Xanthomonas translucens commandeers the host rate-limiting … · 2019. 10. 1. · 1 Supplementary...

1

Supplementary Information for

Xanthomonas translucens commandeers the host rate-limiting step in

ABA biosynthesis for disease susceptibility

Zhao Peng1, Ying Hu2, Junli Zhang1, Jose C. Huguet-Tapia1, Anna K. Block3, Sunghun

Park4, Suraj Sapkota5, Zhaohui Liu5, Sanzhen Liu2, Frank F. White1 1Department of Plant

Pathology, University of Florida, Gainesville, FL 32611, USA; 2Department of Plant

Pathology, Kansas State University, Manhattan, KS 66506, USA; 3 Center for Medical,

Agricultural and Veterinary Entomology, U.S. Department of Agriculture-Agricultural

Research Service, Gainesville, FL, 32608, USA; 4Department of Horticulture and Natural

Resources, Kansas State University, Manhattan, KS 66506, USA; 5Department of Plant

Pathology, North Dakota State University, Fargo, ND, USA

Correspondence to:

Email: [email protected]; [email protected]

This PDF file includes:

Supplementary text Figures S1 to S7 Tables S1 to S4 SI References

www.pnas.org/cgi/doi/10.1073/pnas.1911660116

mailto:[email protected]

2

Supplementary Information Text

Plant Material, Plasmids, and Bacterial Strains.

Seeds of wheat cv. Jagger were provided by KSU Foundation Seed, and the cv. Chinese

Spring were from U.S. Department of Agriculture–Agricultural Research Service National

Small Grains Collection. Plants were grown in Metro-Mix 360 soil (Sungro) and

maintained in growth chamber. Growth condition was set at 24°C and 15-h light at day

time, and 22°C and 9-h dark at night with relative humidity at 75%. The bacterial strains

and plasmids used in this study are listed in Table S1. In brief, mutant strains of XT4699

were previously reported (1) and generated by homologous recombination of a mutant ORF

into the wild type copy using an antibiotic resistance gene maker for the recombinants. The

recombination was validated by PCR.

Virulence assays

Strains of X. translucens pv. undulosa (Xtu) were grown on tryptone sucrose agar (TSA)

for 2 days at 28°C and scraped from plates into sterilized distill water for inoculum with

OD600nm=0.2. The 2nd leaves of 10-day-old or 3rd leaves of 14-day-old wheat plants were

inoculated by needleless syringe infiltration to generate a water-soaking spot without

extension. The lesions were photographed and scored by measuring the lesion length at 7

days post inoculation (DPI). The humidity was set at 75% for standard virulence assays.

Chamber was set to 95% for high humidity infection and 50% for the control treatment.

Plants were grown in a Conviron PRG15 chamber with humidity control. Humidity was

monitored with a Lascar humidity and temperature logger to validate chamber controls.

For the bacterial growth population assay, the whole or sections of leaves were grounded

3

in sterilized water using mortar and pestles. Serially diluted samples were plated onto

TSA plates. Colonies were counted three days after incubation of the plates at 28°C.

RNA-Seq analyses, prediction of candidate S genes for Tal8 and heatmap

constructions

Second leaves of 10-day-old wheat cultivars Jagger and Chinese Spring were infiltrated

with bacterial suspensions (OD600nm=0.5) by needleless syringe. 24 h after inoculation, 5

treated leaves were pooled for RNA extraction using RNeasy Plant Mini Kit (Qiagen).

RNA-seq libraries were prepared at Integrated Genomics Facility at Kansas State

University and sequenced at Genome Sequencing Facility at KU Medical Center. RNA-

Seq reads were subjected to adaptor and quality trimming using Trimmomatic software

(version 0.32) (2). RNA-seq data is deposited at NCBI Short Reads Archive (SRA)

database under project accession PRJNA485724. The remaining high-quality reads were

aligned to the International Wheat Genome Sequencing Consortium (IWGSC) flow-

sorted Chromosome Survey Sequence (CSS) contigs of Chinese Spring using STAR

version 2.5.0b. DESeq2 package were used to perform the differential expression analysis

(3-5) . Multiple test correction was accounted for by converting p-values to q-values (6).

Significantly differentially expressed genes (≥ 8-fold, p-value<0.01) in the highly up-

regulated genes of WT treated leaves from the cultivar Jagger compared to M6 were

selected and considered as candidate S genes. The promoter sequence, 500bp upstream of

translation start site, of all the candidate genes were searched for EBE of Tal8 by Target

finder based on its RVDs (7). The assumption was made that the unusual RVDs, KG, Y*,

and QD in Tal8 were functionally equivalent to NG, N*, and HD of the consensus or

common RVDs of other TALes. All the heatmaps were produced with heatmap.2

4

function of the R package gplots using the log10 transformed reads per million reads

(RPM). All the genes annotated as homologs genes of wheat NPR1 were used for

generating heatmap (8). The wheat PR1 genes used for generating heatmap of TaPR1

were previously reported (9).

Sequence analysis of additional low and high virulence strains of Xtu

Genome sequencing was conducted using SMRT-Pacbio technology. Raw reads were

assembled using Canu software v1.5 (10). Assemblies were polished using the genomic

consensus package with the arrow algorithmic

(https://github.com/PacificBiosciences/GenomicConsensus). Genome comparisons and

alignments were conducted using mummer4 (11). TAL effector sequences analysis was

conducted using QueTALE software (12). Assembled genomes were submitted to the

Genbank with accession numbers: CP043540 for Xtu LW16 and CP043500 for Xtu P3.

dTALe construction and transformation

The promoter of the Tal6 effector in XT4699, which was previously shown to drive gene

expression in wheat, was used for driving the expression of dTALes in strains of Xtu (1) .

The repeat regions of dTALes were constructed using Golden Gate assembly strategies

(13,14). The dTALes driven by tal6 promoter were individually transferred into a broad

host-range vector pHM1 by restriction enzymes. The final constructs were sequenced and

transformed into M6 or LW16 strains by electroporation (Bio-rad).

Quantitative RT-PCR (qRT-PCR) analyses

Xtu strains harboring dTALes were validated by qRT-PCR analyses of targeting host

genes. The leaves were inoculated with Xtu at OD600nm=0.2 and were harvested at 1DPI

for RNA isolation (Trizol reagent, Invitrogen). 1μg total RNA was treated with DNaseI

5

(Invitrogen) first and applied for reverse transcription reaction (Verso cDNA Synthesis

Kit). The 10X diluted cDNA samples were subjected to real-time PCR reactions in the

96-well CFX machine (Bio-rad). The wheat EF-1α gene was used as the control and the

relative gene expression level was calculated using 2-∆∆Ct method (15). Primer sequences

are available in Table S2.

Transient GUS assay in Nicotiana benthamiana

Primers (Table S2) for the predicted EBETal8 of TaNCED_5BS and TaNCED_5DS were

annealed and ligated into upstream of CsLobT. The synthetic promoters were cloned into

pBI101 (Clonetech) to fuse upstream of uidA. Approximately 500 bp of native promoters

of TaNCED_5S were PCR amplified using specific primers (Table S2) and fused with

uidA. The constructs were individually transformed into Agrobacterium LBA4404. The

full length tal8 was PCR amplified using primers (Table S2), fused under 35S promoter

in pBICaMV (16), and then transformed into Agrobacterium strain LBA4404. 1:1 ratio

mixture of Agrobacterium (OD600nm=0.3) with designated promoter::uidA and tal8 was

infiltrated onto young leaves of 1-month old N. benthamiana plants by needleless

syringe. The N. benthamiana plants were maintained at 24°C with 12-h photoperiod in

the lab room. Leaf discs of inoculated areas were punched and collected 3 days later to

test the GUS activity in qualitative and quantitative manners as described (16).

ABA quantification assay

The 2nd leaves of 11-day-old Jagger plants were infiltrated with bacterial suspensions

(OD600nm=0.2) of X. translucens strains by needleless syringe. The inoculated leaves were

collected at 1 DPI, weighed at scale, frozen with liquid nitrogen and stored at -80°C. The

endogenous ABA was extracted following the protocol as described (17). The

6

quantification of ABA was performed with Phytodetek ABA Test Kit following

directions of the manufacturer.

Application of ABA and effect of Tal8 in wheat leaves

The 10-day-old or 14-day-old Jagger plants were sprayed with ABA (100µM) coated

with 0.02 % Silwet L–77. The water-soaking streak phenomenon induced by the sprayed

ABA was observed in one day. At this time, 2nd leaves of 10-day-old or 3rd leaves of 14-

day-old ABA treated plants and control plants (sprayed without ABA) were infiltrated

with bacterial suspension (OD600nm=0.2) to form a water-soaking spot by needleless

syringe. The symptoms were photographed and scored by measuring the lesion length at

7 DPI. The 2nd leaves of 12-day-old Jagger plants were inoculated by bacterial suspension

(OD600nm=0.2) and at 1 DPI the inoculated leaves were placed in the middle of chamber

of LI-COR 6800 for measurement following the manual. GasEx E values were recorded

to determine the transpiration rate. For water loss rate measurement, four infiltrated 2nd

leaves of 12-day-old Jagger at 1DPI were excised, pooled and weighed at time points of

0, 20, 40, 60, 120, 180min. The water loss rate was calculated as percentage of initial

fresh weight (18).

SA measurement and application

The 2nd leaves of 12-day-old Jagger plants were inoculated by X. translucens pv.

undulosa strains (OD600nm=0.2). The inoculated leaves were sampled at 1 DPI, 2 DPI and

3 DPI. To quantify total and free SA a GC-MS method was adapted from Schmelz et.al.,

2004 (19). Briefly tissues were flash frozen in liquid nitrogen and then c.a. 100mg

extracted in 600µl of H2O:1-propanol:HCL (1:2:0.005) spiked with 200ng of d4-SA as an

internal standard. Samples were homogenized using 1g of ceramic beads and partitioned

7

into two equal samples one for free SA and one for total SA. For total SA 2µl of glacial

HCL was added to the sample, and the sample was boiled for 30 min in a water bath and

allowed to cool to 25 °C. Both free and total SA samples were then extracted with 1ml

MeCl2. Following centrifugation, the MeCl2:1-propanol layer was collected and

derivatized with trimethylsilydiazomethane, quenched with acetic acid in hexane and

compounds collected by vapor phase extraction on SuperQ columns. Compounds were

then eluted in MeCl2 and analyzed using GC-MS. One day before inoculation, the 1mM

SA solution coated with 0.02 % Silwet L–77 were sprayed onto leaves or the plant soil

was thoroughly soaked by 1mM SA solution. The 2nd leaves of 10-day-old Jagger were

used for inoculation.

8

Fig. S1

9

Fig. S1. Tal8 is a virulence effector for XT4699. (A and B) M6 mutant shows loss of

virulence compared to wild type XT4699, complemented M6 [M6(tal8)], and mutants

M1-5, M7, and M8 on wheat cultivars cv. Jagger (A) and cv. Chinese Spring(B) as

measured by lesion length. *** indicates reduced virulence of M6 (P-value < 0.001)

compared to XT4699 with Student’s t-test. (C) Representative leaf inoculations on cv.

Chinese Spring at 7DPI with strains indicated above panel. (D and E) Total and surface

bacterial populations were not measurably different between XT4699, M6(tal8) and M6

on cv. Jagger (D) or cv. Chinese Spring (E).

10

Fig. S2

Fig. S2. Tal8 amino acid sequence and repeat variable di-amino acid (RVD)

residues. (A) Tal8 amino acid sequence from NCBI accession AKK68514. The repeat-di-

residues (RVD) of each repeat are in bolded font. (B) Single letter code for the RVD of

Tal8 effector. The lowercase letters g and d were used for indication of unusual di-amino

acid residues KG and QD at the 12th and 13th positions. The RVDs KG, QD, and Y*

were treated as equivalent to NG, HD, and N* for purposes of EBE analysis.

