Defence Mechanisms of a Resistant Monocot Model to Phytophthora Cinnamomi - DocsLib

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Defence mechanisms of a resistant monocot model to Phytophthora cinnamomi

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ACKNOWLEDGEMENTS

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ii

TABLE OF CONTENTS

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7*(>00*/!.9TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT6'

7*(>0( 79TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT6'4

7*(>0 .9?* (*><TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT64'

7*(>0!7*) (*>< .99<( (*><TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT64'''

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CHAPTER 1: INTRODUCTION AND LITERATURE REVIEW ...... 1 STST !)%0-3,"TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTS

STTT>-+7!#2#.* ,2. 2&-%#,1TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTU

STUT(&#/#,31Phytophthora ...... U

STVTPhytophthora cinnamomi. ,"1TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTV

STVTST(&#+-0.&-*-%7 ," 1#63 *1203!230#1-$P. cinnamomiTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTX

STVTTT(&#*'$#!7!*# ,"',$#!2'-,.0-!#11-$P. cinnamomiTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTY STVTUT#-*#!3* 0.&7*-%#,7 ,"'"#,2'$'! 2'-,TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT[

STVTVT(&##$$#!21-$P. cinnamomi-,4#%#2 2'-,TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTSS STVTWT)300#,2!-,20-*+#2&-"1TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTSV

STWT* ,20#1'12 ,!#+#!& ,'1+1TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTSW

STWTST* ,20#1'12 ,!#2-. 2&-%#,15'2&"'$$#0#,2*'$#127*#1 ,"+-"#1-$ ',$#!2'-,TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTSX

STWTTT-12V1.#!'$'!"'1# 1#0#1'12 ,!#TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTSY

STWTUT<-,V&-12"'1# 1#0#1'12 ,!#TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTSZ

STXT.#1'12 ,!#2-Phytophthora 1.#!'#1TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTU

STYT.#1'12 ,!#2-P. cinnamomi ...... TW

STYTST.#1'12 ,!#2-P. cinnamomi 22&#!#**3* 0*#4#*TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTY

STYTTT.#1'12 ,!#2-P. cinnamomi 22&#+-*#!3* 0*#4#*TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTZ

iii

STZT/#,-+'!1 ..0- !-2{"7-$.* ,2VP. cinnamomi',2#0 !2'-,1TTTTTTTTTTTTTTTZ ST[T* ,2+-"#*1712#+1TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT[

ST[TSTZea mays 1 +-,-!-2+-"#*$-00#1'12 ,!#2-P. cinnamomiTTTTTTTTTTTTTTTTTTTTTTTTTTTTTUR STSRT('1- (#!2'4#1TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTUS

CHAPTER 2: CHARACTERISATION OF RESISTANCE RESPONSES TO PHYTOPHTHORA CINNAMOMI IN THE MODEL MONOCOT ZEA MAYS ...... 34

2.1. Introduction ...... 34

TTSTTT#$',',%.* ,20#1.-,1#12-P. cinnamomi ...... UW

2.2. Materials and Methods ...... 39

TTTTST/0-52& ,"+ ',2#, ,!#-$Z. mays ...... U[

TTTTTT>.2'+'1 2'-,-$Z. mays%0-52&1712#+1$-0#6.#0'+#,2 * , *71'1TTTTTTTTTTTTTTTTTTVR TTTTUT/0-52& ,"+ ',2#, ,!#-$#6.#0'+#,2 *.* ,215'2&', 1-'*V$0##.* ,2 %0-52&1712#+TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTVS

TTTTVT#2#0+', 2'-,-$-.2'+3+.* ,2 %#$-0',-!3* 2'-,TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTVU

TTTTWT0'#*"!-**#!2'-,-$1-'*1 +.*#1TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTVV

TTTTXT*1-* 2'-,-$P. cinnamomi$0-+1-'*1 +.*#1TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTVV

TTTTYT*1-* 2'-,-$P. cinnamomi-,1#*#!2'4#+#"' . ,"#12 *'1&+#,2 ,"+ ',2#, ,!#-$5-0)',%!3*230#1TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTVW

TTTTZT)-,$'0+ 2'-,-$. 2&-%#,'"#,2'27 ,".30'27TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTVX

TTTT[T)-,$'0+ 2'-, ,"+ ',2#, ,!#-$4'03*#,!#-$P. cinnamomi'1-* 2#1TTTTTTTTTTTTTVY TTTTSRTH--1.-0#.0-"3!2'-,TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTVZ

TTTTSST#2#0+', 2'-,-$8--1.-0#"#,1'27TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTV[

TTTTSTT*,-!3* 2'-,.0-!#"30#1$-0Z. mays ,"L. angustifolius...... WR TTTTSUT)-,$'0+ 2'-,-$',-!3* 2'-,TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTWR

TTTTSVT*,$#!2'-,2'+#!-301#TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTWS

TTTTSWT.#* 2'4#.* ,2&# *2&.-12V',-!3* 2'-,5'2&P. cinnamomi ...... WT

TTTTSXT 0 +#2#012-"#2#0+',#0#1'12 ,!#2-P. cinnamomi ...... WT

iv

TTTTSYT## 130#+#,2-$.* ,2$0#1&5#'%&2TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTWT

TTTTSZT## 130#+#,2-$0--2*#,%2& ,".#0!#,2 %#0--2*#1'-,*#,%2&TTTTTTTTTTTTTTTTTTTTTTTWU

TTTTS[TI3 ,2'$'! 2'-,-$!&*-0-.&7**5'2&',*# $2'113#TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTWU

TTTTTRT#2#0+', 2'-,-$*#4#*1-$P. cinnamomi5'2&',',-!3* 2#".* ,20--21TTTTTTTWV TTTTTST2 2'12'! * , *71'1-$" 2 TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTWV

TTTTTTT<-02�, *-2 , *71'12-',4#12'% 2#P. cinnamomi !!3+3* 2'-,', ',-!3* 2#"Z. mays0--22'113#TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTWV TTTTTUT('113#1 +.*',%$-0.< #620 !2'-,1TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTWW

TTTTTVT.< #620 !2'-,20' *1 ,"-.2'+'1 2'-,TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTWW

TTTTTWT.< #620 !2'-,$0-+Z. mays0--22'113#TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTWW TTTTTXT.< %#*#*#!20-.&-0#1'1$-0,3!*#'! !'"20 ,1$#0TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTWX

TTTTTYT<-02�,&7 0'"'1 2'-,TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTWY

TTTTTZT<3!*#'! !'"20 ,1$#04' ! .'** 07 *-22',%TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTWY

TTTTT[T0#V&7 0'"'1 2'-,TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTWZ

TTTTURT0- #* #**',% ,".30'$'! 2'-,TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTWZ

TTTTUST##+ 0 ,#5 1&',%TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTW[

TTTTUTT0'*+#6.-130# ,""#4#*-.+#,2TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTXR

2.3. Results...... 61

TTUTST#-0.&-*-%'! *'"#,2'$'! 2'-,-$P. cinnamomi'1-* 2#1TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTXS TTUTTT#-*#!3* 0!-,$'0+ 2'-,-$'1-* 2#1TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTXS

TTUTUT*,$#!2'-,2'+#!-301#TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTXT

TTUTVT* ,2&# *2&$-**-5',%',-!3* 2'-,5'2&P. cinnamomi ...... XU TTUTWT.#120'!2#"*#1'-,.0-"3!2'-, ,"$-0+ 2'-,-$* 2#0 *.0'+ 070--21', P. cinnamomi',-!3* 2#"Z. mays ...... XW TTUTXT* ,2$0#1&5#'%&2.-12',-!3* 2'-,TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTXX

TTUTYT.--2*#,%2&1.-12',-!3* 2'-,TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTXY

TTUTZT7#1'-,*#,%2&1.-12',-!3* 2'-,TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTXZ

v

TTUT[T.#* 2'4#!&*-0-.&7**!-,!#,20 2'-,',*# $2'113#TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTYR

TTUTSRT)-+. 0'1-,-$P. cinnamomi&7.& *%0-52&5'2&',',-!3* 2#"0--2 1#%+#,21TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTYT

TTUTSST 11#11+#,2-$0#1'12 ,!#2-P. cinnamomi',Z. mays ,"L. angustifolius ... YV

TTUTSTTI3 ,2'$'! 2'-,-$P. cinnamomi*#4#*15'2&',',-!3* 2#"Z. mays0--2 2'113#TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTYW

2.4. Discussion ...... 77 TTVTST.#1'12 ,!#0#1.-,1#1 22&#+ !0-1!-.'!*#4#*TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTYY

TTVTTT0-0+ 2'-,-$0#120'!2#"*#1'-,1 ,"* 2#0 *0--21',Z. mays ...... Y[

TTVTUT<-02�, *-2 , *71'1-$P. cinnamomi',-!3* 2#"Z. mays0--22'113# "#+-,120 2#" 0#"3!2'-,',. 2&-%#,*#4#*1-4#02'+#TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTZR

TTVTVT)&*-0-.&7**!-,2#,2'1,-20#"3!#"',P. cinnamomi',-!3* 2#"Z. mays*# $ 2'113#TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTZS

CHAPTER 3: IDENTIFICATION OF CELLULAR COMPOUNDS PRODUCED IN PHYTOPHTHORA CINNAMOMI INOCULATED ZEA MAYS ROOT TISSUE ...... 83

3.1. Introduction ...... 83 UTSTST* ,2!#**5 **1TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTZU

UTSTTT203!230# ,"$3,!2'-,-$.* ,20--22'113#TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTZV

UTSTUT)#**5 **$-02'$'! 2'-,TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTZW

UTSTVT(&#&7.#01#,1'2'4#0#1.-,1#. ,"0# !2'4#-67%#,1.#!'#1.>TTTTTTTTTTTZX

3.2. Materials and Methods ...... 89

UTTTSTZ. mays%0-52& ,"+ ',2#, ,!#TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTZ[

UTTTTTP. cinnamomi+ ',2#, ,!# ,"8--1.-0#.0-"3!2'-,TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTZ[ UTTTUT*,-!3* 2'-,.0-!#"30#1TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTZ[

UTTTVT#2#!2'-,-$ 32-$*3-0#1!#,!#TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT[R

UTTTWT ,'*',# *3#12 ',',%$-0! **-1#"#2#!2'-,TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT[R

UTTTXT&*-0-%*3!',-*V)*12 ',',%$-0"#2#!2'-,-$*'%,',TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT[S

UTTTYTU_U_"' +',- #,8'"',#12 ',',%$-0"#2#!2'-,-$&7"0-%#,.#0-6'"#TTTTTTTTTTTTTTTTT[S

vi

UTTTZTI3 ,2'$'! 2'-,-$*'%,',31',%2&'-%*7!-*'! !'""#0'4'2'1 2'-,TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT[T

UTTT[T*,&' '2'-,-$! **-1#TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT[U

UTTTSRT*,&' '2'-,-$&7"0-%#,.#0-6'"#TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT[U

UTTTSSTI3 ,2'$'! 2'-,-$0#1'12 ,!#0#1.-,1#1$-**-5',%',&' '2-020# 2+#,2TTTTTTTTTT[V

UTTTSTT2 2'12'! * , *71'1-$" 2 TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT[V

UTTTSUT0'+#0"#1'%, ,"-.2'+'1 2'-,TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT[V

UTTTSVT('113#1 +.*',%$-0.< #620 !2'-,1TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT[V

UTTTSWT.< #620 !2'-,$0-+Z. mays0--22'113#31',%(.*8-*TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT[W

UTTTSXT<-02�,&7 0'"'1 2'-, ," , *71'1TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT[X

3.3. Results...... 97 UTUTST. .'"* 2#0 *0--2$-0+ 2'-,4'#5#"3,"#0!?V#.'$*3-0#1!#,!#TTTTTTTTTTTTTTTTTTTTTTTTT[Y

UTUTTT0-"3!2'-,-$ 32-$*3-0#1!#,2+ 2#0' *',',-!3* 2#"0--22'113#TTTTTTTTTTTTTTTTTTTTTT[Y

UTUTUT) **-1#.0-"3!2'-,',',-!3* 2#"0--22'113#TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTSRR

UTUTVT7'%,',.0-"3!2'-,',',-!3* 2#"0--22'113#TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTSRS

UTUTWT7'%,',/3 ,2'$'! 2'-,TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTSRV

UTUTXT7"0-%#,.#0-6'"#.0-"3!2'-,',',-!3* 2#"0--22'113#TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTSRW

UTUTYT7-! *'1#"!#**"# 2&',',-!3* 2#"2'113# 2 22#+.2#".#,#20 2'-,1'2#1TTTTTTSRW

UTUTZT 11#11+#,2-$"#$#,!#0#1.-,1#1$-**-5',%',&' '2-020# 2+#,2TTTTTTTTTTTTTTTTTTTTTSRY

UTUT[T96.0#11'-,-$*'%,', ,"T>TV0#* 2#"%#,#1',P. cinnamomi',-!3* 2#" Z. mays0--22'113#TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTSRZ

3.4. Discussion ...... 110

UTVTST#.-1'2'-,-$ 32-$*3-0#1!#,2+ 2#0' *',P. cinnamomi',-!3* 2#"Z. mays 0--22'113#TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTSSR

UTVTTT7"0-%#,.#0-6'"#T>T !!3+3* 2'-,',P. cinnamomi',-!3* 2#" Z. mays0--22'113#TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTSSS

UTVTUT) **-1#.0-"3!2'-,',P. cinnamomi',-!3* 2#"Z. mays0--22'113#TTTTTTTTTTTTTTTTTSSU

vii

UTVTVT7'%,',"#.-1'2'-,',P. cinnamomi',-!3* 2#"Z. mays0--22'113#TTTTTTTTTTTTTTTTTTTTSSV

UTVTWT96.0#11'-,-$"#$#,!#V0#* 2#"%#,#1',P. cinnamomi',-!3* 2#"Z. mays 0--22'113#TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTSSW

CHAPTER 4: GENE EXPRESSION PROFILING OF ZEA MAYS ROOT TISSUE IN RESPONSE TO PHYTOPHTHORA CINNAMOMI...... 117 4.1. Introduction ...... 117

VTSTSTZea mays 1 +-"#*$-0+-*#!3* 0 , *71'1-$0#1.-,1#12-P. cinnamomi .. SSZ VTSTTT/#,-+'!1$-0.* ,2V. 2&-%#,',2#0 !2'-,1TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTSS[

4.2. Materials and Methods ...... 122 VTTTST* ,2%0-52& ,"+ ',2#, ,!#TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTSTT

VTTTTT 2&-%#,%0-52& ,"+ ',2#, ,!#TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTSTT

VTTTUT*,-!3* 2'-,.0-!#"30#1TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTSTT

VTTTVT96.#0'+#,2 *"#1'%,TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTSTU

VTTTWT9620 !2'-, ,".30'$'! 2'-,-$2-2 *.< TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTSTW

VTTTXT)-,$'0+ 2'-,-$2-2 *.< .30'27TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTSTX

VTTTYT#'!0- 00 7.0-!#11',%* #**',%Q&7 0'"'1 2'-, ,"1! ,,',%TTTTTTTTTTTTTTTTTTTTTTTTTSTY

VTTTZT'-',$-0+ 2'!1 , *71'1TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTST[

VTTT[T'-*-%'! *. 2&5 7 , *71'1TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTSUR

VTTTSRT 2 13 +'11'-,2-.3 *'!0#.-1'2-07TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTSUR

4.3. Results...... 131

VTUTST'$$#0#,2' *%#,##6.0#11'-,',Z. mays0--21$-**-5',%',-!3* 2'-,5'2& P. cinnamomi8--1.-0#1TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTSUS

VTUTTT9 0*7"#$#,!#0#1.-,1#12-P. cinnamomi','2' 2#"',Z. mays 0--21TTTTTTTTTTTTTTTTSUT VTUTUT!.V0#%3* 2'-,-$"#2-6'$7',%%*32 2&'-,#V20 ,1$#0 1#20 ,1!0'.21TTTTTTTTTTTTTTTTSUU

VTUTVT'$$#0#,2' *#6.0#11'-,-$ ,"920#* 2#"%#,#1TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTSUU

VTUTWT*,"3!2'-,-$. 2&-%#,"#%0 "',%#,87+#1Q2-6',1 ,"#,87+#',&' '2-01TTSUV

VTUTXT# # < '-*-%'! *. 2&5 7 , *71'1-.2'+'1#"$-0Z. mays ...... SVU

4.4. Discussion ...... 147

viii

VTVTST9 0*73.V0#%3* 2'-,-$.&72- *#6',1TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTSVZ

VTVTTT(&# 92. 2&5 7TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTSV[

VTVTUT. .'"',"3!2'-,-$%*32 2&'-,#V20 ,$#0 1#1TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTSWT

VTVTVT 2&-%#,#1'1V0#* 2#" ,"2& 3+ 2',V*')#.0-2#',1TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTSWU

VTVTWT7'%,', ,"! **-1#TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTSWV

CHAPTER 5: GENERAL DISCUSSION ...... 159 WTST3++ 07TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTSW[

WTTT)& 0 !2#0'1 2'-,-$Z. mays 1 0#1'12 ,2+-,-!-2+-"#*$-0#6 +',',% "#$#,!#+#!& ,'1+12-P. cinnamomi3,"#0!-,20-**#"!-,"'2'-,1TTTTTTTTTTTTTTTTTTTTTTTTTTTTTSXR WTUT#4#*-.+#,2 ,"-.2'+'1 2'-,-$ ,- (#!2'4#+#2&-"2-"#2#0+',# 0#1'12 ,!#2-P. cinnamomi3,"#0!-,20-**#"!-,"'2'-,1TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTSXR

WTVTI3 ,2'$'! 2'-,-$P. cinnamomi5'2&',Z. mays0--22'113#"#+-,120 2#" 0#"3!2'-,',. 2&-%#,*#4#*1-4#02'+#TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTSXT

WTWT*"#,2'$'! 2'-,-$',"'4'"3 *!#**3* 0!-+.-3,"1.0-"3!#"',Z. mays0--2 2'113#',0#1.-,1#2-P. cinnamomi',-!3* 2'-,0#4# *#" +3*2'V$ !#2#""#$#,!# 0#1.-,1#TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTSXU WTXT/#,##6.0#11'-,.0-$'*',%0#13*2#"',2&##*3!'" 2'-,-$+-*#!3* 00#1.-,1#1 2-P. cinnamomi',Z. mays0--22'113# ,"2&#'"#,2'$'! 2'-,-$)#7"#$#,!#V 0#* 2#"%#,#1TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTSXV

WTXTST )#70-*#$-02� .'"%#,#0 2'-,-$.> ,"/(1',Z. mays0--22'113# ',"#$#,!#2-P. cinnamomiTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTSXX WTYT(&#'+.-02 ,20-*#-$0--22'113#',"#$#,!#0#1.-,1#12-P. cinnamomiTTTTTTTTTSXY WTZT ,',!0# 1#"3,"#012 ,"',%-$"#$#,!#+#!& ,'1+1','2' 2#"', 0#1'12 ,2 .* ,2',0#1.-,1#2-P. cinnamomi',$#!2'-,TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTSXZ WT[T0', *!-,!*31'-,1TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTSYS

REFERENCES...... 173

APPENDICES ...... 197 ..#,"'6STRT0-0+ 2'-,-$0#120'!2#"*#1'-,',2&# 3120 *' ,, 2'4#%0 11 Lomandra filiformisTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTS[Y

..#,"'6STST>,*',#%#,-+'!0#1-30!#1$-0Z. maysTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTS[Z

..#,"'6TTRT* ,2%0-52&20' *1$-0Z. maysTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTS[[

ix

..#,"'6TTST*1-* 2'-,-$P. cinnamomi$0-+$'#*"!-**#!2#"1-'*1 +.*#1TTTTTTTTTTTTTTTTRR

