ,AtCIPK23ÞBƒ AtCBL1ÐO,vw„* K+MG (AKT1)Hë K+ ]I J [8]。 K Á  à L Ò < < ü ý þ ,SOS3 (CBL4)¥ CBL10 €  Ca2+, M 8 ƒ SOS2 (CIPK24)z{„x€ìíz{ç.,ª†
NO¥NÜ2ªu_Çå-z{]´Ô

G…õ#
¯PQ?RS<<üýTU]VW [9]。 †¿À  , M‚RNAibŸXY犗狊犆犐犘犓23¯ZV[¿À âg±]R[10]。 ]^_ú ,CBLCIPKN‡`a bcàƒ#SŠÌŽ1]‚Œ

¿À Os
CIPK15ƒ ¿ À ] d E e ± Å &,Þ ¯ u 7
SnRK1A f  d E ù å Ì g N ‡ ä å[11]。†
CIPK6®¯dh]ié.犮犻狆犽6,j.4ƒjk
4]Sö%lmnµÊR,
oí#p´q|”


Ostuc

vwŽSxy=Södz
[12]。
]^_ú

&2CIPKz{„x†#ùåS üý‚Œ]O{Ŭ}¨]KL

|5}
~

*±E
(reactiveoxygenspecies,ROS)†# â\äå÷Ó&kF]O , F¯Z@÷¦§ € , G…‚ƒ\„’]ë}…†‡ , ˆÞO|} ›N‡ª“Vâ\Ç&.œ]´Ô [13]。 †Á Í ,AtCIPK26¯ZÌ}›NADPHE‹xAtR bohFÐO‰u_­ŠSROS[14]。†¿À,Os CIPK14/15¯‹ÒÓ\„Ç&ª34(microbe associatedmolecularpatterns,MAMP)]“V,Œ OsCIPK14/15]´ÔÒӍƒ,MAMP“VŠS ]ROS{ÒӍƒ,G…Žµ OsCIPK14/15àƒ ¬PAMP“V]â±(PAMPtriggeredimmunity, PTI)[15]。†,CBL10ÌCIPK6¯Zßu_ ¦§IJäå̦§Œ²±‘ 。 †’“ , CIPK6ƒ CBL10]ÐOàƒ ETI(effectortrig geredimmunity)‚ŒROS]ŠS[16]。ÅKL´ µ , ¿qwN‡Mð{ÒÓ¬CIPK]u7。†Á ÂÍ ,NPR1¯Z”CIPK11/PKS5vw‹,G… “V WKY38Ì WKY62]´Ô[17]。 •: , XY&2CIPKz{„x†Süý
]®¯~_–—

KLXYCIPKz{„x†X
YZ[’ÐO.`]®¯á˜™­OšŠÄÅ
kF]›œ

œ^

KLª~˜™¬}›z{„
x.œ犜犪犆犐犘犓16,žqRTPCRbŸ、 ¡¢
£¤bŸ¥y¦§¨©bŸª«¬­®¯°±

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ë}…Ë­†XYZ[’ÐO‚Œ]ªš
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1.1 QRST@RURV
mn5XYZ[’SŠX#|

Z23
‡
’(CYR23)Ì‘Z31‡’(CYR31),XY¤#|

¥¦169’Ì‘¿§11’。XY¨©、Z[’ª«¥
9#¢£à¬Kang=[18]]¢£。TaqDNA ­
x
(Fermentas,®),dNTP、pMD18T(Takara,¯

),
°±g²³ DNA´Jµ¶·(¸ ¹–,),
Top10ºÒ»¦§(Wolsen,®),¼½¾¿·(Bi
oMiga,®),‚ƒ±ÀÁx(Fermentas,®),T4
DNAÂ9x(Fermentas),BIOZOL(BioFlux,¯

