Novel Approaches in Fighting Sudden Death Syndrome in ......Acknowledgements Bhattacharyya Lab...

Madan K. BhattacharyyaAgronomy Department

Iowa State University

Novel Approaches inFighting Sudden

Death Syndrome inSoybean

http://soysdsresistance.ag.iastate.edu/index.html

Dr

Why to enhance sudden death syndrome(SDS) resistance in soybean?



Dr

Why to enhance sudden death syndrome(SDS) resistance in soybean?

v The estimated soybean yield suppression fromSDS in 2010 was 2.1% of total yield, which wasvalued at $0.82 billion.

v Growing SDS resistant cultivars is the majormethod of controlling this disease.

v Unfortunately, SDS resistance encoded by a largenumber QTL; and breeding SDS resistance ischallenging.

v We need novel single-genes encoding SDSresistance.


Two Novel Approaches to EnhanceSDS Resistance in Soybean

1) Understand how the pathogen causesSDS and apply the knowledge to createSDS resistant soybean lines.

2) Identify and express plant genes toenhance SDS resistance.

3/16/16 4


Lead PI: Madan K. Bhattacharyya (ISU)Co-Pis: Liang Dong (ISU)

Jing Yong Ye (UTSA)

SDS symptoms – (i) root necrosis and (ii) foliar SDS

Small chlorotic spotson leaves

Dead interveinal tissue falls off

Leaf drop –followed by flower and pod drop

Young seedlings Mature plants

Pathogen remains in infected roots.


3/16/16 6

v In North America, Fusarium virguliforme causes SDS.v In South America, F. tucumaniae is the major causal

agent of SDS.v Both are haploid organisms.v They produce sickle-shaped conidia for spread of the

disease.v They overwinter as chlamydospores.

v


Express plant genes

Microfluidic Assays

Dong Lab, Iowa State University


3/16/16 8

How quickly conidium germinate?

3/16/16 9

Conidia development

3/16/16 10

Structure of the MAT locus

Hughes et al. 2013: Mycologia 106:686–697.

3/16/16 11

Frequency distributions of alleles at the Mat locusamong the Fusarium spp.

Hughes et al. 2013: Mycologia 106:686–697.

A maximum-likelihood tree of Fusarium species (based on genome-wide SNP data)

BRR

BRR (bean root rot)

SDS

SDS

SDS

SDS

BRR

Huang et al. 2016: PloS One, Under Revision Huang et al. 2016: PloS One, Under Revision

BRR


Phylogenetic tree of F. virguliforme isolates.

13



Phylogenetic tree of F. tucumaniae isolates.

14


Two Approaches to EnhanceSDS Resistance in Soybean

1) Understand how the pathogen causesSDS and apply the knowledge to createSDS resistance.

2) Express plant genes to enhance SDSresistance.

3/16/16 15

FvTox1 – acidic proteincauses foliar SDS

3/16/16 16Brar et al. (2011) MPMI 24: 1179-1188

Pudake et al. (2013) Current Genetics, 59:107-117


Lead PI: Madan K. Bhattacharyya (ISU)Co-PIs: Liang Dong (ISU)

Jing Yong Ye (UTSA)

Brar and Bhattacharyya 2012: MPMI 25:817–824



Dr

Work flow for identifying peptides fromphage display peptides libraries

3/16/16 18Wang et al. (2015) PLoS OneDOI:10.1371/journal.pone.0145156



Dr

Four classes of M13 phage clonesdisplayed FvTox1-interacting peptides.

3/16/16 19

Class (sequence) Clone Number Peptide Library

Class I (SYLPETIYEYRL)

1,2,4,5,6,7,8,10,11,12,13,14,15,16,17,18,19,20,23,28,29,30,36,38,39

Ph.D.-12

Class Ⅱ (VENKTRYHDREV)

3,9,22,25,27,37 Ph.D.-12

Class Ⅲ (HEGAWHNYARSV)

24 Ph.D.-12

ClassⅣ (SNGRVAD)

31 Ph.D.-7



Dr

Nine synthetic genes encodingFvTox1-interacting peptides

20


Dr

21

Transgenic soybean lines carrying FvTox1-interactingPep -1 showed enhanced foliar SDS resistance.


FvTox1-interacting peptides conferenhanced tolerance of roots to FvTox1.

A

B D

CR² = 0.51712

0

10

20

30

40

50

60

0 1 2 3 4 5 6

Roo

t Rot

Sev

erity

(%)

Foliar SDS Score

R² = 0.83681

0

10

20

30

40

50

0 1 2 3 4 5 6

Stem

Rot

Sev

erity

(%)

Foliar SDS Score

R²#=#0.83919#

0

20

40

60

80

100

120

140

0 1 2 3 4 5 6

Deve

lopm

ent o

f New

Ro

ot (

%)

Foliar SDS Score

A

C

B

22Williams 82 Williams 82 + Pep 1


Dr

23

0 0.5

1 1.5

2 2.5

3 3.5

4

MN1

606

Pep

9C -

7

Pep

8C -

9

Pep

1C -

12

Pep

5C -

14

Pep

2C -

8

Pep

5C -

4

Pep

1 - 6

Pep

9 -

25

Pep

9C -

2

Pep

1 -

7

Pep

2C

- 14

Pep

8 -

14

Pep

8C

- 7

Pep

2C

- 6

Pep

5 -

15

Pep

1C

- 6

Pep

9 -

3

Spen

cer

Will

iam

s 82

Pep

8 -

17

Folia

r SDS

Sco

re

0 0.5

1 1.5

2 2.5

3 3.5

4 4.5

5

Folia

r SDS

Sco

re

Resistant Susceptible

02468

1 01 21 4

Num

ber o

f Pla

nts

R e s i s t an tS us c e p t ib l e

B

A

Transgenic soybean lines showed enhancedresistance to F. virguliforme infection.