..#,"'6TTTT##"' ,"1-*32'-,1$-0P. cinnamomi%0-52& ,"+ ',2#, ,!#TTTTTRS

..#,"'6TTUTPhytophthora< '"#,2'$'! 2'-,TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTRU ..#,"'6TTVT(&# ..*'! 2'-,-$=-!&_1.-123* 2#12-.* ,2. 2&-*-%7TTTTTTTTTTTTTTTTTTTTTTRV

..#,"'6TTWT-*32'-,1 ,"5 1&!-,"'2'-,1$-0,-02�,&7 0'"'1 2'-,TTTTTTTTTTTTTTTTRW

..#,"'6UTRTc',# ..*#c.< #620 !2'-,.0-!#"30#TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTRX

..#,"'6VTRT **"'$$#0#,2' **7#6.0#11#"%#,#1',P. cinnamomi',-!3* 2#" Z. mays0--22'113#TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTRZ ..#,"'6VTST?'13 *'1 2'-,2--*1$-0.0#V.0-!#11',%+'!0- 00 7" 2 TTTTTTTTTTTTTTTTTTTTTTTTS[

x

LIST OF FIGURES

CHAPTER 1: Figure 1.1. #-0.&-*-%7-$P. cinnamomi 1#63 *1203!230#1',!3*230#TTTTTTTTTTTTTTTTTTTTTTTTTZ

Figure 1.2. 7'$#!7!*#-$P. cinnamomi ...... SR

Figure 1.3. 9$$#!21-$"'1# 1#! 31#" 7P. cinnamomi',!301'-,', 3120 *' , , 2'4#4#%#2 2'-,TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTST

Figure 1.4. '1# 1#.0-%0#11'-,! 31#" 7P. cinnamomi',2&# 3120 *' ,, 2'4# .* ,2Xanthorrhoea australis ...... SU

Figure 1.5. )300#,2),-5*#"%#-$.* ,20#1'12 ,!#+#!& ,'1+12- '-20-.&1 ," ,#!0-20-.&1TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTS

Figure 1.6.)300#,2),-5*#"%#-$.* ,20#1.-,1#12-P. cinnamomi',2�--2 2'113#-$0#1'12 ,2 ," -0$'#*"0#1'12 ,21.#!'#1TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT[

CHAPTER 2: Figure 2.1. /0-52&-$Z. mays1##"*',%15'2&', 1-'*V$0##%0-52&1712#+TTTTTTTTTTTTTTTTVT

Figure 2.2. (#," 7-*"1##"*',%1-$Z. mays ,"L. angustifolius ...... VU

Figure 2.3.*,-!3* 2'-,-$Z. mays0--21TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTWR

Figure 2.4.#-0.&-*-%'! *'"#,2'$'! 2'-,-$P. cinnamomi'1-* 2#1TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTXS

Figure 2.5.#-*#!3* 0!-,$'0+ 2'-,-$'1-* 2#.30'27TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTXT

Figure 2.6.P. cinnamomi',$#!2'-,2'+#!-301#',Z. mays0--22'113#TTTTTTTTTTTTTTTTTTTTTTTTTXU

Figure 2.7.* ,2&# *2&-$Z. mays ,"L. angustifolius$-**-5',%',-!3* 2'-, 5'2&P. cinnamomi ...... XV

Figure 2.8..#120'!2#"*#1'-,.0-"3!2'-, ,"$-0+ 2'-,-$* 2#0 *0--21', ',-!3* 2#"Z. mays ...... XW

Figure 2.9. 00#1&5#'%&2-$!-,20-* ,"P. cinnamomi',-!3* 2#"Z. mays ," L. angustifolius.* ,21TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTXX

Figure 2.10. )-+. 0'1-,-$0--2%0-52&-$!-,20-* ,"P. cinnamomi',-!3* 2#" Z. mays ,"L. angustifolius ...... XY

Figure 2.11. )-+. 0'1-,-$*#1'-,1-$P. cinnamomi',-!3* 2#"Z. mays ," L. angustifolius 0--21TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTXZ

xi

Figure 2.12. )-+. 0'1-,-$*#1'-,*#,%2&1-$P. cinnamomi',-!3* 2#"Z. mays ,"L. angustifolius 0--21TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTX[

Figure 2.13. 4#0 %#0#* 2'4#!&*-0-.&7**!-,!#,20 2'-,1',*# $2'113#-$ !-,20-* ,"P. cinnamomi',-!3* 2#"Z. mays ,"L. angustifolius ...... YR

Figure 2.14. )-+. 0'1-,-$!&*-0-1'1',*# $2'113#-$P. cinnamomi',-!3* 2#" Z. mays ,"L. angustifolius 0--21TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTYS

Figure 2.15. P. cinnamomi&7.& *%0-52&',0--21#%+#,21-$',-!3* 2#"Z. mays ,"L. angustifolius ...... YU

Figure 2.16. 7#1'-,*#,%2&1',P. cinnamomi',-!3* 2#"L. angustifolius ...... YW

Figure 2.17. <-02�, , *71'1-$ !!3+3* 2'-,-$P. cinnamomi',',-!3* 2#" Z. mays0--22'113#TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTYX

CHAPTER 3: Figure 3.1. , 2-+7-$Z. mays0--21TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTZW

Figure 3.2. -12V#+ 07-,'!* 2#0 *0--2$-0+ 2'-,',P. cinnamomi',-!3* 2#" Z. mays0--21TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT[Y

Figure 3.3. 0-"3!2'-,-$ 32-$*3-0#1!#,!#',#.'"#0+ *2'113#-$ Z. mays 0--21TT[[

Figure 3.4.0-"3!2'-,-$ 32-$*3-0#1!#,!#',4 1!3* 02'113#-$Z. mays0--21TTTTSRR

Figure 3.5. ) **-1#"#.-1'2'-,',P. cinnamomi',-!3* 2#"Z. mays0--21TTTTTTTTTTTTTTTTTSRT

Figure 3.6. 7'%,',"#.-1'2'-, ',P. cinnamomi',-!3* 2#"Z. mays0--21TTTTTTTTTTTTTTTTTTTSRU

Figure 3.7.7'%,', !!3+3* 2'-,',P. cinnamomi',-!3* 2#"Z. mays0--21TTTTTTTTTTTTSRV

Figure 3.8. 7"0-%#,.#0-6'"# !!3+3* 2'-,',P. cinnamomi',-!3* 2#" Z. mays0--21TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTSRX

Figure 3.9.7-! *'1#"!#**"# 2&',2 1!3* 02'113#-$P. cinnamomi ',-!3* 2#"Z. mays 0--21TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTSRY

Figure 3.10. /#,##6.0#11'-,-$*'%,', ,"&7"0-%#,.#0-6'"#V0#* 2#"%#,#1TTTTTSR[

CHAPTER 4: Figure 4.1. 96.#0'+#,2 *"#1'%,$-0+'!0- 00 7 , *71'1-$Z. mays0--22'113#TTTSTV

Figure 4.2. < ,-0-.1! ,-$.< 7'#*" ,".30'27TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTSTY

xii

Figure 4.3. ?-*! ,-.*-25'2&%#,#1 00 ,%#" !!-0"',%2- '-*-%'! * ," 12 2'12'! *1'%,'$'! ,!#TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTST[

Figure 4.4. 03,!2'-, **7 ,,-2 2#"%#,#1"'$$#0#,2' **7#6.0#11#"', P. cinnamomi',-!3* 2#"Z. mays ...... SUT

Figure 4.5. # # <. 2&5 7 , *71'1-$"'$$#0#,2' **7#6.0#11#"%#,#1 '**3120 2',% ,-4#04'#5-$%#,#1',4-*4#"',)#7!#**3* 0$3,!2'-,1TTTTTTTTTTTTTTTTTTTTTTTTTTTSVW

Figure 4.6. # # <. 2&5 7 , *71'1-$"'$$#0#,2' **7#6.0#11#"%#,#1 ',4-*4#"',0#1.-,1#12- '-2'!120#11TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTSVX

Figure 4.7. !&#+ 2'!0#.0#1#,2 2'-,-$2&# '-17,2'1. 2&5 7',!*3"',% %#,#13.V0#%3* 2#"',P. cinnamomi',-!3* 2#"Z. mays0--21TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTSWS

Figure 4.8. # (-0"#$#,!#. 2&5 71Q',!*3"',%)#7%#,#1Q'"#,2'$'#"',2&'1 123"7',P. cinnamomi',-!3* 2#"Z. mays0--21TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTSWY

CHAPTER 5: Figure 5.1. 3++ 07-$2&#+ !0-1!-.'!Q!#**3* 0 ,"+-*#!3* 0"#$#,!# +#!& ,'1+1',"3!#"',P. cinnamomi',-!3* 2#"Z. mays0--22'113#'"#,2'$'#"', 2&'1123"7TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTSYR

APPENDIX: Figure A1T7#1'-,0#120'!2'-,',Lomandra filiformis0--22'113#TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTS[Y

Figure A2. )-+. 0'1-,-$1-'*V%0-5,.* ,21TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTS[[

Figure A3. (&#.0-!#11-$P. cinnamomi'1-* 2'-,$0-+1-'*1 +.*#1TTTTTTTTTTTTTTTTTTTTTTTTTTRR

Figure A4.# 2+ .0#.0#1#,2',%0#* 2'4#%#,##6.0#11'-,4 *3#1TTTTTTTTTTTTTTTTTTTTTTTTTTTTS[

xiii

LIST OF TABLES

CHAPTER 1: Table 1.1. (7.#* ,"(7.#**,-,V&-120#1'12 ,!#'"#,2'$'#"',.* ,21 % ',12 Phytophthora 1.#!'#1TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTV

CHAPTER 2: Table 2.1.)300#,2 ,".0#4'-31!* 11'$'! 2'-,1-$.* ,20#1.-,1#12- P. cinnamomi ...... UY Table 2.2. #/3#,!#-$Phytophthora%#,31V1.#!'$'!.0'+#01#2TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTVX Table 2.3. 11#11+#,2-$0#1.-,1#12-P. cinnamomi',Z. mays ," L. angustifolius0--21TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTYV

CHAPTER 3: Table 3.1. 0'+#01#2131#"',2&'1123"7TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT[W Table 3.2. 11#11+#,2-$0#1.-,1#12-P. cinnamomi',Z. mays0--21$-**-5',% ',&' '2-020# 2+#,21TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTSRZ

CHAPTER 4: Table 4.1. 03,!2'-, **7 ,,-2 2#"%#,#1 3.V0#%3* 2#" 2X&.'',P. cinnamomi ',-!3* 2#"Z. mays0--21TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTSUW Table 4.2. 03,!2'-, **7 ,,-2 2#"%#,#1 3.V0#%3* 2#" 2TV&.'',P. cinnamomi ',-!3* 2#"Z. mays0--21TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTSU[ Table 4.3.03,!2'-, **7 ,,-2 2#"%#,#1 "-5,V0#%3* 2#" 2X&.'',P. cinnamomi',-!3* 2#"Z. mays0--21TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTSVR Table 4.4. 03,!2'-, **7 ,,-2 2#"%#,#1 "-5,V0#%3* 2#" 2TV&.'',P. cinnamomi',-!3* 2#"Z. mays0--21TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTSVS Table 4.5. 03,!2'-, *%0-3.1-$"#$#,!#V0#* 2#"%#,#1"'$$#0#,2' **7#6.0#11#" 2X&.'',P. cinnamomi',-!3* 2#"Z. mays0--21TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTSVT Table 4.6. 03,!2'-, *%0-3.1-$"#$#,!#V0#* 2#"%#,#1"'$$#0#,2' **7#6.0#11#" 2TV&.'',P. cinnamomi',-!3* 2#"Z. mays0--21TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTSVU Table 4.7. /#,#1',2&#.&#,7*.0-. ,-'". 2&5 75&'!&5#0#,-2 "'$$#0#,2' **7#6.0#11#"',P. cinnamomi',-!3* 2#"Z. mays0--21TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTSWW

APPENDIX Table A1. PhytophthoraV1.#!'$'!)...... TRU Table A2. P. cinnamomi0#120'!2'-,"'%#12TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTRU

xiv

Table A3. **%#,#1"'$$#0#,2' **7#6.0#11#"',P. cinnamomi',-!3* 2#"Z. mays 0--22'113# 2X&.'TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTR[

Table A4. **%#,#1"'$$#0#,2' **7#6.0#11#"',P. cinnamomi',-!3* 2#"Z. mays 0--22'113# 2TV&.' ...... TSX

xv

LIST OF ABBREVIATIONS

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xvi

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VUT &-1.&-031UT

# . 2&-%#, 11-!' 2#"+-*#!3* 0. 22#0,

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xvii

PUBLICATIONS AND PRESENTATIONS ARISING FROM THIS RESEARCH

CONFERENCE PRESENTATIONS: )3**3+Q Q/3,,',%Q(=Q.--)#1Q9 ,") &'**Q#TRR[03," +#,2 * !-+.-,#,21-$0#1'12 ,!#2-Phytophthora cinnamomiS31',% +-"#*1712#+ ..0- !&QSY2&'#,,' * 3120 * 1' ,* ,2 2&-*-%7-!'#27)-,$#0#,!#Q <#5! 12*#Q<3

)3**3+Q Q.--)#1Q9 ,") &'**Q#TRRZ)& 0 !2#0'1 2'-,-$!#**3* 0 ," +-*#!3* 0.* ,20#1'12 ,!#+#!& ,'1+12-Phytophthora cinnamomiQ[2& *,2#0, 2'-, *)-,%0#11-$* ,2 2&-*-%7Q(-0',-Q*2 *7

.--)#1Q9Q)3**3+Q Q30'%&2Q#7Q/3,,',%Q(= ,") &'**Q#TRRZ ##!& ,'1+1-$, 230 *0#1'12 ,!#-$.* ,21 % ',12Phytophthora cinnamomi, *,2#0, 2'-, *)-,$#0#,!#-,'-2'!*,2#0 !2'-,1Q0'1 ,#QI7

)3**3+Q ,") &'**Q#TRRY)-+.-,#,21-$2�#1'12 ,!#0#1.-,1#1-$ .* ,212-Phytophthora cinnamomiQSX2&'#,,' * 3120 * 1' ,* ,2 2&-*-%7 -!'#27)-,$#0#,!#Q "#* '"#Q

PUBLICATIONS: ** 0"7!#Q ,") &'**Q#TRSR ,- (#!2'4#Q1#+'V/3 ,2'2 2'4#+#2&-"2- 11#110#1'12 ,!#2-Phytophthora cinnamomi',2&#+-"#*+-,-!-2Zea mays, 13 +'22#"$-0.3 *'! 2'-,

** 0"7!#Q Q.--)#1Q9Q) &'**Q#TRSR(0 ,1!0'.2'-, * , *71'1-$2&# "#$#,!#0#1.-,1#-$Zea mays0--21$-**-5',%',-!3* 2'-,5'2&Phytophthora cinnamomi, ',.0#. 0 2'-,

xviii

ABSTRACT

(&# 1-'*V -0,# --+7!#2#Q Phytophthora cinnamomi . ,"1 '12&#! 31 *

%#,2-$"#4 12 2',%"'1# 1#', -2&, 230 * ," %0'!3*230 *1712#+1

5-0*"5'"#T 1+ ** ,3+ #0 -$ .* ,21 0# 0#1'12 ,2 2- 2&# . 2&-%#,Q &-5#4#0

*'22*#'1),-5, -322&#+#!& ,'1+1-$"#$#,!##+.*-7#"',2#.* ,21T

3&'*#. 120#1# 0!&& 1$-!311#"-,.* ,20#1.-,1#1',2&#$'#*"Q ',. 02'!3* 0

2&-1#-$131!#.2' *#1.#!'#1Q2�#0#+ ',+ (-0% .1',2&#!300#,2),-5*#"%#

-$ .* ,2VP. cinnamomi ',2#0 !2'-,1 ', 0#1'12 ,2 .* ,21 2 -2& 2&# !#**3* 0 ,"

+-*#!3* 0 *#4#*T ""'2'-, **7Q 2�# '1 * !) -$ !-,1#,131 0#% 0"',% 2&#

"#$','2'-,-$.* ,20#1'12 ,!#2- P. cinnamomiT(�#1# 0!&.0#1#,2#"',2&'1

2'1 31#" +3*2'V$ !#2#"Q * -0 2-07V 1#" ..0- !& 5&'!& !-+.0'1#"

20 "'2'-, *.* ,2. 2&-*-%7Q&'12-!&#+'! * , *71'1 ,"20 ,1!0'.2'-, * , *71'1

2- ""0#11 2# )#7 % .1T (&# + (-0 $-!31 #',% 2&# #6 +', 2'-, -$ "#$#,!#

+#!& ,'1+1 !2'4 2#" ', 2&# 0--2 2'113# -$ 0#1'12 ,2 .* ,2 1.#!'#1 $-**-5',%

. 2&-%#,',-!3* 2'-,T

(&#+-,-!-2!0-..* ,2Zea mays5 1!& 0 !2#0'1#" 1 0#1'12 ,2+-"#*Q

2&0-3%& 2&# "#4#*-.+#,2 ," ..*'! 2'-, -$ 0'%-0-31 !0'2#0' 2- - (#!2'4#*7

"#2#0+',# 0#1'12 ,!# 2- 2&# . 2&-%#, 3,"#0 * -0 2-07 !-,"'2'-,1T (#

!0'2#0' ',!*3"#" !-+. 0'1-, -$ .* ,2 $0#1& 5#'%&2Q 0--2 *#,%2&Q *#1'-, *#,%2&Q

," 0#* 2'4# !&*-0-.&7** !-,2#,2T 2 Y " 71 .-12 ',-!3* 2'-,Q 2�# 5 1 ,-

1'%,'$'! ,2"'$$#0#,!# #25##,!-,20-* ,"',-!3* 2#"Z. mays. 0302� , *71'1

-$. 2&-%#,*#4#*15'2&',0--22'113#Q31',% -2&1#*#!2'4#+#"' ,",-02�,

*-2 , *71'1"#+-,120 2#"2& 2 . 2&-%#,*#4#*15#0#0#"3!#" 5'2&',2�--2

2'113#-4#02'+#Q13%%#12',% , !2'4#"#$#,!#+#!& ,'1+#+.*-7#"5'2&',2&#

0#1'12 ,2.* ,2T

xix

(&# 0-*# -$ )#7 !#**3* 0 "#$#,!# +-*#!3*#1Q ',!*3"',% *'%,',Q ! **-1#Q

&7"0-%#,.#0-6'"# ,".&#,-*'!!-+.-3,"1Q5 1#6 +',#"31',%&'12-!&#+'! *

, *71'1 !0-11 STR&-302'+#!-301#T9 !&-$2#!-+.-3,"15 1"#2#!2#"

5'2&', P. cinnamomi ',-!3* 2#" 0--2 2'113#Q &-5#4#0Q !&#+'! * ',&' '2'-, -$

! **-1# ," &7"0-%#, .#0-6'"# TV & .0'-0 2- ',-!3* 2'-, "'" ,-2 0#13*2 ',

',!0# 1#" 131!#.2' '*'27Q 13%%#12',% 2& 2 ',"'4'"3 **7 2# !-+.-3,"1 "- ,-2

!-,20-* 0#1'12 ,!#T<-02�, *-2 , *71'1-$*'%,',0#* 2#"%#,#1CCoA1 ,"