>Tal8 NCBI Accession AKK68514, locus tag FD63_14025 CP008714 REGION:

complement(3256565..3260576) MDPIRSRTPSPARELQAGSQPDAVQPIADRLVSPPAGSPLDGLPARRTMSRTQLPSPPASVPAFSAGSFSDLLRQ

VDSSLFDASFFDSMPAFGAHHAQAATGELDEVQSALRAADDPQPPVRVAVTAARPPRAKPAQRPRRAAQTSDASP

AADVDLSTFGYSQQQQEKIKPTVRSTVAQHHAALVGHGFTHAHIVELSKHPAALGTIAARYSEMIAALPEATHED

IVGVGKQWSGARALEALLMVAEELRAPPLQLVTGQLLKIAKRGGVTAVEAVHASRNALTGAPLH LTPDQVVAIVSNNGGKQALETVQRLLPVLCKPPYG LTPEQVVAIASNGGGKPALETVQRLLPVLCQEYG LTPNQVVAIASHDGAKQALETVQRLLPVLCQPPHP LTPNQVVAIASHDGGKPALETVQRLLPVLCKPPYG LTPEQVVAIANHDGAKQALETVQRLLPVLCQPPYG LTPEQVVAIASKGGGKQALETVQRLLPVLCQPPYG LTPDQVVTIASNNGGKPALETVRRLLPVLCKPPYG LTPKQVVAIASY*GGKQSLETVQRLLPVLCKPPYG LTPEQVVAIASNGGGKPALETVQRLLPVLCQEYG LTPNQVVAIASHDGAKPALETVQRLLPVLCQEYG LTPNQVVAIASHDGAKQALETVQRLLPVLCKPPYG LTPEQVVAIASQDGGKQSLETVQRLLPVLCKPPYG LTPEQVVAIASHNGGTQALESIFAQLSSPDPALAALTNGRLVALACIGGRPALDAVKKGLPHAPALITRVHNRVP

EGTAHLVADLAQVVRVLSFFQCHSHPAQAFDEAMRQFGMSRHGLLQLFRRVGVTELEAISGTLPPASQRWDRMLQ

ASGRKGAKPSSASAQTPSQESVDAFADSLERELDAHSPMHQAGQTLASSRKRSRSESSVNRSSAQQAAEVFVPEQ

RDAPPLLPLSSWGVKRRRTRIGGLPDPGTPTHGDLAASSAAFLEQDADPFAGAAEDFPAFDQEEIAWLKELLAH

A

Single letter code for RVD of Xtu 4699 Tal8 NN NG HD HD HD KG NN Y* HG HD HD QD HN N G D D D g N * G D D d N

B

11

Fig. S3

12

Fig. S3. Promoter sequences and predicted EBETal8 of TaNCED and TaERF. (A) The

predicted alignments are shown for RVDs of Tal8 and EBEs of TaNCED_5BS,

TaNCED_5DS and TaNCED_5AS. Nonconsensus nucleotide in the RVD pairings are

highlighted in yellow. The alignment of EBEs for all three genes is shown above the

sequences for the respective promoter regions. The start codon ATG is highlighted in

green. Concensus nucleotide matches in EBEs are highlighted in turquoise. The putative

TATA box is underlined. (B) The alignment of RVD of Tal8 with EBEs in the promoter

regions of TaERF_1BL, TaERF_1DL and TaERF_1AL. Highlighting as in A.

13

Fig. S4

Fig. S4. Sequence analysis of X. translucens pv. undulosa strains P3 and LW16. (A)

TAL effector genes (arrow) are located in same positions of the genomes in LW16, P3,

and XT4699. (B) Dot plot analysis of LW16 (y-axis) versus P3 (x-axis). Red dots

represent forward alignments. Blue dots represent reverse alignments (inversions).

0

500000

1000000

1500000

2000000

2500000

3000000

3500000

4000000

4500000

0 500000 1000000 1500000 2000000 2500000 3000000 3500000 4000000 4500000

lw16r1

tig00000001|arrow|join_near_2309204|pilon

P3

Lw1

6

B

1 2 3 4 5 6 7 8

1 2 3 4 5 6 7 8

LW16

P3

A

1 2 3 4 5 6 7 8 XT4699

14

Fig. S5

Fig. S5. Alignment of RVDs of TAL effectors of XT4699, LW16 and P3 strains of X.

translucens pv. undulosa. LW16-Tal8 RVDs are marked with red and do not align with

RVDs from Tal8 of P3 and XT4699. Other TAL effectors were highly similar among

three strains except variable RVDs, which were marked with purple highlight. The

missing 13th amino acid in the repeat was represented by *.

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 XT4699-Tal1 HD YD NI NG NG NN YK NG HD NG NG ND NG QD NH HD P3-Tal1 HD YD NI NG NG NN YK NG HD NG NG ND NG QD NH HD LW16-Tal1 HD YD NI NG NG NN YK NG HD NG NG ND NG QD NH QD XT4699-Tal2 NN HD NG NN HN KG NI HD NI NN HD HN HD HD NI HN HD QD P3-Tal2 NN HD NG NN HN KG NI HD NI NN HD HN HD HD NI HN HD QD LW16-Tal2 NN HD NG NN HN KG NI HD NI HN HD HN HD HD NI HN HN HD XT4699-Tal3 NN HD NG HD HD HN NF NI NH HD HD HD HN HN HD P3-Tal3 NN HD NG HD HD HN NF NI NN NN HD HD HN HN HD LW16-Tal3 NN HD KG HD HD HN NF NI NN HD HD HD HN HN HD XT4699-Tal4 NH NN HD NN HD NH HD YK NG NH Y* HD NN NI NG QD P3-Tal4 NH NN HD NN HD NH HD YK NG NH Y* HD NN NI NG QD LW16-Tal4 NH NN HD NN HD NH HD YK NG NH Y* HD NN NI NG QD XT4699-Tal5 NN HD NG NN HN HN NI NI NI NH NN HD NN NH HD HD P3-Tal5 NN HD NG NN HN HN NI NI NI NH NN HD NN NH HD HD LW16-Tal5 NN HD NG NN HN HN NN NI NI NH NN HD NN NH HD HD XT4699-Tal6 HD HN HN HD NH NH HG HD KG NN Y* NG HD HD HN P3-Tal6 HD HN HN HD NH NH HG HD KG NN Y* NG HD HD HN LW16-Tal6 HD HN HN HD NH NH HG HD KG NN Y* NG HD HD HN XT4699-Tal7 HN HD HD HD NI NI NI HN HD HD NN NN NI NN HD P3-Tal7 HN HD HD HD NI NI NI HN HD HD NN NN NI NN HD LW16-Tal7 NN HD HD HD NI NI NI HN HD HD NN NN NI NN HD XT4699-Tal8 NN NG HD HD HD KG NN Y* NG HD HD QD HN P3-Tal8 NN NG HD HD HD KG NN Y* NG HD HD QD HN LW16-Tal8 NN HD NG NN HN NG NI HD NI NN HD HD NN NN NI HN HD

15

Fig. S6

Fig. S6 TaNCED_5BS expression is associated with enhanced virulence of M6. (A)

Empty vector (ev) and dTALes, dERF101 and dNCED63, designed specifically for

16

TaERF_1BL and TaNCED_5BS, were transformed into the tal8 mutant strain M6.

M6(dERF1011) and M6(dERF1012) are two independent transformants of dERF101.

qRT-PCR results revealed the specific inductions of gene by corresponding dTALes. (B)

Lesion length measurements at 7 DPI showed only the dNCED63 transformants

enhanced the virulence for M6 on the 2nd leaves of 10-day-old Jagger plants. *** -

indicates values differ in the significance at the p-value < 0.001 relative to lesion lengths

of M6(ev) with Student’s t test. ns – values non-significant difference relative to lesion

lengths of M6(ev).

17

Fig. S7

18

Fig. S7. Phylogenetic analysis of NCED and presence and effects of salicylic acid in

wheat leaves after infection. (A) Protein sequences of NCED from Arabidopsis, rice,

maize and wheats were analyzed and aligned. Phylogenetic tree was generated by

Neighbor-joining method in the Geneious software with AtCCD1 sequence as outgroup.

Upregulated wheat genes are indicated by red line. Genbank accession numbers of

proteins are: AtCCD1 (NP_191911),AtNCED2 (NP_193569), AtNCED3(NP_188062),

AtNCED4(NP_193652), AtNCED5(NP_174302), AtNCED6(NP_189064),

AtNCED9(NP_177960), OsNCED1(XP_015626662),

OsNCED2(XP_015619611),OsNCED3(XP_015631538), OsNCED4(XP_015645858),

OsNCED5(XP_015618707), ZmVP14(AAB62181). Protein sequences are provided in

Table S4. (B and C) Free SA (B)and total SA (C) from inoculated wheat leaf samples

were quantified by GC-MS. Bacterial strains LW16(ev), LW16(dNCED63) and

LW16(tal8) were inoculated onto 2nd leaves of 12-day-old Jagger. Inoculated samples

with bacterial suspensions (OD600nm=0.2) were harvested at 1 DPI, 2 PDI and 3 DPI.

Values indicate mean ± standard error. Values with the same lowercase letters do not

differ significantly (p-value < 0.05) using Tukey statistics and ANOVA analysis. Details

for SA measurements are described in Methods and Materials. (D and E) 9-day-old

Jagger plants were sprayed (D) with 1mM SA or soil-drenched (E) with 1mM SA,

respectively. Control plants were not treated with SA. The 2nd leaves of Jagger plants

were inoculated one day after treatment. Each bar signifies the range of mean ± standard

deviation. The lower-case letters show significantly different groups (p-value < 0.05) in

the Tukey statistic and ANOVA analysis.

19

Table S1

Table S1. Bacterial strains and plasmids used in this study.

Xtu strains Notes

XT4699 WT strain (2)

LW16 WT strain (2)

XT130 WT strain(2)




LB10 WT strain (2)

LG48 WT strain (2)

P3 WT strain (2) M1 XT4699 tal2 mutant (2)

M2 XT4699 tal6 mutant (2)







M6 (ev) M6 strain harboring empty broad host range cloning vector pHM1

M6 (tal8) M6 strain harboring tal8 in pHM1

M6 (dNCED63) M6 harboring dNCED63 in the pHM1 M6 (dERF1011) harboring dERF101-1 in pHM1, independent transformant of M6 M6 (dERF1012) harboring dERF101 -2 in pHM1, independent transformant of M6 LW16 (ev) LW16 harboring empty vector pHM1

LW16 (tal8) LW16 harboring tal8 gene in pHM1

LW16 (dNCED41) LW16 harboring dNCED41 in pHM1




LW16 (dERF3) LW16 harboring dERF3 in pHM1




Agrobacterium

LBA4404 WT strain from Dr. Sunghun Park at Kansas State University

LBA4404 (tal8) LBA4404 strain with tal8 coding sequence fused to 35S CaMV promoter

LBA4404 (pthA4) LBA4404 strain with overexpressed pthA4 (Hu et al., 2014)

LBA4404 (NCEDB) LBA4404 strain with native promoter of TaNCED_5BS fused upstream of uidA

gene in the pBI101

LBA4404 (NCEDD) LBA4404 strain with native promoter of TaNCED_5DS fused upstream of uidA

gene in pBI101

LBA4404 (CsLob) LBA4404 strain with native promoter CsLob1 fused upstream of uidA gene in

pBI101

LBA4404 (CsLobT) LBA4404 strain with truncated promoter CsLobT fused upstream of uidA gene

in pBI101

LBA4404 (EBEB-

CsLobT)

LBA4404 strain with synthetic promoters consisting of EBETal8 of TaNCED_5BS

placed upstream of CsLobT, and then fused upstream of uidA gene in pBI101

LBA4404 (EBED-

CsLobT)

LBA4404 strain with synthetic promoters consisting of EBETal8 of

TaNCED_5DS placed upstream of CsLobT, and tfused upstream of uidA gene in

pBI101

20

Plasmids

pHM1 Broad host-range vector for Gram-negative bacteria (White lab)

pBICaMv Modified vector from pBI121, suitable for overexpressing genes under CaMV

35S promotor (Hu et al., 2015)

pBI101 Clonetech vector

pBI101-CsLobT Truncated CsLob1 promoter inserted upstream of uidA gene in the pBI101 vector

(Hu et al., 2014)

Peng, Z. et al., (2016) BMC Genomics, 17, 21.

Hu, Y. et al., (2014) Proc Natl Acad Sci U S A, 111, E521-9.

Hu, Y. et al., (2015) Horticulture Research, 2.

