Ossds1, a Negative Regulator, Significantly Reduces Salt and Drought Tolerance in Transgenic

OsSDS1, a negative regulator, significantly reduces salt and drought tolerance in transgenic Arabidopsis

Guoyun Xu · Mingjuan Li · Mei Huang · Yanchun Cui · Manling Wang · Xinjie Xia

OsSDS1 (1) --MAAYGKPQPP----LNDAYYGPPIPPPPAAAYYGAA------APPPAPRRSGAHRLFCCLFRV

BAB63813 (1) -----MANNHEHKVDHLDQPFYGPPVLPPVEPLSAAAAR------RRCVADPYALCCRAIRV

NP001044894 (1) MGDHAMANNHEHKVDHLDQPFYGPPVLPPVEPLSAAAAR------RRCVADPYALCCRAIRV

AAB97367 (1) ------MGSASRAVS------CLCCPCKCLACGLFSCLCSI

NP001054317 (1) ------MGSASRAVS------CLCCPCKCLACGLFSCLCSI

NP001147044 (1) ------MGSASRAIS------C-LCCPCRCLFSCLCSI

AAF88022 (1) ------MGSKQP--YLNGAYYGPSIPPPPKAHRSYNSPGFG-CCCFSCLGSCLRCCGCCILSLICNI

AF264698 (1) ------MGSKQP--YLNGAYYGPSIPPPPKAHRSYNSPGFG-CCCFSCLGSCLRCCGCCILSLICNI

AAF88023 (1) ------MADLNGAYYGPSIPPPKKVSHSHGRR----GGGCGCLGDCLGCCGCCILSVIFNI

NP196250 (1) ------MADLNGAYYGPSIPPPKKVSHSHGRR----GGGCGCLGDCLGCCGCCILSVIFNI

ACH63239 (1) ------MTSYQKQPQLNGAYYGPAIPPPEPAKSSYRPGRGGGGRGCCCLFDCLCDCGCCLLSCVFKI

BAG15858 (1) ------MAEKQP--HLNGAYYGPAIPPPKN---YHRP------GGRSG--SGCGCCLLSFLFKL

AF212183 (1) ------MPESN----LNGAYYGPSIPPPAKSYHRHGR------GSSCNPCSCLFGCLCNCFFKI

BAC15623 (1) ------MPESN----LNGAYYGPSIPPPAKSYHRHGR------GSSCNPCSCLFGCLCNCFFKI

BAD22534 (1) ------MPESN----LNGAYYGPSIPPPAKSYHRHGR------GSSCNPCSCLFGCLCNCFFKI

BAD22533 (1) ------MAESH----LNGAYYGPSIPPSSKTYHHHGR------GSSCNPCSCLFGCLCNCIFQI

AAS67334 (1) ------MPEPN----LNGAYYGPSIPPPTKTYHRSCG------GSSCNPCSCLFNCLCTCICQI

AAN52931 (1) ------MSN----LNGAYYGPSIPPPSKSYHRHGR------GSSCNPCSCLFGCLCNCIFQI

BAD11071 (1) ------MPESN----LNGAYYGPSIPPPSKSYHRHGR------GSSCNPCSCLFGCLCNCIFQI

OsSDS1 (54) LAVAVIALGTAVLVLWLIYRPSVVKAYADTAALSRFDLTNGGS-----LLVYNLTVGMRVRNPN-RFG

BAB63813 (52) LTIVVIAVGVVALVLWLVSLPNALKAYVDSAELTRFELGGSDGAKRGQLLRYNLTVAVSIRNPNRDQA

NP001044894 (57) LTIVVIAVGVVALVLWLVSLPNALKAYVDSAELTRFELGGSDGAKRGQLLRYNLTVAVSIRNPNRDQA

AAB97367 (30) LISLLVTLGVLALIFYLIFRPHMIAATVDSAALTQFTLSTNSA------LAYSLTVDLTVRNPNKRVG

NP001054317 (30) LISLLVTLGVLALIFYLIFRPHMIAATVDSAALTQFTLSTNSA------LAYSLTVDLTVRNPNKRVG

NP001147044 (26) LVSLAVTLGLLALALYLILKPRMIAATVDSASLSQFALSASST------LAYDLNVTMTVRNPNTRVG

AAF88022 (59) LIAVAVILGVAALILWLIFRPNAVKFYVADANLNRFSFDPNNN------LHYSLDLNFTIRNPNQRVG

AF264698 (59) LIAVAVILGVAALILWLIFRPNAVKFYVADANLNRFSFDPNNN------LHYSLDLNFTIRNPNQRVG

AAF88023 (52) LITIAVLLGIAALIIWLIFRPNAIKFHVTDAKLTEFTLDPTNN------LRYNLDLNFTIRNPNRRIG

NP196250 (52) LITIAVLLGIAALIIWLIFRPNAIKFHVTDAKLTEFTLDPTNN------LRYNLDLNFTIRNPNRRIG

ACH63239 (62) ICSIIVAIGILILVVWLIIHPHEPKFHVTDASITTFNYS-GNQ------LSYNLAVNFTIRNSNHRIG