CCoA2 15#** 12&#-6'" 2'4# 3012+ 0)#0%#,#gst11&-5#",-',"3!2'-,',

',-!3* 2#"0--22'113#!-+. 0#"2-!-,20-*1T

(- ""0#112&#+ (-0% .1',2&#),-5*#"%#-$+-*#!3* 00#1.-,1#12-

2&#. 2&-%#,Q%#,-+#V5'"# , *71'131',%2&# $$7+#20'6+ '8#/#,#)&'. 00 7

5 1!-,"3!2#"',!-,20-* ,"P. cinnamomi',-!3* 2#"Z. mays0--22'113# 2X

," TV & .-12 ',-!3* 2'-,T >4#0 URR %#,#1 5#0# "'$$#0#,2' **7 #6.0#11#" ',

',-!3* 2#"0--21!-+. 0#"2-!-,20-*1T>$2.V0#%3* 2#"%#,#15&'!&5#0#

$3,!2'-, **7 ,,-2 2#"Q %0# 2#0 2& , XR$ 5#0# '+.*'! 2#" ', .* ,2 "#$#,!#

0#1.-,1#1 2- '-2'! 120#11Q ',!*3"',% .&72- *#6',1Q %*32 2&'-,# 1V20 ,1$#0 1#1Q

1#0',#V.0-2# 1# ',&' '2-01Q ( 1+-,'! !'" #2&7*#,# '-17,2'1 %#,#1 ,"

%#,#0 *"#$#,!#V0#* 2#".0-2#',1T

(&'1 0#1# 0!& & 1 !-,20' 32#" 2- %0# 2*7 '+.0-4#" 3,"#012 ,"',% -$

2&#+#!& ,'1+1-$"#$#,!#2-P. cinnamomi',2�--22'113#-$ 0#1'12 ,2.* ,2

22&#+ !0-1!-.'!Q!#**3* 0 ,"+-*#!3* 0*#4#*T

xx

CHAPTER 1: Introduction and literature review

Chapter 1: Introduction and Literature Review

1.1. Background

(&#--+7!#2#QPhytophthora cinnamomi . ,"1, =',%"-+S20 +#,-.'* R

&7*3+S >-+7!-2 Q '1 1-'*V -0,# 0--2 . 2&-%#, -$ #!-,-+'! ,"

#,4'0-,+#,2 *1'%,'$'! ,!#5-0*"5'"#T*2'12&#+-125'"#*7"'120' 32#"-$ **

Phytophthora 1.#!'#1 5'2& &-12 0 ,%# %0# 2#0 2& , UWRR 1.#!'#1Q 5&'!&

',!*3"#1 !0-.1 13!& 1 .',# ..*#Q 4-! "-Q + ! " +' ," .# !&Q 1 5#** 1

-0, +#,2 *1 13!& 1 ) +#**' ," .&-"-"#,"0-, 0"& + TRRWR 0"& +

,"* !)+ ,TRSRT*, 3120 *' Q"'1# 1#! 31#" 7P. cinnamomi& 10#13*2#"

',"#4 12 2',%*-11#1', -2& %0'!3*230 * ,", 230 *1712#+1', **12 2#1 ,"

2#00'2-0'#1 07!# et al. TRRTT *, S[[[Q '2 5 1 *'12#" 1 )#7 2&0# 2#,',%

.0-!#11 3,"#0 2&# )-++-,5# *2& %-4#0,+#,2_1 9,4'0-,+#,2 0-2#!2'-, ,"

'-"'4#01'27 )-,1#04 2'-, !2Q ," 2&0# 2 2#+#,2 .* , ( 5 1

"#4#*-.#" ', TRRS 9,4'0-,+#,2 3120 *' TRRST )300#,2 *#4#*1 -$

P. cinnamomi',$#12 2'-,',, 2'4# 3120 *' ,$-0#121 0##12'+ 2#"2- #1#4#0 *

&3,"0#" 2&-31 ," &#!2 0#1Q .0-+.2',% 2&# 0#!#,2 0#*# 1# -$ 0#4'1#" (

#. 02+#,2 -$ 312 ', '*'27Q 9,4'0-,+#,2Q 3 2#0Q -.3* 2'-,1 ,"

)-++3,'2'#1TRR[T#1.'2##62#,1'4#0#1# 0!&-4#02&#. 121#4#0 *"#! "#1Q

,-#$$#!2'4#+#2&-"1& 4# ##,"#4#*-.#"2-#0 "'! 2# 2&#. 2&-%#,',#'2�

, 2'4# -0 %0'!3*230 * 1712#+1Q 5'2&-32 ! 31',% 1'%,'$'! ,2 4#%#2 2'-, " + %#

," *-11 0"7 et al.TRRST3&'*122&'1!& .2#0$-!31#1-,2&##$$#!21-$2&#

. 2&-%#,', 3120 *' Q'2'1 !),-5*#"%#"�#2& 2"'1# 1#1! 31#" 7

Phytophthora1.#!'#1 0#-$+ (-0!-,!#0,5-0*"5'"#T

S CHAPTER 1: Introduction and literature review

3&'*#+3!&.0-%0#11& 1 ##,+ "#2-5 0"13,"#012 ,"',%.* ,2V

. 2&-%#,',2#0 !2'-,15'2&$-*' 0. 2&-%#,1Q2&#+#!& ,'1+1-$.* ,20#1'12 ,!#

2- 0--2 . 2&-%#,1 0#+ ', .--0*7 3,"#012--"T '1# 1#1 ! 31#" 7 $-*' 0

. 2&-%#,12#,"2- #.-*7!7!*'!Q5'2&.* ,2V2-V.* ,21.0# "-$* 0%# +-3,21-$

1.-0# ',-!3*3+Q 5�# 1 0--2 . 2&-%#,1 ! 31# +-,-!7!*'! "'1# 1# 5'2& *'22*#

20 ,1$#0 -$ ',-!3*3+ $0-+ .* ,2 2- .* ,2 >)3 0 ," 3*'28 TRRWT >,*7

1+ **,3+ #0-$"#$#,!#. 2&5 71 ,"0#1'12 ,!#+#!& ,'1+1"#1!0' #"$-0

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V CHAPTER 1: Introduction and literature review

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W CHAPTER 1: Introduction and literature review

1.4.1. The morphology and asexual structures of P. cinnamomi

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X CHAPTER 1: Introduction and literature review

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Figure 1.2. P. cinnamomi life cycle. In response to environmental cues, zoospores are released from the sporangium and swim towards plant roots. Zoospores form cysts on the root surface, which subsequently germinate and penetrate the epidermal tissue, growth of mycelium then occurs within the plant root. In the absence of a suitable host, the pathogen can remain dormant within the soil as chlamydospores until conditions are favourable. The sexual structures consist of a female oogonium and a male antheridium which fertilises the oogonium, resulting in a diploid oospore. (Schematic not to scale).

SR CHAPTER 1: Introduction and literature review

1.4.4. The effects of P. cinnamomi on vegetation

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Figure 1.3. Effects of disease caused by P. cinnamomi incursion on the abundance and distribution of vegetation in the Anglesea Heathlands, south-west Victoria. (a) Healthy, uninfected vegetation with an intact over-storey of Eucalypt species and mid-understorey of X. australis and B. marginata (b) Disease spread down-slope resulting in collapse and death of understorey species (c) Older diseased site, with some recolonisation of grass species (blue arrowhead), rotted stumps of X. australis have left depressions in the ground (white arrowheads). Photographs taken April, 2008.

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Figure 1.4. Disease progression in the susceptible Australian native Xanthorrhoea australis (a) Healthy X. australis with dark green leaves, this plant is at least 200 years old and ~ 2 m tall (b) Diseased individual with advanced leaf chlorosis (white arrow) (c) Complete crown collapse which results in (d) a rotted stump (blue arrow) and disintegration of the plant. Photographs taken 2008-2009, Great Otway National Park, Victoria.

SU CHAPTER 1: Introduction and literature review

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Pathogen Non-host Plant Tissue Symptoms Reference resistance produced

P. infestans Type I Nicotiana Leaf None Kamoun et al. clevelandii 1999 N. tabacum cv. Leaf None xanthi

Type II N.benthamiana Leaf HR Kamoun et al. N. rustica Leaf HR 1999

Type II Solanum nigrum- Leaf HR Vleeshouwers et SN18 al. 2003

P. sojae Type II Arabidopsis Leaf HR Kamoun 2001

P. Type II A. paradoxa Root HR Tippett and cinnamomi Malajczuk 1979

Unknown Z. mays Root Variable Hinch et al. 1985

Type I Arabidopsis Root None Rookes et al. 2008

Type II Arabidopsis Leaf HR Rookes et al. 2008

Table 1.1. Type I and type II non-host responses in plants against Phytophthora species.

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TY CHAPTER 1: Introduction and literature review

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Cellwallfortification: Synthesisofnewroots Callose P.cinnamomi Restrictedlesions Lignin Absenceofdisease Phenolics Cellwallfortification Resistant Productionofcallose, plant GenerationofROS? lignin,phenolics roots H2O2? essentialforresistance?

Molecularresponses: Molecularresponses: Molecularresponses: JA/Etpathway? Productionofplant Pathogenrecognition? Phytoalexins? derivedtoxins? ROS? Defencerelatedgenes? PRproteins?

Figure1.6. Schematichighlightingthecurrentgapsintheknowledgeofdefenceresponses toP.cinnamomiinroottissueofresistant,and/orfieldresistantspecies.Responseswhich have been observed previously in several plant species are indicated in green boxes, unknownand/orunexaminedcellularresponsesareindicatedinpinkboxes,unknownand /orunexaminedmolecularresponsesare indicatedinblueboxes.

1.9.Plantmodelsystems

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Chapter 2: Characterisation of resistance responses to Phytophthora cinnamomi in the model monocot Zea mays

2.1. Introduction

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36 CHAPTER 2: Characterisation of resistance responses to Phytophthora cinnamomi

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Classification of Description of host response References host response Byrt and Holland 1978; Extensive root necrosis Whitfield et al. 1981; Cahill Highly Water-soaked tissue et al. 1989; Podger and susceptible/Fully Leaf wilt/chlorosis Brown 1989; Aberton et susceptible Plant death al.2001; Reiter et al. 2004; Cahill et al. 2008 Cahill et al.1989; Podger and Extensive root necrosis Brown 1989; Hüberli et Water-soaked tissue al.2002; Krebs and Wilson Susceptible Leaf wilt/chlorosis 2002; Reiter et al. 2004; Plant death Cahill et al. 2008

Pathogen reproduction Byrt and Holland 1978; Moderately Root necrosis Podger and Brown 1989; susceptible Considerable plant death Duncan and Keane 1996; Cahill et al. 2008 Slightly susceptible Some disease symptoms Podger and Brown 1989

Host penetration Hinch and Weste 1979; Tolerant No disease symptoms Cahill and Weste 1982; Formation of callose papillae Rookes et al. 2008 Restricted root lesions Phenylpropanoid biosynthesis Phillips and Weste 1984; Cahill and McComb 1992; Field resistant Production of lignin Formation of new roots Cahill et al. 1992; Cahill et al. No disease symptoms 1993; Wilson et al. 2000

Restricted root lesions Hinch and Weste 1979; Phenylpropanoid biosynthesis Tippett and Maljczuk 1979; Production of lignin Hinch and Clarke 1982; Resistant Formation of new roots Cahill and Weste 1983; No disease symptoms Cahill et al. 1989; Podger Formation of callose papillae and Brown 1989; Hüberli et Hypersensitive cell death al. 2002; Cahill et al. 2008 Highly Cahill et al. 1989; Rookes et resistant/Fully No disease symptoms al. 2008 resistant Table 2.1. Current and past classifications of host responses to P. cinnamomi have applied more than one term to describe an identical response.

37 CHAPTER 2: Characterisation of resistance responses to Phytophthora cinnamomi

*,2&#!300#,2123"7Q',-0"#02-!* 11'$7 .* ,21.#!'#1 10#1'12 ,22-

P. cinnamomi 3,"#0 !-,20-**#" !-,"'2'-,1Q ',-!3* 2#" .* ,21 & " 2- +##2 $'4#

1.#!'$'! 12 ," 0"'1#" !0'2#0' Q 5&'!& 5#0# 11#11#" 2 1#4#, " 71 $-**-5',%

',-!3* 2'-,T (&# !0'2#0' 31#" 2- #12 *'1& 2# 20 '21 5#0# 1#" -,

+# 130 *#Q+ !0-1!-.'!- 1#04 2'-,15&'!&$-0+2&# 1'1-$.* ,2. 2&-*-%7

"'1# 1# 11#11+#,21 7 0%#S[XXR/ 3,2S[[WR<'*11-, et al.S[[WQ5&'!&! ,

#12 2'12'! **7 , *71#"T(#!0'2#0' ',!*3"#.* ,2$0#1&5#'%&2Q0--2%0-52&Q

2&#! *!3* 2'-,-$.#0!#,2 %#-$2-2 *0--2,#!0-1'1 7+# 130',%*#1'-,*#,%2&

1 .#0!#,2 %# -$ 2-2 * 0--2 *#,%2&Q *# $ !&*-0-1'1 1 , 4#0 %# -$ 2-2 *

!&*-0-.&7** !-,!#,20 2'-,Q ," 2&# .#0!#,2 %# -$ . 2&-%#, %0-52& -, 0--2

1#!2'-,1.* 2#"-,2-PhytophthoraV1#*#!2'4#+#"' T ,3+#0'! *1! *#5 12&#,

..*'#"2-2&#.* ,21T(&'1 11#11+#,25 1,-2',2#,"#"$-00 .'"1!0##,',%-$

* 0%# ,3+ #0 -$ "'$$#0#,2 .* ,2 1.#!'#1R '2 5 1 120',%#,2 2#12 2- !-,$'0+ 2&#

0#1.-,1#1 -$ 131.#!2#" 0#1'12 ,2 -0 131!#.2' *# .* ,2 2- 0--2 . 2&-%#,

3,"#0!-,20-**#"!-,"'2'-,1T

(&'1 11#11+#,2'1.32$-05 0"�# 1 +#2&-"2- !!30 2#*7!* 11'$7

.* ,20#1.-,1#12-P. cinnamomiT(&# ..*'! 2'-,-$2# 11#11+#,2!0'2#0' Q

1#"-,+# 130#+#,21-$+ !0-1!-.'!17+.2-+1Q5 131#"2-$'0+*7#12 *'1&

2&# +-,-!-2 Z. mays 1 1-3," 0#1'12 ,2 +-"#* $-0 .* ,2VP. cinnamomi

',2#0 !2'-,1T(&'1#,130#"2& 22�#1.-,1#1- 1#04#"',"-5,120# + , *71#1

2 -2&2&#!#**3* 0 ,"+-*#!3* 0*#4#*5#0#-!!300',%', 203*70#1'12 ,2.* ,2

," .0-4'"#1 0- 312 , *71'1 1712#+ 5&'!& ! , # ..*'#" 2- -2� .* ,2V

P. cinnamomi',2#0 !2'-,1T

38 CHAPTER 2: Characterisation of resistance responses to Phytophthora cinnamomi

2.2. Materials and Methods

2.2.1. Growth and maintenance of Z. mays

,','2' *12-!)-$UR1##"1-$Z. maysYU 31(.)0URVZXTQ5#0#)',"*7

"-, 2#" 70#2#07 50#,!#$0-+2&# 3120 *' ,(0-.'! *)0-.1 ,"0-0 %#1

/#0+.* 1+ )-**#!2'-,Q #. 02+#,2 -$0'+ 07*,"3120'#1Q '*-#* Q I3##,1* ,"T

1-,*7 *'+'2#",3+ #0-$ YU1##"1!-3*" #- 2 ',#"Q'2 5 1 ,#!#11 072-

3*)3.1##"12-!)1$-0$302�#6.#0'+#,2 2'-,T##"1-$Z. mays9 0*77# +',%

9"#, ##"1Q 7-5#0 ##!&+-,2Q I7Q 3120 *' Q VTSS 5#0# 31#" ',

#6.#0'+#,2 * 5-0) ', !-,(3,!2'-, 5'2& YUT *, .0#*'+', 07 123"'#1Q ,-

1'%,'$'! ,2"'$$#0#,!#5 1- 1#04#" #25##,2&#+ !0-1!-.'!0#1.-,1#1-$YU

,"9 0*77# +',%*',#1$-**-5',%',-!3* 2'-,5'2&P. cinnamomi.

##"1 5#0# 20 ,1$#00#" ',2- TW !+ "' +#2#0 .-21 -,# 1##" .#0 .-2

$'**#" 5'2& .-22',% +'6 >1+-!-2# #3*2' 30.-1# #'6Q !-221 3120 *' Q

3*)& +'**1Q<3T(K#0#2&#,%0-5,5'2&', 2#+.#0 230#!-,20-**#"

! ',#2 3,"#0 U 6 XRRV5 22 &'%&V.0#1130# 1-"'3+ *'%&21 5'2& SX & Z &

.&-2-.#0'-" 2 TS5) 2 TRR =+-* +VT1VS(�+-*',#7&#Q+'2&$'#*"Q<3Q

+-"#* /TSWTSZT[ (T 2 X 5##)1 -$ %#Q 5&#, 2 5#0# 2-- 2 **

..0-6'+ 2#*7S+$-02&#! ',#21Q.* ,215#0#20 ,1$#00#"2- %0-52&0--+

,".* !#"',$0-,2-$ 1-"'3+*'%&2 2SWRVSYW=+-*+VT1VST# 230#!- 15#0#

& 04#12#" $-0 1##" 12-!)Q .* !#" 5'2&', . .#0 % ," "0'#" 5'2&', 5 0+

0--+TT5) + '#,22#+.#0 230#$-03.2-Y" 71R1##"15#0#2&#,0#+-4#"

$0-+2&#!- 1 ,"12-0#"',. .#0 %1 20--+2#+.#0 230#T

39 CHAPTER 2: Characterisation of resistance responses to Phytophthora cinnamomi

2.2.2. Optimisation of Z. mays growth systems for experimental analysis

(&#+ (-0%- *-$2&'1-.2'+'1 2'-,5 12- !!#110--22'113#$-0"'0#!2

',-!3* 2'-,5'2&-32"'1230 ',%-0" + %',%0--22'113#Q ,"2-- 2 ',!*# ,0--2

+ 2#0' *5'2&+','+ * 22 !&#". 02'!3* 2#+ 22#0T ,3+ #0-$%0-52&+#"' Q

',!*3"',%1-'*Q4#0+'!3*'2# ,"1-'*V$0##5#0#20' **#"2-"#2#0+',#2&#-.2'+3+

%0-52& !-,"'2'-,1 ," +#"' $-0 Z. mays .* ,21 0#/3'0#" $-0 "-5,120# +

#6.#0'+#,2 * , *71'1T ** .* ,21 5#0# ','2' **7 %0-5, ', 20 71 -$ 1-'* -0

4#0+'!3*'2# ..#,"'6TTRT$'%30# S Q&-5#4#0Q2&'15 1"'1! 0"#""3#2-2&#

+-3,2 -$ . 02'!3* 2# + 22#0 2& 2 "�#" 2- 2&# 0--21 ," 2&# "'$$'!3*27 -$

!!#11',%0--21$-0"'0#!2',-!3* 2'-,T

)*# 0V$0-,2#" .-*7! 0 -, 2# 0--2 - 1#04 2'-, -6#1 ..#,"'6 TTRT

$'%30# S "'+#,1'-,1SZS!+6VS!+6X!+5#0#.30.-1#V 3'*22-$ !'*'2 2#

0--2 ',-!3* 2'-, ," - 1#04 2'-,1 -$ "'1# 1# .0-%0#11'-, 5'2& +','+ *

"'1230 ,!# 2- 2&# 0--2 2'113# ', 2&# 1-'* #,4'0-,+#,2 90 * 12'!1Q /##*-,%Q

?'!2-0' T (&# #01.#6 $0-,2 5 1 &',%#" ," **-5#" $-0 # 17 !!#11 2- 2&#

0--21 5'2& +','+ * "'1230 ,!# 2- 2&# .* ,21T (&# 3,'21 5#0# $'**#" 5'2& 1-'*Q

* '"$* 2 ,"$'4#V" 7V-*"1##"*',%1-$Z. mays5#0#.* !#"5'2&',2-.* 7#0-$

1-'* 5&'!& #,!-30 %#" 0--2 %0-52& 2-5 0"1 2&# !*# 0 #01.#6 $0-,2T (&#

1##"*',%15#0# 2&#,5 2#0#" 2-03,-$$ ," 1&##2-$ * !).* 12'!5 12 .#"