21

Table S2

Table S2. Primers used in this study. Primers used in qRT-PCR

TaNCED-qF1 TCAAGAAGCCGTACCTCAAG

TaNCED-qR1 ACGAAGTTCTCGGTGATGG

TaERF-qF ATGGCGACAATAACGACAAC

TaERF-qR CATACTCCTGGTTCTTGAATGG

TaEF-1α-qF1 CAGATTGGCAACGGCTACG

TaEF-1α-qR1 CGGACAGCAAAACGACCAAG

TaNPR1-F GAGATGCCTGGAGATAGTAGTC

TaNPR1-R CCAAGAGTTATCCGTGAATCAAT

Primers used in transient GUS assay

NCED-pF1 AATCGCATGCCCCCCACATGAGCATACATA

NCED-pF2 AATCGCATGCGAAAGAAAAGAAGAGTGGAGAG

NCED-pR ATCATCTAGAGCTGATCCTYGGAAGAAGAA

NCED5B-FOR AGCTCTGTCCCTTCTCCCCCCCTCCCAAA

NCED5B-REV CTAGTTTGGGAGGGGGGGAGAAGGGACAG

NCED5D-FOR AGCTCTGTCCCGTCTCCCCCCCTCCCAAA

NCED5D-REV CTAGTTTGGGAGGGGGGGAGACGGGACAG

Tal8-OX-F actaTCTAGAATGGATCCCATTCGTTCTCG

Tal8-OX-R atgaGAATTCTCAATGAGCCAATAGCTCCTTC

22

Table S3

Table S3. TAL effectors of Xtu P3 (high virulence) and LW16 (low virulence)a

P3-Tal1

MDPIRSRTPSPARELLAGSQPDGVQPTADPRVSPPAGSPLDGLPARRTMSRTQLPPPPASGPAFSAGSFSD

LLRQVDSSLFNASLFDSMPAFGAHHAQAATGELDEAQSALRAVDDPQPSASAAITAAPPRTKAAARRRSAQ

TLDALPAADVDISTFGYSQQQQEKIKPKVRSTVAQHHAALVGHGFTHAHIVELSKHPPALGTIAARYSEMI

AALPEATHEDIVGVGKHCAGARTLEVLLMVVQELRAPPLQLVTSQLLKIAKRGGVTAVEAVHASRNALTGA

PLHLLPDQVVAIVSHDGGKQSLETVQRLLPVLRQPPYGLTPNQVVAIASYDGGKQSLETVERLLPVLCQPP

YGLTPNQVVAIASNIGGKPALETVQRLLPVLCQPPYGLTPNQVVAIASNGGGKQALETVQRLLPVLCQEYG

LIPEQVVAIASNGGGKQALETVQRLLPVLCKEYGLTPEQVVAIASNNGGKLALETVERLLPVLCQPPYGLT

PKQVVAIASYKGANQALGTVQRLLPVLCKPPYGLTPDQVVAIASNGGAKQALETVERLLPVLCKPPYGLTP

DQVVAIASHDGGRQALETVQRLLPVLCQPPYGLTPNQVVAIASNGGGKQALETVQRLLPVLCQEYGLTRQQ

VVAIASNGGGKQSLETVQRLLPVLCKPPYNLTPDQVVAIASNDGGKQALETVQRLLPVLCKPPYNLTPEQV

VAIASNGGGKQALETVQRLLPVLCKEYGLTPEQVVAIASQDGGKQSLETVQRLLPVLCQLPYGLTPNQVVA

IASNHGGKQSLETVQRLLPVLCQPPYGLTPNQVVAIASHDGGTQALESIFAQLSSPDPALAALTNDRLVAL

ACIGGRPALDAVKKGLPHAPALITRVHNRVPEGTAHLVADHAQVVRVLSFFQCHSQRGQVFHEAMKRFEMS

REGLLQLFRRVGVTELEAISGTLPPASQRWDRMLQASGRKGAKPPSASAQTQGQESLDAFADSLERELDAP

SPMHQAGQALASSRKRSRSASSVNRSSAQQAAEVFVPEQRDAPPLLPLSSWGVKRRRTRIGGLPDPGTPTH

GDLAASSAAFLEQDADPFAGAAEDFPAFDQEEIAWLKELLAH

P3-Tal2

MDPIRSRTPSPARELQAGSQPDGVQPTADPRVSPPAGSPLDGLPARRTMSRTQLPPPPASVPAFSAGSFSD

LLRQVDSSLFDASFFDSMPAFGAHHAQAATGELDEVQSALRAADDPQPPVRVAVTAARPPRAKPAQRPRRA

AQTSDASPAADVDLSTFGYSQQQQEKIKPTVRSSVAQHHAALVGHGFTHAHIVELSKHPAALGTIAARYSE

MIAALPEATHEDIVGVGKQWSGARALEALLMVAEELRAPPLQLVTGQLLKIAKRGGVTAVEAVHASRNALT

GAPLHLTPDQVVAIVSNNGGKQALETVQRLLPVLCQPPYGLTPEQVVAIASHDGAKPALETVQRLLPVLCQ

PPYGLTPEQVVAIASNGGGKQALETVQRLLPVLCKEYGLTPEQVVAIASNNGGKLALETVERLLPVLCQPP

YGLTPKQVVAIASHNGGKQALETVQRLLPVLCQPPYGLTPEQVVAIASKGGGKQALETVQRLLPVLCQPPY

GLTPDQVVTIANNIGAKQALETVQRLLPVLCQPPYGLTPNQVVAIASHDGGKQALETVQRLLPVLCQEYGL

TPGQVVAIANNIGGKPALETVQRLLPVLCQPPYNLTPNQVVAIASNNGGKQALETVQRLLPVLCKPPHPLT

PNQVVAIASHDGAKQALETVQRLLPVLCKSPYGLTPDQVVVIASHNGGKQALETVQRLLPVLCKPPYGLTP

EQVVAIASHDGGKPALETVQRLLPVLCQEYGLTPNQVVAIASHDGGKQALETVQRLLPVLCQDHGLTPGQV

VAIANNIGGKPALETVQRLLPVLCKEYGLTPNQVVAIASHNGGKQALETVQRLLPVLCQPPYGLTPEQVVA

IASHDGGKPALETVQRLLPVLCQPPYGLTPNQVVAIASQDGGTQALESIFAQLSSPDPALAALTNDRLVAL

ACIGGRPALDAVKKGLPHAPALITRVHNRVPEGTAHLVADHAQVVRVLSFFQCHSQRGQVFHEAMKRFEMS

REGLLQLFRRVGVTELEAISGTLPPASQRWDRMLQASGRKGAKPPSASAQTQGQESLDAFADSLERELDAP

SPMHQAGQTLASNRKRSRSESSVNRSSAQQAAEVFVPEQRDAPPLLPLSSWGVKRRRTRIGGLPDPGTPTH


P3-Tal3

MDPIRSRTPSPARELLAGSQPDGVQPTADPRVSPPAGSPLDGLPARRTMSRTQLPPPPASGPSFSAGSFSD

LLRQVDSSLFDASFFDSMPAFGAHHAQAATGELDEAQSALRAVDDPQPSASAAITAAPPRTKAAARRRSAQ

TSDALPAADVDLSTFGYSQQQQEKIKPKVRSTVAQHHAALVGHGFTHAHIVELSKHPPALGTIAARYSEMI

AALPEATHEDIVGVGKQWSGARALEALLMVAEELRAPPLQLVTGQLLKIAKRGGVTAVEAVHASRNALTGA

PLHLTPDQVVAIVSNNGGKQALETVQRLLPVLCQPPYNLTPEQVVAIASHDGAKPALETVQRLLPVLCQPP

YGLTPEQVVAIASNGGKQALEAVQRLLPVLCKEYGLTPEQVVAIASHDGAKQALETVQRLLLVLCQEYGLT

PNQVVAIASHDGAKQALETVQRLLPVLCQPPYGLTPNQVVAIASHNGGKQALETVQRLLPVLCQPPYGLTP

EQVVTIANNFGGKPALETVQRLLPVLCQPPYGLTPNQVVAIANNIGAKPALETVQRLLPVLCQPPYGLTPE

QVVAIAGNNGAKQALETVQRLLPVLCQPPYGLTPEQVVAIAGNNGAKQALETVQRLLPVLCQPPHPLTPNQ

VVAIASHDGAKQALETVQRLLPVLCKPPYGLTPDQVVAIASHDGAKQALETVQRLLPVLCQPPYGLTPDQV

VAIASHNGGKQALETVQRLLPVLCQPSYGLTRNQVVAIASHNGGKQALETVQRLLPVLCKEYGLTPEQVVA

IASHDGGTQALESIFAQLSSPDPALAALTNDRLVALACIGGRPALDAVKKGLPHAPALITRVHNRVPEGTA

HLVADHAQVVRVLSFFQCHSQRGQVFHEAMKRFEMSREGLLQLFRRVGVTELEAISGTLPPASQRWDRMLQ

ASGRKGAKPPSASAQTQGQESLDAFADSLERELDAPSPMHQAGQTLASSRKRSRSESSVNRSSAQQAAEVF

23

VPEQRDAPPLLPLSSWGVKRRRTRIGGLPDPGTPTHGDLAASSAAFLEQDADSFAGAAEDFPAFDQEEIAW

LKELLAH

P3-Tal4

MEPIRSRTPSTARELQAGSQPDAVQPIADRLVSTPASSPLDGLPARRMVSRTSPPPSPARSPAFSAGSLSG

LLRQQIDPSLFAGSPFDSLPSFGAARAESAPGEGDEVQSGLRAVDDPQPSASAAITAAPPRTKAAARRRSA

QTSDALPAAHVDLGTFGYSQQQQEKIKPKVRSTLAQHHEALVGHGFTHAHIVELSKHPPALGTIAARYSEM

IAALPEATHEDIVGVGKQKSGARALEALLTVAEELRAPPLQLVTGQLLKIAKRGGVNAVEAVHASRNALTG

APLHLLPDQVVAIVSNHGSKLALGTVQRLLPVLCKPPYGLTRNQVVAIVNNNGGKQALETVHRLLPVLCQP

PYGLTPEQVVAIASHDGGRQALETVHRLLPVLRQPPYGLTLEQVVAIASNNGGKQALETVQRLLPVLCQPP

YGLTPDQVVTIASHDGGRQALETVQRLLPVLCQPPYGLTPNQVVAIASNHGGKQALETVQRLLPVLCQPLH

GLTPDQVVTIASHDGGRQALETVQRLLPVLCQPPYGLTPKQVVAIANYKGAKQALETVQRLLPLLCKPPYG

LTPDQVIAIVSNGGGKPALETVRRLLPVLCKHPYGLTPKQVVAIVSNHGGKPALETVRRLLPVLCEHPYGL

TPKQVVAIASYGGKPALETVQRLLPVLCQPPYGLTPNQVVAIASHDGGRQALETVQRLLPVLRQPPYGLTP

DQVVAIASNNGGKQALETVQRLLPVLRQPPYSLIPDQVVAIASNIGAKQSLETVQRLLPVLCQPPYGLTTD

QVIAIASNGGSKQALETVQRLLPVLCQPPYGLTPDQVVAIASQDGGKQALESIFAQLSSPDPALAALTNDR

LVALACLGGRPALDAVKKGLPHAPALVTRTHNRVPEGTAHLVADHAQVVRVLGFFQCHSHPAQAFHEAMTR

FEMSREGLLQLFRRVGVTELEAISGTLPPASQRWHRILQALGVKGAKPSSASAQTPSQESVDAFADSLERE

LDAPSPIHEADRARASNRKRSRSESSVNRSSAQQAAEVFVPEQRDAPPLLPLSSWGVKRQRTRIGGLPDPG

MPTDGELAASSAAFLEQDADPFAGAAEDFPVFDQEEIAWLMTLLPH

P3-Tal5

MDPIRSRTPSPARELQAGSQPDAVQPIADRLVSPPAGSPLDGLPARRTMSRTQLPSPPASVPAFSAGSFSD


AQTSDASPAADVDLSTFGYSQQQQEKIKPTVRSTVAQHHAALVGHGFTHAHIVELSKHPPALGTIAARYSE


GAPLHLTPDQVVAIVSNNGGKQALETVERLLPVLCQPPYGLTPEQVVAIASHDGAKPALETVQRLLPVLCQ

PPYGLTPEQVVAIASNGGGKPALETVQRLLPVLCKEYGLTPEQVVAIASNNGGKLALETVERLLPVLCQPP