),RNaseinhibitor(Promega,®),MMLV ä
Ãĵ¶·
(Fermentas,®),SYBRGreen Ì
RoxreferenceÅ¡(Invitrogen,®)。
1.2 W X
1.2.1 犜犪犆犐犘犓16,<=>? †TAIRv10(ht
tp://www.arabidopsis.org/)Ì RGAPv7 (ht
tp://rice.plantbiology.msu.edu/)ÆÇȍ™Ó
ÁÂÃÌ¿À]CIPK²³,†É.œÊÆÇ
4701 ! " #  % &                   36Ë È (http://wheaturgi.versailes.inra.fr/Seq Repository/Genesannotations)™ÓXYcDNAÆ ÇÈ , B¿ÀÌÁÂÃ]CIPK²³|#²³, àÌNCBI“BLAST+usermanual”]¢£,áO Í]XYcDNAÆÇÈëÎNBLASTÏÐ,™ ÓXYÑÒ]CIPK²³,M‚ MEGA5.0½Ó ë‹ÔµÕƒOsCIPK16Ö×&`ØÙ]²³,Ú |ÑÒ]TaCIPK16。wDz³ª«™Ó]ÑÒ. œ²³Û)@TaCIPK16F/R(´ 1)。¾¿¿§
11”•Ü RNAäÃÄícDNA,Ä.¢£àÌ
FermentsReverAidFirstStrandcDNAsynthesis
µ¶·ÝOŽµ

BcDNA|3Þ,PCR†ßŒ²
|95℃àé±5min,95℃ é±30s,55℃ áâ
30s,72℃xã1min,ä35›åê;72℃ xã
10min。PCRŠæ‹8Â9ÓpGEMTEasy Í. , çcèé’Ë8¾¿¼½êë² 。 1.2.2 YZCD ìmn–—ë²]€ÝÌM‚ àë™Ó]ÑÒ.œ²³ëÎÇȱíá 。 õ ExPAsyîï]†ðñòComputepI/Mwtool(ht tp://web.expasy.org/compute_pi/)ëÎ)ó¶· z{]=ôl̪Ñ ; ž DNASTAR ñò, BLAST(http://www.ncbi.nlm.nih.gov/blast/) ÌORFFinder(http://www.ncbi.nlm.nih.gov/ gorf/gorf.html)ëÎcDNA ²³]ª«。ž ClustalW2 (http://www.ebi.ac.uk/Tools/msa/ muscle/)Ì DNAMAN7.0 ñò (version7.0; LynnonBiosoft,USA)ëβ³í᪫。õ MEGA5software (version5.0)ñòBõ9£ (NeighborJoiningmethod)ëΕ]z{ƒ­ö§ z{]èk²³íá¥ë‹Ôª« 。 õInter ProScan(http://www.ebi.ac.uk/Tools/pfa/ipr scan/)ÌPROSITEScan(http://prosite.expasy. org/scanprosite/)Uàëz{]d»z{€½¾。 1.2.3 [\]^_CD Û)í TaCBL4 (GenBank÷ć| KU736850)Ì TaCBL9(Gen Bank÷ć| KU736852) ¡¢£¤–—@
TaCBL4BDF/R、TaCBL9BDF/R(´ 1),–—
pGBKT7Í.Ü,Û)TaCIPK16 ¡¢£¤–
—@ TaCIPK16ADF/R (´ 1), – —  pGADT7Í.Ü。øÌ ¡¢£¤ùú(Clontech, Japan),ì TaCBL4ƒ TaCIPK16?P TaCBL9ƒ TaCIPK16äà¡’pAH109,ª†dz
±¨©.SDLeucineTryptophan(LT)ÌSDAde
nineHistidineLeucineTryptophan(º20μg·mL
-1
]
`1 ab`
Table1 Primersusedinthisstudy
@ûüPrimer @²³Primersequence(5′→3′)
TaCIPK16F GAGGAGTGGTAGTGATTAGTGA
TaCIPK16R CTTCAAGTTGCTCTGCACGTTAC
TaCBL4BDF CATGCCATGGGCTGCGTGTTGTCATC
TaCBL4BDR GGCTGCAGTTATTTGCTGATTCCGCTGTAA
TaCBL9BDF CATGCCATGGCCTCACGCTTCAGCTT
TaCBL9BDR GGCTGCAGTTAGTCTTCAACCGCGGTATTG
TaCIPK16ADF CCGGAATTCATGTCCGCGAGCCCGG
TaCIPK16ADR AAAACTGCAGGCTACGTCAAGATGTACCAGAAGA
TaCIPK16163F AAGCTTATGGCGAGAGGGCGGGAGGG
TaCIPK16163R GGATCCCGCGACGGCGCCCGG
TaCIPK16qRTF1 GCCGCCGCAGGCCGCGCGGCTGC
TaCIPK16qRTR1 CCTTCTCCCGGTCGAGCACCTTGAT
TaEF1F TGGTGTCATCAAGCCTGGTATGGT
TaEF1R ACTCATGGTGCATCTCAACGGACT
XαGal)(LTHA+ Xαgal)ÜëΨ©。¨©
4d8,žýþ)ÆÞì ¡¦§ÿ™Ó!"#
¿ºÅ107、106、105Ì104›¦§,lLTHA+
Xαgal¨©.ܨ©,24h8ÌÇ。
1.2.4 cdefg ˆÇpCaMV35S:GFPè
–—©lN , Û)íTaCIPK16y¦§¨© – — @ TaCIPK16163F/R (´ 1), ì
TaCIPK16–— pCaMV35S:GFP ,àÌ Li
=
[19]
]¢£