!0-11 2&# #01.#6 2- #6!*3"# *'%&2 $0-+ 2&# 0--21T * ,21 5#0# 2&#, %0-5,

5'2&',2#+.#0 230#!-,20-**#"! ',#21 1.0#4'-31*7"#1!0' #"1#!2'-,TTTTSTT

3&'*#2&'1+#2&-"5 1'"# *$-00--2',-!3* 2'-,Q ,""'",-2

!-+.0-+'1# 2&# ',2#%0'27 -$ 0--2 2'113# -0 .* ,2 &# *2&Q 2&# ,3+ #0 -$ .* ,21

5'2&',# !&3,'25 1*'+'2#"2- + 6'+3+-$#'%&2T(#'%&2SW)%5&#,

$'**#"5'2&1-'* ,".* ,21 ,"1'8#-$2,'21 *1-.0-4#"'+.0 !2'! *$-0*-,%V

40 CHAPTER 2: Characterisation of resistance responses to Phytophthora cinnamomi

2#0+#6.#0'+#,2 2'-,-, * 0%#1! *#Q 1-,*71'63,'21!-3*" #.* !#"5'2&',

! ',#2 2 ,7-,#2'+#T(&'1+#2&-"5 12�#$-0#"'1!-,2',3#"',$ 4-30-$

2+ **#0Q1-'*V$0##%0-52&20 713,'21T

*2&-3%&# !&-$2&0##+#2&-"1.0-"3!#"&# *2&7.* ,21Q 1-'*V$0##

.* ,2 %0-52& 1712#+ 0'%30# TTS 5 1 1#*#!2#" 1 2&# +-12 13'2 *#

+#"'3+$-0- 2 ',',%!*# ,0--2+ 2#0' *$0##-$1-'*-0-2�. 02'!3* 2#+ 22#0

/3,,',% ,") &'**TRR[T(&'1& 1 ,3+ #0-$ "4 ,2 %#1$-0"-5,120# +

..*'! 2'-,1',!*3"',%+'!0-1!-.7-0 '-!&#+'! * ,"+-*#!3* 0 , *71'1T.--2

+ 2#0' *$0##-$1-'*'1*#11*')#*72- #13 (#!2#"2-+#!& ,'! *" + %#Q! 31#"

7 2&# 0#+-4 * -$ .* ,21 $0-+ 2&# 1-'*Q ," '2 0#"3!#1 2&# *')#*'&--" -$

!-,2 +', 2'-,$0-+2-'*T

2.2.3. Growth and maintenance of experimental plants within a soil-free plant growth system (SPS)

96.#0'+#,2 *.* ,215#0#%0-5,31',% +#2&-"+-"'$'#"$0-+/3,,',%

,") &'**TRR[T##"1-$Z. mays5#0#1- )#"', # )#0-$2 .5 2#0$-0TV&

.0'-02-1##"',%T (K#0#2&#, 20 ,1$#00#"2- 12#0'*'1 2'-,1-*32'-,-$S$

4 41-"'3+&7.-!&*-0'2#$-0T+',32#1 #$-0# #',%'++#"' 2#*70',1#"5'2&

2 . 5 2#0$-0 $302�T+',32#1T##"15#0#.* !#"-,$-30 * 7#01-$. .#0

2-5#*',.0#. 0 2'-,$-01##"',%T(&# 1#-$# !&.-*7! 0 -, 2#3,'25 1$'**#"

5'2& ZRR +7 -$ S U 120#,%2& (-2 * -02'!3*230 * )-,!#,20 2# () ,320'#,2

1-*32'-, ()Q 96!#* '120' 32-01Q .#1#04-'0Q 3120 *' T (&# $* 2 !) -$ # !&

20 75 1!-4#0#"5'2& 1/3 0#-$$'*2#0. .#03& 2+ ,<-TS=#,2Q9,%* ,"

," 2&# . .#0 1 230 2#" 5'2& 2 . 5 2#0 31',% .0#1130'1#" 1.0 7 -22*#T '0

3 *#15#0#0#+-4#" 7%#,2*7*'$2',% ,"0#.* !',%2&#$'*2#0. .#0T(&0##

41 CHAPTER 2: Characterisation of resistance responses to Phytophthora cinnamomi

!-22-, 5--* 1/3 0# . "1 5'11.#01Q 3120 *' Q !32 ', & *$ *#,%2&5'1#Q 5#0#

.* !#" !0-112-.-$2&#$'*2#0. .#0 ,"1 230 2#"5'2&2 .5 2#0T(&#$0-,2

," !)1#!2'-,1-$2 715#0#2&#,!*'..#"2-%#2� 5'2&$-*"V !)!*'.1

(3"-0Q .#0*',KQ0#12-,Q 3120 *' ,"0#.* !#"5'2&',2&# 1#3,'2T##"1

5#0#.* !#" *-,%2-.-$2 75'2&2&#&'*'3+$ !',%"-5,5 0"1$'%30#

TTSQ ,"!-4#0#"5'2&1&##21-$!-22-,5--*0'..*#1Q5&'!&5#0#2&#,1 230 2#"

5'2&2 .5 2#0$0-+ 1.0 7 -22*#',-0"#02-)##.2##"1+-'12T(##"1

5#0#20 ,1$#00#"2-2#+.#0 230#!-,20-**#"%0-52&! ',#213,"#0U6XRRV5 22

&'%&V.0#1130#1-"'3+*'%&215'2& SX& Z&.&-2-.#0'-" 2TS5) 2TRR =+-*+VT

1VST ##"1 5#0# +'12#" " '*7 5'2& 1.0 7 -22*# !-,2 ',',% S U 120#,%2& ()

,320'#,21-*32'-,T $2#0 U " 71Q 2&# !-22-,5--* 5 10#+-4#"$0-+ 2##"1

," ,71##"15&'!&& "$ '*#"2-%#0+', 2#5#0#"'1! 0"#"T (&#$'*2#0. .#0

,",320'#,21-*32'-,Q5'2&',2&# 1#-$2,'2Q5#0#0#.* !#"#4#07W" 71T

##"1 -$ L. angustifolius 4 0T 3-,% 5#0# - 2 ',#" $0-+ < 00 !--02#

##"1Q< 00 !--02#Q-32& 3120 *' Q ,"%0-5,', 1"#1!0' #"$-0

Z. maysT##"15#0#1- )#"',2 .5 2#0$-0VVX&.0'-02-1##"',%T ,71##"1

5&'!&& ",-215#**#" $2#02&'12'+#5#0#"'1! 0"#".0'-02-.* ,2',%T

Figure 2.1. Z. mays plants grown within the SPS (a) Three day old seedlings of Z. mays within the SPS Scale bar = 2 cm (b) Eight day old seedlings of Z. mays within the SPS. Scale bar = 2.5 cm

42 CHAPTER 2: Characterisation of resistance responses to Phytophthora cinnamomi

2.2.4. Determination of optimum plant age for inoculation

*,-0"#02-"#2#0+',#2&#-.2'+3+.* ,2 %#$-0',-!3* 2'-,Q ,3+ #0-$

',-!3* 2'-, 2'+#V.-',21 5#0# ','2' **7 20' **#" ', 2&'1 123"7 " 2 ,-2 1&-5,T

(#," 715 11#*#!2#" 12&#-.2'+3+',-!3* 2'-, %#$-0 **.* ,21%0-5,

5'2&',2&#1712#+ ,"5 1!-,2',3#"2&0-3%&-32 **!#**3* 0 ,"+-*#!3* 0

5-0)T(&'12'+#.-',2 **-5#"$-02&#$'0121#2-$203#*# 4#12-"#4#*-.',2&#

1##"*',%1Q ," $-0 %--" 0--2 "#4#*-.+#,2T * ,21 ',-!3* 2#" 2 2#, " 71

0#+ ',#"&# *2&75'2&',2&#1712#+$-03.2-SV" 71 $2#0',-!3* 2'-,T(&#

.0'+ 070--22'.1-$# !&.* ,25#0##6!'1#" 2W" 712-#,!-30 %#$0#1&* 2#0 *

0--2%0-52&$-0',-!3* 2'-,$'%30#TTTT

ab

Figure 2.2. Ten day old seedlings of (a) Z. mays and (b) L. angustifolius within the SPS developed fresh lateral root growth for inoculation 5 days after excision of the primary root tip (arrowhead). Scale bar = 2.5 cm.

43 CHAPTER 2: Characterisation of resistance responses to Phytophthora cinnamomi

2.2.5. Field collection of soil samples

** '1-* 2#1 -$ P. cinnamomi31#"', 2&'1123"7 5#0#- 2 ',#"$0-+1-'*

2 )#,$0-+2&# 1#-$Xanthorrhoea australis %0-5',% 2 "'1# 1#"1'2# 2 *"

'**1 .- "Q ,%*#1# Q ?'!2-0' T -'* 1 +.*#1 5#0# !-**#!2#" $0-+ 1#4#0 *

X. australis "'1.* 7',% # 0*7 17+.2-+1 -$ "'1# 1# ..#,"'6 TTSTQ $'%30# T T

(-.SR!+-$1-'*5 1!-**#!2#"5'2& 20-5#*$0-+2&0## 0# 1WRVSRR!+

$0-+2&# 1#-$# !&.* ,2 ,".* !#"', 8'.V*-!).* 12'! %T(-5#*5 1

130$ !# 12#0'*'1#" 5'2& ZR$ 4 4 #2& ,-* #25##, # !& .* ,2T (- .0#4#,2

"#1'!! 2'-,Q **'1-* 2#15#0#12-0#"', 1# *#"!-,2 ',#0 ,"20 ,1.-02#"2-2&#

* -0 2-07$-0'1-* 2'-,-$P. cinnamomi.

2.2.6. Isolation of P. cinnamomi from soil samples

*1-* 2'-, -$ P. cinnamomi$0-+1-'*1 +.*#15 1!-,"3!2#"31',%

+#2&-" +-"'$'#" $0-+ # 0)1 ," = 11 7 S[YV ..#,"'6 TTSTT

..0-6'+ 2#*7 WR % -$ 1-'* $0-+ # !& 1 +.*# % 5 1 .* !#" ',2- ',"'4'"3 *

.* 12'!!3.1/#,$ !* 12'!1Q?*)Q5&'!&5#0#2&#,$'**#"2-S!+$0-+2-.

5'2&2 .5 2#0T(-'* ,"5 2#05#0#+'6#"2- 1*30075'2&%#,2*#12'00',%

31',% 5--"#, 12'00#0T P. cinnamomi 5 1 2&#, '2#" 31',% TRV" 7V-*"

!-27*#"-,1 -$ Eucalpytus sieberi, 5&'!& & " ##, %0-5, ', 3,'21 5'2&',

2#+.#0 230# !-,20-**#" ! ',#21 1 "#1!0' #" ', 1#!2'-, TTSTWT , E. sieberi

!-27*#"-,1##"*',%5 1* '" !0-112-.-$2-'*1*3007T(&#!3.15#0#2&#,

',!3 2#"$-0YT&',2&#" 0)5'2&', 2#+.#0 230#!-,20-**#"%0-52&! ',#2

1"#1!0' #"',1#!2'-,TTTTST

44 CHAPTER 2: Characterisation of resistance responses to Phytophthora cinnamomi

2.2.7. Isolation of P. cinnamomi on selective media (PARPH) and establishment and maintenance of working cultures

##"*',%1-$E. sieberi5#0#0#+-4#"$0-+2-'*1*300731',%12#0'*'1#"

$-0!#.1Q ,"2�--21#6!'1#"T(�--215#0#130$ !#12#0'*'1#"',ZR$4 4

#2& ,-*$-0VW1#!-,"12-0#+-4#2&#. 2&-%#,$0-+2‚$ !#Q2&31#,130',%

2& 2 ,7%0-52&-,2&#+#"'3+#+ , 2#"$0-+',1'"#2&#.* ,20--2T.--21

5#0#0',1#"',1"T>$-0XR1#!-,"1 ,".* !#"-,12#0'*#$'*2#0. .#0$-0XRV[R

1#!-,"1 2- "07T 3'2&', * +', 0 $*-5 5-0)12 2'-,Q 0--21 5#0# 2&#, !32 ',2-

SR ++ 1#!2'-,1 5'2& 1! *.#*T !1',% $-0!#.1Q 0--2 1#!2'-,1 5#0# * '"

1#/3#,2' **7 -,2- 2&# 130$ !# -$ PhytophthoraV1#*#!2'4# +#"' !-,2 ',',% 2&#

,2' '-2'!1 '+ 0'!',Q +.'!'**',Q .'$ +.'!',Q #,2 !&*-0-,'20- #,8#,# ,"

7+#6 8-* . ..#,"'6TTT=#** + ,")-$$#7S[ZWT(�--215#0#

.0#11#" ',2- 2&# 130$ !# 1*'%&2*7 2- #,130# 2&# . 2&-%#, %0#5 -32 -$ 2&# 0--2

2'113# ,"',2-2&#+#"'3+T(&#.* 2#15#0#1# *#"5'2& 0 $'*+#Q

',!3 2#"',2&#" 0) 2TV ) ," +-,'2-0#" #4#07 TV & $-0 2&# .0#1#,!# -$

P. cinnamomi %0-52& ..#,"'6 TTTTQ $'%30# T!T $2#0 U " 71Q U ++T.*3%1

$0-+2&##"%#-$2&#P. cinnamomi!-*-,75#0# 1#.2'! **720 ,1$#00#"-,2-SR$

!* 0'$'#"?Z % 0.* 2#1SR$)?Z ,"',!3 2#"',2&#" 0) 2TV5) ..#,"'6

TTTT$'%30# T"T $2#0W" 712&#!3*230#5 1#12 *'1&#" 1 5-0)',%!3*230#

," 5 1 + ',2 ',#" ', 2&# " 0) 2 TV5)T 9 !& '1-* 2# 5 1 2&#, 13 !3*230#"

#4#07YVSR" 71-,2-SR$)?Z % 02-+ ',2 ', $0#1&5-0)',%!3*230#T(#

'1-* 2#15#0# ""#"2-2&## )',!,'4#01'27!3*230#!-**#!2'-,Q ,"# !&,#5

'1-* 2#5 1 11'%,#" ,3+#0'! *'"#,2'$'! 2'-,!-"#5'2&2&#.0#$'6!T

45 CHAPTER 2: Characterisation of resistance responses to Phytophthora cinnamomi

2.2.8. Confirmation of pathogen identity and purity

0'-0 2- 2&# !-++#,!#+#,2 -$ 2&# !& 0 !2#0'1 2'-, -$ .* ,2V. 2&-%#,

',2#0 !2'-,1QP. cinnamomi5 1'1-* 2#"$0-+1-'*1 +.*#12 )#,$0-+2&#$'#*"

,"'21'"#,2'27!-,$'0+#" 7'"#,2'$7',%!& 0 !2#0'12'!+-0.&-*-%'! *20 '21 ,"

72#-$PhytophthoraV1#*#!2'4# +#"' ," .07 , *71'1 0#,2& et al.

TRRXT (&'1 #,130#" 2& 2 2&# '1-* 2#1 31#" 5#0# .30#

P. cinnamomiQ$0##$0-+ !2#0' *!-,2 +', 2'-, ," -02&#.0#1#,!#-$-2�

Phytophthora 1.#!'#1T(&'1! , # !-,$-3,"',%4 0' *#',$'#*"123"'#15�#

2&# .0#1#,!# -$ -2� . 2&-%#,1Q ',!*3"',% -2� Phytophthora1.#!'#1! ,

0#"3!#2'03*#,!#-$P. cinnamomi *1 **S[ZTT

(&#'"#,2'27 ,".30'27-$2&#P. cinnamomi!3*230#131#"',2&'10#1# 0!&

5#0# ','2' **7 "#2#0+',#" 7 - 1#04 2'-, -$ !& 0 !2#0'12'! +-0.&-*-%'! *

$# 230#13,"#0*'%&2+'!0-1!-.7T(#$# 230#1',!*3"#!-0 **-'"Q&7 *',#

+7!#*'3+ 5'2& 3," ,2 &7.& * 15#**',%1T (&# '1-* 2#1 5#0# 2&#, $302�

!-,$'0+#" 31',% < V 1#" '"#,2'$'! 2'-, +#2&-" 0#,2& et al. TRRXT (-

!-,$'0+2& 22&#'1-* 2# #*-,%#"2-2&#%#,31PhytophthoraQ%#,-+'!< 5 1

+.*'$'#" 7).31',%2&#Phytophthora%#,31V1.#!'$'!.0'+#01#21( *#TTT

0#,2&et al. TRRXT

Primer Oligonucleotide sequence (5’ – 3’)

A2_forward ACT TTC CAC GTG AAC CGT TTC AA

I2_reverse GAT ATC AGG TCC AAT TGA GAT GC

Table 2.2. Sequence of Phytophthora genus-specific primer set.

(&# ). 5 1 .#0$-0+#" ', UR 70# !2'-, ..#,"'6TTUT/#,-+'!

< 5 1- 2 ',#"$0-+ , 6#,'!!3*230#-$P. cinnamomi 71!0 .',% 1+ **

+-3,2-$+7!#*'3+$0-+ SR$)?Z % 0.* 2#31',% .'.#22#2'.T(&#.'.#22#

46 CHAPTER 2: Characterisation of resistance responses to Phytophthora cinnamomi

2'.5 1"'..#"',2- STW+79..#,"-0$23 #!-,2 ',',%2&#+ 12#0+'6 ,"

12'00#"%#,2*7$-0W1#!-,"12-0#*# 1#2&#%< T< 5 1 +.*'$'#"',

2�+ * !7!*#0 # .#1# 0!& ()VTRRQ # Q ! 5'2& , ','2' * "#, 230 2'-,

12#. 2[V5)$-0U+',32#1Q$-**-5#" 7VR!7!*#1"#, 230 2'-, 2[V5)$-0UR

1#!-,"1Q ,,# *',% 2WT5)$-0VW1#!-,"1 ,"#62#,1'-, 2YT5)$-0UR1#!-,"1T

1+ ** +-3,2-$2&# +.*'$'#".0-"3!2W75 103,-, RTZ$ % 0-1#%#*',

(0'1V-0 2#V9( (9 3$$#0 % ',12 S) * ""#07.#0* ""#0*Q'-*',#T

(&##6.#!2#"$0 %+#,21'8#5 1ZT[ .T(&0##0#.*'! 2#15#0#03,# !&2'+#

*-,%1'"# 1',%*#,#% 2'4#,-%#,-+'!< !-,20-*T

(-!-,$'0+2&#'1-* 2#2-1.#!'#1*#4#*Q2&0##"'$$#0#,20#120'!2'-,

#,87+#1Q Msp*, Rsa* ," Taq*Q5#0#31#"T9 !&"'%#12'-,5 1!-,"3!2#"',

2-2 *4-*3+#-$SR7!-+.0'1',%X7-$2&#)..0-"3!2 ..#,"'6TTUTT(&#

1 +.*#15#0#',!3 2#"$-0STW& 2UY5)$-0Msp* ,"Rsa* ," 2XW5)$-0Taq*T

00 %+#,215#0#03,-, U$ % 0-1#%#*',(0'1V-0 2#V9( (9 3$$#0

% ',12 SRR .* ""#07.#0* ""#0*?Q'-*',# ,"2&# ,"',%. 22#0, -$

# !&5 1!-+. 0#"2-2&-1#"#1!0' #"$-0P. cinnamomi',0#,2&et al.TRRXT

2.2.9. Confirmation and maintenance of virulence of P. cinnamomi isolates

P. cinnamomi'1-* 2#1Q1#*#!2#"$-031# 1',-!3*3+Q5#0#. 11#"2&0-3%&

2�--21-$ L. angustifolius 1##"*',%1 ..0-6'+ 2#*7 #4#07 ST 2- SX 5##)1Q 2-

.0#4#,2 ,7 *-11 -$ 4'03*#,!# "3# 2- .0-*-,%#" 13 !3*230',% 905', S[ZUT

L. angustifolius1##"*',%15#0#%0-5,$-0W" 71 1"#1!0' #"',1#!2'-,TTTTUT

*1-* 2#1-$P. cinnamomi5#0#2 )#,2&0-3%&2-2--1.-0#.0-"3!2'-,12 %#

1"#1!0' #"',1#!2'-,TTTTSRT(�--21-$L. angustifolius5#0#',-!3* 2#"5'2&

W 6 TR =7 "0-.*#21 -$ P. cinnamomi 8--1.-0#1 2 !-,!#,20 2'-, -$ S 6 SRW

47 CHAPTER 2: Characterisation of resistance responses to Phytophthora cinnamomi

8--1.-0#1+7VS 1"#1!0' #"',1#!2'-,TTTTSTTQ ,"2�--215#0#& 04#12#" 2

YT&T(�--215#0#2&#,.* 2#"-,2- .+#"'3+ 1"#1!0' #"',1#!2'-,

TTTTYT $2#0YT&Q25-W++T.*3%1Q$0-+2&#-32#0%0-52&#"%#-$# !& .