YGLTPNQVVAIASHNGGKQSLETVQRLLPVLCQPPYGLTPEQVVAIASHNGGKQALETVQRLLPVLCQPPY

GLTPEQVVTIANNIGGKQALETVERLLPVLCQPPYGLTPNQVVAIASNIGAKPALETVQRLLPVLCQPPYG

LTPEQVVTIANNIGGKPALETVQRLLPVLRKPPYGLTPEQVVAIASNHGGKQALETVQRLLPVLCKPPYGL

TPNQVVAIASNNGAKQALETVQRLLPVLCKPPHPLTPNQVVAIASHDGAKQALETVQRLLPVLCQPPYGLT

PEQVVVIASNNGGKQALETVQRLLPVLCQPSYGLTPEQVVAIASNHGGKQALETVQRLLPVLCQPPYGLTP

NQVVAIASHDGGKPALETVQRLLPVLCQPPYGLTPNQVVAIASHDGGTQALESIFAQLSSPDPALAALTND

RLVALACIGGRPALDAVKKGLPHAPALITRVHNRVPEGTAHLVADLAQVVRVLSFFQCHSHPAQAFDEAMR

QFGMSRHGLLQLFRRVGVTELEAISGTLPPASQRWDRMLQASGRKGAKPSSASAQTPSQESVDAFADSLER

ELDAPSPMHQAGQTLASSRKRSRSESSVNRSSAQQAAEVFVPEQRDAPPLLPLSSWGVKRRRTRIGGLPDP

GTPTHGDLAASSAAFLEQDADPFAGAAEDFPAFDQEEIAWLKELLAH

P3-Tal6



AQTSDASPAADVDLSTFGYSQQQQEKIKPTVRSTVAQHHAALVGHGFTHAHIVELSKHPAALGTIAARYSE

MIAALPEATHEDIVGVGKQWSGARALETLLMVAEELRAPPLQLVTGQLLKIAKGGGVTAVEAVHASRNALT

GAPLHLTPDQVVAIVSHDGGKQSLETVQRLLPVLCQPPYGLTPNQVVAIASHNGGKQALETVQRLLPVLCQ

EYGLTPEQVVAIASHNGGKQALETVQRLLPVLCQEYGLTPNQVVAIASHDGAKQALETVQRLLPVLCQPPY

GLTPEQVVAIASNHGGKLALETVERLLPVLCQPPYGLTPNQVVAIASNHGGKQALETVQPLLPVLCQPPYG

LTPEQVVAIASHGGAKQALKTVQRLLPVLCQDHGLTPEQVVAIANHDGAKQALETVQRLLPVLCQPPYGLT

PEQVVAIASKGGGKQALETVQRLLPVLCQPPYGLTPDQVVTIASNNGGKPALETVRRLLPVLCKPPYGLTP

KQVVAIASYGGKQALETVQRLLPVLCKPPYGLTPEQVVAIASNGGGKPALETVQRLLPVLCQEYGLTPNQV

VAIASHDGAKPALETVQRLLPVLCQEYGLTPNQVVAIASHDGGKPALETVQRLLPVLCQPPYGLTPEQVVA

IASHNGGTQALESIFAQLSSPDPALAALTNDRLVALACIGGRPALDAVKKGLPHAPALITRVHNRVPEGTA

HLVADLAQVVRVLSFFQCHSHPAQAFDEAMRQFGMSRHGLLQLFRRVGVTELEAISGTLPPASQRWDRMLQ


VPEQRDAPPLLPLSSWGVKRRRTRIGGLPDPGTPTHGDLAASSAAFLEQDADPFAGAAEDFPAFDQEEIAW

LKELLAH

24

P3-Tal7

MDPIRSRTPSPARELLAGSQPDGVQPTADPRVSPPAGSPLDGLPARRTMSRTQLPPPSASGPAFSAGSFSD

LLRQVDSSLFNASLFDSMPAFGAHHAQAATGELDEAQSALRAVDDPQSSASAAITAAPPRTKAAARRRSAQ

TLDASPAADVDLSTFGYSQQQQEKIKPTVRSSVAQHHAALVGHGFTHAHIVELSKHPAALGTIAARYSEMI


PLHLTPDQVVAIVSHNGGKQALETVQRLLPVLCQPPYNLTPEQVVAIASHDGGKQALETVQRLLLVLCQEC

GLTPNQVVAIASHDGAKPALETVHRLLPVLCQPPYGLTPNQVVAIASHDGAKQALETVQRLLPVLCQDHGL

TPGQVVAIADNIGGKQALETVQRLLPVLCKPPYGLTPEQVVTIANNIGGKPALETVQRLLPVLCQPPYGLT

PEQVVTIANNIGAKPALETVHRLLPVLRKPPYGLTPEQVVAIASHNGAKPALETVQRLLPVLCQPPYGLTP

NQVVAIASHDGAKQALETVQRLLPVLCKPPHPLTPNQVVAIASHDGAKQALETVQRLLPVLCKPPYGLTPD

QVVVIASNNGGKQALETVQRLLPVLCQPPYGLTPEQVVAIASNNGGKPALETVQRLLPVLCKPPYDLTPDQ

VVAIANNIGAKPALETVQRLLPVLCQPPYGLTPNQVVAIASNNGGKQALETVQRLLPVLCQPPYGLTPYQV

VAIASHDGGTQALESIFAQLSSPDPALATLTNDRLVALACIGGRPALDAVKKGLPHAPALITRVHNRVPEG

TAHLVADHAQVVRVLGFFQCHSQRGQVFHEAMKRFEMSREGLLQLFRRVGVTELEAISGTLPPASQRWDRM

LQASGRKGAKPPSASAQTQGQESLDAFADSLERELDAPSPMHQAGQTLASSRKRSRSESSVNRSSAQQAAE

VFVPEQRDAPPLLPLSSWGVKRRRTRIGGLPDPGMPTDGELAASSAAFCEQDADPFAGAAEDFPAFDQEEI

AWLRELLAH

P3-Tal8





GAPLHLTPDQVVAIVSNNGGKQALETVQRLLPVLCKPPYGLTPEQVVAIASNGGGKPALETVQRLLPVLCQ

EYGLTPNQVVAIASHDGAKQALETVQRLLPVLCQPPHPLTPNQVVAIASHDGGKPALETVQRLLPVLCKPP

YGLTPEQVVAIANHDGAKQALETVQRLLPVLCQPPYGLTPEQVVAIASKGGGKQALETVQRLLPVLCQPPY

GLTPDQVVTIASNNGGKPALETVRRLLPVLCKPPYGLTPKQVVAIASYGGKQSLETVQRLLPVLCKPPYGL

TPEQVVAIASNGGGKPALETVQRLLPVLCQEYGLTPNQVVAIASHDGAKPALETVQRLLPVLCQEYGLTPN

QVVAIASHDGGKPALETVQRLLPVLCKPPYGLTPEQVVAIASQDGGKQSLETVQRLLPVLCKPPYGLTPEQ

VVAIASHNGGTQALESIFAQLSSPDPALAALTNGRLVALACIGGRPALDAVKKGLPHAPALITRVHNRVPE

GTAHLVADLAQVVRVLSFFQCHSHPAQAFDEAMRQFGMSRHGLLQLFRRVGVTELEAISGTLPPASQRWDR

MLQASGRKGAKPSSASAQTPSQESVDAFADSLERELDAHSPMHQAGQTLASSRKRSRSESSVNRSSAQQAA

EVFVPEQRDAPPLLPLSSWGVKRRRTRIGGLPDPGTPTHGDLAASSAAFLEQDADPFAGAAEDFPAFDQDE

LAWLKELLAH

LW16-Tal1



TLDALPAADVDLSTFGYSQQQQEKIKPKVRSTVAQHHAALVGHGFTHAHIVELSKHPPALGTIAARYSEMI

AALPEATHEDIVGVGKHCAGARTLEVLLMVVQELRAPPLQLVTSQLLKIAKRGGVTAVEAVHASRNALTGA

PLHLLPDQVVAIVSHDGGKQSLETVQRLLPVLRQPPYGLTPNQVVAIASYDGGKQSLETVERLLPVLCQPP

YGLTPNQVVAIASNIGGKPALETVQRLLPVLCQPPYGLTPNQVVAIASNGGGKQALETVQRLLPVLCQEYG

LIPEQVVAIASNGGGKQALETVQRLLPVLCKEYGLTPEQVVAIASNNGGKLALETVERLLPVLCQPPYGMT

PKQVVAIASYKGANQALGTVQRLLPVLCKPPYGLTPDQVVAIASNGGAKQALETVERLLPVLCKPPYGLTP

DQVVAIASHDGGRQALETVQRLLPVLCQPPYGLTPNQVVAIASNGGGKQALETVQRLLPVLCQEYGLTRQQ

VVAIASNGGGKQSLETVQRLLPVLCKPPYNLTPDQVVAIASNDGGKQALETVQRLLPVLCKPPYNLTPEQV

VAIASNGGGKQALETVQRLLPVLCKEYGLTPEQVVAIASQDGGKQSLETVQRLLPVLCQLPYGLTPNQVVA

IASNHGGKQSLETVQRLLPVLCQPPYGLTPNQVVAIASQDGGKQALESIFAQLSSPDPALAALTNDRLVAL

ACIGGRPALDAVKKGLPHAPALITRVHNRVPEGTAHLVADLAQVVRVLSFFQCHSHPAQAFDEAMRQFGMS

RHGLLQLFRRVGVTELEAISGTLPPASQRWDRMLQASGRKGAKPPSASAQTQGQESLDAFADSLERELDAP

SPMHQAGQTLASSRKRSRSESSVNRSSAQQAAEVFVPEQRDAPPLLPLSSWGVKRRRTRIGGLPDPGTPTH


LW16-Tal2



TLDASPAADVDLSTFGYSQQQQEKIKPTVRSTVAQHHAALVGHGFTHAHIVELSKHPAALGTIAARYSEMI

25


PLHLTPDQVVAIVSNNGGKPALETVQRLLPVLCQPPYGLTPNQVVAIASHDGAKPALETVQRLLPVLCQPP

YGLTPEQVVAIASNGGGKQALETVQRLLPVLCKEYGLTPEQVVAIASNNGGKLALETVERLLPVLCQPPYG

LTPKQVVAIASHNGGKQALETVQRLLPVLCQPPYGLTPEQVVAIASKGGGKQALETVQRLLPVLCQPPYGL

TPDQVVTIANNIGAKQALETVQRLLPVLCQPPYGLTPNQVVAIASHDGAKQALETVQRLLPVLCQEYGLTP

GQVVAIANNIGGKPALETVQRLLPVLCQPPYNLTPNQVVAIASHNGGKQALETVQRLLPVLCKPPHPLTPN

QVVAIASHDGAKQALETVQRLLPVLCKSPYGLTPDQVVVIASHNGGKQALETVQRLLPVLCKPPYGLTPNQ

VVAIASHDGGKPALETVQRLLPVLCQPPYGLTPDQVVAIASHDGGKQALETVQRLLPVLCQDHGLTPGQVV

AIANNIGGKPALETVQRLLPVLCKPPYGLTPDQVVAIASHNGGKQALETVQRLLPVLCQPSYGLTRNQVVA

IASHNGGKQALETVQRLLPVLCKEYGLTPEQVVAIASHDGGTQALESIFAQLSSPDPALAALTNDRLVALA

CIGGRPALDAVKKGLPHAPALITRVHNRVPEGTAHLVADHAQVVRVLGFFQCHSQRGQVFHEAMKRFEMSR

EGLLQLFRRVGVTELEAISGTLPPASQRWHRILQALGVKGAKPSSASAQTQGQESLDAFADSLERELDAPS

PMHQAGQTLASSRKRSRSESSVNRSSAQQAAEVFVPEQRDAPPLLPLSSWGVKRRRTRIGGLPDPGTPTHG

DLAASSAAFLEQDADPFAGAAEDFPAFDQEEIAWLKELLAH

LW16-Tal3

MDPIRSRTPSPARELQAGSQPDAVQPIADRLVSPAAGSPLDGLPARRTMSRTQLPSPPASVPAFSAGSFSD



MIAALPEAAHEDIVGVGKQWSGARALEALLMVAEELRAPPLQLVTGQLLKIAKRGGVTAVEAVHASRNALT

GAPLHLTPDQVVAIVSNNGGKQALETVQRLLPVLCQPPYNLTPEQVVAIASHDGAKPALETVQRLLPVLCQ

PPYGLTPEQVVAIASKGGGKQALETVQRLLPVLCQPPYGLTPEQVVAIASHDGAKQALETVQRLLLVLCQE

YGLTPNQVVAIASHDGAKQALETVQRLLPVLCQPPYGLTPNQVVAIASHNGGKQALETVQRLLPVLCQPPY