ƒ%XY„S¼.

à̑¢£ì
rÍ.pCaMV35S:TaCIPK16∶GFP¥&Í.
pCaMV35S∶GFPÃXY„S¼.,¨ ©18h
8

M‚ä­Ê()ëÎ*+ÌÇ

1.2.5 狇犚犜犘犆犚CD wǖ—,U]犜犪犆犐犘犓16
.œÌÀà.œXYxãœ犜犪犈犉1α(GenBank÷
ć Q03033)ªÛ)°Ø¨Ñ@ TaCIPK16
qRTF1/R1¥TaEF1F/R(´ 1)。-.9#Z[
’80、12、18、24、72Ì120hþ/l]”•¾¿
RNA,äÃÄ|cDNA,B^cDNA|3Þ,qRT
PCRä匲|95℃àé±5min,95℃é±,
10s,55℃ áâ20s,72℃xã40s,ä40›åê。
qRTPCR]äå.`¥Œ²àÌ Feng=]¢
£
[20],
Ø8-2-ΔΔCt£)ó•0.œáÀà.œ
]Çá´ÔÑ

2 €Ýƒª«
2.1 45犜犪犆犐犘犓16,<=>?@YZCD
M‚RTPCRbŸ–—™Ó]cDNA²³Ä
57016;        MYOTHWIN,=:XYCBL€z{„x.œ犜犪犆犐犘犓16]–—¥°1ª«
Å1606›23w(41),¶·1›447¸.w]z
{

z{ªÑ|49.521kD,=ôl|9.09。õ
InterProScanñòª«´µ,­N6¹º}›z
{„x€½¾
,C6ºÅ}›u_€½¾,†u7
¾¹º¯ZƒCBL°Ø€]NAF€½¾¥ƒ
PP2CÇÐO]PPI€½¾(42)。žÁÂÃ
ƒ¿À5Å]CIPK²³¸.w²½Óë‹ÔŽ
q‘z{ƒ¿À] OsCIPK16(LOC_Os09g25090)
¥ÁÂÍ] AtCIPK5(AAF86504)、AtCIPK16
(AAK50348)¥AtCIPK25(AF44226)ö1}›ª
:

