Host plant species determines symbiotic bacterial community mediating suppression of plant defenses

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1 Host plnt species determines symiotic cteril community mediting suppression of plnt defenses Seung Ho Chung 1, Erin D. Scully 2, Michelle Peiffer 3, Scott M. Gei 4, Cristin Ros 5, Kelli Hoover 3, Gry W. Felton 3 1 Deprtment of Entomology, Cornell University, Ithc, NY 14853, USA 2 Stored Product Insect nd Engineering Reserch Unit, USDA-Agriculturl Reserch Service, Center for Grin nd Animl Helth Reserch, Mnhttn, KS 6652, USA 3 Deprtment of Entomology, Pennsylvni Stte University, University Prk, PA 1682, USA 4 Tropicl Crop nd Commodity Protection Reserch Unit, USDA-ARS Dniel K. Inouye Pcific Bsin Agriculturl Reserch Center, Hilo, HI 9672, USA 5 Deprtment of Plnt Pthology nd Environmentl Microiology, Pennsylvni Stte University, University Prk, PA 1682, USA Correspondence nd requests for mterils should e ddressed to G.W.F. (emil: gwf1@psu.edu) Supplementry Informtion:

2 PPO ctivity mod/min/mg protein c c Tomto PPO ctivity mod/min/mg protein Potto PPO ctivity mod/min/mg protein Eggplnt PPO ctivity mod/min/mg protein Bufflour PPO ctivity mod/min/mg protein Horsenettle PPO ctivity mod/min/mg protein Nightshde Fig. S1. Polyphenol oxidse (PPO) ctivity in plnts spryed with methyl jsmonte (MJ), slicylic cid (SA), oth elicitors (Both), or the ethnol (control). PPO ctivity ws mesured 48 h fter tretment. Vlues re mens ± SEM. Different letters ove the rs represent significnt differences (ANOVA, P <.5, N = 5-1).

3 Lrvl mss (mg) c 6 Tomto Lrvl mss (mg) 6 Potto Lrvl mss (mg) Eggplnt Lrvl mss (mg) Bufflour Lrvl mss (mg) c 6 Horsenettle 5 Lrvl mss (mg) Nightshde Fig. S2. Weight of lrve fed on plnts spryed with methyl jsmonte (MJ), slicylic cid (SA), oth elicitors (Both), or the ethnol (control). Neontes were llowed to feed on excised leflets for 5 dys nd lrvl mss ws determined. Vlues re mens ± SEM. Different letters ove the rs represent significnt differences (ANOVA, P <.5, N = 2-3).

4 Orl secretions (nl/cm) Tomto ** ** *** Potto Eggplnt Bufflour Horsenettle Nightshde Fig. S3. The mount of orl secretions (OS) secreted y Colordo potto eetle lrve tht were rered on different host plnts. Vlues re mens ± SEM (N = 5-1). Asterisks indicte significnt differences compred with tomto. ** P <.1; *** P <.1.

5 12 Numer of OTUs oserved Numer of sequences Tomto-1 Tomto-2 Tomto-3 Potto-1 Potto-2 Potto-3 Eggplnt-1 Eggplnt-2 Eggplnt-3 Bufflour-1 Bufflour-2 Bufflour-3 Horsenettle-1 Horsenettle-2 Horsenettle-3 Nightshde-1 Nightshde-2 Nightshde-3 Fig. S4. Rrefction curves for 16S gut cteril communities of Colordo potto eetle lrve tht were rered on different host plnts.

6 Color Key Row Z Score N3 N1 N2 E2 E3 H3 H2 E1 H1 P3 P2 P1 B3 B2 B1 T3 T2 T1 Otu1 Otu6 Otu8 Otu2 Otu13 Otu16 Otu223 Otu13 Otu4 Otu154 Otu172 Otu189 Otu499 Otu652 Otu169 Otu755 Otu123 Otu144 Otu293 Otu1397 Otu1482 Otu1552 Otu61 Otu444 Otu276 Otu62 Otu5 Otu36 Otu134 Otu342 Otu18 Otu55 Otu175 Otu232 Otu258 Otu285 Otu448 Otu47 Otu72 Otu723 Otu1235 Otu1513 Otu1578 Otu158 Otu46 Otu44 Otu138 Otu3 Otu9 Otu42 Otu11 Otu29 Otu3 Otu4 Otu21 Otu14 Otu6 Otu28 Otu38 Otu17 Otu23 Fig. S5. 2D hierchil clustering dendrogrm nd hetmp showing reltive udnce of OTUs over 1% of totl reds from Colordo potto eetle lrve tht were rered on different host plnts (T, tomto; P, potto; E, eggplnt; B, ufflour; H, horsenettle; N, nightshde-fed lrve). OTU tle ws presented in Tle S4.

7 .8 T1.4 N3 P1 P3 NMDS2. P2 E2 N1 H3 N2 B1 B2 T3 E3 H2 H1 E1.4 B3 T NMDS1 Fig. S6. Non-metric multi-dimensionl scling (NMDS) plot showing similrities etween gut cteril communities from Colordo potto eetle lrve tht were rered on different host plnts (T, tomto; P, potto; E, eggplnt; B, ufflour; H, horsenettle; N, nightshde-fed lrve). Bry-Curtis dissimilrity mtrix with singletons ws used to generte NMDS coordintes (Stress:.141, R 2 :.92).