GLTPEQVVTIANNFGAKPALETVQRLLPVLCQPPYGLTPNQVVAIANNIGAKPALETVQRLLPVLCQPPYG

LTPEQVVAIAGNNGAKQALETVQRLLPVLCQPPYGLTPNQVVAIASHDGAKQALETVQRLLPVLCKPPHPL

TPNQVVAIASHDGAKQALETVQRLLPVLCKPPYGLTPDQVVAIASHDGAKQALETVQRLLPVLCKPPYGLT

PDQVVAIASHNGGKQALETVQRLLPVLCQPSYGLTRNQVVAIASHNGGKQALETVQRLLPVLCKEYGLTPE

QVVAIASHDGGTQALESIFAQLSSPDPALAALTNDRLVALACIGGRPALDAVKKGLPHAPALITRVHNRVP

EGTAHLVADLAQVVRVLSFFQCHSHPAQAFDEAMRQFGMSRHGLLQLFRRVGVTELEAISGTLPPASQRWD

RMLQASGRKGAKPPSASAQTQGQESLDAFADSLERELDAPSPMHQAGQTLASSRKRSRSESSVNRSSAQQA

AEVFVPEQRDAPPLLPLSSWGVKRRRTRIGGLPDPGTPTHGDLAASSAAFLEQDADPFAGAAEDFPAFDQE

EIAWLKELLAH

LW16-Tal4

MEPIRSRTPSTARELQAGSQPDAVQPIADRLVSTPASSPLDGLPARRMVSRTSPPPSPARSPAFSAGSLSG

LLRQQIDPSLFAGSPFDSLPSFGAARAESAPGEGDEVQSGLRAVDDPQPSASAAITAAPPRTKAAARRRSA

QTSDALPAAHVDLGTFGYSQQQQEKIKPKVRSTLAQHHEALVGHGFTHAHIVELSKHPPALGTIAARYSEM

IAALPEATHEDIVGVGKQKSGARALEALLTVAEELRAPPLQLVTGQLLKIAKRGGVNAVEAVHASRNALTG

APLHLLPDQVVAIVSNHGSKLALGTVQRLLPVLCKPPYGLTRNQVVAIVNNNGGKQALETVHRLLPVLCQP

PYGLTPEQVVAIASHDGGRQALETVHRLLPVLRQPPYGLTLEQVVAIASNNGGKQALETVQRLLPVLCQPP

YGLTPDQVVTIASHDGGRQALETVQRLLPVLCQPPYGLTPNQVVAIASNHGGKQALETVQRLLPVLCQPLH

GLTPDQVVTIASHDGGRQALETVQRLLPVLCQPPYGLTPKQVVAIANYKGAKQALETVQRLLPLLCKPPYG

LTPDQVIAIVSNGGGKPALETVRRLLPVLCKHPYGLTPKQVVAIVSNHGGKPALETVRRLLPVLCKHPYGL

TPKQVVAIASYGGKPALETVQRLLPVLCQPPYGLTPNQVVAIASHDGGRQALETVQRLLPVLRQPPYGLTP

DQVVAIASNNGGKQALETVQRLLPVLCQPPYSLIPDQVVAIASNIGAKQALETVQRLLPVLCQPPYGLTTD

QVIAIASNGGSKQALETVERLLPVLCQPPYGLTPDQVVAIASQDGGKQALESIFAQLSSPDPALAALTNDR

LVALACLGGRPALDAVKKGLPHAPALVTRTHNRVPEGTAHLVADHAQVVRVLGFFQCHSHPAQAFHEAMTR

FEMSREGLLQLFRRVGVTELEAISGTLPPASQRWHRILQALGVKGAKPSSASAQTPSQESVDAFADSLERE

LDAPSPIHEADRARASNRKRSRSESSVNRSSAQQAAEVFVPEQRDAPPLLRLSSWGVKRQRTRIGGLPDPG

MPTDGELAASSAAFCEQDADPFAGAAEDFPVFDQEEIAWLMTLLPH

LW16-Tal5

MDPIRSRTPSPARELQAGSQPDAVQPIADRLVSPAAGSPLDGLPARRTMSRTQLPSPPASVPAFSAGSFSD

LLRQVDSSLFDASFFDSMPAFGAHHAQAATGELDEVQSALRAADDPQPSASAAITAAPPRTKAAARRRSAQ


AALPEATHEDIVGVGKQWSGARALETLLMVAEELRAPPLQLVTGQLLKIAKRGGVTAVEAVHASRNALTGA

PLHLTPDQVVAIVSNNGGKQALETVERLLPVLCQPPYGLTPNQVVAIASHDGAKPALETVQRLLPVLCQPP

26

YGLTPEQVVAIASNGGGKPALETVQRLLPVLCKEYGLTPEQVVAIASNNGGKLSLETVERLLPVLCQPPYG

LTPNQVVAIASHNGAKQALETVQRLLPVLCQPPYGLTPEQVVAIASHNGGKQALETVQRLLPVLCQPPYGL

TPEQVVAIVSNNGGKPALETVERLLPVLCQPPYGLTPNQVVAIASNIGAKPALETVQRLLPVLCQPPYGLT

PEQVVTIANNIGGKQALETVQRLLPVLRKPPYGLTPEQVVAIASNHGGKQALETVERLLPVLRKPPYGLTP

NQVVAIASNNGAKQALETVQRLLPVLCKPPHPLTPNQVVAIASHDGAKQALETVQRLLPVLCQPPYGLTPE

QVVVIASNNGGKQALETVQRLLPVLCQPSYGLTPEQVVAIASNHGGKQALETVQRLLPVLCQPPYGLTPNQ

VVAIASHDGGKPALETVQRLLPVLCQPPYGLTPNQVVAIASHDGGTQALESIFAQLSSPDPALAALTNDRL

VALACIGGRPALDAVKKGLPHAPALITRVHNRVPEGTAHLVADLAQVVRVLSFFQCHSHPAQAFDEAMRQF

GMSRHGLLQLFRRVGVTELEAISGTLPPASQRWDRMLQASGRKGAKPSSASAQTPSQESVDAFADSLEREL

DAPSPMHQAGQTLASSRKRSRSESSVNRSSAQQAAEVFVPEQRDAPPLLPLSSWGVKRRRTRIGGLPDPGT

PTHGDLAASSGAFLEQDADPFAGAAEDFPAFDQEEIAWLKELLAH

LW16-Tal6



AQTSDASPAADVDLSTFGYSQQQQEKIKPTVRSTVSQHHAALVGHGFTHAHIVELSKHPAALGTIAARYSE

MIAALPEATHEDIVGVGKQWSGARALETLLMVAEELRAPPLQLVTGQLLKIAKGGGVTAVEAVHASRNALT

GAPLHLTPDQVVAIVSHDGGKQSLETVQRLLPVLCQPPYGLTPNQVVAIASHNGGKQALETVQRLLPVLCQ

EYGLTPEQVVAIASHNGGKQALETVQRLLPVLCQEYGLTPNQVVAIASHDGAKQALETVQRLLPVLCQPPY

GLTPEQVVAIASNHGGKLALETVERLLPVLCQPPYGLTPNQVVAIASNHGGKQALETVQPLLPVLCQPPYG

LTPEQVVAIASHGGAKQALKTVQRLLPVLCQDHGLTPEQVVAIANHDGAKQALETVQRLLPVLCQPPYGLT

PEQVVAIASKGGGKQALETVQRLLPVLCQPPYGLTPDQVVTIASNNGGKPALETVRRLLPVLCKPPYGLTP

KQVVAIASYGGKQALETVQRLLPVLCKPPYGLTPEQVVAIASNGGGKPALETVQRLLPVLCQEYGLTPNQV

VAIASHDGAKPALETVQRLLPVLCQEYGLTPNQVVAIASHDGGKPALETVQRLLPVLCQPPYGLTPEQVVA

IASHNGGTQALESIFAQLSSPDPALAALTNDRLVALACIGGRPALDAVKKGLPHAPALITRVHNRVPEGTA

HLVADLAQVVRVLSFFQCHSHPAQAFDEAMRQFGMSRHGLLQLFRRVGVTELEAISGTLPPASQRWDRMLQ


VPEQRDAPPLLPLSSWGVKRRRTRIGGLPDPGTPTHGDLAASSGAFLEQDADPFAGAAEDFPAFDQEEIAW

LKELLAH

LW16-Tal7

MDPIRSRTPSPARELLAGSQPDGVQPTADPRVSPPAGSPLDGLPARRTMSRTQLPPPPASVPAFSAGSFSD

LLRQVDSSLFDASFFDSMPAFGAHHAEAATGELDEAQSALRAVDDPQPSASAAITAAPPRTKAAARRRSAQ



PLHLTPDQVVAIVSNNGGKQALETVQRLLPVLCQPPYNLTPEQVVAIASHDGGKQALETVQRLLLVLCQEC

GLTPNQVVAIASHDGAKPALETVHRLLPVLCQPPYGLTPNQVVAIASHDGAKQALETVQRLLPVLCQDHGL

TPGQVVAIADNIGGKQALETVQRLLPVLCKPPYGLTPEQVVTIANNIGGKPALETVQQLLPVLCQPPYGLT

PEQVVTIANNIGAKPALETVHRLLPVLRKPPYGLTPEQVVAIASHNGAKPALETVQRLLPVLCQPPYGLTP

NQVVAIASHDGAKQALETVQRLLPVLCKPPHPLTPNQVVAIASHDGAKQALETVQRLLPVLCQPPYGLTPD

QVVVIASNNGGKQALETVQRLLPVLCKPPYGLTPEQVVAIASNNGGKPALETVQRLLPVLCQPPYDLTPDQ

VVAIANNIGAKPALETVQRLLPVLCQPPYGLTPEQVVAIASNNGGKQALETVQRLLPVLCQPPYGLTPYQV

VAIASHDGGTQALESIFAQLSSPDPALATLTNDRLVALACIGGRPALDAVKKGLPHAPELITRVHNRVPEG

TAHLVADHAQVVRVLGFFQCHSQRGQVFHEAMKRFEMSREGLLQLFRRVGVTELEAISGTLPPASQRWDRM

LQASGRKGAKPPSASAQTQGQESLDAFADSLERELDAPSPMHQAGQTLASSRKRSRSESSVNRSSAQQAAE

VFVPEQRDAPLLLPLSSWGVKRRRTRIGGLPDTGMPTDGELAASSAAFLEQDADPFAGAAEDFPAFDQEEI

AWLRELLAH

LW16-Tal8




MIAALPEATHEDIVGVGKQWSGARALETLLMVAEELRAPPLQLVTGQLLKIAKRGGVTAVEAVHASRNALT

GAPLHLTPDQVVAIVSNNGGKQALETVQRLLPVLCQPPYGLTPEQVVAIASHDGAKPALETVQRLLPVLCQ

PPYGLTPEQVVAIASNGGGKQALETVQRLLPVLCKEYGLTPEQVVAIASNNGGKLALETVERLLPVLCQPP

YGLTPKQVVAIASHNGGKQALETVQRLLPVLCQPPYGLTPEQVVAIACNGGGKQALETVQRLLPVLCQPPY

GLTPDQVVTIANNIGAKQALETVQRLLPVLCQPPYGLTPNQVVAIASHDGAKQALETVQRLLPVLCQEYGL

27

TPGQVVAIANNIGGKPALETVQRLLPVLCQPPYNLTPNQVVAIASNNGAKPALETVQRLLPVLCQPPYGLT

PNQVVAIASHDGAKQALETVQRLLPVLCKPPHPLTPNQVVAIASHDGAKQALETVQRLLPVLCQPPYGLTP

DQVVVIASNNGGKQALETVQRLLPVLCQPPYGLTPEQVVAIASNNGGKPALETVQRLLPVLCKPPYDLTPD

QVVAIANNIGAKPALETVQRLLPVLCQPPYGLTPEQVVAIASHNGGKQALETVQRLLPVLCQPPYGLTPYQ

VVAIASHDGGTQALESIFAQLSSPDPALATLTNDRLVALACIGGRPALDAVKKGLPHAPALITRVHNRVPE

GTAHLVADLAQVVRVLSFFQCHSHPAQAFDEAMRQFGMSRHGLLQLFHRVGVTELEARSGTLPPASQRWDR

MLQASGRKGAKPPSASAQTQGQESLDAFADSLERELDAPSPMHQAGQTLASSRKRSRSESSVNRSSAQQAA

EVFVPEQRDAPPLLPLSSWGVKRRRTRIGGLPDPGTPTHGDLAASSAAFLEQDADPFAGAAEDFPAFDQEE

IAWLKELLAH

Table S4

Table S4. NCED proteins of selected species and wheat.