.* 2#Q5#0#20 ,1$#00#"-,2-SR$)?Z % 0.* 2#1 ,"',!3 2#"',2&#" 0) 2

TV5)T

2.2.10. Zoospore production

H--1.-0#15#0#.0-"3!#"31',% +#2&-" 1#"-,2& 2-$702 ,"

/0 ,2 S[Y[T ',%*# ',-!3*3+ .*3%1 W ++T -$ P. cinnamomi5#0#.* !#"

',4#02#"-,2-2&#!#,20#-$ [!+ .#20'"'1!-,2 ',',%SR$)?Z % 0Q ,"

+','+3+-$$-300#.*'! 2#.* 2#15 1.0-"3!#"# !&2'+#T* 2#15#0#1# *#"

5'2&. 0 $'*+ ,"',!3 2#"',2&#" 0) 2TV5)$-0U" 71T'6 % 0.*3%15#0#

2&#,20 ,1$#00#"$0-+2&#-32#0%0-52&#"%#-$2&'1!-*-,7-,2-.0#V12#0'*'1#"

+'0 !*-2&"'1!1) * '-!&#+Q7 -** Q) -,SR$)?Z % 0.* 2#1T(#.* 2#1

5#0#1# *#" ,"',!3 2#"',2&#" 0) 2TV5)$-0W" 71T

#'0 !*-2& "'1!1 5#0# 1#. 0 2#" $0-+ 2&# % 0 31',% 12#0'*'1#" $-0!#.1

,".* !#"',2-12#0'*#TWR+790*#,+#7#0$* 1)1!-,2 ',',%SRR+7-$W$)?Z

0-2& ..#,"'6 TTTTT 0* 1)1 5#0# 1# *#" ," .* !#" -, , -0 '2 * 1& )#0

*,$-01 /Q-22+',%#, 2SSR0.+3,"#0U6URV5 22$*3-0#1!#,2*'%&21', ,

',!3 2-0 2TV5)$-0SX&T(&#W$)?Z 0-2&5 1"#! ,2#" ,"2&#+'0 !*-2&

0',1#"',2&#$* 1)5'2&SRR+7-$#',#0 * *2-*32'-,# ..#,"'6TTTU

2'+#1 2SWV+',32#',2#04 *12-',"3!#1.-0 ,%' *$-0+ 2'-,T $', *SRR+7-$

#5 1 ""#" #$-0#2&#$* 1)15#0#0#230,#"2-2&#-0 '2 *1& )#0',2&#

',!3 2-0$-0 $302�TR&T

48 CHAPTER 2: Characterisation of resistance responses to Phytophthora cinnamomi

(&# # 5 1 2&#, "#! ,2#" ," 2&# +'0 !*-2& 0',1#" 5'2&', 2&# $* 1)

5'2& $302�WR+7-$#$-0SW+',32#1T(&#+'0 !*-2&5 12&#,20 ,1$#00#"

2- $0#1&12#0'*#[!+.#20'"'1& ,"0',1#"-,!#5'2&TR+71"T>.0#V!&'**#"

2-V5)T $302�TR+71"T>.0#V!&'**#"2-V5)5 1 ""#" #$-0#2&#.* 2#1

5#0#1# *#"5'2&. 0 $'*+ ,"',!3 2#"',2&#" 0) 2V5)$-0VWVXR+',32#12-

**-5 $-0 2&# ','2' * 0#*# 1# -$ 8--1.-0#1 "3# 2- !-*" 1&-!)T .-0 ,%' *

.0-"3!2'-, 5 1 !-,$'0+#" 2 2&'1 12 %# 7 4'#5',% # !& .* 2# 3,"#0 *'%&2

+'!0-1!-.# 2 *-5 .-5#0T (&# .* 2#1 5#0# 0#230,#" 2- 0--+ 2#+.#0 230# 2-

$ !'*'2 2# !-,2',3#" 8--1.-0# 0#*# 1# ," +-,'2-0#" $-0 0#*# 1# 2 UR +',32#

',2#04 *1TH--1.-0#131.#,1'-,0#+ ',#"4' *#$-031# 1',-!3*3+$-0U&T

2.2.11. Determination of zoospore density

(&#"#,1'27-$8--1.-0#1',2"T> 5 1 "#2#0+',#" 7 20 ,1$#00',%

2&# 131.#,1'-, 2- WR +7 !#,20'$3%# 23 # ," **-5',% '2 2- 1#22*# $-0 SW

+',32#1T S +7 "0-. -$ 8--1.-0# 131.#,1'-, 5 1 0#+-4#" $0-+ 2&# 3..#0

* 7#0-$2&#!#,20'$3%#23 # ,".* !#"', STW+7!#,20'$3%#23 #T( #

5 14-02#6#"$-0XR1#!-,"12-',"3!#8--1.-0##,!712+#,2 ,"',4#02#"TVU

2'+#1 #$-0# SR 7 "0-. 5 1 .* !#" -, # !& !-3,2',% %0'" -$ 0'%&2 *',#

& #+-!72-+#2#01*'"# 311#0!'#,2'$'!)-+. ,7Q-01& +Q T(&#,3+ #0

-$ #,!712#" 8--1.-0#1 5 1 !-3,2#"Q 5'2& $', * !-3,21 8--1.-0#1 +7VS

0#.0#1#,2',%2&#+# ,-$ **[1/3 0#1',# !&%0'"T(--1.-0#131.#,1'-,

5 1"'*32#"2- !-,!#,20 2'-,-$S6SRW1.-0#1+7VS5'2&1"T>.0'-02-31# 1

',-!3*3+T

49 CHAPTER 2: Characterisation of resistance responses to Phytophthora cinnamomi

2.2.12. Inoculation procedures for Z. mays and L. angustifolius

0'-0 2- ',-!3* 2'-, -$ 0--2 2'113#Q # !& 3,'2 5 1 * '" $* 2 ," 2&#

.#01.#6$0-,21#!2'-,15#0#0#+-4#"Q ," 120'.-$ 0 $'*+##!&',#7

* 12'!1Q***',-'1Q! 5 1.* !#" #,# 2&# !&0--22- #',-!3* 2#"T00#1&0--2

2'113# -$ Z. mays5 1',-!3* 2#"5'2&TR=7"0-.1-$8--1.-0#131.#,1'-,Q

.* !#" *-,%2&#*#,%2&-$* 2#0 *0--21(312 #&',"2�--22'.2- + 6'+3+-$

SRR=7.#0.* ,20'%30#TTUT(�--21-$# !&.* ,25#0#',-!3* 2#" ,"2&#

#01.#6$0-,21#!2'-,1-$2&# 3,'215#0#2&#, * '" !0-112&#.* ,21 2 ,

,%*#2-.0#4#,2!-,2 !25'2&2&#',-!3*3+"0-.1 ,"2-+ ',2 ', &3+'"

#,4'0-,+#,2T (0 71 5#0# *#$2 *7',% $* 2 $-0 S & 2- **-5 $-0 8--1.-0#

#,!712+#,2Q 2&# $0-,2 #01.#6 1#!2'-,1 5#0# 2&#, !*'..#" !) ',2- .* !# ,"

2 710#230,#"2-2&#%0-52&!-,"'2'-,1 1"#1!0' #"',1#!2'-,TTTTUT.--21

-$L. angustifolius5#0#',-!3* 2#" 1"#1!0' #"$-0Z. maysT

Figure 2.3. Inoculation of Z. mays roots. Five x 20L droplets (black arrow) containing 1 x 5 -1 10 zoospores mL (maximum of 100 uL per plant) were placed along each experimental plant root from ~0.5 cm behind the root cap with a maximum spread of 2.5 cm. Control plants were mock inoculated with the same number of droplets of sdH20. Scale bar = 5 mm.

2.2.13. Confirmation of inoculation

(-!-,$'0+2&#.0#1#,!#-$2&#. 2&-%#,5'2&',',-!3* 2#".* ,21Q0--21

5#0# 1#!2'-,#" ," .* 2#" -,2- 2&# PhytophthoraV1#*#!2'4# +#"' .

50 CHAPTER 2: Characterisation of resistance responses to Phytophthora cinnamomi

5&'!& !-,1'121 -$ $'4# "'$$#0#,2 ,2' '-2'!1Q 2- 5&'!&

P. cinnamomi '1 0#1'12 ,2 #$$#01 ," # 02', S[ZXR 905', ," .' #'0- S[[XT

.--215#0##6!'1#"$0-+# !&.* ,2Q 2TV&',2#04 *1$-0Y" 71.-12',-!3* 2'-,T

.--215#0##6!'1#" ,"'++#"' 2#*7130$ !#12#0'*'1#"',ZR$4 4#2& ,-*$-0

VW1#!-,"12-#,130#2& 2 ,7%0-52&-,2&#+#"'3+#+ , 2#"$0-+',1'"#2&#

.* ,20--2T(K#0#2&#,0',1#"',1"T>$-0S+',32# ,".* !#"-,12#0'*#

$'*2#0. .#0$-0S+',32#2-"07T.--215#0#.* !#"-, !*# ,!#0 +'!2'*# ,"

!3220 ,14#01#*7',2-W++1#!2'-,15'2& 1! *.#*T.--21#!2'-,15#0#.* !#"

1#/3#,2' **7-,2-2‚$ !#-$ ..* 2#1Q2�--215#0#.0#11#"',2-2&#

130$ !#1*'%&2*72-#,130#2&#. 2&-%#,%0#5-32-$2�--22'113# ,"',2-2&#

+#"'3+T (&# .* 2#1 5#0# 1# *#" 5'2& 0 $'*+ ," ',!3 2#" ', 2&# " 0) 2

TV ) $-0 YT &T (&# %0-52& -$ P. cinnamomi *-,%2�--21#!2'-,15 1

+-,'2-0#" ,"0#!-0"#"', **.* ,21T(&#. 2&-%#,5 1 *#2- #13 1#/3#,2*7

0#V'1-* 2#"$0-+2&# .+#"' Q5&'!&1 2'1$'#"=-!&_1.-123* 2#1 ..#,"'6

TTVT

2.2.14. Infection time course

2'+#!-301#5 1!-,"3!2#"-4#0SZR+',32#1Q2-#6 +',#2.##"

5'2& 5&'!& 2&# 8--1.-0#1 5#0# 220 !2#" 2- 2&# 0--2 130$ !# ," .#,#20 2#"

0--22'113#Q ,"2-- 1#04#2�--28-,#2-5&'!&2K#0#+-12 220 !2#"T(-

- 1#04# 2&# ',$#!2'-, .0-!#11 -$ P. cinnamomi 8--1.-0#1Q 0--21 -$

L. angustifolius ,"Z. mays 5#0# #6!'1#" ," .* !#" -, +'!0-1!-.# 1*'"#T

TR=7"0-.-$8--1.-0#131.#,1'-,5 1.* !#"-,2*'"# ,"2&#',$#!2'-,

.0-!#11 5 1 - 1#04#" $-0 SZR +',32#1 3,"#0 *'%&2 +'!0-1!-.# 6'-1)-. T

+-2 .*31Q H#'11Q /ö22',%#,Q UYRURQ /#0+ ,7T &-2-1 -$ 2&# ',2#0 !2'-, 5#0#

51 CHAPTER 2: Characterisation of resistance responses to Phytophthora cinnamomi

2 )#,#4#07UR+',32#131',% ! +#0 +-3,2#" 2-2&#+'!0-1!-.#>(.(

*'"#0Q ' %,-12'! *,1203+#,21 *,!T *+ %#1 5#0# !/3'0#" 31',% >( .(

-$25 0#4UTRT

2.2.15. Relative plant health post-inoculation with P. cinnamomi

(- ','2' **7 "#2#0+',# 0#1.-,1#1 2- 2&# . 2&-%#, 5'2&', !-,20-**#"

!-,"'2'-,1Q +','+3+-$UR.* ,21-$Z. mays ,"UR.* ,21-$L. angustifolius

5#0# ',-!3* 2#" 5'2& P. cinnamomi 8--1.-0#1 2 SR " 71 ," - 1#04#" $-0

$302� SV " 71T .#!-0"1 -$ %#,#0 * .* ,2 &# *2& 5#0# 0#!-0"#" 2 " '*7

',2#04 *1 $-0 Y ".' $-**-5',% 4'13 * 11#11+#,21 ," +# 130#+#,21Q ," 2&#

#6.#0'+#,25 10#.# 2#"25'!#T

2.2.16. Parameters to determine resistance to P. cinnamomi

* ,2$0#1&5#'%&2Q0--2*#,%2&Q0--2*#1'-,*#,%2&Q*# $!&*-0-1'1 ,"2&#%0-52&

-$ P. cinnamomi 5'2&', 0--2 1#%+#,21 .* 2#" -,2- . 1 "#1!0' #" ',

1#!2'-,TTTTYT5#0#2&#W. 0 +#2#0131#"2- 11#112&#&# *2&-$Z. mays ,"

L. angustifolius .* ,21 $-**-5',% ',-!3* 2'-, 5'2& 2&# . 2&-%#,T * ,21 5#0#

11'%,#" + 6'+3+1!-0#-$S$-0# !&!0'2#0' T +','+3+-$[R!-,20-* ,"

[R ',-!3* 2#" .* ,21 -$ # !& 1.#!'#1 5#0# 31#" $-0 # !& #6.#0'+#,2 ," **

#6.#0'+#,215#0#0#.# 2#"2&0##2'+#1T

2.2.17. Measurement of plant fresh weight

* ,2 $0#1& 5#'%&2 5 1 +# 130#" #4#07 TV & $-0 SXZ & ', !-,20-* ,"

P. cinnamomi ',-!3* 2#"Z. mays ,"L. angustifoliusT* ,215#0#0#+-4#"$0-+

52 CHAPTER 2: Characterisation of resistance responses to Phytophthora cinnamomi

2&#1712#+1Q.* !#"', 5#'%&20 7 ,"2#'%&20#!-0"#"Q2K#0#2&#,

'++#"' 2#*70#230,#"2-2&#1712#+T

2.2.18. Measurements of root length and percentage root lesion length

>,.* ,21',-!3* 2#" 2W" 71-*"Q0--2*#,%2&5 1+# 130#"#4#07TV&

$-0 [X & ', !-,20-* ," P. cinnamomi ',-!3* 2#" .* ,21T .--2 %0-52& 5 1

+# 130#"3,2'*2�--21#62#,"#"2-2&# -22-+-$2&#20 715&'!&Q-,

4#0 %#Q5 1V" 71.-12',-!3* 2'-,Q $2#02&'12'+#Q2&#+ (-0'27-$0--2%0-52&

5 1* 2#0 *0--2$-0+ 2'-,T7#1'-,*#,%2&1$-0# !&0--25#0#+# 130#"#4#07

TV&$-0[X&31',% ?#0,'#0! **'.#0 ,"2�#13*2 ,24 *3#5 1!-,4#02#"2-

.#0!#,2 %#-$2�--2*#,%2&-$# !&.* ,2T>,.* ,21',-!3* 2#" 2SR" 71-*"Q

0--2*#,%2&5 1+# 130#"#4#07TV&$-0SXZ& ,"#6.0#11#" 1%0-52&!+

TV&0VST7#1'-,*#,%2&15#0#+# 130#" 1"#1!0' #"$-0W" 7-*".* ,21T

2.2.19. Quantification of chlorophyll within leaf tissue

(&# 0#* 2'4# +-3,2 -$ !&*-0-.&7** ', *# $ 2'113# -$ !-,20-* ,"

',-!3* 2#"*# 4#1$0-+RSXZ&.'5 1+# 130#"#4#07TV&31',% -'** ,2

, *71'1 #4#*-.+#,2 +#2#0 VWRTQ #',-*2 )-TQ 72"T (&0##

1#. 0 2# +# 130#+#,21 5#0# 2 )#, $0-+ 2&# +'"V4#', -$ # !& *# $ -$ #4#07

.* ,2 ," +','+3+ -$UR+# 130#+#,215#0#- 2 ',#"$0-+ # !&%0-3. 2

# !& 2'+# .-',2T (&# ! *!3* 2#1 2&# -.2'! * "#,1'27 -$ *'%&2 20 ,1+'22#"

2&0-3%&2&#*# $ ,"2&'15 1#6.0#11#"Q31',% ,3+#0'! *4 *3# #25##,S ,"

SRRQ 1 0#* 2'4# !&*-0-.&7** !-,!#,20 2'-,T (&# 4#0 %# 4 *3# - 2 ',#" $0-+

2#+# 130#+#,215 1#6.0#11#" 10#* 2'4#!&*-0-.&7**!-,!#,20 2'-,T

53 CHAPTER 2: Characterisation of resistance responses to Phytophthora cinnamomi

2.2.20. Determination of levels of P. cinnamomi within inoculated plant roots (- "#2#0+',# 2&# *#4#* -$ P. cinnamomi 5'2&', ',-!3* 2#" .* ,21Q 0--21

5#0# #6!'1#" $0-+ # !& .* ,2 2 TV & ',2#04 *1 $-0 Y ".'Q 2 5#0# 2&#,

1#!2'-,#" ," .* 2#" 1#/3#,2' **7 -,2- 2&# Phytophthora 1#*#!2'4# .

