Figure S1 Morphological features of 58 endophytic strains (Red box: SH-1.2-R-15).

72

68

74

69

54

65 66

62 53

47

38

44 43 42 41

51

40 39

49 50

27

30

28

36 34

29 21

31

26

20

25

15 16

14

18 17

6 4 5 3 2 1

46 52

75

59 60 58

7 9 10 11 13

19

29

70

71 73

67

Table S1 16S rRNA analysis of 58 endophytic strains.

NO. Strains Host plant Parts Closest sequences by BlastN Percent

Identity

Genbank accession

1 SH-1.1-R-1 Dendrobium

officinale -

one year

old-1

Root Bacillus sp. (in: Bacteria) strain 12D6 97.14% MN784185

2 SH-1.1-R-2 Root Stenotrophomonas rhizophila strain PgBE58 99.72% MN784186

3 SH-1.1-R-3 Root Bacillus sp. BS98 99.90% MN784187

4 SH-1.1-R-4 Root Streptomyces sp. strain SHP 1-2 99.80% MN784188

5 SH-1.1-R-5 Root Stenotrophomonas sp. strain H9 100.00% MN784189

6 SH-1.1-R-6 Root Streptomyces sp.SR3-82 gene 99.85% MN784190

7 SH-1.1-R-7 Root Bacillus thuringiensis strain ZLynn1000-39 97.59% MN784191

8 SH-1.1-R-8 Root Same as 5 - -

9 SH-1.1-R-9 Root Aerococcus viridans strain NBRC 12219 99.78% MN784192

10 SH-1.1-L-1 Leaf Bacillus cereus strain BC2 99.90% MN784193

11 SH-1.1-L-2 Leaf Bacillus thuringiensis strain GA-A07 99.90% MN784194

12 SH-1.1-L-3 Leaf Same as 58 - -

13 SH-1.1-S-1 Stem Stenotrophomonas sp. Strain FB14 99.93% MN784195

14 SH-1.2-R-1 Dendrobium

officinale -

one year

old-2

Root Streptomyces sp. NK04104 96.58% MN784196

15 SH-1.2-R-2 Root Streptomyces sp. CdTB01 97.13% MN784197

16 SH-1.2-R-3 Root Pseudomonas sp. BMS12 99.33% MN784198

17 SH-1.2-R-4 Root Streptomyces sp. g2b 99.79% MN784199

18 SH-1.2-R-5 Root Streptomyces sp. HF-2 96.82% MN784200

19 SH-1.2-R-6 Root Pseudomonas sp. Strain IAE244 99.92% MN784201

20 SH-1.2-R-7 Root Streptomyces sp. LCB 0297 97.94% MN784202

21 SH-1.2-R-8 Root Streptomyces sp. strain QJSt8 97.47% MN784203

22 SH-1.2-R-9 Root Same as 7 - -

23 SH-1.2-R-10 Root Same as 8 - -

24 SH-1.2-R-11 Root Same as 21 - -

25 SH-1.2-R-12 Root Bacillus cereus strain ULT15 99.60% MN784204

26 SH-1.2-R-13 Root Streptomyces sp. TM-74 97.51% MN784205

27 SH-1.2-R-14 Root Streptomyces fulvissimus strain DSM 40593 97.64% MN784206

28 SH-1.2-R-15 Root Streptomyces chartreusis strain ISP 5085 99.21% MN784207

29 SH-1.2-R-16 Root Streptomyces variabilis strain NRRL B-3984 97.52% MN784208

30 SH-1.2-R-17 Root Streptomyces sp. strain TM-A158 99.86% MN784209

31 SH-1.2-R-18 Root Bacillus thuringiensis strain GA-A07 chromosome

99.93% MN784210

32 SH-1.2-R-19 Root Same as 58 - -

33 SH-1.2-L-1 Leaf Micromonospora aurantiaca strain IMB16-201 99.92% MN784211

34 SH-3.1-R-1 Dendrobium

officinale -

three years

old-1

Root Bacillus sp. (in: Bacteria) strain 12D6 98.67% MN784212

35 SH-3.1-R-2 Root Same as 36 - -

36 SH-3.1-R-3 Root Streptomyces sp. GSENDO-0578 96.80% MN784213

37 SH-3.1-R-4 Root Same as 50 - -

38 SH-3.1-R-5 Root Streptomyces sp. 13-22 96.92% MN784214