43)。5Z²³í᪫´µ,‘z{²³ƒ
1~2.PCR†ßŠ;M.Marker 41 犜犪犆犐犘犓16.œcDNA]PCR†ß•5 1-2.PCRproduct;M.Marker Fig.1 Electrophoresisof犜犪犆犐犘犓16cDNAfragment At.ÁÂÃCIPK²³;Os.¿ÀCIPK²³;678,mn –—]TaCIPK16²³;7lÜ]Æ9´ËBootstrapnÉ .21000:kç‘7l]ÞNÆ 43 TaCIPK16ƒ¿À、ÁÂÍCIPK(š]ë‹Ôª« At,CIPKsequencesof犃狉犪犫犻犱狅狆狊犻狊downloadfromTAIRv10; Os,CIPKsequencesofricedownloadfromRGAPv7;Thearrow indicatesTaCIPK16;Thenumbersatthenodesrepresentsthe reliabilitybasedon1000replicates Fig.3 PhylogeneticanalysisofTaCIPK16withCIPK familyfromriceand犃狉犪犫犻犱狅狆狊犻狊 ;ð´ËN6z{„x€½¾;<ð´ËC6u_€½¾;<=´ËNAF€½¾;;ð=´ËPPI€½¾ 42 TaCIPK16ƒ¿À、ÁÂÃö§.œ]è²³íá Theconservedproteinkinasedomainandregulatorydomainaremarkedwithblackfullineanddashline,respectively. TheconservedNAFandPPImotifsaremarkedwithblackboxanddashbox Fig.2 MultisequencealignmentofTaCIPK16withhomologousproteinsinriceand犃狉犪犫犻犱狅狆狊犻狊 6701 ! " #  % &                   36Ë
OsCIPK16ÇÈÆÔÓ88.04%,ƒÁÂÃ(犃狉犪犫犻
犱狅狆狊犻狊狋犺犪犾犻犪狀犪) AtCIPK16 Ç È Æ Ô Ó
76.43%,>?û|TaCIPK16。
2.2 [\]^CD
|¬nÉ TaCIPK16ÌTaCBL4?TaCBL9ñ@
ÅÇÐO

ìTaCIPK16ƒTaCBL4?PTaCIPK16
ƒTaCBL9äöࡍ。†AÒ¨©.SD
LeucineTryptophanÜ ¡¯ZßBSö,Žµ2
#r¼½í®ÃC ¡

44,A)。†AÒ¨
©.SDAdenineHistidineLeucineTryptophan(º20
ºÅkʼ½]AH109 ¡’p†ÒD¨©.SDLeucineTryptophan(LT)ÌSDAdenineHistidineLeucineTryptophan(E20μg/mL
]Xαgal)¨ ©;A、B.F±áÌ,Takala ¡µ¶·]SV40largeTantigenÌmurinep53ÐO€Ý:C、D.TaCIPK16ÌTaCBL4]ÐO
€Ý
;E、F.TaCIPK16ÌTaCBL9]ÐO€Ý;G、H.G±áÌ,Takala ¡µ¶·SV40largeTantigenƒhumanlamin]ÐO€Ý
44 TaCIPK16ƒTaCBL4、TaCBL9]ÐOª«
CelsofyeaststrainAH109harboringtheindicatedplasmidcombinationsweregrownonselectivemediaSDLeucineTryptophan
(LT)andSDAdenineHistidineLeucineTryptophan(containing20μg/mLXαgal);A,B.Positivecontrol,theinteraction
betweenSV40largeTantigenandmurinep53;C,D.TheinteractionofTaCIPK16andTaCBL4:E,F.Theinteractionbetween
TaCIPK16andTaCBL9;G,H.Negativecontrol,theinteractionbetweenSV40largeTantigenandhumanlaminC.
Fig.4 AnalysisofinteractionsbetweenTaCIPK16andTaCBL4/TaCBL9
A~D.F±áÌ,pCaMV35S:GFPË]XY„S¼¦§;E~H.pCaMV35S:TaCIPK16:GFPË]XY„S¼¦§;A、E.Ò
„Ž]GFPHI̔J.+ŽHIʁ€Ý;B、F.”J.+ŽHI;C、G.҄Ž]GFPHI;D、H.{IO]„S¼¦§
45 XY„S¼.TaCIPK16y¦§¨©ª«(Bar=5μm)
A~D.Positivecontrol,pCaMV35S:GFP;E~H.pCaMV35S:TaCIPK16:GFP;A,E.overlay;B,F.Chloroplast;C,G.GFP;
D,H.Brightfield.
Fig.5 SubcelularlocalizationofTaCIPK16inwheatprotoplasts
77016;        MYOTHWIN,=:XYCBL€z{„x.œ犜犪犆犐犘犓16]–—¥°1ª«
46 犜犪犆犐犘犓16.œ†XYZ[’ÐO‚Œ]
ÃÄ´Ô°1
Fig.6 Expressionpatternof犜犪犆犐犘犓16duringthe
compatibleandincompatibleinteractionbetweenwheatandPst
μg·mL
-1XαGal) ¡¯ZSöˆíKL,ƒF
± á Ì Ç È