>O49505.1 RecName: Full=9-cis-epoxycarotenoid dioxygenase NCED2,

chloroplastic; Short=AtNCED2; Flags: Precursor

MVSLLTMPMSGGIKTWPQAQIDLGFRPIKRQPKVIKCTVQIDVTELTKKRQLFTPRTTATPPQHNPLRLNI

FQKAAAIAIDAAERALISHEQDSPLPKTADPRVQIAGNYSPVPESSVRRNLTVEGTIPDCIDGVYIRNGAN

PMFEPTAGHHLFDGDGMVHAVKITNGSASYACRFTKTERLVQEKRLGRPVFPKAIGELHGHSGIARLMLFY

ARGLCGLINNQNGVGVANAGLVYFNNRLLAMSEDDLPYQLKITQTGDLQTVGRYDFDGQLKSAMIAHPKLD

PVTKELHALSYDVVKKPYLKYFRFSPDGVKSPELEIPLETPTMIHDFAITENFVVIPDQQVVFKLGEMISG

KSPVVFDGEKVSRLGIMPKDATEASQIIWVNSPETFCFHLWNAWESPETEEIVVIGSCMSPADSIFNERDE

SLRSVLSEIRINLRTRKTTRRSLLVNEDVNLEIGMVNRNRLGRKTRFAFLAIAYPWPKVSGFAKVDLCTGE

MKKYIYGGEKYGGEPFFLPGNSGNGEENEDDGYIFCHVHDEETKTSELQIINAVNLKLEATIKLPSRVPYG

FHGTFVDSNELVDQL

>Q9LRR7.1 RecName: Full=9-cis-epoxycarotenoid dioxygenase NCED3,

chloroplastic; Short=AtNCED3; AltName: Full=Protein SALT TOLERANT 1;

Flags: Precursor

MASFTATAAVSGRWLGGNHTQPPLSSSQSSDLSYCSSLPMASRVTRKLNVSSALHTPPALHFPKQSSNSPA

IVVKPKAKESNTKQMNLFQRAAAAALDAAEGFLVSHEKLHPLPKTADPSVQIAGNFAPVNEQPVRRNLPVV

GKLPDSIKGVYVRNGANPLHEPVTGHHFFDGDGMVHAVKFEHGSASYACRFTQTNRFVQERQLGRPVFPKA

IGELHGHTGIARLMLFYARAAAGIVDPAHGTGVANAGLVYFNGRLLAMSEDDLPYQVQITPNGDLKTVGRF

DFDGQLESTMIAHPKVDPESGELFALSYDVVSKPYLKYFRFSPDGTKSPDVEIQLDQPTMMHDFAITENFV

VVPDQQVVFKLPEMIRGGSPVVYDKNKVARFGILDKYAEDSSNIKWIDAPDCFCFHLWNAWEEPETDEVVV

IGSCMTPPDSIFNESDENLKSVLSEIRLNLKTGESTRRPIISNEDQQVNLEAGMVNRNMLGRKTKFAYLAL

AEPWPKVSGFAKVDLTTGEVKKHLYGDNRYGGEPLFLPGEGGEEDEGYILCFVHDEKTWKSELQIVNAVSL

EVEATVKLPSRVPYGFHGTFIGADDLAKQVV

>Q9C6Z1.1 RecName: Full=Probable 9-cis-epoxycarotenoid dioxygenase

NCED5, chloroplastic; Short=AtNCED5; Flags: Precursor

MACSYILTPNPTKLNLSFAPSDLDAPSPSSSVSFTNTKPRRRKLSANSVSDTPNLLNFPNYPSPNPIIPEK

DTSRWNPLQRAASAALDFAETALLRRERSKPLPKTVDPRHQISGNYAPVPEQSVKSSLSVDGKIPDCIDGV

YLRNGANPLFEPVSGHHLFDGDGMVHAVKITNGDASYSCRFTETERLVQEKQLGSPIFPKAIGELHGHSGI

ARLMLFYARGLFGLLNHKNGTGVANAGLVYFHDRLLAMSEDDLPYQVRVTDNGDLETIGRFDFDGQLSSAM

IAHPKIDPVTKELFALSYDVVKKPYLKYFKFSPEGEKSPDVEIPLASPTMMHDFAITENFVVIPDQQVVFK

LSDMFLGKSPVKYDGEKISRFGILPRNAKDASEMVWVESPETFCFHLWNAWESPETDEVVVIGSCMTPADS

IFNECDEQLNSVLSEIRLNLKTGKSTRRTIIPGSVQMNLEAGMVNRNLLGRKTRYAYLAIAEPWPKVSGFA

KVDLSTGEVKNHFYGGKKYGGEPFFLPRGLESDGEDDGYIMSFVHDEESWESELHIVNAVTLELEATVKLP

SRVPYGFHGTFVNSADMLNQA

>Q9LRM7.1 RecName: Full=9-cis-epoxycarotenoid dioxygenase NCED6,


MQHSLRSDLLPTKTSPRSHLLPQPKNANISRRILINPFKIPTLPDLTSPVPSPVKLKPTYPNLNLLQKLAA

TMLDKIESSIVIPMEQNRPLPKPTDPAVQLSGNFAPVNECPVQNGLEVVGQIPSCLKGVYIRNGANPMFPP

28

LAGHHLFDGDGMIHAVSIGFDNQVSYSCRYTKTNRLVQETALGRSVFPKPIGELHGHSGLARLALFTARAG

IGLVDGTRGMGVANAGVVFFNGRLLAMSEDDLPYQVKIDGQGDLETIGRFGFDDQIDSSVIAHPKVDATTG

DLHTLSYNVLKKPHLRYLKFNTCGKKTRDVEITLPEPTMIHDFAITENFVVIPDQQMVFKLSEMIRGGSPV

IYVKEKMARFGVLSKQDLTGSDINWVDVPDCFCFHLWNAWEERTEEGDPVIVVIGSCMSPPDTIFSESGEP

TRVELSEIRLNMRTKESNRKVIVTGVNLEAGHINRSYVGRKSQFVYIAIADPWPKCSGIAKVDIQNGTVSE

FNYGPSRFGGEPCFVPEGEGEEDKGYVMGFVRDEEKDESEFVVVDATDMKQVAAVRLPERVPYGFHGTFVS

ENQLKEQVF

>Q9M9F5.1 RecName: Full=9-cis-epoxycarotenoid dioxygenase NCED9,


MTIITIISGMYIYSLLSQDAHHSQYGQNTNLVLKKPIPKPQTAAFNQESTMASTTLLPSTSTQFLDRTFST

SSSSSRPKLQSLSFSSTLRNKKLVVPCYVSSSVNKKSSVSSSLQSPTFKPPSWKKLCNDVTNLIPKTTNQN

PKLNPVQRTAAMVLDAVENAMISHERRRHPHPKTADPAVQIAGNFFPVPEKPVVHNLPVTGTVPECIQGVY

VRNGANPLHKPVSGHHLFDGDGMVHAVRFDNGSVSYACRFTETNRLVQERECGRPVFPKAIGELHGHLGIA

KLMLFNTRGLFGLVDPTGGLGVANAGLVYFNGHLLAMSEDDLPYHVKVTQTGDLETSGRYDFDGQLKSTMI

AHPKIDPETRELFALSYDVVSKPYLKYFRFTSDGEKSPDVEIPLDQPTMIHDFAITENFVVIPDQQVVFRL

PEMIRGGSPVVYDEKKKSRFGILNKNAKDASSIQWIEVPDCFCFHLWNSWEEPETDEVVVIGSCMTPPDSI

FNEHDETLQSVLSEIRLNLKTGESTRRPVISEQVNLEAGMVNRNLLGRKTRYAYLALTEPWPKVSGFAKVD

LSTGEIRKYIYGEGKYGGEPLFLPSGDGEEDGGYIMVFVHDEEKVKSELQLINAVNMKLEATVTLPSRVPY

GFHGTFISKEDLSKQALC

>O65572.2 RecName: Full=Carotenoid 9,10(9',10')-cleavage dioxygenase 1;

AltName: Full=AtCCD1; AltName: Full=Neoxanthin cleavage enzyme NC1;

