wel come

Non-host Resistance in Plants:

New insights

Jayappa PALB4255

JayappaID.No: 4255Sr. M.Sc (Agri)

OUTLINE OF SEMINAR

Introduction

Components

Types

Applications

Case studies

Conclusion

INTRODUCTION

“Whatever the plant defence or resistance, it is controlled by its genes”

(Agrios., 2007 )

higher

,

Non Host response of the cell/ nucleus

against the pathogen . Ultimately

no disease to the plants

Host response of the cell/ nucleus against

the pathogen and cause disease

Non Host response of the cell/ nucleus

against the pathogen . Ultimately no disease

to the plants

Host response of the cell/ nucleus against the pathogen as R

Gene mediated resistance develops

and cause no disease

Nucleus response

NH pathogen signals

NH

pathogen

Avr

path

ogen

Vir

pathog

en Infection/

diseaseIndu

ced

defe

nce

&

PTI

Prefo

rmed

defen

c &

phys

ical

barr

iers

R- gene m

ediated im

munity

( Shamim et al., 2013)

No

dise

ase

No disease

Interaction and disease development phenomenon in plant

Is a broad spectrum resistance Resistance of an entire plant species to all isolates of a microbial species Preformed barriers such as cell wall, cuticle, phytoanticipins Induced defense responses such as lignin accumulation, production of

antimicrobials like phytoalexins, HR response, induction of pathogenesis-related (PR) proteins

Eg., Barley is typically susceptible to P. hordei, to Which wheat is a nonhost. The reverse is true for P. triticina

Non host resistance Plant Innate Immunity, Species Resistance, Durable resistance,

Basal defense and Nonspecific resistance

Components of Non host resistance

Preformed or passive defence mechanisms

Inducible plant defence mechanisms

Plant defense signalling

Broad-spectrum disease resistance genes

Preformed or passive defense mechanisms

Sheath around a hypha

Cork layer Phenolic compounds

T, Tyloses in xylem vessels

Abscission layer

Necrotic defense reaction in a cell

N, nucleus; PS, protoplasmic strands; Z, zoospore; H, hypha; G, granular material; NC, necrotic cell

( Tomiyama et al., 1999 )

Resistant potato variety infected by Phytophthora infestans.

Plant Defense Signalling

(Yang et al., 1999)

Non host pathogen

Gene/protein Function in non-host resistance (NHR) References

PAMPsPep-13 Induces defense responses in non-host plants

like PotatoNurnberger et al. (1994)

Harpin (Hrp Z) Elicits HR-like cell death and defense responses in various plants

He et al. (1993)

Genes PEN1(Penetration1)/ROR2

This gene is involved in timely deposition of papillae during non-host interactions.

Collins et al. (2003)

NHO1 Required for NHR of Arabidopsis against Pseudomonas syringae pv. phaseolicola

Kang et al. (2003)

R-gene mediated genes SGT1 Silencing of SGT1 in N. benthamiana

compromises NHR against P. syringae pv. maculicola and Xanthomonas axonopodis pv. Vesicator

Peart et al.(2002)

Heat-shock proteins (Hsps)

Silencing of Hsp90 and Hsp70 in N. benthamiana individually compromised NHR against P. cichorii.

Kanzaki et al. (2003)

Genes involved in NHR

Types of Non host resistance

( Mysore et al.,2004)

Examples of type I

( Mysore et al.,2004)

Pathogen Strain NH plants Symptoms

P. s. pv. syringe B76 Arabidopsis None

P.s. pv. phaseolicola S2 N. tabacum None

P.s.pv.delphini PDDCC529 Arabidopsis None

P. infestans 88069 N. clevalandi None

P. infestans 88069 N. tabacum cv. xanthi None

Cont..

( Mysore et al.,2004)

Type II non host resistance

Pathogen Strain NH plants Symptoms

P. s.pv. tomato DC3000 N. tabacum HR

P. s. pv. glycinea PG4180 N. tabacum HR

P. s. pv. pisi ATCC 11055 N. tabacum HR

P. s. pv. syringae 61 N. tabacum HR

P. cichorii 83-1 Arabidopsis HR

Application of NHR in Agriculture

Application References

1. Successful use of a PRR gene, EFR, from Arabidopsis in N. benthamiana and tomato to reduce the growth of their respective host pathogens

(Lacombe et al., 2010)

2. R gene, Bs2, from pepper has been shown to impart resistance to X. campestris pv. vesicatoria in tomato (Tai et al., 1999)

3. Resistance to Erwinia carotovora, the causal agent of potato (Solanum tuberosum) tuber soft rot, was achieved by replacing chromosome 8 of potato with the corresponding chromosome from its wild relative Solanum brevidens

(Tek et al.,2004)

CASE STUDIES

Objective : To know the function of MPKs in NHR to M.oryzae in A.thaliana

Quantitative analysis of NHR to M. oryzae in A. thaliana

Mean frequency of M. oryzae penetration into A. thaliana mutants @ 48hpi (Expressed as a total no. of infection sites.)