Ž µ TaCIPK16 ƒ TaCBL4 Ì
TaCBL9n͆ÐO(44,B)。ì ¡¨©Mÿ™
Ó!"#ºÅ104› ¡¦§þ,N¯*+Ó ¡
]Sö
,´
µTaCIPK16ƒTaCBL4ÌTaCBL9Ä
ÅÎÏ]ÐO

2.3 cdefgCD
|nÉTaCIPK16†XY¦§]ªOPQ,ì
rÍ.pCaMV35S∶TaCIPK16∶GFPB¥&Í.
pCaMV35S∶GFPÃRXY„S¼.。€Ý´
µ
,TaCIPK16†XY¦§ªOƒF±áÌGFP]
PQÇö

†f›XY¦§nŪO

45)。
2.4 45犜犪犆犐犘犓16,<=hifj 犚犜犘犆犚
CD
  mþ¨ÑRTPCRª«€Ý´µ,犜犪犆犐犘犓16
ÒÓZ[’“V´Ô

†SÖÌÐO
(CYR23),
犜犪犆犐犘犓16ÃÄ¿ÿ†9#86~36hC20h´
Ô¿ÿ

ÕOu´Ô

KÓ48h´ÔѵÊÜu,
ÔÓT9

Çá´ÔÑ|10.8),†B8]þ/l,
´ÔÑUçÓVW¿ÿ

4 6)。†ÖÌÐO
(CYR31),犜犪犆犐犘犓16]Çá´ÔÙڃ†SÖ
̍]´ÔÙÚÇö

X†9’48h8S(]Üu
´ÔÑ

Çá´ÔÑ|2)ÊÖC2†SÖÌÐO
´ÔÑ

46)。Y^Z[,犜犪犆犐犘犓16Þ¯àƒ¬
XYáZ[’]â\^ãäå

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8701 ! " #  % &                   36Ë kl!M : [1] ZHUJK.Saltanddroughtstresssignaltransductioninplants [J].犃狀狀狌犪犾犚犲狏犻犲狑狅犳犘犾犪狀狋犅犻狅犾狅犵狔.2002,53:247. [2] ZHAOJ,SUNZ,ZHENGJ,犲狋犪犾.Cloningandcharacteriza tionofanovelCBLinteractingproteinkinasefrommaize[J]. 犘犾犪狀狋犕狅犾犲犮狌犾犪狉犅犻狅犾狅犵狔,2009,69(6):661674. [3] LUANS,KUDLAJ,RODRIGUEZCONCEPCION M,犲狋 犪犾.CalmodulinsandcalcineurinBlikeproteinscalciumsensors forspecificsignalresponsecouplinginplants[J].犜犺犲犘犾犪狀狋 犆犲犾犾,2002,14:S389S400. [4] KOLUKISAOGLU,WEINLS,BLAZEVICD,犲狋犪犾.Calci umsensorsandtheirinteractingproteinkinases:genomicsof the犃狉犪犫犻犱狅狆狊犻狊andriceCBLCIPKsignalingnetworks[J]. 犘犾犪狀狋犘犺狔狊犻狅犾狅犵狔,2004,134(1):4358. [5] ALBRECHTV,RITZO,LINDERS,犲狋犪犾.TheNAFdo maindefinesanovelproteinproteininteractionmodulecon servedinCa2+regulatedkinases[J].犜犺犲犈犕犅犗犑狅狌狉狀犪犾, 2001,20(5):10511063. [6] LEESC,LANWZ,KIMBG,犲狋犪犾.Aproteinphosphoryla tion/dephosphorylationnetworkregulatesaplantpotassium channel[J].犘狉狅犮犲犲犱犻狀犵狊狅犳狋犺犲犖犪狋犻狅狀犪犾犃犮犪犱犲犿狔狅犳犛犮犻 犲狀犮犲狊,2007,104(40):1595915964. [7] LIR,ZHANGJ,WEIJ,犲狋犪犾.Functionsandmechanismsof theCBLCIPKsignalingsysteminplantresponsetoabiotic stress[J].