39 SH-3.1-R-6 Root Streptomyces sp. MI02-7b 99.58% MN784215

40 SH-3.1-R-7 Root Streptomyces anulatus strain 174456 97.06% MN784216

41 SH-3.1-R-8 Root Same as 38 - -

42 SH-3.1-R-9 Root Streptomyces sp. DSM 40835 clone K12 99.70% MN784217

43 SH-3.1-R-10 Root Bacillus thuringiensis strain JW-1 99.81% MN784218

44 SH-3.1-R-11 Root Bacillus thuringiensis strain GA-A07 100.00% MN784219

45 SH-3.1-R-12 Root Same as 66 - -

46 SH-3.1-R-13 Root Bacillus sp. JAS24-2 chromosome 100.00% MN784220

47 SH-3.1-R-14 Root Streptomyces sp. strain MUSC11 98.46% MN784221

48 SH-3.1-R-15 Root Same as 66 - -

49 SH-3.1-R-16 Root Streptomyces sp. strain BOR09 93.69% MN784222

50 SH-3.1-R-17 Root Viridibacillus sp. strain TM-B117 99.69% MN784223

51 SH-3.1-R-19 Root Streptomyces sp. SR3-82 96.17% MN784224

52 SH-3.1-R-20 Root Bacillus thuringiensis strain GCU1 99.79% MN784225

53 SH-3.1-R-21 Root Streptomyces prunicolor NBRC 13075 99.80% MN784226

54 SH-3.1-R-22 Root Streptomyces sp. strain GDMCC 60254 96.51% MN784227

55 SH-3.1-L-1 Leaf Same as 59 - -

56 SH-3.1-L-2 Leaf Same as 65 - -

57 SH-3.1-L-3 Leaf Same as 60 - -

58 SH-3.1-S-1 Stem Streptomyces sp. strain GDMCC 60254 96.92% MN784228

59 SH-3.1-S-2 Stem Bacillus cereus strain NRRL B-23957 98.73% MN784229

60 SH-3.1-S-3 Stem Streptomyces sp. SM17 99.31% MN784230

61 SH-3.1-S-4 Stem Same as 65 - -

62 SH-3.2-R-1 Dendrobium

officinale -

three years

old-2

Root Bacillus sp. HT-Z74-B2 98.55% MN784231

63 SH-3.2-R-2 Root Same as 50 - -

64 SH-3.2-R-3 Root Same as 60 - -

65 SH-3.2-R-4 Root Bacillus cereus strain GE16 98.78% MN784232

66 SH-3.2-R-5 Root Streptomyces sp. GKU 867 100.00% MN784233

67 SH-3.2-R-6 Root Bacillus thuringiensis strain QZL38 chromosome

100.00% MN784234

68 SH-3.2-R-7 Root Streptomyces rochei 7434AN4 DNA 99.79% MN784235

69 SH-3.2-R-8 Root Streptomyces sp. Endophyte N2 chromosome 100.00% MN784236

70 SH-3.2-R-9 Root Bacillus cereus strain Sneb2000 100.00% MN784237

71 SH-3.2-R-10 Root Brevundimonas sp. strain 7002-176 100.00% MN784238

72 SH-3.2-R-11 Root Corynebacterium sp. strain ABYHD3-2 99.85% MN784239

73 SH-3.2-R-12 Root Streptomyces sp. TJ-27 97.98% MN784240

74 SH-3.2-R-13 Root Bacillus sp. AR4-2 chromosome 100.00% MN784241

75 SH-3.2-L-1 Leaf Bacillus thuringiensis strain GA-A07 100.00% MN784242

Figure S2 Scatterplot of 75 cultured endophytic strains screened in triplicate at 25 or 50 µg/mL in Hep3B2.1-

7 cell viability assay. Data are shown normalized to percent max response of the high control (Medium only

group). Each data point represents the mean and standard deviation of three replicates in 96 wells. The hit

cutoff calculated as the average percent inhibition plus three times the standard deviation of the low control

wells, is shown as the black line at 11.7%.

Table S2. Strains showed inhibition against Hep3B2.1-7 cell more than the hit cutoff (>11.7%).