+#"'3+ 1"#1!0' #"',1#!2'-,TTTT[T!.2-V',"'4'"3 *0--215#0#.* !#"-,

1',%*# .* 2#T * 2#1 5#0# ',!3 2#" 2 TV5) $-0 YT & 2- **-5 $-0 . 2&-%#,

%0-52& ,"2&#%0-52&-$P. cinnamomi *-,%2�--21#%+#,215 10#!-0"#"

,".&-2-%0 .&#"T(&##6.#0'+#,25 10#.# 2#"2&0##2'+#1T(-2 *. 2&-%#,

%0-52&-,# !&0--25 1#6.0#11#" 1 .#0!#,2 %#-$2-2 *,3+ #0-$0--2

1#%+#,21.#0.* ,2T

2.2.21. Statistical analysis of data

2 2'12'! * , *71'15 1!-,"3!2#"31',%'%+ *-24#01'-,SS712 2

-$25 0#Q ,-1#Q) Q! T(&#+# ,1$-0# !&2'+#V.-',25#0#!-+. 0#"

12 2'12'! **7 31',% tV2#12T /0-3.1 2& 2 5#0# ,-2 ,-0+ **7 "'120' 32#" 5#0#

13 (#!2#"2- # ,,V3&'2,#7. ,)3+(#12T

2.2.22. Northern blot analysis to investigate P. cinnamomi accumulation in inoculated Z. mays root tissue

(- $302� ',4#12'% 2# *#4#*1 -$ P. cinnamomi5'2&',Z. mays0--21Q

,-02�,&7 0'"'1 2'-,31',% 0 "'-V* #**#"P. cinnamomi.0- #5 1

!-,"3!2#"T(&##6.#0'+#,25 10#.# 2#"2&0##2'+#1T

54 CHAPTER 2: Characterisation of resistance responses to Phytophthora cinnamomi

2.2.23. Tissue sampling for RNA extractions

.--22'113#$0-+!-,20-* ,"',-!3* 2#".* ,215 1& 04#12#" 2RQTVQVZQ

,"YT&.'T **2'113#5 1'++#"' 2#*7$0-8#,',*'/3'",'20-%#, ,"12-0#" 2

7ZR5)3,2'*0#/3'0#"T +','+3+ -$SW0--21 -$# !&20# 2+#,2 2# !&2'+#V

.-',25 1.--*#"2-+','+'1# '-*-%'! *4 0' 2'-,1 #25##,.* ,21T

2.2.24. RNA extraction trials and optimisation

#4#0 * .< #620 !2'-, +#2&-"1 5#0# 20' **#" ', -0"#0 2- - 2 ', 2&#

%0# 2#127'#*" ,".30'27$0-+2�--22'113#Q',!*3"',%!-++#0!' *.<

#620 !2'-, )'21 ? (-2 * .< *1-* 2'-, 712#+Q ) 2 *-%3# 3HUSRRQ 0-+#% Q

3'1!-,1',Q ! Q , #620 !2'-, +#2&-" 2 '*-0#" $-0 &'%& ! 0 -&7"0 2#V

!-,2 ',',% 2'113# ..#,"'6 UTR) 88-,#**' et al. S[[Z ," (.*8-* 1#"

+#2&-"T<-.< !-3*" #- 2 ',#"$0-+2&#$'012+#2&-" ,"-,*7 1+ **7'#*"

0RTS=% =75 1- 2 ',#"$0-+2#!-,"T(&#(.*8-*+#2&-"#, *#"&'%&

7'#*"11S=% =7 ,"#6!#**#,2.30'27-$STZTTTT

/--"/3 *'27.< '14'2 *$-0"-5,120# ++-*#!3* 0 , *71'131',% -2&

<-02�, *-22',% ," < +'!0- 00 71T 3&'*12 2&# (.*8-* +#2&-" '1 +-0#

2'+#!-,13+',%2& ,31',% !-++#0!' *.< #620 !2'-,)'2Q'2'1 0#*' *#

+#2&-"2-#620 !2 !-,1'12#,2*7%--"7'#*" ,"/3 *'27 .< $0-+Z. mays0--2

2'113#T

2.2.25. RNA extraction from Z. mays root tissue

#25##,TRRVTWR+%-$$0-8#,0--22'113#5 1%0-3,"2- $',#.-5"#0

',*'/3'",'20-%#,5'2&', .0#V!&'**#"+-02 0QT+7-$(.*8-*S+7 SRR+%

2'113#5 1 ""#" ,"2 +.*#5 1%0-3,"2- *'/3'"T(&#&-+-%#, 2#5 1

55 CHAPTER 2: Characterisation of resistance responses to Phytophthora cinnamomi

2&#,20 ,1$#00#"2-SR+7.< 1#V$0##!#,20'$3%#23 #1 ,"',!3 2#" 20--+

2#+.#0 230#.($-0W+',32#1T( +.*#5 1!#,20'$3%#" 2URRR6%$-0UR

+',32#1 ,"2&# /3#-31.& 1#5 120 ,1$#00#"2- $0#1&.< 1#V$0##23 #1T*2

5 12&#,.0#!'.'2 2#" 72&# ""'2'-,-$S+7'1-.0-.7* *!-&-*RTW+7 S+7

(.*8-*Q +'6#" 5#** 7 ',4#01'-, ," ',!3 2#" 2 .( $-0 SR +',32#1T (&#

.0#!'.'2 2# 5 1 !#,20'$3%#" 2 URRR 6 % $-0 UR +',32#1 ," 2&# 13.#0, 2 ,2

"'1! 0"#" 7"#! ,2',%Q ,70#+ ',',%13.#0, 2 ,25 1! 0#$3**70#+-4#"5'2&

.'.#22#T (&# .#**#2 5 1 5 1&#" 5'2& RTW +7 -$ YW$ #2& ,-* + "# 5'2&

.< 1#V$0##5 2#0 ,"!#,20'$3%#" 2URRR6%$-0TR+',32#1T( .#0, 2 ,2

5 1 % ',"'1! 0"#" 7"#! ,2',% ,"2 #',4#02#"-,2- 1-0 #,2. .#0

='+5'.#Q='+ #0*#7V)* 0)Q#'*1-,1-',2Q<32-"0 5-$$ ,70#1'"3 **'/3'"T

( #5 1 **-5#"2- '0V"07$-0SW+',32#1Q2-#,130# **2&##2& ,-*& "

#4 .-0 2#"$0-+2 **1-$2 #Q.0'-02-2&# ""'2'-,-$WR=7.< 1#V$0##

5 2#0T(&#.#**#25 10#131.#,"#" 7.'.#22',%3. ,""-5,TRVUR2'+#1T

(&# .< 5 1 2&#, /3 ,2'$'#" 31',% < ,-0-. .#!20-.&-2-+#2#0

<VSRRRQ 3'*+',%2-,Q 9Q S[ZSRQ ! T S 7 "0-. -$ # !& 1 +.*# 5 1

.* !#" -, 2&# +# 130#+#,2 .#"#12 * ,"2&#< ,-0-.1#22-.< VVRT(&#

$', * !-,!#,20 2'-, -$ 2&# 1 +.*#Q 1#" -, 2&# 1-0 ,!# 2 TXR ,+Q 5 1

#6.0#11#" 1,% 72-2 *.< T(� 2'--$1 +.*# 1-0 ,!# 2TXR TZR5 1

31#"2-"#2#0+',#2&#.30'27-$2 +.*#T

2.2.26. RNA gel electrophoresis for nucleic acid transfer

.< %#* #*#!20-.&-0#1'1 $-0 ,3!*#'! !'" 20 ,1$#0 4' ! .'** 07 *-22',%

5 1 !-,"3!2#" 31',% +-"'$'#" +#2&-" $0-+ + 0--) ," .311#**Q TRRST

0'#$*7Q.< 5 1"#, 230#" 2XW5)$-0W+',32#1 #$-0#SR=%-$# !&1 +.*#

56 CHAPTER 2: Characterisation of resistance responses to Phytophthora cinnamomi

5 103,-, STW$ % 0-1#%#*',RTW$(0'1-0 2#9( (9 3$$#0 2SRR

4-*21 $-0 WR +',32#1T /#*1 5#0# 2&#, 12 ',#" 5'2& #2&'"'3+ 0-+'"# RTW

=% +7$-0W+',32#1 ,""#V12 ',#"',"T>$-0 $302�W+',32#1.0'-02-

4'13 *'1 2'-,3,"#03*20 V4'-*#2*'%&2T

2.2.27. Northern hybridisation

<-02�, &7 0'"'1 2'-,1 "#1!0' #" #*-5 5#0# !-,"3!2#" 31',%

+#2&-" +-"'$'#" $0-+ + 0--) ," .311#** TRRST (&# #6.#0'+#,2 5 1

0#.# 2#"2&0##2'+#1T

2.2.28. Nucleic acid transfer via capillary blotting

%* 11 13..-02 ," 5'!)Q !-+.0'1',% 25- * 7#01 -$ U## $'*2#0 . .#0

3& 2+ ,U##. .#0Q) 2 *-%3#3SRRS[SYQ# '"12-,#Q9,%* ,"Q5#0#1#23.

!0-11 .* 12'!"'1&$'**#"5'2&20 ,1$#0 3$$#0TRK)Q ..#,"'6UTS ,"2&#

5'!)1- )#"',SRK)T(&#.< %#*5 1.* !#" !0-112'!) ,"

.-1'2'4#*7 !& 0%#" ,7*-, +#+ 0 ,# +#01& + 7 -,"V<&Q /9 # *2&! 0#Q

3!)',%& +1&'0#Q!=5 1!322-1'8# ,"* '" !0-112&#%#*Q '0 3 *#15#0#

0#+-4#" 70-**',% . 12#30.'.#22# !0-112‚$ !#T(&#+#+ 0 ,#5 1

!-4#0#"5'2& * 7#0-$.0#V!32U##$'*2#0. .#0 ,"5#25'2&SRK)Q2&'15 1

0#.# 2#"5'2& ,-2�W* 7#01-$. .#0T 2&'!)+','+3+W!+&'%&12 !)-$

. .#02-5#*5 1.* !#" !0-112-.Q2-..#"5'2& %* 11.* 2# ," UWRVVRR%

5#'%&25 11#2',.* !#Q#,130',%#4#,.0#1130# !0-112&#+#+ 0 ,#T(&#

.. 0 2315 1*#$2-4#0,'%&2 2.($-0.< 20 ,1$#02-2&#,7*-,+#+ 0 ,#T

(0 ,1$#0#$$'!'#,!75 1!&#!)#" 74'13 *'1',%2&#.< %#*3,"#0!?*'%&2T.<

5 12&#,$'6#"2-2&#+#+ 0 ,# 7.* !',%2&#+#+ 0 ,#', &7 0'"'1 2'-,

57 CHAPTER 2: Characterisation of resistance responses to Phytophthora cinnamomi

-4#, 2ZR5)$-0T&-301T(&#+#+ 0 ,#5 112-0#"5'2&', 8'.V*-!) % 2.(

3,2'*0#/3'0#"T

2.2.29. Pre-hybridisation

(&# +#+ 0 ,# 5 1 .* !#" 5'2&', %* 11 &7 0'"'1 2'-, -22*# 5'2& 2&#

*-22#" 1'"# $ !',% ',5 0"1 ," SR +7 -$ )&30!& 3$$#0 ..#,"'6 TTW 5 1

""#"2-2&# -22*#T(&# -22*#!-,2 ',',%2&#+#+ 0 ,#5 1.* !#"5'2&',

.0#V&# 2#"&7 0'"'1 2'-,-4#,XW5)$'22#" 5'2& 0-2 2',% !7*',"#05&##*$-0

..0-6'+ 2#*7S&-30T

2.2.30. Probe labelling and purification

< .0- #1* #**#"5'2&JV#+'22',%Q0 "'- !2'4#&-1.&-031VUTVUT

9 172'"#1")(Q VUT#0)',9*+#0Q/*#,3 4#0*#7Q?'!2-0' USWR5#0#31#"

$-0 **&7 0'"'1 2'-,#6.#0'+#,21T3'2&', 0 "' 2'-,0--+Q2&#VUT5 1.* !#"

5'2&', .#01.#6 -62-2& 5$-0S&-30T0- #* #**',%5 1!-,"3!2#"31',%

+#01& + .#"'.0'+# ** . ,"-+ 0'+#0 7 #**',% 712#+ /9 # *2&! 0#Q

3120 *' Q !!-0"',% 2- 2&# + ,3$ !230#0_1 ',1203!2'-,1T 0'#$*7Q 2&# <

1#!2'-,TTTTZT5 1"'*32#"2-TTWVTW,%',VW=7-$SR+#(9 3$$#0Q'25 12&#,

"#, 230#" 7&# 2',%2-[W5)$-0W+',32#1 ,"1, .!--*#"-,'!#$-0 $302�

W+',32#1T(&#"#, 230#"< 5 12&#, ""#"2-2 ..*'#"0# !2'-,23 #

,"W=7-$.#"'43#UT")(5 1 ""#"2-2 # ,"+'6#" 7.'.#22',%

ST2'+#1T( #5 12&#,',!3 2#" 2.($-0UR+',32#1T

(&#.0- #5 1.30'$'#"31',%2&#/9***3120 0- #I3 ,2/VWR#'!0-

)-*3+, 1.#0+ ,3$ !230#0_1',1203!2'-,1T0'#$*7Q2&#!-*3+,5 1.0#. 0#"

74-02#6',%2-0#131.#,"2�#1',Q2&#! .5 1*--1#,#"-,#V/3 02#0230, ,"

58 CHAPTER 2: Characterisation of resistance responses to Phytophthora cinnamomi

2&# -22-+!*-130#1, ..#"-$$T(&#!-*3+,5 1.* !#"5'2&', !-**#!2'-,23 #

,"!#,20'$3%#"$-0S+',32# 2YUW6%T(&#!-*3+,5 12&#,20 ,1$#00#"2-

$0#1&1!0#52-.STW+723 # ,"WR=7-$1 +.*#5 1 ..*'#"2-2&#!#,20#-$

2�#1', #"T( +.*#5 1#*32#" 7!#,20'$3%',%$-0T+',32#1 2YUW6%T

(&#.30'$'#".0- #5 1"#, 230#" 2[W5)$-0W+',32#1 ,"1, .!--*#"

-,'!#$-0W+',32#1TWR=7-$2&#.30'$'#"Q0 "'-V* #**#".0- #5 12&#, ""#"

2- 2&# &7 0'"'1 2'-, -22*# !-,2 ',',% 2&# +#+ 0 ,# ," 2&# +#+ 0 ,# 5 1

&7 0'"'1#"5'2&%#,2*# %'2 2'-,-4#0,'%&2 2XW5)T

2.2.31. Membrane washing

(&# +#+ 0 ,# 5 1 !&#!)#" $-0 JV#+'22',% . 02'!*#1 31',% /#'%#0

!-3,2#0Q ,"2x',%#,!7-$+#+ 0 ,#5 1 1"#2#0+',#" 1#"-,2&#

!-3,2- 2 ',#" ..#,"'6TTWTT(7.'! *5 1&',%!-,!#,20 2'-,1 0#"#1!0' #"

�#T (&# )&30!& 3$$#0Q !-,2 ',',% 2&# .0- #Q 5 1 #+.2'#" $0-+ 2&#

&7 0'"'1 2'-, -22*# ,"2&#+#+ 0 ,# 0'#$*75 1&#"5'2& 1+ **4-*3+#-$

TK) RTS$ ..#,"'6UTSQ #$-0#2&# -22*#5 1$'**#"5'2&VRR+7-$TK

) RTS$2-!-4#02&#+#+ 0 ,#T(&# -22*#5 1',!3 2#" 2.($-0SW

+',32#1T(&#+#+ 0 ,#2&#,20 ,1$#00#"2- .* 12'!!-,2 ',#0 ,"!-4#0#"

5'2& 1-*32'-, -$ TK ) RTS$ .0#V&# 2#" 2- WW5) ," 2&# !-,2 ',#0

.* !#"-, .* 2$-0+0-!)#05'2&', &7 0'"'1 2'-,-4#, 2WW5)$-0SW+',32#1

..#,"'6 UTST (&# 1-*32'-, 5 1 0#.* !#" 5'2& RTTK ) RTS$ ," 2&#

!-,2 ',#00#230,#"2-2&#&7 0'"'1 2'-,-4#,$-0 $302�SW+',32#1

..#,"'6TTWT( 1&#"+#+ 0 ,#5 1"0 ',#"-,U##$'*2#0. .#0 ,"

! 0#$3**7!-4#0#"5'2& 1',%*#* 7#0-$!*# 0.* 12'!50 ./7 Q 3120 *' T*2

5 1 2&#, 1! ,,#" 5'2& 2&# /#'%#0 !-3,2#0 2- "#2#0+',# 1'%, * ',2#,1'27 -$ JV

59 CHAPTER 2: Characterisation of resistance responses to Phytophthora cinnamomi

#+'22',%. 02'!*#1 ,"2�#/3'0#"#6.-130#2'+#5 1#12'+ 2#" 1#"-,2&#

*#4#*-$1'%, *- 2 ',#"Q 27.'! *#6.-130#2'+#5 1TVVVZ&Q 1#"-, 1'%, *

*#4#*-$SRVSW!-3,21.#01#!-,"!.1T

2.2.32. Film exposure and development

(&#+#+ 0 ,#5 1.* !#"5'2&', ! 11#22# ,"20 ,1$#00#"2- " 0)0--+T

!,"#0 0#" *'%&2 2&# ! 11#22# 5 1 -.#,#" ," 2&# +#+ 0 ,# !-4#0#" 5'2&

1',%*#1&##2-$$'*+ +#01& +7.#0$'*+#Q/9# *2&! 0#Q

3!)',%& +1&'0#Q!=T(&#! 11#22#5 1.* !#"5'2&', $0##8#0 27ZR5)$-02&#

0#/3'0#" #6.-130# 2'+#T 0-**-5',% #6.-130#Q 2&# ! 11#22# 5 1 0#+-4#" $0-+

2&#$0##8#0 ,".* !#"5'2&', " 0)0--+2-2& 5T(&#$'*+5 10#+-4#" ,"

20 ,1$#00#" 2- 2&# "#4#*-.#0 1-*32'-, **$-0" &#,'1-* '%& )-,20 12 0'*+

#4#*-.#0$-0W+',32#1Q0',1#" 0'#$*7',1"T> ,"20 ,1$#00#"2-2&#$'6 2'4#

1-*32'-,**$-0"7. +. .'" .#0 ,"0'*+0'6#0$-0T+',32#1T

60 CHAPTER 2: Characterisation of resistance responses to Phytophthora cinnamomi

2.3. Results

2.3.1. Morphological identification of isolates

** '1-* 2#1 31#" ', #6.#0'+#,2 * 5-0) 5#0# ','2' **7 '"#,2'$'#" 7

#6 +',',%2&#+-0.&-*-%7-$+7!#*'3+%0-5',%-,SR$)?Z % 0.* 2#13,"#0

*'%&2 +'!0-1!-.# 2 *-5 ," &'%& .-5#0 0'%30# TTV T *1-* 2#1 "'1.* 7',%

27.'! * +-0.&-*-%'! * !& 0 !2#0'12'!1 -$ P. cinnamomi$'%30#TTV ',!*3"',%

!-00 *-'" +7!#*'3+ 5'2& &7.& * 15#**',%1 5#0# 1#*#!2#" $-0 $302�

!-,$'0+ 2'-,31',%< '"#,2'$'! 2'-,T a b

Figure 2.4. (a) Isolate DU067 of P. cinnamomi growing on 10% CV8 agar after 72 h on the media. Scale bar = 1 cm. (b) Isolate DU067 of P. cinnamomi showing distinct hyphal swellings. Scale bar = 10 m.

2.3.2. Molecular confirmation of isolates

).-$# !&1 +.*#.0-"3!#" .0-"3!2-$ZT[ .5&'!&!-,$'0+#"'21

'"#,2'27 1 #*-,%',% 2- 2&# %#,31 Phytophthora 0'%30# TTW T 3 1#/3#,2

.#120'!2'-, 00 %+#,2 7#,%2& -*7+-0.&'1+ .07 , *71'1 -$ 2&# +.*'!-,

31',% 2&# 0#120'!2'-, #,87+#1 MspI, RsaI ,"TaqI .0-"3!#" $0 %+#,21

!-,1'12#,25'2&2&-1#$-0P. cinnamomi 0'%30#TTW Q2&31!-,$'0+',%2& 22&#

'1-* 2#5 1P. cinnamomi.

61 CHAPTER 2: Characterisation of resistance responses to Phytophthora cinnamomi

a) 1 2 3 4 5 6 b) 1 2 3 4

300 bp 800 bp 100 bp

Figure 2.5. (a) Lane 1: 1 kb ladder, Lanes 2-6: PCR identification using the genus specific primer pair A2-I2 of isolate DU067 resulted in a consistent 829 bp fragment indicative of the Phytophthora genus. (b) Subsequent RFLP analysis of the amplicon confirmed the isolate to be P. cinnamomi. Lane 1: 100 bp ladder, Lane 2: restriction enzyme MsPI produced fragments of 339, 221, 147, 122 bp Lane 3: RsaI produced fragments of 377, 170, 165, 106 bp Lane 4: TaqI produced fragments of 194, 181, 150 and 90 bp.