犘狉狅犵狉犲狊狊犻狀 犖犪狋狌狉犪犾犛犮犻犲狀犮犲,2009,19(6): 667676. [8] LIL,KIM BG,CHEONGY H,犲狋犪犾.ACa2+ signaling pathwayregulatesaK+channelforlowKresponsein犃狉犪犫犻 犱狅狆狊犻狊[J].犘狉狅犮犲犲犱犻狀犵狊狅犳狋犺犲犖犪狋犻狅狀犪犾犃犮犪犱犲犿狔狅犳犛犮犻犲狀犮犲狊, 2006,103(33):1262512630. [9] XUJ,LIHD,CHENLQ,犲狋犪犾.Aproteinkinase,interac tingwithtwocalcineurinBlikeproteins,regulatesK+trans porterAKT1in犃狉犪犫犻犱狅狆狊犻狊[J].犆犲犾犾,2006,125(7):1 3471360. [10] YANG W,KONGZ,OMOIKERODAHE,犲狋犪犾.Calci neurinBlikeinteractingproteinkinaseOsCIPK23functions inpolinationanddroughtstressresponsesinrice(犗狉狔狕犪狊犪 狋犻狏犪犔.)[J].犑狅狌狉狀犪犾狅犳犌犲狀犲狋犻犮狊犪狀犱犌犲狀狅犿犻犮狊,2008,35 (9):531543. [11] LEEKW,CHENPW,LUCA,犲狋犪犾.Coordinatedrespon sestooxygenandsugardeficiencyalowriceseedlingstotol erateflooding[J].犛犮犻犲狀犮犲犛犻犵狀犪犾犻狀犵,2009,2(91):ra61. [12] TRIPATHIV,PARASURAMANB,LAXMIA,犲狋犪犾. CIPK6,aCBLinteractingproteinkinaseisrequiredforde velopmentandsalttoleranceinplants[J].犜犺犲犘犾犪狀狋犑狅狌狉 狀犪犾,2009,58(5):778790. [13] MITTLERR,VANDERAUWERAS,SUZUKIN,犲狋犪犾. ROSsignaling:thenewwave?[J].犜狉犲狀犱狊犻狀犘犾犪狀狋犛犮犻 犲狀犮犲,2011,16(6):300309. [14] KIMURAS,KAWARAZAKIT,NIBORIH,犲狋犪犾.The CBLinteractingproteinkinaseCIPK26isanovelinteractor of犃狉犪犫犻犱狅狆狊犻狊NADPH oxidaseAtRbohFthatnegatively modulatesitsROSproducingactivityinaheterologousex pressionsystem[J].犑狅狌狉狀犪犾狅犳犅犻狅犮犺犲犿犻狊狋狉狔,2013,153 (2):191195. [15] KURUSUT,HAMADAJ,NOKAJIMAH,犲狋犪犾.Regula tionofmicrobeassociatedmolecularpatterninducedhyper sensitivecel death,phytoalexinproduction,anddefense geneexpressionbycalcineurinBlikeproteininteractingpro teinkinases,OsCIPK14/15,inriceculturedcels[J].犘犾犪狀狋 犘犺狔狊犻狅犾狅犵狔,2010,153(2):678692. [16] DELATORREF,GUTI?RREZBELTR?NE,PAREJA JAIMEY,犲狋犪犾.ThetomatocalciumsensorCbl10andits interactingproteinkinaseCipk6defineasignalingpathwayin plantimmunity[J].