No. Endophytic strains %Inhibition No. Endophytic strains %Inhibition

2 SH-1.1-R-2 22.14±4.46 54 SH-3.1-R-22 35.74±2.31

4 SH-1.1-R-4 44.24±1.39 56 SH-3.1-L-2 18.64±1.16

16 SH-1.2-R-3 14.67±1.99 58 SH-3.1-S-1 14.87±3.17

18 SH-1.2-R-5 97.24±0.38 59 SH-3.1-S-2 47.80±0.99

19 SH-1.2-R-6 89.27±9.58 61 SH-3.1-S-4 12.00±1.38

24 SH-1.2-R-11 23.97±4.63 62 SH-3.2-R-1 33.77±2.22

27 SH-1.2-R-14 98.44±0.36 63 SH-3.2-R-2 41.40±1.71

28 SH-1.2-R-15 20.30±3.76 64 SH-3.2-R-3 31.27±4.01

31 SH-1.2-R-18 34.47±2.74 65 SH-3.2-R-4 12.30±3.31

33 SH-1.2-L-1 14.47±1.28 66 SH-3.2-R-5 47.07±1.86

42 SH-3.1-R-9 23.14±1.77 67 SH-3.2-R-6 62.20±6.76

43 SH-3.1-R-10 26.37±2.21 68 SH-3.2-R-7 98.67±0.62

44 SH-3.1-R-11 12.17±4.47 69 SH-3.2-R-8 89.50±2.46

47 SH-3.1-R-14 45.34±1.71 73 SH-3.2-R-12 49.97±1.41

Figure S3 Scatterplot of 75 cultured endophytic strains screened in triplicate at 25 or 50 µg/mL in S. aureus

inhibitory assay. Data are shown normalized to percent max response of the high control (Bacterial treated

with 100 µg/mL Ampicillin). Each data point represents the mean and standard deviation of three replicates

in 96 wells. The hit cutoff calculated as the average percent inhibition plus three times the standard deviation

of the low control wells, is shown as the black line at 36.2%.

Table S3 Strains showed inhibition against S. aureus more than the hit cutoff (>36.2%).

No. Endophytic strains %Inhibition No. Endophytic strains %Inhibition

12 SH-1.1-L-3 41.86±3.50 37 SH-3.1-R-4 51.50±1.96

13 SH-1.1-S-1 50.13±5.59 44 SH-3.1-R-11 41.81±8.64

14 SH-1.2-R-1 36.70±6.74 48 SH-3.1-R-15 40.45±21.15

15 SH-1.2-R-2 40.17±5.94 50 SH-3.1-R-17 48.24±15.86

18 SH-1.2-R-5 95.12±3.34 52 SH-3.1-R-20 47.21±7.18

23 SH-1.2-R-10 54.04±7.21 56 SH-3.1-L-2 46.37±3.41

24 SH-1.2-R-11 72.75±6.14 57 SH-3.1-L-3 38.59±3.27

26 SH-1.2-R-13 51.38±3.27 68 SH-3.2-R-7 100.00±0.24

27 SH-1.2-R-14 99.66±0.58 69 SH-3.2-R-8 99.32±0.53

28 SH-1.2-R-15 100.12±0.09 70 SH-3.2-R-9 51.01±6.67

29 SH-1.2-R-16 40.21±5.97

Figure S4 Scatterplot of 75 samples screened in triplicate at 25 or 50 µg/mL in E. coli inhibitory assay. Data

are shown normalized to percent max response of the high control (Bacterial treated with 100 µg/mL

Ampicillin). Each data point represents the mean and standard deviation of three replicates in 96 wells. The

hit cutoff calculated as the average percent inhibition plus three times the standard deviation of the low

control wells, is shown as the black line at 26.4%.

Table S4 Strains showed inhibition against E. coli more than the hit cutoff (>26.4%).

No. Endophytic strains %Inhibition

59 SH-3.1-S-2 31.49±6.86

Scheme S1 Work-up scheme for the Streptomyces sp. SH-1.2-R-15.

Figure S5. (+) and (‒)-ESI-MS of compound 1.