8 Figure S7. Reltive undnce of cteril communities t the order level from Colordo potto eetle lrve tht were rered on different host plnts. Tx with reltive undnce less thn.1 were ssigned to other.

9 Figure S8. Reltive undnce of cteril communities t the order level in guts nd OS from Colordo potto eetle lrve tht were rered on tomto. Tx with reltive undnce less thn.1 were ssigned to other.

10 Tle S1. The threshold cycle (Ct) of rpod copy numer deposited on plnts y Colordo potto eetle lrve. rpod copy numers were mesured to estimte the mount of Pseudomons sp. Deposited on leves fter 2 h of insect feeding. Vlues re untrnsformed mens ± SEM (N = 5-6). Con, undmged plnts; AB(-), plnts dmged y untreted lrve; AB(+), plnts dmged y AB-treted lrve. rpod on potto leves dmged y tomto fed lrve Tretment Ct* Con 39.8 ±.2 rpod on potto leves dmged y potto fed lrve Tretment Ct Con 39.7 ±.2 rpod on ufflour dmged y ufflour fed lrve Tretment Ct Con 39.5 ±.3 AB(-) 39.8 ±.2 AB(+) 39.7 ±.2 * Ct vlue of 4. represent tht zero copy of rpod.

11 Tle S2. Alph diversity indexes with singletons from Colordo potto eetle lrve tht were rered on different host plnts. Host Coverge OTUs Cho1 Shnnon 1/Simpson Tomto Tomto Tomto (men ± SEM) (82.3 ± 12.) (13.8 ± 27.4) (1.86 ±.52) (5.16 ± 2.16) Potto Potto Potto (men ± SEM) (53.3 ± 7.2) (1.2 ± 2.1) (.61 ±.15) (1.5 ±.2) Eggplnt Eggplnt Eggplnt (men ± SEM) (4.3 ± 9.) (56.37 ± 9.3) (.46 ±.15) (1.46 ±.22) Bufflour Bufflour Bufflour (men ± SEM) (63.7 ± 4.9) (153.9 ± 38.3) (1.6 ±.6) (2.57 ±.26) Horsenettle Horsenettle Horsenettle (men ± SEM) (19. ± 1.5) (31.3 ± 3.2) (.25 ±.3) (1.14 ±.3) Nightshde Nightshde Nightshde (men ± SEM) (46.3 ± 8.4) (73.9 ± 26.7) (.56 ±.22) (1.44 ±.19)

12 Tle S3. Reltive undnce of the 1 most undnt OTUs from lrve tht were rered on different host plnts. Enterocter Lctococcus Stenotrophomons Lctocillus Serrti Enterococcus Pseudomons Rlstoni Sphingomons Spiroplsm Host OTU1 OTU2 OTU3 OTU5 OTU6 OTU8 OTU9 OTU11 OTU14 OTU17 Tomto Tomto Tomto (men) (9.56) (2.16) (29.32) (1.72) (1.4) (.55) (1.5) (4.6) (2.18) (7.49) Potto Potto Potto (men) (8.81) (5.11) (1.65) - (.42) (11.79) - (.1) (.3) - Eggplnt Eggplnt Eggplnt (men) (18.88) (8.71) (.4) - (.6) - - (.8) (.1) - Bufflour Bufflour Bufflour (men) (22.82) (23.75) - (52.39) (.28) (.9) - (.6) - (.38) Horsenettle Horsenettle Horsenettle (men) (5.69) (93.79) - (.1) (.43) Nightshde Nightshde Nightshde (men) (32.2) (57.46) (.6) - (.9) - (.3) (.39) (.2) (8.15)