2.3.3 Infection time course

H--1.-0#15#0#.0#$#0#,2' **7 220 !2#"2-2-,#-$#*-,% 2'-,-$2&#

0--21 ," & " #%3, 2- %%0#% 2# 0-3," 2&'1 8-,# 5'2&', SW V UR +',32#1

$'%30#TTX T $2#0 ..0-6'+ 2#*7VW+',32#1Q1YR$-$2--1.-0#1& "1&#"

2&#'0$* %#** ,"#,!712#"Q13 1#/3#,2*71WR$-$ **!7121$-0+#"%#0+23 #1

$'%30#TTX Q +','+3+-$WR1.-0#15#0#!-3,2#"$-0# !&- 1#04 2'-,T7XR

V[R+',32#1Q2&##,!712#"8--1.-0#1& "#62#,"#"2&#'0%#0+23 #12-$ !'*'2 2#

.#,#20 2'-, -$ 2&# #.'"#0+ * 130$ !# -$ 2&# 3,13 #0'8#" .0'+ 07 0--2 2'113#

$'%30#TTX T 2SZR+',32#1Q%#0+23 #1& " #%3,2-.#,#20 2#2&##.'"#0+'1

-$2�--22'113# 22-,#-$#*-,% 2'-,T(&#. 22#0,-$','2' *0--2 220 !2'-,Q

',!*3"',% 2&# 1.##" -$ 8--1.-0# %%0#% 2'-, ," #,!712+#,2 "'" ,-2 "'$$#0

1'%,'$'! ,2*7 #25##,0--21-$Z. mays, -0L. angustifolius " 2 ,-21&-5,.

62 CHAPTER 2: Characterisation of resistance responses to Phytophthora cinnamomi

a

GT CY

b

GT

CY

EC

Figure 2.6. Z. mays root tissue 180 minutes post inoculation with 10 L of P. cinnamomi

zoospores (a) zoospores formed walled cysts (CY) which subsequently adhered to the root tissue of Z. mays. Many of the cysts (>50%) extended germ tubes (GT) towards the root and attempted penetration of the epidermis. Scale bar = 20 m (b) Adherence of walled cyst (CY) to an epidermal cell (EC) in the zone of elongation. Scale bar = 5 m

2.3.4. Plant health following inoculation with P. cinnamomi

,','2' *4'13 * 11#11+#,2-$5&-*#.* ,21-4#0 2'+#!-301#-$SV" 71

1&-5#" *'22*# "'$$#0#,!# #25##, !-,20-* +-!) ',-!3* 2#" ," ',-!3* 2#"

Z. maysT $2#0Y" 71.-12',-!3* 2'-,".'Q.* ,210#+ ',#"&# *2&7 ,"4' *#

," 0--21 !-,2',3#" 2- %0-5 $'%30# TTY T $2#0 SV ".'Q ** ',-!3* 2#" .* ,21

0#+ ',#" &# *2&7T *, !-,20 12Q 2&# &# *2& ," %0-52& -$ ',-!3* 2#"

L. angustifolius.* ,215 11'%,'$'! ,2*7!-+.0-+'1#" ,"0--2 ,"*# $%0-52&

63 CHAPTER 2: Characterisation of resistance responses to Phytophthora cinnamomi

& " !# 1#" 7 #25##, W ," Y ".' $'%30# TTY T 7# 4#1 -$ ',-!3* 2#"

L. angustifolius5#0#0#"3!#"', -2&1'8# ,",3+ #0Q!-+. 0#"2-!-,20-*

.* ,21 ,"& " 1+ **0SR$2-2 **# $130$ !# +-3,2-$',2#04#', *!&*-0-1'1T

7SV".'Q1YW$-$ **',-!3* 2#"L. angustifolius5#0#"# "Q ,"2&#&# *2&-$

1304'4',%.* ,215 11'%,'$'! ,2*7!-+.0-+'1#"T

ab

Figure 2.7. (a) Z. mays plants 7 dpi with P. cinnamomi zoospores (right) exhibit no disease symptoms, the plants remain healthy and root growth is not attenuated, small hypersensitive lesions (arrowhead) are visible at the points of inoculation. (b) L. angustifolius plants 7 dpi with P. cinnamomi zoospores (right) are visibly diseased, with curled leaves, and dark, water soaked lesions present on >75% of the total root area. Plant growth is also significantly compromised in all experimental plants with experimental plants considerably (>50%) smaller than control plants. Control (mock inoculated) plants are shown on the left. Plants shown are representative of 30 plants per species per treatment group. Scale bar = 1.5 cm

64 CHAPTER 2: Characterisation of resistance responses to Phytophthora cinnamomi

2.3.5. Restricted lesion production and formation of lateral primary roots in P. cinnamomi inoculated Z. mays

.--21-$Z. mays',-!3* 2#"5'2&P. cinnamomi8--1.-0#11&-5#" 0 .'"

.0-"3!2'-,-$0#120'!2#"*#1'-,10VZ&.'5&'!&5#0#!-,$',#"2-1+ ** 0# 1

"'0#!2*7 #,# 2&2'2#-$',-!3* 2'-,$'%30#TTZ T 2YT&.'Q+3*2'.*#* 2#0 *

0--21 & " $-0+#" 0-3," 2&# *#1'-,Q 1 5#** 1 "'12 **7 -4# 2&# 1'2# -$

',-!3* 2'-,$'%30#TTZ T*,!-,20 12Q0--21-$L. angustifoliusQ',-!3* 2#"5'2&

2 +#!-,!#,20 2'-,-$P. cinnamomi8--1.-0#1Q"#+-,120 2#"5 2#0V1- )#"

*#1'-,15&'!>#,"#". 122&#','2' *.-',2-$',-!3* 2'-, ,"0#13*2#"',

2'113#!-** .1#Q* 2#0 *0--2%0-52&5 1 1#,2$'%30#TTZ T

a

b

Figure 2.8. (a) Z. mays root 72 h post inoculation (hpi) with P. cinnamomi zoospores, showing restricted lesions beneath the point of inoculation (arrowhead). (b) L. angustifolius root 72 hpi, a spreading lesion beneath the point of inoculation (white arrowhead) root tissue is water soaked and has collapsed (black arrowhead). Pictures representative of 3 independent experiments. Scale bar = 0.5 cm

65 CHAPTER 2: Characterisation of resistance responses to Phytophthora cinnamomi

2.3.6. Plant fresh weight post inoculation (�#5 1,-1'%,'$'! ,2"'$$#0#,!# #25##,2&#$0#1&5#'%&2-$!-,20-*

,"',-!3* 2#"Z. mays.* ,21$0-+VZVSXZ&.'$'%30#TT[ T 2SXZ&.'Q2&#

+# ,$0#1&5#'%&2-$',-!3* 2#"Z. mays 5 1 STRY%Q ( RTRT +# , ( 9#

!-+. 0#"2-2&#+# ,-$!-,20-*.* ,21 2STSR%(RTRTT-5#4#0Q2�#5 1

1'%,'$'! ,2 "'$$#0#,!# #25##, 2&# $0#1& 5#'%&2 -$ !-,20-* ," ',-!3* 2#"

L. angustifolius $0-+VZVSXZ&.'$'%30#TT[ -RTRSU 2VZ&.'Q0RTRRS **

13 1#/3#,2 2'+#V.-',21T 7 SXZ &.'Q 2&# +# , $0#1& 5#'%&2 -$ ',-!3* 2#"

L. angustifolius5 1RTXZ%(RTRUQ *+-12& *$2& 2-$!-,20-*1 2STS[%(RTRWT

1.20 a) Control 1.00 Inoculated

0.80

0.60

Fresh weight (g)Fresh weight 0.40

Z. mays 0.20

0.00 48 72 120 168 Time post inoculation (h)

b) 1.40 1.20

1.00 0.80 * * * Fresh weight (g)Fresh weight * 0.60

0.40

0.20 L. angustifolius angustifolius L. 0.00 48 72 120 168 Time post inoculation (h)

Figure 2.9. Fresh weight of whole plants over 168 h following inoculation with P. cinnamomi zoospores or mock inoculated with dH O (a) Fresh weight of inoculated Z. mays plants (b) 2 Fresh weight of inoculated L. angustifolius plants * denotes means that were significantly different to controls. Data shown is the mean of 3 independent experiments, bars represent standard error of the mean (SEM), n = 30 plants per species per treatment group.

66 CHAPTER 2: Characterisation of resistance responses to Phytophthora cinnamomi

2.3.7. Root lengths post inoculation

(�# 5 1 ,- 1'%,'$'! ,2 "'$$#0#,!# 1RTRW #25##, 2&# %0-52& -$

!-,20-* ,"P. cinnamomi',-!3* 2#"Z. mays0--21.#0TV&-4#0 SXZ&2'+#

!-301#$'%30#TTSR T-5#4#0Q2�#5 1 1'%,'$'! ,20#"3!2'-,0RTRRS',

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0--2 #62#,1'-, !# 1',% 7 STR &.' $'%30# TTSR T .--2 %0-52& 1*-5#" $0-+

STRVSXZ&', -2&!-,20-* ,"',-!3* 2#"Z. mays ,"2,V',-!3* 2#"!-,20-*1

-$L. angustifolius 12�--210# !&#"2&# -22-+-$2 73,'21T

4.50 a) Control 4.00 Inoculated 3.50

3.00 2.50 2.00 1.50 1.00 0.50 Average growth (cm) per per (cm) 24h growth Average 0.00 24h 48h 72h 120h 168h

Hours post inoculation

b) 5.00 4.50 4.00 3.50 3.00 2.50 2.00 * 1.50 1.00 0.50 *

Average growth Average growth (cm) per 24h * 0.00 * 24h 48h 72h 120h 168h

Hours post inoculation

Figure 2.10. Root growth over 168 hpi (a) Growth of control and inoculated Z. mays roots (b) Growth of P. cinnamomi inoculated L. angustifolius roots compared to controls. * denotes means which are significantly different to controls. Data shown is mean of 3 independent experiments, bars represent SEM, n = 30 plants per species, per treatment group.

67 CHAPTER 2: Characterisation of resistance responses to Phytophthora cinnamomi

2.3.8. Lesion lengths post inoculation

7#1'-, *#,%2&1 #6.0#11#" 1 .#0!#,2 %# -$ 2-2 * 0--2 *#,%2& 5#0#

1'%,'$'! ,2*7"'$$#0#,20RTRW #25##,P. cinnamomi',-!3* 2#"Z. mays ,"

L. angustifolius -4#02'+#T(&#+# ,*#1'-,*#,%2&1-$',-!3* 2#"Z. mays5#0#

RTV!+-00TTW$-$2-2 *0--2*#,%2& 2VZ&.'Q0# !&',% + 6'+3+-$STU!+-0

YTW$ -$ 2-2 * 0--2 *#,%2& 2 STR V SXZ &.' $'%30# TTSS Q TTST !Q ," 2#

0#+ ',#" 1 2'%&2*7 !-,20-**#"Q ,#!0-2'! $*#!)1 "'0#!2*7 #*-5 2&# .-',21 -$

',-!3* 2'-,T *, !-,20 12Q 2&# +# , *#1'-, *#,%2&1 -$ ',-!3* 2#" L. angustifolius

',!0# 1#"$0-+RTYW!+-00TW$-$2-2 *0--2*#,%2& 2VZ&.'Q2-XTY!+-0WR

YW$-$2-2 *0--2*#,%2& 2STR&.'$'%30#TTSS QTTST ! ," 7SXZ&.'5#0#

ZT[!+-01YW$-$2-2 *0--2*#,%2&$'%30#TTSS QTTST !T

a b

Figure 2.11. Lesions on roots of Z. mays and L. angustifolius at 120 hpi with P. cinnamomi zoospores. (a) Restricted lesions on roots of Z. mays. (b) Water-soaked spreading lesions on L. angustifolius at 120 hpi. Plants shown were inoculated at 5 days old and are representative of 3 separate experiments, n = 30 plants per species, per time-point. Scale bars = 1.5 cm. Several lesions are highlighted with black brackets.

68 CHAPTER 2: Characterisation of resistance responses to Phytophthora cinnamomi

25 a) Root length

20 Lesion length

15

10

5 Growth per 24h (cm) 24h per Growth

0 24h 48h 72h 120h 168h Hours post inoculation

b) 14 12 10 8 6

4

(cm) 24h per Growth 2 0

24h 48h 72h 120h 168h Hours post inoculation c) 90 * 80 Z. mays 70 L. angustifolius 60 * 50 40 30 * 20 * 10 * Lesion lengthrootLesion length) (% 0 24h 48h 72h 120h 168h

Hours post inoculation

Figure 2.12. Lesion lengths of (a) Z. mays and (b) L. angustifolius in comparison to root length. (c) Mean lesion lengths for each species (expressed as a percentage of total root length) were compared over a 168 h time course. *denotes means which are significantly different. Data shown is the mean of 3 independent experiments, bars represent SEM, n = 30 plants per species, per treatment group.

69 CHAPTER 2: Characterisation of resistance responses to Phytophthora cinnamomi

2.3.9. Relative chlorophyll concentration in leaf tissue

(�#* 2'4# +-3,2-$!&*-0-.&7**.0#1#,2',*# $2'113#-$ Z. mays ,"

L. angustifolius5 1#6.0#11#" 1 4 *3# #25##,S ,"SRR 72&# +#2#0T

>4#0Y" 71Q2�#5 1,-1'%,'$'! ,2"'$$#0#,!#1RTRW #25##,2&# +-3,2

-$!&*-0-.&7**',*# 4#1-$!-,20-* ," P. cinnamomi',-!3* 2#"Z. mays $'%30#

TTSU T*,L. angustifolius2�#5 1 1'%,'$'! ,2"'$$#0#,!#0RTRRS #25##,

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*# 4#1-$',-!3* 2#".* ,21$0-+RYT&.'1RTRWT

a) 36 Control 35 Inoculated 34 33 32

31 30 29 28 Relative chlorophyll concentration 27 0 24 48 72 96 120 168 Time post inoculation (h) b) 61

59

57 55 * 53 *

51 * 49 47 Relative chlorophyll concentration 0h 24h 48h 72h 96h 120h 168h Time post inoculation (h)

Figure 2.13. Average relative chlorophyll concentration over time in control and inoculated leaves over 168 h using the Soil Plant Analysis Development (SPAD) meter. (a) Control and inoculated leaves of Z. mays. (b) Control and inoculated leaves of L. angustifolius. *denotes means which are statistically different to controls. Data shown is means of 3 independent experiments, bars represent SEM, n=30 plants per species, per treatment group.

70 CHAPTER 2: Characterisation of resistance responses to Phytophthora cinnamomi

7# 4#1-$',-!3* 2#".* ,215#0#4'13 **7 11#11#"$-0!&*-0-1'1-4#0 SV

" 7.#0'-"T 2SV".'Q2�#5 1,-"'$$#0#,!# #25##,!-,20-* ,"',-!3* 2#"

*# 4#1 -$ 2&# 0#1'12 ,2 Z. mays$'%30#TTSV T*,!-,20 12Q*# 4#1-$

L. angustifolius5#0#4'1' *7!&*-0-2'!Q ,"2&'17#**-5',%5 1.0#1#,2',1WR$

-$2‚$ !#-$2&#*# 4#1$'%30#TTSV T

a

b

Figure 2.14. Leaves of control (left) and inoculated (right) of Z. mays and L. angustifolius at 14 dpi. (a) No visible chlorosis in the leaves of inoculated Z. mays. (b) Inoculated

L. angustifolius show significant levels of chlorosis. Plants shown are representative of 3 independent experiments, n = 30 plants per species, per treatment group. Scale bar = 1.5 cm

71 CHAPTER 2: Characterisation of resistance responses to Phytophthora cinnamomi

2.3.10. Comparison of P. cinnamomi hyphal growth in inoculated root segments

.--21 -$ Z. mays .* 2#" -,2- . +#"'3+ 2 TV SXZ &.' 1&-5#"

"#!0# 1',%*#4#*1-$P. cinnamomi',$#!2'-,-4#02'+#T*,0--21& 04#12#" 2TVV

&.'Q$-**-5',%U" 71',!3 2'-,Q 1+ ** +-3,2-$%0-52&& "1.0# " 0-3,"

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5 1 0#120'!2#" 2- *#11 2& , TW$ -$ ** 0--2 1#%+#,21 $'%30# TTSWT T *, 0--21

& 04#12#" 2VZYT&.'Q2&#*#4#*-$. 2&-%#,%0-52&-,2&#.* 2#& "1.0# "

2-TWVR$-$2-2 *.* 2# 0# ,"!-4#0#" ,3+ #0-$ "( !#,20--2

1#%+#,21$'%30#TTSW!T7STRVSXZ&.'Q. 2&-%#,%0-52&5 10#120'!2#"2-

2&#.-',21-$',-!3* 2'-,-,*7R2&'15 1*#112& ,TR$-$2-2 *-$ **.* 2#"

0--2 1#%+#,21 $'%30# TTSWT#T (&'1 "#+-,120 2#1 2& 2 *#4#*1 -$ P. cinnamomi

',$#!2'-, 0#1'%,'$'! ,2*7 22#,3 2#"5'2&',Z. mays-4#02'+#T

.--21 -$ L. angustifolius .* 2#" -,2- . +#"'3+ 2 TV& V SXZ &.'

"#+-,120 2# 0 .'" ',!0# 1# -$ P. cinnamomi ',$#!2'-, -4#0 2'+#T >, 0--21

& 04#12#" 2TV&.'Q. 2&-%#,%0-52&& "!-4#0#"+-0#2& ,[R$-$ **.* 2#"

0--21#%+#,21$'%30#TTSW Q ," $2#0YT&.'P. cinnamomi+7!#*'3+!-4#0#"

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%0-52&& "1.0# "2-2&#-32#0#"%#-$2&#.* 2# ,"!-4#0#"SRR$-$ **0--2

1#%+#,21$'%30#TTSW$T

72 CHAPTER 2: Characterisation of resistance responses to Phytophthora cinnamomi

Z. mays PARPH L. angustifolius PARPH +72 h incubation +72 h incubation a bb

24 hpi

c d

72 hpi

e f

168 hpi

Figure 2.15. Root sections inoculated with P. cinnamomi zoospores plated sequentially onto Phytophthora-selective media (PARPH). (a,c,e) Z. mays root sections harvested at 24 hpi, 72hpi (4 roots plate-1) and 168 hpi (3 roots plate-1), and incubated for 72 h at 24°C to allow for pathogen growth. Scale bar = 1.25 cm. (b,d,f) L. angustifolius root sections harvested at the same time-points after 72 h incubation. Arrowheads indicate point of inoculation. Images shown are representative of 3 replicate plates and the experiment was repeated twice. Scale bar = 1.5 cm.