Figure S6. HRESI-MS spectrum of compound 1.

m / z1 0 0 2 0 0 3 0 0 4 0 0 5 0 0 6 0 0

0

1 0

2 0

3 0

4 0

5 0

6 0

7 0

*M S D 1 S P C , tim e= 5 .0 0 0 o f D :\u s er1 \d a ta \Z H A O H U IM IN \Z H A O H U IM IN 2 0 1 9 -0 4 -2 3 1 5 -1 0 -0 2 \D J 6 0 -7 L H 7 P 3 .D E S -A P I, P os , S c an , F r

M a x : 1 0 3 4 0 0

431.2

292.2

410.2

409.2

m / z1 0 0 2 0 0 3 0 0 4 0 0 5 0 0 6 0 0

0

2 0

4 0

6 0

8 0

1 0 0

*M S D 2 S P C , tim e= 5 .0 1 0 o f D :\u s er1 \d a ta \Z H A O H U IM IN \Z H A O H U IM IN 2 0 1 9 -0 4 -2 3 1 5 -1 0 -0 2 \D J 6 0 -7 L H 7 P 3 .D E S -A P I, N eg , S c an , F r

M a x : 1 4 4 1 2 8

407.2

249.0

227.2

113.2

200 250 300 350 400 450 500 550 600 650 700

m/z

0

10

20

30

40

50

60

70

80

90

100

0

10

20

30

40

50

60

70

80

90

100

Re

lative

Ab

un

da

nce

409.2444

431.2260274.2740

318.3000230.2478197.0921 447.1993362.3262 475.2771 552.3053

391.2328

504.2688 638.4754574.2767

407.2294

507.1544284.0769 473.2618255.1341 341.1707 439.2177394.2227 550.2842

421.2029

719.4003663.3711215.0092176.0700 599.3121

NL: 3.03E9

60-7LH7P3#549 RT: 3.66 AV: 1 T: FTMS + p ESI Full ms [100.0000-1500.0000]

NL: 1.18E9

60-7LH7P3#548 RT: 3.65 AV: 1 T: FTMS - p ESI Full ms [100.0000-1500.0000]

Figure S7. 1H NMR (500 MHz, DMSO-d6) spectrum of compound 1.

Figure S8. 1H NMR (500 MHz, CDCl3) spectrum of compound 1.

Figure S9. 13C NMR (125 MHz, CDCl3) spectrum of compound 1.

Figure S10. 1H-1H COSY (500 MHz, CDCl3) spectrum of compound 1.

Figure S11. HSQC (500 MHz, CDCl3) spectrum of compound 1.

Figure S12. HMBC (500 MHz, CDCl3) spectrum of compound 1.

Figure S13. ROESY (500 MHz, CDCl3) spectrum of compound 1.

Figure S14. (+) and (‒)-ESI-MS of compound 2.

m / z1 0 0 2 0 0 3 0 0 4 0 0 5 0 0

0

2 0

4 0

6 0

8 0

1 0 0

*M S D 1 S P C , tim e= 5 .1 7 1 o f D :\u s er1 \d a ta \Z H A O H U IM IN \Z H A O H U IM IN 2 0 1 9 -0 4 -2 3 1 5 -1 0 -0 2 \D J 6 0 -7 L H 7 P 4 .D E S -A P I, P os , S c an , F r

M a x : 3 0 5 2 8 0

425.2

445.2

306.2 4

24.2

423.2

m / z1 0 0 2 0 0 3 0 0 4 0 0 5 0 0

0

1 0

2 0

3 0

4 0

*M S D 2 S P C , tim e= 5 .1 6 2 o f D :\u s er1 \d a ta \Z H A O H U IM IN \Z H A O H U IM IN 2 0 1 9 -0 4 -2 3 1 5 -1 0 -0 2 \D J 6 0 -7 L H 7 P 4 .D E S -A P I, N eg , S c an , F r

M a x : 1 3 6 8 3 2

249.0

227.2

422.4

113.2

Figure S15. HRESI-MS spectrum of compound 2.

Figure S16. 1H NMR (500 MHz, DMSO-d6) spectrum of compound 2.

200 250 300 350 400 450 500 550 600 650 700

m/z

0

10

20

30

40

50

60

70

80

90

100

0

10

20

30

40

50

60

70

80

90

100

Re

lative

Ab

un

da

nce

423.2597

445.2412306.1807278.1851

211.1075 461.2160331.1863240.0871 646.3412545.3363 567.3140405.2542356.2169 481.1740 668.3198

421.2451

521.1702407.2104

484.2415339.1921 375.1832286.0928255.1342 457.2207226.9763 557.2133439.2525

644.3322616.3126

NL: 4.19E9

60-7LH7P4#565 RT: 3.77 AV: 1 T: FTMS + p ESI Full ms [100.0000-1500.0000]

NL: 1.68E9

60-7LH7P4#568 RT: 3.79 AV: 1 T: FTMS - p ESI Full ms [100.0000-1500.0000]

Figure S17. 1H NMR (500 MHz, CDCl3) spectrum of compound 2.