Mean length of infection hyphae measured @48hpi

(Okawa and Ishikawa, 2013)

Quantitative analysis of Post penetration resistance to M.oryzae in A.thaliana

I. Cell wall penetrationII. Establishment of infection

hyphaeIII. Elongation of infection

hyphaeIV. Branch formation on

infection hyphae

Microscopic views of infection sites in Arabidopsis mutants

Light microscopic view @ 48hpi

Fluorescence microscopic

view @ 48hpi

pen2

Pen2 mpk6

48hpi

(Okawa and Ishikawa, 2013)

Objective: Detection of DNAse and their activity in Non host resistance

Non-host to Fusarium solani f.sp. phaseoli (Fsph) Host to Fusarium solani f.sp. pisi (Fspi )

Pea

,2013

Effect of VdDNase enzyme concentrations on the non-host resistance (to Fsph) and susceptibility (to Fspi) responses of pea endocarp tissue.

Ng, no growthDg, spores degradedDt, spores deteriorated+ hyphal growth equalent to length of the spore++ two length+++ 3length

VdDNase dilution series applied to the pea endocarp surface10 min prior to applying Fsph or Fspi spore suspensions

(Hadwiger et al., 2013)

VdDNase induces pisatin accumulation

Vd DNase assay and pisatin eliciting activity:(A) Pod halves (0.5 g fr. wt.) were treated

with 2 ml of superdex 75 fractions. (B) Superdex 75 fractions (2.0 ml) were incubated in the standard DNase buffer for 10 min wit 0.2micro gram of plasmid DNA.

DNase activity

24 h

(Hadwiger et al., 2013)

A northern blot analysis of the expression of PR gene, DRR49

Treatments: 1 = 02 = 5000 Units VdDNase, 3 = 2500 units VdDNase, 4 = 1250 units VdDNase 5 = 06 = 5000 units VdDNase7 = 2500 units VdDNase 8 = 1250 units VdDNase

4Hr

7Hr

mRNA levels

(Hadwiger et al., 2013)

Objective : To confirm that PRR activity is retained after its transfer between two plant families

EFR(Elongation factor receptor, PRR) Elf18( Elongation factor 18aa)

EF-TU(Elongation factor thermo unstable)EF-Tu is highly conserved in all Phytopathogenic bacteria

2010

Transgenic expression of EFR in N. benthamiana and tomato

N. benthamiana plants S. lycopersicum plants

Oxidative Burst triggered by 100 nM elf18 or flg22

Four-week-old Four-week-old

Oxidative Burst triggered by 100 nM elf18 or flg22

(Lacombe et al., 2010)RLU= measures cleanliness of leaf surface

Gene expression of marker genes determined by RT-PCR

(Lacombe et al., 2010)

Transgenic expression of EFR in N. benthamiana

a. Infection with P. syringae pv. syringae (Pss) B728ab. Infection with P. syringae pv. tabaci (Pta) 11528

(Lacombe et al., 2010)

N. benthamiana infection with A. tumefaciens A281

(Lacombe et al., 2010)

Transgenic expression of EFR in tomato

(Lacombe et al., 2010)

Objective: 1. Identification of non-adapted P. cucumerina isolates 2. To find out the role of different trp-derived compounds.

Plectosphaerella cucumerina isolates Pc1187 and Pc2127 are non-adapted necrotrophic fungi on Arabidopsis wild-

type plants

Lactophenol Trypan Blue staining of

inoculated leaves at 12 and 20 h post

inoculation

Relative quantification of fungal DNA

Average disease rating

(Sanchez- Vallet et al., 2010)

Quantitative real-time PCR quantification of fungal DNA (Pcb-tubulin)

Average disease rating

Lactophenol Trypan Blue staining of inoculated leaves

(Sanchez- Vallet et al., 2010)

Objective: 1. Inoculation with a Xoo results in induction of HR and

nonhost resistance2. Exogenous supply of accelerated Xoo induced HR

,2015

Effect of the Xoo inoculum concentration on HR induction.

Xoo at 1×, cfuconcentrations was inoculated in fully expanded N. benthamianaleaves. The inoculated plants were grown at 28 °C.

24h

(Li et al., 2015)

is essential for HR induction during the interaction between Xoo and N. benthamiana

Effect of exogenous on Xoo-induced HR.

Effect of exogenous catalase on Xoo-induced HR

DAB staining for detection

O

O

(Li et al., 2015)

Ability of three Xoo-derived T3SS gene mutants to induce HR and accumulation in N. benthamiana

DAB staining for detection

(Li et al., 2015)

Bacterial numbers were counted in catalase treated leaves inoculated with Xoo and the Δhpa1 mutant

(Li et al., 2015)

CONCLUSION