BAG15858 (46) IITAVVIIGIAILVIWLVLRPNKIKFHVTDATLTQFNITANNT------LQYNLALNLTVRNPNKRIG

AF212183 (49) IFTLLIILGVIALVLWLVLRPNKVKFYVTDATLTQFDLSTTNN-----TIFYDLALNMTIRNPNKRIG

BAC15623 (49) IFTLLIILGVIALVLWLVLRPNKVKFYVTDATLTQFDLSTTNN-----TIFYDLALNMTIRNPNKRIG

BAD22534 (49) IFTLLIILGVIALVLWLVLRPNKVKFYVTDATLTQFDLSNTNN-----TIFYDLALNMTIRNPNKRIG

BAD22533 (49) FFTILVIIGVAALVLWLVLRPNKVKFYVTDATLTQFDLSTTNN-----TINYDLALNMTIRNPNKRIG

AAS67334 (49) IFTLLIALGVIALVLWLVLRPNKVKFYVTDATLTQFDLSTANN-----TLYYDLALNMTIRNPNKRVG

AAN52931 (47) IFTILIVLGVIALVLWLVLRPNKVNFYVTDATLTQFDYSTTNN-----TLYYDLALNMTIRNPNKRIG

BAD11071 (49) IFTILIVLGVIALVLWLVLRPNKVKFYVTDATLTQFDLSTANN-----TLYYDLALNMTIRNPNKRIG

OsSDS1 (116) INFRSVDAQASYDGDRFGYAP-LQPLYVGRKSDARFDVTLS-GSAAIDDRDVERT-YRRETAQGSYEV

BAB63813 (120) VLYRRLEAVVLYGGERFGYVD-FPRTRQGRKSTMVIRPSFVG--QGVLAGAAAFG---REKEEGFFNI

NP001044894 (125) VLYRRLEAVVLYGGERFGYVD-FPRTRQGRKSTMVIRPSFVG--QGVLAGAAAFG---REKEEGFFNI

AAB97367 (92) LYYDNVESLALFDGQRFGYAP-LDSFYQSTEASTKLSPAFK--GQQPLQGDITAANFRSQQTAGKFDI

NP001054317 (92) LYYDNVESLALFDGQRFGYAP-LDSFYQSTEASTKLSPAFK--GQQPLQGDITAANFRSQQTAGKFDI

NP001147044 (88) LYYDDVQALASYEGQRFGYAP-LDAFYQGTGAATKLAPGFR--GTQLLQGDVAAGQFRQQQSAGRFDV

AAF88022 (121) VYYDEFSVSGYYGDQRFGSAN-VSSFYQGHKNTTVILTKIE-GQNLVVLGDGARTDLKDDEKSGIYRI

AF264698 (121) VYYDEFSVSGYYGDQRFGSAN-VSSFYQGHKNTTVILTKIE-GQNLVVLGDGARTDLKDDEKSGIYRI

AAF88023 (114) VYY-EIEVRGYYGDQRFGMSNNISKFYQGHKNTTVVGTKLV-GQQLVLLDGGERKDLNEDVNSQIYRI

NP196250 (114) VYYDEIEVRGYYGDQRFGMSNNISKFYQGHKNTTVVGTKLV-GQQLVLLDGGERKDLNEDVNSQIYRI

ACH63239 (123) IYYDAFEANTFYQDQRFGMVE-VDPFFQGKKNTTEIGPVVFKGQSGITLGSS--SDFESQKNGGKFDI

BAG15858 (108) IYYDKIDAAAFYEGNRFGHAL-LTPFYQGHKNTSVLSHVLQ-GQHLLLLGTDELTQFNQEKTAGLYSI

AF212183 (112) IYYDSIEARALYQGERFDSTN-LEPFYQGHKNTSSLRPVFK-GQSLVLLGDREKSNYNNEKNLGVYEM

BAC15623 (112) IYYDSIEARALYQGERFDSTN-LEPFYQGHKNTSSLRPVFK-GQSLVLLGDREKSNYNNEKNLGVYEM

BAD22534 (112) IYYDSIEARAMYQGERFHSTN-LKPFYQGHKNTSSLHPVFK-GQSLVLLGDREKSNYNNEKNLGVYEM

BAD22533 (112) IYYDSIEARAMYQGERFASQN-LEPFYQGHKNTSSLHPVFK-GQSLVLLGDREKTNYNNEKNAGVYEM

AAS67334 (112) IYYDSIEARAMYQGQRFASHN-LEPFYQGHKNTSNLHPVFK-GQSLVLLGDREKSNYNNEKNSGVYDI

AAN52931 (110) IYYDSIEARGMYQGQRFASQN-LERFYQGHKNTSYLHPVFK-GQNLVLLGDREKTNYNNEKNLGIYEM

BAD11071 (112) IYYDSIEARGMYKGERFASQN-LEPFYQGHKNTSSLHPVLK-GQSLVLLGDREKSNYNNEKNSGVYDI

OsSDS1 (181) KVRVYARQGFKVRGFRLN--NKSKFTCTLNLPAPNSGNGTASGTPTTVFTRKQPKCDVDY--

BAB63813 (182) NVKLHMRVRLKVMVFVDSVEYRPDVDCYIRVPDP------SNATAVAMGFTATRCRVDDFM

NP001044894 (187) NVKLHMRVRLKVMVFVDSVEYRPDVDCYIRVPDP------SNATAVAMGFTATRCRVDDFM

AAB97367 (157) EVKLNAKLRVKVWAFKVPG-PKPKISCPITVPAS------APNAPAFQRTDCKVWF--

NP001054317 (157) EVKLNAKLRVKVWAFKVPG-PKAKISCPITVPAS------APNAPAFQRTDCKVWF--