Figure S18. 13C NMR (125 MHz, CDCl3) spectrum of compound 2.

Figure S19. 1H-1H COSY (500 MHz, CDCl3) spectrum of compound 2.

Figure S20. HSQC (500 MHz, CDCl3) spectrum of compound 2.

Figure S21. HMBC (500 MHz, CDCl3) spectrum of compound 2.

Figure S22. ROESY (500 MHz, CDCl3) spectrum of compound 2.

Figure S23. (+) and (‒)-ESI-MS of compound 3.

Figure S24. HRESI-MS spectrum of compound 3.

m / z2 0 0 4 0 0 6 0 0 8 0 0

0

2 0

4 0

6 0

8 0

1 0 0

*M S D 1 S P C , tim e= 4 .6 4 3 o f D :\u s er1 \d a ta \Z H A O H U IM IN \Z H A O H U IM IN 2 0 1 9 -0 3 -1 4 2 0 -1 6 -3 5 \D J -1 0 0 -4 .D E S -A P I, P os , S c an , F rag :

M a x : 3 6 0 0 8

101.2

213.2

660.0

980.0

360.6

271.2

143.2

335.2

m / z2 0 0 4 0 0 6 0 0 8 0 0

0

2 .5

5

7 .5

1 0

1 2 .5

1 5

1 7 .5

*M S D 2 S P C , tim e= 4 .6 3 3 o f D :\u s er1 \d a ta \Z H A O H U IM IN \Z H A O H U IM IN 2 0 1 9 -0 3 -1 4 2 0 -1 6 -3 5 \D J -1 0 0 -4 .D E S -A P I, N eg , S c an , F rag :

M a x : 6 6 8 5

147.0

671.0

113.2

640.0

639.2

400 450 500 550 600 650 700 750 800 850 900 950 1000 1050 1100 1150 1200

m/z

0

10

20

30

40

50

60

70

80

90

100

0

10

20

30

40

50

60

70

80

90

100

Re

lative

Ab

un

da

nce

663.1688

989.2375384.3091

417.1764

870.4260

679.1422

641.4619431.1870 475.3294 818.2929555.5335 914.4601760.5887

969.3007

1143.2715

702.4931

639.1683

702.1633

655.1626

757.1034401.0848 789.0935491.1156 843.1236

NL: 4.39E7

CHARESINms640#535 RT: 3.66 AV: 1 T: FTMS + p ESI Full ms [100.0000-1500.0000]

NL: 7.52E8

CHARESINms640#526 RT: 3.60 AV: 1 T: FTMS - p ESI Full ms [100.0000-1500.0000]

Figure S25. 1H NMR (500 MHz, DMSO-d6) spectrum of compound 3.

Figure S26. 13C NMR (125 MHz, DMSO-d6) spectrum of compound 3.

Figure S27. 1H-1H COSY (500 MHz, DMSO-d6) spectrum of compound 3.

Figure S28. HSQC (500 MHz, DMSO-d6) spectrum of compound 3.

Figure S29. HMBC (500 MHz, DMSO-d6) spectrum of compound 3.

Figure S30. (+) and (‒)-ESI-MS of compound 4.

Figure S31. HRESI-MS spectrum of compound 4.

Figure S32. 1H NMR (500 MHz, DMSO-d6) spectrum of compound 4.

Figure S33. 13C NMR (125 MHz, DMSO-d6) spectrum of compound 4.

Figure S34. (+) and (‒)-ESI-MS of compound 5.

Figure S35. HRESI-MS spectrum of compound 5.

Figure S36. 1H NMR (500 MHz, CDOD3) spectrum of compound 5.

Figure S37. 13C NMR (125 MHz, CDOD3) spectrum of compound 5.

Figure S38. (+) and (‒)-ESI-MS of compound 6.

Figure S39. HRESI-MS spectrum of compound 6.

Figure S40. 1H NMR (500 MHz, CDOD3) spectrum of compound 6.

Figure S41. 13C NMR (125 MHz, CDOD3) spectrum of compound 6.

Figure S42. (+) and (‒)-ESI-MS of compound 7.

Figure S43. HRESI-MS spectrum of compound 7.

Figure S44. 1H NMR (500 MHz, DMSO-d6) spectrum of compound 7.

Figure S45. 13C NMR (125 MHz, DMSO-d6) spectrum of compound 7.