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1、作物学报 ACTA AGRONOMICA SINICA 2017, 43(5): 708717 http:/zwxb.chinacrops.org/ ISSN 0496-3490; CODEN TSHPA9 E-mail: SE1S (31400583), ig$-9B (cstc2014jcyjA80011)n=UaH , =sM, =sc10-11; rV9Y Ml , CMV , 7V?/ly8-12b 8%=0i1 4e , sY ZIP (ZRT-IRT-like protein)aNramp (natural resistance-associated macrophage pr
2、otein)aHMA (P1B-ATPase or CPx-type ATPase) CDF (cation diffusion fa-cilitator)13-14b CDF EB 400 F , V|%=% , 9V|%0% , Pui , V7%bo CDF , c 46t , N C Y%= ; YcBFu , 4 5 tu N C 15-16b- , 3 CDFEB$XHbd?sV , Vs Zn-CDFaFe/Zn-CDF Mn-CDF EB17b , CDF $0s(metal tolerance protein, MTP)byFV , E2yFc 12 I MTP y , At
3、MTP1aAtMTP3aAtMTP4aAtMTP8aAtMTP1 AtMTP11 118-19; OsMTP1 OsMTP8a PtMTP1a( CuMTP8 9$0M113, 20-23bE2 AtMTP1 AtMTP3 sY$A , (0eV24-25; AtMTP11 $A-8Fs , 126; ShMTP1$A , V|%0A , V7%i , r0srT27b -111c#3?VY , (33Z , #s0ebXH0sy CsMTP11, isy? , wV?1Tb 1 材料与方法 1.1 k 2 M3 108 ! , 2vkbk41 EASYspin Plus p RNA4|k4
4、, 13S/K ; EasyScript One-Step gDNA Removal and cDNA Synthesis SuperMixaTransStart Taq DNA Polymerase, 1T3/K ; SMARTer RACE 5/3 KitaXH8 pMD18-T v)JM109 s% , 1 TaKaRa (v ); DNAMlk4a Golden Easy PCR SystemaDNA Marker 1?3S/ ( )K ; Sso-FastEvaGreen A1 Bio-Rad b YNB !#1 Sigma ; y (Z )Kb 1.2 9 RNA 4|# cDNA
5、 sY|aa=a=aaaT2F 2 g, tz , A9RNA 4| , 4|v EASYspin Plus p RNA 4|k4 , DNase I (RNase free)h4|9 RNAyF DNA, 710 T 43 B9 RNA sY MTP11 yXHVrsb MTP11y9Vrs9RNA vT EasyScript One-Step gDNA Removal and cDNA Synthesis SuperMix k4yQc8 , 42 1 h; 85 5 sQc cDNATb 1.3 CsMTP11 yXH V NCBIo/X MTP11y , i1 , yo , iN!9ei
6、 MTP11-280-L: TYCCYGGAA TGTCARAGGARGA MTP11-1150-R: TCRTAATC DAGATGVACAAARGC, F9 RNAQc cDNA , XHyo, iN!99 5 3y+s GSP1- MTP11: CCAATGGCTGCATGCGTTTCTTTCC GSP2-MTP11: GTTGGCTAGAAGTGAAACAACAG CC; v SMARTer RACE 5 /3 Kitk4 , sY 5-RACE-Ready cDNA 3-RACE-Ready cDNABT , sYY UPMy+s MTP11-GSP1 MTP11-GSP2sY9y
7、5 3 , P9 94 3 min; 94 30 s, 68 30 s, 72, 2 min; 34 , 72 5 min, j , Ml , pMD18-T8 , sT , y 5 3 , i Seqman qy , CsMTP11y cDNA , iN!9 MTP11- F: ATGGTGGAGCCGGTGGAGGGTACGA MTP11- R: CTAAGAGTGTGACTGTGCGTGCTC, PCR9 CsMTP11y cDNAb | PCRMl , pMD18-T 8 , v)JM109, ) PCRk 3XHb 1.4 3s Seqman q CsMTP11 y , Editse
8、qq ORF ; NCBIosY Blast N Blast Xs ; ClustalX1.8aDNAMAN MEGA4.0 1iydbTMHMMServer v.2.0tb 1.5 CsMTP11 yVrTs 71 (2015 M 10 ), VvlB=M3 108 ! , sY|B=aaa=a , htz , A , 80bQi , qRT-PCR kbI)H , |vl3B=M3 108! , MS! 1 , sYF 10 mmol L1MnSO4 (T1) ) 1 mmol L1CoCl2(T2) ), vF (CK)B?! 3 d , |M= 1 g TqRT-PCR , A , 80bQib;Q8 SYBR Premix 10