Short=AtNCED1

MAEKLSDGSSIISVHPRPSKGFSSKLLDLLERLVVKLMHDASLPLHYLSGNFAPIRDETPPVKDLPVHGFL

PECLNGEFVRVGPNPKFDAVAGYHWFDGDGMIHGVRIKDGKATYVSRYVKTSRLKQEEFFGAAKFMKIGDL

KGFFGLLMVNVQQLRTKLKILDNTYGNGTANTALVYHHGKLLALQEADKPYVIKVLEDGDLQTLGIIDYDK

RLTHSFTAHPKVDPVTGEMFTFGYSHTPPYLTYRVISKDGIMHDPVPITISEPIMMHDFAITETYAIFMDL

PMHFRPKEMVKEKKMIYSFDPTKKARFGVLPRYAKDELMIRWFELPNCFIFHNANAWEEEDEVVLITCRLE

NPDLDMVSGKVKEKLENFGNELYEMRFNMKTGSASQKKLSASAVDFPRINECYTGKKQRYVYGTILDSIAK

VTGIIKFDLHAEAETGKRMLEVGGNIKGIYDLGEGRYGSEAIYVPRETAEEDDGYLIFFVHDENTGKSCVT

VIDAKTMSAEPVAVVELPHRVPYGFHALFVTEEQLQEQTLI

>O49675.1 RecName: Full=Probable carotenoid cleavage dioxygenase 4,

chloroplastic; Short=AtCCD4; AltName: Full=AtNCED4; Flags: Precursor

MDSVSSSSFLSSTFSLHHSLLRRRSSSPTLLRINSAVVEERSPITNPSDNNDRRNKPKTLHNRTNHTLVSS

PPKLRPEMTLATALFTTVEDVINTFIDPPSRPSVDPKHVLSDNFAPVLDELPPTDCEIIHGTLPLSLNGAY

IRNGPNPQFLPRGPYHLFDGDGMLHAIKIHNGKATLCSRYVKTYKYNVEKQTGAPVMPNVFSGFNGVTASV

ARGALTAARVLTGQYNPVNGIGLANTSLAFFSNRLFALGESDLPYAVRLTESGDIETIGRYDFDGKLAMSM

TAHPKTDPITGETFAFRYGPVPPFLTYFRFDSAGKKQRDVPIFSMTSPSFLHDFAITKRHAIFAEIQLGMR

MNMLDLVLEGGSPVGTDNGKTPRLGVIPKYAGDESEMKWFEVPGFNIIHAINAWDEDDGNSVVLIAPNIMS

IEHTLERMDLVHALVEKVKIDLVTGIVRRHPISARNLDFAVINPAFLGRCSRYVYAAIGDPMPKISGVVKL

DVSKGDRDDCTVARRMYGSGCYGGEPFFVARDPGNPEAEEDDGYVVTYVHDEVTGESKFLVMDAKSPELEI

VAAVRLPRRVPYGFHGLFVKESDLNKL

>Q6YVJ0.1 RecName: Full=9-cis-epoxycarotenoid dioxygenase NCED1,

chloroplastic; Short=OsNCED1; Flags: Precursor

MQRICPAHCSVTHSLTMKSMRLSYIPPAASAAPQSPSYGRKKNASAAPPSAAASTTVLTSPLVTTTRTPKQ

TEQEDEQLVAKTKTTRTVIATTNGRAAPSQSRPRRRPAPAAAASAASLPMTFCNALEEVINTFIDPPALRP

AVDPRNVLTSNFVPVDELPPTPCPVVRGAIPRCLAGGAYIRNGPNPQHLPRGPHHLFDGDGMLHSLLLPSP

ASSGDDPVLCSRYVQTYKYLVERDAGAPVLPNVFSGFHGVAGMARGAVVAARVLTGQMNPLEGVGLANTSL

AYFAGRLYALGESDLPYAVRVHPDTGEVTTHGRCDFGGRLVMGMTAHPKKDPVTGELFAFRYGPVPPFVTY

FRFDPAGNKGADVPIFSVQQPSFLHDFAITERYAIFPEIQIVMKPMDMVVGGGSPVGSDPGKVPRLGVIPR

YATDESEMRWFEVPGFNIMHSVNAWEEAGGEELVLVAPNVLSIEHALEHMELVHSCVEKVRINLRTGVVTR

TPLAAGNFDFPVINPAFLGRRNRYGYFGVGDPAPKIGGVAKLDFDRAGEGDCTVAQRDFGPGCFAGEPFFV

ADDVEGNGNEDDGYLVCYVHDEATGENRFVVMDARSPDLEIVAEVQLPGRVPYGFHGLFVTQAELQSQHQ

29

>Q5MBR6.1 RecName: Full=9-cis-epoxycarotenoid dioxygenase NCED2,


MEVPIAAMTFAHPANVMTLASRQPKSKRSHISPATTAHRNLQTRLAHHHHATPASLPMAICNTVDKVINRF

IDLPEQRPTVDPRRVLSGNFAPVDELPPTSCHVIRGSIPSCLAGGVYIRNGPNPQHRLPQRTHHLFDGDGM

LHSLLIPSASSTLLSEPVLCSRYVHTYKYLLERETGGPVLPNFFAGFHGVAGLARAVVMIARVLAGQINLN

KGFGLANTSITLFADCLYALCESDLPYSMHINPANGEVTTLGRCDFGGDLSFRMTAHPKKDPVTMELFAFR

YNVFQPFITYFWFDRAGSKVADVPILSLQKPSVMHDFAITERYAIFPESQLIVNPMDMVMRGSSLVGLDRT

MVPRIGVLPRYAKDESDMRWFEVPRFNMLHTTNGWEEADGEEIVLVAPNILSIEHMLGNMELMRARVDMVR

INLCTGDVSCTALSPESLEFGVIHQGYVGRKNRYGYFGVSGPLPKIKGIRKLDFDLVGSGDCTVGRRDFGL

GCFAGEPFFVPDNIDGYGNEDSGYVVCYTHEEDTGESWFVVMDAKSPELDIVAEVQLPSRIPYGFHGIFVK

QAELLAQQ



MATITTPGYAHIQRQHGRCSTTAGRRGASNSVRFSARAVSSVPHAAAASSAPAFLPVPFVPGADAPSPSGK

SAIGVPKAPRKGEEGKRLNFFQRAAAMALDAFEEGFVANVLERPHGLPSTADPAVQIAGNFAPVGETPPAR

ALPVSGRIPPFINGVYARNGANPHFDPVAGHHLFDGDGMVHAVRIRNGAAESYACRFTETARLRQERAMGR

PMFPKAIGELHGHSGIARLALFYARAACGLLDPSHGTGVANAGLIYFNGRLLAMSEDDLPYQVRVTADGDL

ETVGRYDFDGQLGCAMIAHPKLDPATGELHALSYDVIKKPYLKYFYFAPDGTKSADVEIPLDQPTMIHDFA

ITENYVVVPDHQVVFKLQEMLRGGSPVVLDKEKTSRFGVLPKHAADASEMVWVDVPDCFCFHLWNAWEEAD

TDEVVVIGSCMTPADSIFNESDDRLESVLTEIRLNTRTGESTRRAILPPSSQVNLEVGMVNRNLLGRKTRY

AYLAVAEPWPKVSGFAKVDLATGELTKFEYGEGRFGGEPCFVPMDAAAATPRGEDDGYILSFVHDERAGTS

ELLVVNAADMRLEATVQLPSRVPYGFHGTFITGDELTTQA

>Q69NX5.1 RecName: Full=9-cis-epoxycarotenoid dioxygenase NCED4,


MASSAPSAPGLAPVAKPPPPPSKVKVATATVPTNGKIKQGARPMRVSAPPVEPRRRMNPLQRLAAAAIDAV

EEGLVAGLLERGHALPRTADPAVQIAGNYAPVGERPPVRGLPVSGRLPACLDGVYVRNGANPLHAPRAGHH

LFDGDGMLHAVRLAGGRAESYACRFTETARLRQEREMGRPVFPKAIGELHGHSGVARLLLFGSRALCGVLD

ASRGIGVANAGLVYHDGRLLAMSEDDLPYHVRVTHDGDLETVGRYDFHGQLDADGTMIAHPKLDPVTGELF

ALSYNVVSKPYLKYFYFTADGRKSRDVDIPVGAPTMIHDFAVTENYAVVPDQQIVFKLQEMVRGGSPVVYD

REKASRFGVLPKRAADASELRWVEVPGCFCFHLWNAWEDDATGEIVVIGSCMTPPDAVFNEPSQSPEEESF

RSVLSEIRLDPRTGVSRRRDVLRDAAEQVNLEAGMVNRQLLGRKTRYAYLAIAEPWPRVSGFAKVDLESGT

AEKFIYGEGRYGGEPCFVPRAGAAAEDDGHVLCFVHDEERGTSELVVVDAGSEAMEEVAAVKLPGRVPYGL

HGTFIGANELQRQA



MPTTFTPNSPASSCSIHHRASPSRGARNSVRFTRPRAAAAATNSVLSAPSSVPPAYVPPPPPPPTKMFPEA

GDAAAAKAAARRCGKKKDGLNFFQRAAAVALDAFEEGFITNVLERPHALPRTADPAVQIAGNFAPVGEQPP

VRSLPVSGRIPPFINGVYARNGANPHFEPTAGHHLFDGDGMVHAVRIRNGAAESYACRFTETARLGQERAL

GRAVFPKAIGELHGHSGIARLALFYARGLCGLVDPSHGTGVANAGLVYFNGRLLAMSEDDLPYQVRVTADG

DLETVGRYDFDGQLGCAMIAHPKLDPVSGELFALSYDVIKKPYLKYFYFDADGTKSPDVEIELEQPTMIHD

FAITENFVVVPDHQVVFKLGEMFRGGSPVVLDREKTSRFGVLPKHATSSLEMVWVDVPDCFCFHLWNAWEE

AESGEVVVVGSCMTPADSIFNESDEHLESVLTEIRLNTRTGESTRRAVLPPAAQVNLEVGMVNRAMLGRKT

RYAYLAVAEPWPKVSGFAKVDLATGELTKFEYGEGRFGGEPCFVPMGGAGAAASPARGEDDGYILSFVRDE

AAGTSELLVVNAADMRLEATVQLPSRVPYGFHGTFINAGELATQA

>O24592.2 RecName: Full=9-cis-epoxycarotenoid dioxygenase 1,

chloroplastic; AltName: Full=Protein VIVIPAROUS14; Short=VP-14;

Short=VP14; Flags: Precursor

MQGLAPPTSVSIHRHLPARSRARASNSVRFSPRAVSSVPPAECLQAPFHKPVADLPAPSRKPAAIAVPGHA

AAPRKAEGGKKQLNLFQRAAAAALDAFEEGFVANVLERPHGLPSTADPAVQIAGNFAPVGERPPVHELPVS

GRIPPFIDGVYARNGANPCFDPVAGHHLFDGDGMVHALRIRNGAAESYACRFTETARLRQERAIGRPVFPK

AIGELHGHSGIARLALFYARAACGLVDPSAGTGVANAGLVYFNGRLLAMSEDDLPYHVRVADDGDLETVGR

YDFDGQLGCAMIAHPKLDPATGELHALSYDVIKRPYLKYFYFRPDGTKSDDVEIPLEQPTMIHDFAITENL

VVVPDHQVVFKLQEMLRGGSPVVLDKEKTSRFGVLPKHAADASEMAWVDVPDCFCFHLWNAWEDEATGEVV

30

VIGSCMTPADSIFNESDERLESVLTEIRLDARTGRSTRRAVLPPSQQVNLEVGMVNRNLLGRETRYAYLAV

AEPWPKVSGFAKVDLSTGELTKFEYGEGRFGGEPCFVPMDPAAAHPRGEDDGYVLTFVHDERAGTSELLVV

NAADMRLEATVQLPSRVPFGFHGTFITGQELEAQAA

>TaNCED 5DL (BAS30382)

MQTLTASTSVSSIQRHRPRPAGRSSSVSFTARAVSSVPRAPSGAARAPAPSQFVRGADAKPLIAVPKAPAV

ERQEKKLNFFQRAAATALDAFEEGFVANVLERPHGLSRTVDPAVQIAGNFAPVGETPPVHALPVTGRIPPF

INGVYARNGANPRFDPVAGHHLFDGDGMVHALRIRNGVAETYASRFTETERLQQERALGRPMFPKAIGELH

GHSGIARLALFYARAACGLIDPSRGTGVANAGLVYFNGHLLAMSEDDIPYHVRVTDDGDLQTVGRYDFDGQ

LECPMIAHPKLDPATGELHALSYDVIKKPYLKYFYFAADGTKSADVEIPLDQPTMIHDFAITENYVVVPDH

QVVFKLQEMLRGGSPVVLDKEKTSRFGVLPKCAADASEMVWVDVPDCFCFHLWNAWEEEETEEVVVIGSCM

TPADSIFNESDECLESVLTEIRLNTRTGESTRRPILALSEQVNLEVGMVNSNLLGRKTRYAYLAVAEPWPK

VSGFAKVDLATGELTKFEYGEGRFGGEPCFVPMDPAASRGEDGYILTFVHDEAAGTSELLVVNAADMRLEA

TIQLPSRVPYGFHGTFVTGKELESQA

>TaNCED 5AL (AOE46767)

MQTLTASTSVSSIQRHTPRRTGRSGSLSFSARAVSSPPRAPAPSRFVRGADAAPAKPLIAVPKAPAVERQE

KKLNFFQRAAATALDAFEEGFVANVLERPHGLSRTVDPAVQIAGNFAPVGETPPVQALPVTGRIPPFINGV

YARNGANPHFDPVAGHHLFDGDGMVHALRIRNGVAETYASRFTETERLQQERALGRPMFPKAIGELHGHSG

IARLALFYARAACGLIDPSRGTGVANAGLVYFNGHLLAMSEDDIPYHVRVTDDGDLQTVGRYDFDGQLECP

MIAHPKLDPATGELHALSYDVIKKPYLKYFYFKADGTKSADVEIPLDQPTMIHDFAITENYVVVPDHQVVF

KLQEMLRGGSPVVLDKEKTSRFGVLPKCAADASEMVWVDVPDCFCFHLWNAWEEEESDEVVVIGSCMTPAD

SIFNESDECLESVLTEIRLNTRTGESTRRPILALSEQVNLEVGMVNSNLLGRKTRYAYLAVAEPWPKVSGF

AKVDLATGELTKFEYGEGRFGGEPCFVPMDPAASRGEDDGYILTFVHDEAAGTSELLVVNAADMRLEATIQ

LPSRVPYGFHGTFVTGKELESQA

>TaNCED 5BL (referred from TGAVv1 genome)

MQTLTASTSVSSIQRHRPRPAGRSSFSARAVSSAPRAPSGAARVPAPSRFVRGADAAPAKPLIAVPKAPAV

ERQEKKLNFFQRAAATALDAFEEGFVANVLERPHGLSRTVDPAVQIAGNFAPVGETPPVHALPVTGRIPPF

INGVYARNGANPHFDPVAGHHLFDGDGMVHALRIRNGVAETYASRFTETERLQQERALGRPMFPKAIGELH

GHSGIARLALFYARAACGLIDPSRGTGVANAGLVYFNGHLLAMSEDDIPYHVRVTDDGDLQTVGRYDFDGQ

LECPMIAHPKLDPATGELHALSYDVIKKPYLKYFYFKADGTKSADVEIPLDQPTMIHDFAITENYVVVPDH

QVVFKLQEMLRGGSPVVLDKEKTSRFGVLPKCAADSSEMVWVDVPDCFCFHLWNAWEEEETDEVVVIGSCM

TPADSIFNESDECLESVLTEIRLNTRTGESTRRPILALSEQVNLEVGMVNSNLLGRKTRYAYLAVAEPWPK

VSGFAKVDLATGELTKFEYGEGRFGGEPCFVPMDPATSRGEDDGYILTFVHDEAAGTSELLVVNAADMRLE

ATIQLPSRVPYGFHGTFVTGKELESQA

>TaNCED 5DS (referred from TGAVv1 genome)

MQTLSAQPLASSSASSSIQHHHGRRRGASSVRFAPCAAAAAADSVLIAASTSTARTPAYVSSPSTRKVPGY

EQSGPPAIASPHKQGSSSGKDGQSLNFFQRAAAAALDAFEEGFIHNVLERPHALPRTADPAVQIAGNFAPV

GEQAPVRALPVSGRIPPFINGVYARNGANPCFEPTAGHHLFDGDGMVHAIRIRNGAAESYTCRFTETARLS

QERAVGKPVFPKTIGELHGHSGIARLALFYARGACGLVDPSHGTGVANAGLVYFNGRLLAMSEDDLPYQVR

VTADGDLETVGRYDFDGQLDCAMIAHPKLDPVSGELFALSYDVIKKPYLKYFYFHADGTKSADVEIELDQP

TMIHDFAITENFVVVPDHQMVFKLAEMFRGGSPVMLDKEKTSRFGVLPKYAKHSSEMMWVDVPDCFCFHLW

NSWEEPETDEVVVIGSCMTPADSIFNDTDDHLESVLTEIRLNTRTGESTRRAILPLESQVNLEVGMVNRNM

LGRKTRYAYLAVAEPWPKVSGFAKVDLVTGELTKFEYGEGRFGGEPCFVPMDGEHARPGAEDDGYVLSFVR

DEAAGTSELLVVNAADMRLEATVQLPSRVPYGFHGTFIGAADLDAQH

>TaNCED 5AS (referred from TGAVv1 genome)