13 Tle S4. OTU tle of cteril communities from Colordo potto eetle lrve tht were rered on different host plnts. OTUs over 1% of totl reds were included. OTU ID Domin Phylum Clss Order Fmily Genus 1 Bcteri Proteocteri Gmmproteocteri Enterocteriles Enteroctericee Enterocter 2 Bcteri Firmicutes Bcilli Lctocillles Streptococccee Lctococcus 3 Bcteri Proteocteri Gmmproteocteri Xnthomondles Xnthomondcee Stenotrophomons 4 Bcteri Proteocteri Gmmproteocteri Enterocteriles Enteroctericee unclssified 5 Bcteri Firmicutes Bcilli Lctocillles Lctocillcee Lctocillus 6 Bcteri Proteocteri Gmmproteocteri Enterocteriles Enteroctericee Serrti 8 Bcteri Firmicutes Bcilli Lctocillles Enterococccee Enterococcus 9 Bcteri Proteocteri Gmmproteocteri Pseudomondles Pseudomondcee Pseudomons 11 Bcteri Proteocteri Betproteocteri Burkholderiles Burkholdericee Rlstoni 13 Bcteri Proteocteri Gmmproteocteri Pseudomondles Morxellcee Acinetocter 14 Bcteri Proteocteri Alphproteocteri Sphingomondles Sphingomondcee Sphingomons 17 Bcteri Tenericutes Mollicutes Entomoplsmtles Spiroplsmtcee Spiroplsm 18 Bcteri Proteocteri Gmmproteocteri Pseudomondles Pseudomondcee unclssified 21 Bcteri Proteocteri Betproteocteri Burkholderiles Burkholdericee Rlstoni 23 Bcteri Proteocteri Betproteocteri Burkholderiles Burkholdericee Burkholderi 28 Bcteri Firmicutes Bcilli Bcillles Stphylococccee Stphylococcus 29 Bcteri unclssified unclssified unclssified unclssified unclssified 3 Bcteri Proteocteri Alphproteocteri Rhizoiles Brdyrhizoicee Brdyrhizoium 36 Bcteri unclssified unclssified unclssified unclssified unclssified 38 Bcteri Proteocteri Alphproteocteri Culocterles Culoctercee Phenylocterium 4 Bcteri Proteocteri Gmmproteocteri Pseudomondles Pseudomondcee unclssified 42 Bcteri Proteocteri Betproteocteri Burkholderiles Commondcee Curvicter 44 Bcteri Proteocteri Alphproteocteri Rhodocterles Rhodoctercee Prcoccus 55 Bcteri Proteocteri Betproteocteri Neisseriles Neissericee unclssified 6 Bcteri Firmicutes Bcilli Lctocillles Streptococccee Streptococcus 61 Bcteri Proteocteri Alphproteocteri Sphingomondles Sphingomondcee Sphingomons 62 Bcteri Proteocteri Gmmproteocteri Enterocteriles Enteroctericee unclssified 13 Bcteri Actinocteri Actinocteri Actinomycetles Streptomycetcee unclssified 123 Bcteri Tenericutes Mollicutes Entomoplsmtles Spiroplsmtcee Spiroplsm 134 Bcteri Proteocteri Betproteocteri Burkholderiles Oxloctercee Undicterium

14 138 Bcteri Firmicutes Bcilli Lctocillles Streptococccee Streptococcus 144 Bcteri Actinocteri Actinocteri Actinomycetles Dermctercee Brchycterium 154 Bcteri Bcteroidetes Sphingocteri Sphingocteriles Chitinophgcee Hydrotle 16 Bcteri Proteocteri Gmmproteocteri Pseudomondles Morxellcee Acinetocter 169 Bcteri Bcteroidetes Sphingocteri Sphingocteriles Chitinophgcee unclssified 172 Bcteri Bcteroidetes unclssified unclssified unclssified unclssified 175 Bcteri Firmicutes Bcilli Bcillles Bcillles_Incerte_Sedis_XI Gemell 189 Bcteri Acidocteri Acidocteri unclssified unclssified unclssified 223 Bcteri Proteocteri Alphproteocteri Rhizoiles Phylloctericee Phyllocterium 232 Bcteri Actinocteri Actinocteri Actinomycetles Mycoctericee Mycocterium 258 Bcteri Firmicutes Clostridi Clostridiles Ruminococccee Ruminococcus 276 Bcteri Proteocteri Gmmproteocteri Psteurellles Psteurellcee Hemophilus 285 Bcteri Firmicutes Clostridi Clostridiles Lchnospircee unclssified 293 Bcteri Actinocteri Actinocteri Actinomycetles Corynectericee Corynecterium 342 Bcteri Proteocteri Alphproteocteri Rhodospirillles Acetoctercee Acidocell 46 Bcteri Actinocteri Actinocteri Actinomycetles unclssified unclssified 444 Bcteri Acidocteri Acidocteri unclssified unclssified unclssified 448 Bcteri Proteocteri Alphproteocteri Culocterles Culoctercee Brevundimons 47 Bcteri Firmicutes Clostridi Clostridiles Lchnospircee Clostridium 499 Bcteri Bcteroidetes Flvocteri Flvocteriles Flvoctericee unclssified 652 Bcteri Proteocteri Betproteocteri Neisseriles Neissericee Neisseri 72 Bcteri Proteocteri Deltproteocteri Bdellovirionles Bdellovirioncee Bdellovirio 723 Bcteri Firmicutes Bcilli Lctocillles Aerococccee Aiotrophi 755 Bcteri Bcteroidetes Sphingocteri Sphingocteriles Chitinophgcee Sediminicterium 1235 Bcteri Firmicutes Bcilli Lctocillles Crnoctericee Dolosigrnulum 1397 Bcteri Acidocteri Acidocteri unclssified unclssified unclssified 1482 Bcteri unclssified unclssified unclssified unclssified unclssified 1513 Bcteri Firmicutes Bcilli Lctocillles Streptococccee Lctococcus 1552 Bcteri Bcteroidetes Bcteroidi Bcteroidles Prevotellcee unclssified 1578 Bcteri Firmicutes Clostridi Clostridiles Lchnospircee unclssified 158 Bcteri Actinocteri Actinocteri Actinomycetles Corynectericee Corynecterium

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