B. Wang and M.K. Bhattacharyya, unpublished


Express plant genes

Summary

1. Demonstrated that FvTox1-interacting Pep-1enhances foliar SDS resistance.

2. FvTox1-interacting peptides conferringenhanced foliar SDS resistance also showedimproved tolerance of roots to FvTox1.

3. FvTox1 most likely causes suppression of rootgrowth and root necrosis in addition to foliarSDS.

24





1. Arabidopsis nonhost resistance genes2. Altered expression of soybean genes

3/16/16 25

Phytophthora sojae susceptible (pss) mutants

pen1-1 pss1

Mutant Family Mutant Family Mutant Family

pss1 0.2B17I9 pss11 0.25B8C1 pss21 0.25B9D7

pss2 0.2B3D2 pss12 0.25 B9A1 pss22 0.25B9F1

pss3 0.3B6A7 pss13 0.25B8H5 pss23 0.2B20I2

pss4 0.2B21F1 pss14 0.3B6E8 pss24 0.2B9G4

pss5 0.2B15I6 pss15 0.3B6H8 pss25 0.2B8B7

pss6 0.2B4G5 pss16 0.25B7F8 pss26 0.3B1C7

pss7 0.2B21B9 pss17 0.25B7G4 pss27 0.3B8B8

pss8 0.2B4A5 pss18 0.25B9C3 pss28 0.25B9E3

pss9 0.2B21D6 pss19 0.3B6E4 pss29 0.2B1A9

pss10 0.25B7I5 pss20 0.3B6C8 pss30 0.25B7D2

Identified 30 pss mutants.


Dr

Fourteen pss mutants are alsosusceptible to F. virguliforme.

pss12

pss15

pss16

pss1

pss2

pss5

pss3

pss10

pss18

pss19

pss20


Dr

Field evaluation of transgenic soybean plantscarrying Arabidopsis nonhost resistance Pss1 gene

0 20 40 60 80

100

SDS

Res

ista

nt P

lant

s (%

)

30-Sep 15-Oct

29B. Wang et al. unpublished


Dr

30

0

20

40

60

80

100

SDS

Resi

stan

t Pla

nts

(%)

Field evaluation of transgenic soybean plantscarrying Arabidopsis nonhost resistance

Pss30 gene193-16.2 35S-Pss30-16.2 232-2 Prom2-Pss30-2

232-5 Prom2-Pss30-5 Williams 82

A

C

B

D

193-16.2 35S-Pss30-16.2 232-2 Prom2-Pss30-2

232-5 Prom2-Pss30-5 Williams 82

A

C

B

D

S. Kambakam et al., unpublished





1. Arabidopsis nonhost resistance genes2. Altered expression of soybean genes

3/16/16 31


Soybean genes suppressed duringF. virguliforme infection.

32



Expression of GmSAMT1 enhances SDS resistancein transgenic soybean plants under field conditions

33



34

Expression of GmDS1 enhances SDS resistance intransgenic soybean plants under field conditions


Current Status

Gene Name Growth Chamber Field EvaluationPss1 Resistant ResistantPss30 Resistant ResistantPss6 Resistant Not YetPss21 Resistant Not YetPss25 Partially Resistant Not YetGmSAMT1 Resistant ResistantGmDS1 Resistant ResistantGmARP1 Resistant ResistantGmSEQ1 Resistant Not Yet

3/16/16 35

Acknowledgements

Bhattacharyya Lab

Sekhar Kambakam, Ph.D.

Micheline Ngaki, Ph.D.

Bing Wang, Ph.D.

Siva Swaminathan, Ph.D.

Jill Heinrich

Previous Scientists

Hargeet Brar

Ramesh Pudake, Ph.D.

Devinder Sandhu, Ph.D.

Min Xu, Ph.D.

Binod Sahu, Ph.D.

Rishi Sumit, M.S.

Prashant Singh, Ph.D.

Program Grant no. 2013-68004-20374 11:25:52 AM 36

Dong LabXuan Qiao

Huang LabAnindya Das

Ye Lab (UT San Antonio)Bailin Zhang, Ph.D.

Teresa J. HughesKerry O’Donnell

USDA-ARS

ISU Plant Transformation FacilityIowa Soybean Association

CPBRUnited Soybean Board

3/16/16 37

Slide 1Slide 2Slide 3Two Novel Approaches to Enhance SDS Resistance in SoybeanSlide 5Slide 6Slide 7Slide 8Slide 9Slide 10Slide 11Slide 12Slide 13Slide 14Two Approaches to Enhance SDS Resistance in SoybeanFvTox1 – acidic protein causes foliar SDSSlide 17Slide 18Slide 19Slide 20Slide 21Slide 22Slide 23Slide 24Two Approaches to Enhance SDS Resistance in SoybeanSlide 26Slide 27Slide 28Slide 29Slide 30Two Approaches to Enhance SDS Resistance in SoybeanSlide 32Slide 33Slide 34Current StatusAcknowledgementsSlide 37

Novel Approaches in Fighting Sudden Death Syndrome in ......Acknowledgements Bhattacharyya Lab...

Documents

Transcript of Novel Approaches in Fighting Sudden Death Syndrome in ......Acknowledgements Bhattacharyya Lab...