MQTLSAQPLASSSSSIQRQNGRRRGTSSVRFAPRAAAAAADSVLIAASTSTARPPAYLPSPSTRKVPGYEQ

SAPPAIASPQKQGSSKDGQSKGGLNFFQRAAAAALDAFEEGFIHNVLERPHALPRTADPAVQIAGNFAPVG

EQAPVRALPVSGRIPPFINGVYARNGANPCFEPTAGHHLFDGDGMVHAIRIRNGAAESYACRYTETARLSQ

ERAVGRPIFPKTIGELHGHSGIARLALFYARGACGLVDPSHGTGVANAGLVYFNGRLLAMSEDDLPYQVRV

TAGGDLETVGRYDFDGQLDCAMIAHPKLDPVSGELFALSYDVIKKPYLKYFYFHADGTKSADVEIELDQPT

MIHDFAITENFVVVPDHQMVFKLAEMFRGGSPVMLDKEKTSRFGVLPKYAKDSSEMMWVDVPDCFCFHLWN

SWEEPETDEVVVIGSCMTPADSIFNDTDDHLESVLTEIRLNTRTGESTRRTILPLESQVNLEVGMVNRNML

31

GRKTRYAYLAVAEPWPKVSGFAKVDLVTGELTKFEYGEGRFGGEPCFVPMDGEHARPGAEDDGYVLSFVRD

EAAGTSELLVVNAADMRLEATVQLPSRVPYGFHGTFIGAADLDAQH

>TaNCED 5BS (referred from TGAVv1 genome)

MQTLSAQPLASSSASSSIQHHHGRRRGASSVRFSPRAAAAAADSVLIAASTSTARTPAYVSSPSTRKVPGY

EQSALPAIASPQKQSGSGKDGQSLNFFQRAAAAALDAFEEGFTHNVLERPHALPRTADPAVQIAGNFAPVG

EQAPVRALPVSGRIPPFINGVYARNGANPCFEPTAGHHLFDGDGMVHAIRIRNGAAESYACRFTETARLSQ

ERAVGRPVFPKTIGELHGHSGIARLALFYARGACGLVDPSHGTGVANAGLVYFNGRLLAMSEDDLPYQVRV

TAAGDLETVGRYDFDGQLDCAMIAHPKLDPVSGELFALSYDVIKKPYLKYFYFHADGTKSADVEIELDQPT

MIHDFAITENFVVVPDHQMVFKLAEMFRGGSPVMLDKEKTSRFGVLPKYAKESSEMMWVDVPDCFCFHLWN

SWEEPETDEVVVIGSCMTPADSIFNDTDDHLESVLTEIRLNTRTGESTRRAILPLESQVNLEVGMVNRNML

GRKTRYAYLAVAEPWPKVSGFAKVDLVTGELTKFEYGEGRFGGEPCFVPMDGEHARPGAEDDGYVLSFVRD

EAAGTSELLVVNAADMRLEATVQLPSRVPYGFHGTFIGAADLDAQH

>TaNCED 2BL (referred from TGAVv1 genome)

MASSITAPAAAPGSVVRPPAKVRPRPPPSRFNPNDPRKNADGSAVVTWKGPMRAAPARKKWPNPFQRMVAA

ALNAVEDRLVAGILERKHPLPRTADPAVQIAGNYAPVGELPPAGPEDLPVVSGRVPACLQGVYVRNGANPL

HAPRAGHHLFDGDGMVHAVRLGGGRAESYACRFTKTARLRQEREIGRPIFPKAIGELHGHSGVARLALFGA

RSLCGVVDASQGIGVANAGLVYHDGRLLAMSEDDLPYHVRVTPDGDLETVGRYDFNGQLDGAMIAHPKLDP

ATGELFALSYNVVSKPYLKYFYFTADGHKSHDVDIPVDEPTMIHDFAVTENYAVVPDQQIVFKLREMVLGG

SPVVYDKSKTARFGVVPKRAANASELRWVEVPDCFCFHLWNAWEDEATGEIVVIGSCMTPPDAVFNESPDQ

DQSFRSVLSEIRLDPRTGTSRRRAVLREADQVNLEAGMVNRQLVGRKTRYAYLSIAEPWPRVSGFAKVDLE

SGTVEKFTC

>TaNCED 2DL (SPT17866)

MASSITAPAAAPGSVVRPPAKVAPNDPRKNADGSAVVTWKGPMRAAPARKKWPNPFQRMVAAALDAVEDRL

VAGILERKHPLPRTVDPAVQIAGNYAPVGELPPAGPEDLPVVSGRVPACLEGVYVRNGANPLHAPRAGHHL

FDGDGMLHAVRLGGGRAESYACRFTETARLRQEREIGRPIFPKAIGELHGHSGVARLALFGARSLCGVVDA

SQGIGVANAGLVYHDGRLLAMSEDDLPYHVRVTPDGDLETVGRYDFNGQLDGAMIAHPKLDPATGELFALS

YNVVSKPYLKYLYFTADGHKSHDVDIPVDEPPMIHDFAVTENYAVVPDQQIVFKLQEMVLGGSPVVYDKNK

TARFGVLPKRAANALELRWVEVPDCFCFHLWNAWGGEATGEIVVIGSCMTPPDAVFNESPDQEQSFRSVLS

EIRLDPRTGTSRRRAVLREADQVNLEAGMVNRQLLGRKTRYAYLAIAEPWPRVSGFAKVDLESGTAEKFTY

GEGRYGGEPCFVPHADGSGAEDDGYVLCFVHDESRGSADGTSSELLVVNARDLRSEATVELPGRVPYGFHG

TFITATELQRQAQL

>TaNCED 2AL (referred from TGAVv1 genome)

MASSITAPAAAPGSVVRPPAKVRPRPPSSRFNPNDPRRNADGSAVVTWKGPMRAAPARKKWPNPFQRMVAA

ALDAVEDRLVVGILERKHPLPRTADPAVQISGNYAPVGELPPAGPEDLPVVSGRVPACLEGVYVRNGANPL

HAPRAGHHLFDGDGMLHAVRLGGGRAESYACRFTETARLRQEREIGRPIFPKAIGELHGHSGVARLALFGA

RSICGVVDASQGIGVANAGLVYHDGRLLAMSEDDLPYHVRVTPDGDLETVGRYDFNGQLDGAMIAHPKLDP

ATGELFALSYNVVSKPYLKYFYFTADGHKSHDVDIPVDEPTMIHDFAVSENYAIVPDQQIVFKLQEMVLGG

SPVVYDKNKTARFGVLPKRAANASDLRWVEVPGCFCFHLWNAWEDEATGEIVVIGSCMTPPDAVFNESPDQ

EQSFRSVLSEIRLDPRTGTSRRRAVLREADQVNLEAGMVNRQLLGRKTRYAYLAISEPWPRVSGFAKVDLE

SGTVEKFTYGEGRYGGEPCFVPHADGSGAEDDGYVLCFVHDESRGNADGTSSELLVVNARDLRSEATVKLP

GRVPRISRHVHHRHGAAATSPAIAS

32

Supplementary Information References.

1. Peng Z, Hu Y, Xie J, Potnis N, Akhunova A, Jones J, Liu Z, White FF, Liu S (2016)

Long read and single molecule DNA sequencing simplifies genome assembly and

TAL effector gene analysis of Xanthomonas translucens. BMC Genomics. 17, 9.

2. Bolger AM, Lohse M, Usadel B (2014) Trimmomatic: a flexible trimmer for Illumina

sequence data. Bioinformatics. 30, 2114-2120.

3. International Wheat Genome Sequencing Consortium, (IWGSC) (2014) A

chromosome-based draft sequence of the hexaploid bread wheat (Triticum aestivum)

genome. Science. 345, 1251788.

4. Dobin A, Davis CA, Schlesinger F, Drenkow J, Zaleski C, Jha S, Batut P, Chaisson

M, Gingeras TR (2013) STAR: Ultrafast universal RNA-seq aligner. Bioinformatics.

29, 15-21.

5. Love M, Huber W, Anders S (2014) Moderated estimation of fold change and

dispersion for RNA-seq data with DESeq2. Genome Biol. 15, 8.

6. Benjamini Y, Hochberg Y (1995) Controlling the false discovery rate - a practical and

powerful approach to multiple testing. Journal of the Royal Statistical Society Series

B-Methodological 57, 289-300.

7. Doyle EL, Booher NJ, Standage DS, Voytas DF, Brendel VP, Vandyk JK,

Bogdanove AJ (2112) TAL Effector-Nucleotide Targeter (TALE-NT) 2.0: tools for

TAL effector design and target prediction. Nucleic Acids Res. 40, 117.

8. Wang X, Yang B, Li K, Kang Z, Cantu D, Dubcovsky J (2016) A conserved Puccinia

striiformis protein interacts with wheat NPR1 and reduces induction of pathogenesis-

related genes in response to pathogens. Mol. Plant Microbe Interact. 29, 977-989.

33

9. Lu S, Friesen TL, Faris JD (2011) Molecular characterization and genomic mapping

of the pathogenesis-related protein 1 (PR-1) gene family in hexaploid wheat (Triticum

aestivum L.). Mol. Genet. Genomics. 285, 485-503.

10. Koren S, Walenz BP, Berlin K, Miller JR, Bergman NH, Phillippy AM (2017) Canu:

scalable and accurate long-read assembly via adaptive k-mer weighting and repeat

separation. Genome Res. 27(5):722-736.

11. Marçais G, Delcher AL, Phillippy AM, Coston R, Salzberg SL, Zimin A (2018)

MUMmer4: A fast and versatile genome alignment system. PLoS Comput Biol.

14(1):e1005944.

12. Pérez-Quintero AL, Lamy L, Gordon JL, Escalon A, Cunnac S, Szurek B, Gagnevin

L (2015) QueTAL: A suite of tools to classify and compare TAL effectors

functionally and phylogenetically. Front Plant Sci. 6:545.

13. T. Cermak et al., Efficient design and assembly of custom TALEN and other TAL

effector-based constructs for DNA targeting. Nucleic Acids Res 39, e82 (2011).

14. Li T, Huang S, Zhao X, Wright DA, Carpenter S, Spalding MH, Weeks DP, Yang B

(2011) Modularly assembled designer TAL effector nucleases for targeted gene

knockout and gene replacement in eukaryotes. Nucleic Acids Res. 39, 6315-6325.

15. Livak KJ1, Schmittgen TD (2001) Analysis of relative gene expression data using

real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods 25, 402-

408.

16. Hu Y, Zhang J, Jia H, Sosso D, Li T, Frommer WB, Yang B, White FF, Wang N,

Jones JB (2014) Lateral organ boundaries 1 is a disease susceptibility gene for citrus

bacterial canker disease. Proc. Natl. Acad. Sci. U. S. A. 111, 521.

34

17. Liu N, Ding Y, Fromm M, Avramova Z (2014) Endogenous ABA extraction and

measurement from Arabidopsis leaves. Bio Protoc. 4.

18. Hu Y, Wu Q, Peng Z, Sprague SA, Wang W, Park J, Akhunov E, Jagadish KSV,

Nakata PA, Cheng N, Hirschi KD, White FF, Park S (2017) Silencing of OsGRXS17

in rice improves drought stress tolerance by modulating ROS accumulation and

stomatal closure. Sci. Rep. 7, 7.

19. Schmelz EA, Engelberth J, Tumlinson JH, Block A, Alborn HT. (2004) The use of

vapor phase extraction in metabolic profiling of phytohormones and other

metabolites. Plant J. 39:790-808.

Xanthomonas translucens commandeers the host rate-limiting … · 2019. 10. 1. · 1 Supplementary...

Documents

Transcript of Xanthomonas translucens commandeers the host rate-limiting … · 2019. 10. 1. · 1 Supplementary...