NP001147044 (153) DVDLNAKLRVKFWAFKAHG-PNAKITCTLSVPAA------ASASPFQPTDCKVWF--

AAF88022 (187) DAKLRLSVRFKFWFIKSWK-LKPKIKCDDLKIPLG------SSNSTGGFKFQPVQCDFDLS-

AF264698 (187) DAKLRLSVRFKFWFIKSWK-LKPKIKCDDLKIPLG------SSNSTGGFKFQPVQCDFDLS-

AAF88023 (180) DAKLRLKIRFKFGLIKSWR-FKPKIKCDLKVPLT------SNSTSGFVFQPTKCDVDF--

NP196250 (181) DAKLRLKIRFKFGLIKSWR-FKPKIKCDLKVPLT------SNSTSGFVFQPTKCDVDF--

ACH63239 (188) LVRLHLKVRFKLGAFKTGK-WKPKVKCDLTVPLS------SSTNTFESTKCRYYY--

BAG15858 (174) DVKLYMRIRLKVGRFKVEK-SSPRLSAT------

AF212183 (178) EVKLYMRIRLKVGWIKTHK-IKPKIECDFKVPLG------SNGRSSSANFEETRCHLDW--

BAC15623 (178) EVKLYMRIRLKVGWIKTHK-IKPKIECDFKVPLG------SNGRSSSANFEETRCHLDW--

BAD22534 (178) EVKLYMRIRLKAGWIKTHK-IKPKIECDFKVPLE------SNGRSSSANFEETRCHLDW--

BAD22533 (178) EVKLSMRIRLKFGWIKTRK-IKPKIECDFKVPLE------SNGRSSSANFEETRCHLDW--

AAS67334 (178) EMKLYMRIRL------

AAN52931 (176) EMKLYMRIRLKIGWIKTHK-IKPRIECDFKVPLE------SSGRSVNFEETRCHLDW--

BAD11071 (178) EMKLYMRIRLKIGWIKTHK-IKPKIECDIKVPLE------SNGRSSANFEETRCHLDW--

Fig. S1 Analysis of the protein sequence of OsSDS1. a Alignment of the sequence of OsSDS1 with other harpin-induced proteins. Identical and similar residues (similarity value > 50%) are shaded in gray. The accession numbers in genbank: BAB63813, NP001044894, AAB97367, NP001054317 (Oryza sativa); NP001054317 (Zea mays); AAF88022, AF264698, AAF88023, NP196250 (Arabidopsis thaliana); ACH63239 (Rheum australe); BAG15858 (Bruguiera gymnorhiza), AAS67334 and AAN52931 (Solanum tuberosum), BAD11071 (Capsicum chinense), AF212183, BAD22534, BAD22533, BAC15623 (Nicotiana tabacum). b Phylogenetic relationship of OsSDS1 with other harpin-induced proteins. Sequence alignment and phylogenetic tree were conducted by Vector NTI 10

Fig. S2 qRT-PCR analysis of OsSDS1 expression in transgenic lines and wild type plants. Total RNAs were extracted from different transgenic lines of T3 generation and wild type plants. Error bars represent SD for three independent experiments. WT, wild type Arabidopsis; H3, H7, H8, H15, H21 and H23, transgenic lines 3, 7, 8, 15, 21 and 23

Supplementary Table 1 Primer pairs used in this paper

Gene Sense primer (5′-3′) Antisense primer (5′-3′)
OsSDS1 TCTAGACATCCATCACTCGTTCGATCTCTCC CACGTGTGTGTCGTGTCGTTCCCAGCC
OsSDS1-GFP AAGCTTCATCCATCACTCGTTCGATCT CCATGGAATAGTCCACGTCGCACTTCG
RT-OsSDS1 ACGAGGAAGCAGCCGAAGTG CGTTCCCAGCCGAGGAGTTC
P5CS1 AAGGCTTGTGATACGGATATGG ATGCACAAGAAGGGTTTCCA
RD29A ACGTCGAGACCCCGATAAC CAATCTCCGGTACTCCTCCA
RD29B GGTACTCCAGCTTCTCCACCT AAAACGTTTGGAGGAAGATCC
AtSOS1 TCGTTTCAGCCAAATCAGAAAGT TTTGCCTTGTGCTGCTTTCC
AtNHX1 CCGTGCATTACTACTGGAGACAAT GTACAAAGCCACGACCTCCAA
18s AAACGGCTACCACATCCAAG CCTCCAATGGATCCTCGTTA