犜犺犲犘犾犪狀狋犆犲犾犾,2013,25(7):2748 2764. [17] XIEC,ZHOUX,DENGX,犲狋犪犾.PKS5,aSNF1related kinase,interactswithandphosphorylatesNPR1,andmodu latesexpressionofWRKY38andWRKY62[J].犑狅狌狉狀犪犾狅犳 犌犲狀犲狋犻犮狊犪狀犱犌犲狀狅犿犻犮狊,2010,37(6):359369. [18] KANGZS,LIZQ.DiscoveryofanormalT.typenew pathogenicstraintoLovrin10[J].犃犮狋犪犆犾犾犲犵犻犻犛犲狆狋犲狀狋狉犻狅狀犪 犾犻犗犮犮犻犱犲狀狋犪犾犻犃犵狉犻犮狌犾狋狌狉犪犲,1984,4(1):1828. [19] LIC,LIN H,DUBCOVSKYJ.Factorialcombinationsof proteininteractionsgenerateamultiplicityofflorigenactiva tioncomplexesinwheatandbarley[J].犜犺犲犘犾犪狀狋犑狅狌狉狀犪犾, 2015,84(1):7082. [20] FENGH,WANGX,SUNY,犲狋犪犾.Cloningandcharacter izationofacalciumbindingEFhandproteingene犜犪犆犪犫1 fromwheatanditsexpressioninresponseto犘狌犮犮犻狀犻犪狊狋狉犻 犻犳狅狉犿犻狊f.sp.狋狉犻狋犻犮犻andabioticstresses[J].犕狅犾犲犮狌犾犪狉犅犻 狅犾狅犵狔犚犲狆狅狉狋狊,2011,38(6):38573866. [21] THODAYKENNEDYEL,JACOBSAK,ROYSJ.The roleoftheCBLCIPKcalciumsignalingnetworkinregula tingiontransportinresponsetoabioticstress[J].犘犾犪狀狋 犌狉狅狑狋犺犚犲犵狌犾犪狋犻狅狀,2015,76(1):312. [22] BATISTICˇO,WAADTR,STEINHORSTL,犲狋犪犾.CBL mediatedtargetingofCIPKsfacilitatesthedecodingofcalci umsignalsemanatingfromdistinctcelularstores[J].犜犺犲 犘犾犪狀狋犑狅狌狉狀犪犾,2010,61(2):211222. [23] WANGCF,HUANGLL,BUCHENAUERH,犲狋犪犾.His tochemicalstudiesontheaccumulationofreactiveoxygen species(O2-andH2O2)intheincompatibleandcompatible interactionofwheat犘狌犮犮犻狀犻犪狊狋狉犻犻犳狅狉犿犻狊f.sp.狋狉犻狋犻犮犻[J]. 犘犺狔狊犻狅犾狅犵犻犮犪犾犪狀犱 犕狅犾犲犮狌犾犪狉犘犾犪狀狋犘犪狋犺狅犾狅犵狔,2007,71 (4):230239. [24] DENGX,HU W,WEIS,犲狋犪犾.犜犪犆犐犘犓29,aCBLinter actingproteinkinasegenefrom wheat,conferssaltstress toleranceintransgenictobacco[J].犘犔狅犛犗犖犈,2013,8 (7):e69881. ( !" : #%
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