73 CHAPTER 2: Characterisation of resistance responses to Phytophthora cinnamomi

2.3.11. Assessment of resistance to P. cinnamomi in Z. mays and L. angustifolius $2#0Y".'Q $', * 11#11+#,2-$2&#*#4#*-$0#1'12 ,!#5 1 11'%,#"$-0

Z. mays 1#"-,2&#W. 0 +#2#01.0#4'-31*7-32*',#"TZ. mays.* ,211 2'1$'#"

** W !0'2#0' ," 5#0# 2�#$-0# !* 11'$'#" 1 0#1'12 ,2 2-

P. cinnamomi ," 11'%,#" 1!-0#-$W ( *#TTUT(-$302�.322#4 *3#1

',2- !-,2#62Q 2 5#0# !-+. 0#" 2- 2&-1# -$ 2&# 131!#.2' *#

L. angustifolius 5&'!& $ '*#" 2- . 11 ,7 -$ 2&# 11#11#" !0'2#0' 2 Y ".' ,"

5#0# 11'%,#" 1!-0#-$R( *#TTUQ$'%30#TTSX#T

Response post Resistance Resistance Z. mays L. angustifolius inoculation rating rating Not increased Increased (no (significantly Root length difference to 1 0 different to controls) controls) >75% Lesion length (% of <10% 1 root necrosis and 0 total root length) tissue collapse Decreased Increased (no (significantly Plant fresh weight difference to 1 0 different to controls) controls) Growth on Uncontrolled. Minimal, Phytophthora Pathogen growth restricted to site 1 0 selective media in 100% of root of inoculation after 72 h segments Reduced Not reduced (no Relative chlorophyll (significantly difference to 1 0 amount different to controls) controls) Final resistance Resistant 5 Susceptible 0 assessment

Table 2.3. Responses of Z. mays and L. angustifolius to inoculation with P. cinnamomi zoospores assessed at 7 dpi. Plants were assigned 1 for each criteria they passed, and plants which failed to pass a criteria were assigned 0. These were totalled and each plant was assigned a final resistance rating out of 5. Z. mays plants passed all criteria were given a final resistance assessment of 5, L. angustifolius failed each criteria and were assigned a resistance assessment of 0.

74 CHAPTER 2: Characterisation of resistance responses to Phytophthora cinnamomi

(-'**3120 2#&-52&'11! *#5-0)1', 131!#.2' *#.* ,2Q2&#*#1'-,*#,%2&

. 0 +#2#0'11&-5, #*-5',L. angustifolius $'%30#TTSX 1$', * 11#11+#,21

-$ *#1'-, *#,%2& 5#0# 3,"#02 )#, 2 Y ".'Q ** L. angustifolius.* ,215#0#

11'%,#" 1!-0#-$RT

a b c d e

Control = 1 10-25% = 0.75 25-50% = 0.50 50-75% = 0.25 >75% = 0

Figure 2.16. Measurement of lesion length as a percentage of total root length in L. angustifolius at 7 dpi. (a) Control plants with healthy white roots and no sign of lesion development. (b) A score of 0.75 was assigned to plants with lesion development on 10- 25% of the total surface of the root. (c) A score of 0.50 was assigned to plants with lesions on 25-50% of the root (d) A score of 0.25 was assigned to plants with lesions on 50 – 75%

of the root (e) A score of 0 was assigned to plants with necrotic lesion development on >75% of the total root. Square brackets designate the lesioned area. Scale bars = 0.5 cm.

2.3.12. Quantification of P. cinnamomi levels within inoculated Z. mays root tissue P. cinnamomi 5 1 "#2#!2#" 5'2&', ',-!3* 2#" Z. mays 0--2 2'113# 7 ST

&.'Q ,"0# !&#" + 6'+ **#4#*-$#6.0#11'-, 2VZ&.'$'%30#TTSYT7YT

&.'Q 2&# *#4#* -$ 2&# . 2&-%#, 5 1 0#"3!#" 2- *#11 2& , 2& 2 -$ ST &.'T <-

75 CHAPTER 2: Characterisation of resistance responses to Phytophthora cinnamomi

P. cinnamomi 5 1 "#2#!2#" ', ,7 -$ 2&# !-,20-* 1 +.*#1 5&'!& 5#0# +-!)

',-!3* 2#"5'2& ,#/3'4 *#,24-*3+#-$1"T>T

Control Inoculated 0 12 24 48 72 0 12 24 48 72

P. cinnamomi

rRNA

Figure 2.17. Accumulation of P. cinnamomi within inoculated Z. mays root tissue over 72 h. rRNA demonstrates equal loading of samples, representative of three repeats.

76 CHAPTER 2: Characterisation of resistance responses to Phytophthora cinnamomi

2.4. Discussion

(&'1123"7"#1!0' #12&#"#4#*-.+#,2-$ 1#+'V/3 ,2'2 2'4# 11#11+#,2Q

31',% 1.#!'$'! . 0 +#2#01Q 5&'!& #, *# "#2#0+', 2'-, ," !* 11'$'! 2'-, -$

.* ,20#1'12 ,!#-0131!#.2' '*'272- P. cinnamomi3,"#0!-,20-**#"!-,"'2'-,1T

(&# ..*'! 2'-, -$ 2&# . 0 +#2#01 -32*',#" ', 2&'1 5-0) & 1 *1- #12 *'1&#"

Z. mays 1 0#*' *#Q0#1'12 ,2+-"#*$-0.* ,2VP. cinnamomi',2#0 !2'-,1Q5&'!&

! ,2&#, #31#"2-#6 +',#)#7+#!& ,'1+1-$0#1'12 ,!# 22&#!#**3* 0 ,"

+-*#!3* 0*#4#*T3&'*12P. cinnamomi8--1.-0#1! ,13!!#11$3**7.#,#20 2#2&#

#.'"#0+'1-$Z. mays0--21Q2&#. 2&-%#,'10#120'!2#"2-*-! *'1#" 0# 1 22&#

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5'2&',2&#.* ,20--21 0#1'%,'$'! ,2*7 22#,3 2#"-4#02'+#Q5&'!&120-,%*7

13%%#121 , !2'4#"#$#,!#+#!& ,'1+2& 2'1',"3!#"5'2&',2�#1'12 ,2.* ,2T

2.4.1. Resistance responses at the macroscopic level

.#1'12 ,!#Q ', 2&'1 123"7Q 5 1 "#$',#" 1 2&# 1#,!# -$ "'1# 1#

17+.2-+1 ,"2&# !2'4 2'-,-$"#$#,!#0#1.-,1#15&'!&0#13*2', 0#"3!2'-,

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0#1.-,1#1',Z. mays, 11-!' 2#"5'2&0#1'12 ,!#2- P. cinnamomi, ',!*3"#" 32

0# ,-2 *'+'2#" 2-R 0 .'" * 2#0 * 0--2 $-0+ 2'-, ," 2&# .0-"3!2'-, -$ 2'%&2*7

!-,20-**#"Q 0#120'!2#" *#1'-,1 0-3," 2&# 1'2# -$ ',-!3* 2'-,T (&# 1#,!# -$

"#20'+#,2 * #$$#!21 -, 2&# .* ,2 .-12 ',-!3* 2'-, ',!*3"#1 !-,2',3#" 0--2

%0-52&Q ,- 1'%,'$'! ,2 "'$$#0#,!# ', 0#* 2'4# *# $ !&*-0-.&7** !-,2#,2Q ," ,-

0#"3!2'-,',.* ,2$0#1&5#'%&2T(#0#1'12 ,20#1.-,1#15#0#0#!-0"#"', **

Z. mays.* ,21-4#0 Y" 7.#0'-"$-**-5',%',-!3* 2'-,5'2&P. cinnamomi

77 CHAPTER 2: Characterisation of resistance responses to Phytophthora cinnamomi

8--1.-0#1 ', SR " 7 -*" .* ,21T '1# 1# 17+.2-+1 5#0# !* 11'$'#" 1 ,7

!& ,%#15&'!&!-+.0-+'1#.* ,2&# *2&-00#2 0".* ,2%0-52&Q0#13*2',%',2&#

*-11-$.&-2-17,2'! ,"0#.0-"3!2'4#! . '*'2'#1Q ,"2&##4#,23 *"# 2&-$

2&#.* ,2T

(.##"-$8--1.-0# %%0#% 2'-, ,"#,!712+#,2"'",-2 ..# 02-

"'$$#0 #25##, 0--21 -$ 2&# &'%&*7 0#1'12 ,2 Z. mays, -0 2&# &'%&*7 131!#.2' *#

L. angustifolius " 2 ,-21&-5,. (&'1'1!-,1'12#,25'2&.0#4'-31$',"',%1',

2�--21-$ 3120 *' ,, 2'4#.* ,21.#!'#1',!& ,"3#12#S[Y[T-5#4#0Q

2.##"-$ !23 *0--2.#,#20 2'-,+ 7"'$$#0 #25##,2-1.#!'#1T (&'1

+ 70#"3!#-00#2 0". 2&-%#,',%0#11 ,"!-,1#/3#,2*72&#*#4#*-$',-!3*3+

5'2&', 2&# 0--2 2'113#T 0302�+-0#Q 2&'1 5-3*" **-5 2&# .* ,2 2'+# 2-

0#!-%,'1# 2&# . 2&-%#, ," +-3,2 , #$$#!2'4# ," 0 .'" "#$#,!# 0#1.-,1#

3'2#+ et al. TRRUT

<-1'%,'$'! ,2"'$$#0#,!#5 1- 1#04#"+ !0-1!-.'! **7 #25##,!-,20-*

,"P. cinnamomi',-!3* 2#"Z. mays .* ,21 !0-11 SXZ&2'+#V.-',2Q5'2&2&#

#6!#.2'-, -$ 0#120'!2#" *#1'-,1 ', ',-!3* 2#" 0--21 -$ Z. maysT <- "'1# 1#

17+.2-+15#0#- 1#04#"', ,7-$2&#',-!3* 2#".* ,21 ,"0--2%0-52&5 1

,-2 0#"3!#"T (&'1 '1 ', !-,20 12 2- 2&# 0#1.-,1# -$ ',-!3* 2#"

L. angustifolius5&'!&',!*3"#"!#11 2'-,-$0--2%0-52& ,"0 .'"#6. ,1'-,-$

,#!0-2'! *#1'-,1T (�# '1 1-+# #4'"#,!# 2- 13%%#12 2& 2 0--2 %0-52& '1

12'+3* 2#" 7 . 2&-%#, ',$#!2'-, ', 0#1'12 ,2 1.#!'#1Q 5&'!&#$$#!2'4#*7 **-51

2&# .* ,2 2- -32%0-5 2&# . 2&-%#, &'**'.1 ," 3#12# S[ZVT -5#4#0Q 2&'1

"-#1 ,-2 1##+ 2- # 2&# ! 1# ', 2&# ',2#0 !2'-, #25##, Z. mays ,"

P. cinnamomi 12�#5 1,-1'%,'$'! ,2"'$$#0#,!# #25##,2�--2%0-52&-$

!-,20-* ,"',-!3* 2#".* ,21T

78 CHAPTER 2: Characterisation of resistance responses to Phytophthora cinnamomi

2.4.2. Formation of restricted lesions and lateral roots in Z. mays

7 TV &.'Q 0#120'!2#" *#1'-,1 $-0+#" "'0#!2*7 #,# 2& 2&# 1'2# -$

',-!3* 2'-,Q ," 2# 5#0#- 1#04#" 1 1#0'#1 -$ 2'%&2*7 !-,20-**#" ,#!0-2'!

$*#!)1T (&'1 0#1.-,1# '1 !& 0 !2#0'12'! -$ 27.# ** ,-,V&-12 0#1'12 ,!# 2-

--+7!#2#1Q 1.0-.-1#" 7#71-0# ,".73TRRVT3&'*#2&#.'1,-2

!-++-,*70#% 0"#" 1 #',%',4-*4#"',.* ,20#1.-,1#12-P. cinnamomi

) &'** et al. TRRZQ '2 & 1 ##, "#1!0' #" .0#4'-31*7 ', Acacia pulchella, ,

3120 *' ,, 2'4#.* ,25&'!&5 1!* 11'$'#" 10#1'12 ,22-2&#. 2&-%#,3,"#0

!-,20-**#"!-,"'2'-,1('..#22 ,"# * (!83)S[Y[R) &'**et al. S[Z[T *-5

*#4#* -$ .V*')# !#** "# 2& 5 1 *1- 0#.-02#" ', 0#1.-,1# 2- P. cinnamomi',

0 '"-.1'1*# $ 2'113#Q&-5#4#0Q'2"'",-2-!!30',0--2 2'113#.--)#1 et al.

TRRZT.#120'!2#"*#1'-,"#4#*-.+#,2 ,"2&#$-0+ 2'-,-$* 2#0 *0--21"'0#!2*7

-4#2'2#-$',-!3* 2'-,5#0#!-,1'12#,2*7- 1#04#"',',-!3* 2#" Z. maysT

(# 0#1.-,1#1 & 4# *1- ##, - 1#04#" ', 1#4#0 * 3120 *' , , 2'4# .* ,21

!* 11'$'#"#'2� 1$'#*"0#1'12 ,2-0$3**70#1'12 ,23,"#0!-,20-**#"!-,"'2'-,1

) &'**et al. S[Z[T

3&'*# . 2&-%#, 0#1'12 ,!# '1 -$2#, #/3 2#" 5'2& *#1'-, 0#120'!2'-,Q ,"

!!-+. ,7',% &7.#01#,1'2'4# !#** "# 2&Q 2&# !-,2',3#" #62#,1'-, -$ ,#!0-2'!

*#1'-,1 ," 13 1#/3#,2 2'113# !-** .1# '1 !& 0 !2#0'12'! -$ 131!#.2' '*'27

= +-3, et al. S[[[R ?*##1&-35#01 et al. TRRRR 3'2#+ et al. TRRUR >)3 0

," 3*'28TRRWT*,+ ,7',2#0 !2'-,15'2&0--2. 2&-%#,1Q2&#+# 130#+#,2

-$*#1'-,*#,%2&#6.0#11#" 1 .#0!#,2 %#-$0--2*#,%2&'131#" 1 +#2&-"

$-0/3 ,2'$7',%.* ,2"'1# 1#>_0'#, ,"4 ,03%%#,S[[TR?*##1&-35#01 et al. TRRRR? , 0*#,et al. TRRVT(&# '*'27Q-0', '*'27Q-$P. cinnamomi2-

0#.0-"3!# ," ',!0# 1# *#4#*1 -$ ',-!3*3+ 2&0-3%&-32 2&# .* ,2 '1 ,-2�

79 CHAPTER 2: Characterisation of resistance responses to Phytophthora cinnamomi

. 0 +#2#05&'!&& 1 ##,31#".0#4'-31*72- 11#110#1'12 ,!#-0131!#.2' '*'27

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. 2&-%#,5 1.0#1#,2', **1#%+#,21-$2�--2T

2.4.3. Northern blot analysis of P. cinnamomi inoculated Z. mays root tissue demonstrated a reduction in pathogen levels over time

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!-,20-**#"!-,"'2'-,1T

80 CHAPTER 2: Characterisation of resistance responses to Phytophthora cinnamomi

2.4.4. Chlorophyll content is not reduced in P. cinnamomi inoculated Z. mays leaf tissue

(�# 5 1 ,- 1'%,'$'! ,2 "'$$#0#,!# #25##, 2&# 0#* 2'4# !&*-0-.&7**

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81 CHAPTER 2: Characterisation of resistance responses to Phytophthora cinnamomi

#12 *'1&+#,2 -$ 1-*'" 0#1'12 ,2 +-"#*1 '1 0#/3'0#" 2- ',!0# 1# 2&#

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#6 +',',%.* ,215&'!&' '2203#Q"30 *#0#1'12 ,!#2-P. cinnamomiT

(&'1 123"7 & 1 ""0#11#" 2&# 1&-02$ ** ', 2&# 4 '* '*'27 -$ !-,1'12#,2

," 5#**V"#$',#" . 0 +#2#01 2- "#1!0' # .* ,2 ',2#0 !2'-,1 5'2& P. cinnamomi

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+-*#!3* 0 +#!& ,'1+1 -$ 0#1'12 ,!# 2- P. cinnamomiT Z. mays ,"

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82 CHAPTER 3: Identification of cellular defence responses to P. cinnamomi in Z. mays roots

Chapter 3: Identification of cellular defence responses to Phytophthora cinnamomi in Zea mays root tissue

3.1. Introduction

3&'*# 2&# ',"3!2'-, -$ 1#4#0 * !#**3* 0 !-+.-3,"1Q 13!& 1 .&#,-*'!1Q

*'%,', ,"! **-1#Q& 4# ##,'"#,2'$'#".0#4'-31*7',"#$#,!#2-Phytophthora cinnamomiQ2�#'1*'+'2#"!-,1#,131 12-2�-*# ,"'+.-02 ,!#-$# !&',

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-$),-5*#"%#-$0#1'12 ,!#+#!& ,'1+12-P. cinnamomiT

3.1.1. Plant cell walls

-2&2&# -4# ," #*-5%0-3,"',2#0, *-0% ,1-$.* ,21 0#.0-2#!2#"

$0-+ '-2'! ," '-2'! 120#11 7 $-02'$'#" !#** 5 **T 3'2&', 2&'1 1203!230 **7

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.* ,2 !#** 5 ** '1 &'%&*7 "7, +'! 1203!230#Q 5&'!& 0#1.-,"1 2- -2&

"#4#*-.+#,2 * ,"#,4'0-,+#,2 *!3#1 ,"3,"#0%-#11'%,'$'! ,2+-"'$'! 2'-,

1 0#13*2M+ 27et al. TRR[T(&#!#**5 **.-*7+#01!32', ,"13 #0', 0#

83 CHAPTER 3: Identification of cellular defence responses to P. cinnamomi in Z. mays roots

!-+.-1#" -$ &7"0-.&- '! .-*7#12#01 -$ $ 227 !'" "#0'4 2'4#1 2& 2 !-,20-* 2&#

',$*36 ,"#$$*36-$5 2#0 ,"1-*32#1*#',et al. TRRTT#1.'2#2&#'03 '/3'27Q

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3.1.2. Structure and function of plant root tissue

* ,20--210#.0#1#,22&#$'012*',#-$"#$#,!#',2&#.* ,2 1-'*',2#0$ !#

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84 CHAPTER 3: Identification of cellular defence responses to P. cinnamomi in Z. mays roots

-$0--21"'$$#01 #25##,#3"'!-21 ,"+-,-!-21 ,"! , 4 07 #25##,1.#!'#1

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a b

RC ZD ZE ZM

Figure 3.1. The anatomical structure of a newly emerging primary Z. mays root includes (a) a highly mucilaginous root cap (b) and overlapping zones of division with varying cell types. RC = root cap, ZD = zone of differentiation, ZE = zone of elongation, ZM = zone of maturation. Scale bar = 2.5 mm.

3.1.3. Cell wall fortification

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85 CHAPTER 3: Identification of cellular defence responses to P. cinnamomi in Z. mays roots

SQUVV%*3! , 2& 2 '1 17,2'1#" 7 .* ,21 ', 0#1.-,1# 2- '-2'! ," '-2'!

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3.1.4. The hypersensitive response (HR) and reactive oxygen species

(ROS)

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86 CHAPTER 3: Identification of cellular defence responses to P. cinnamomi in Z. mays roots

. 2&-%#,',%0#11T-2&2'+',% ,"2&##62#,2-$.! ,4 07 !!-0"',%2-

2&# , 230# -$ 2&# ',$#!2',% . 2&-%#, ," 2&# . 02'!3* 0 &-12 = +-3, et al.

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'+.-02 ,!#-$# !&',2�#1'12 ,!#0#1.-,1#T(#',!*3"#"!&#+'! *

',&' '2'-, -$ ! **-1# .0-"3!2'-,Q 2&# 31# -$ 1! 4#,%#0 2- 0#+-4# T>TQ ,"

,-02�, *-2 , *71'1-$*'%,',V0#* 2#"%#,#1 ," T>T0#* 2#"%#,#T

88 CHAPTER 3: Identification of cellular defence responses to P. cinnamomi in Z. mays roots

3.2. Materials and Methods

3.2.1 Z. mays growth and maintenance

##"1-$Z. maysYU ,"9 0*77# +',%5#0#%0-5,', 1-'*V$0##1712#+

1 "#1!0' #" ', 1#!2'-, TTTTUTQ ," + ',2 ',#" 5'2&', 2#+.#0 230#

!-,20-**#".* ,2%0-52&! ',#23,"#0U6XRRV5 22&'%&.0#1130#1-"'3+*'%&21

5'2& SX& Z&.&-2-.#0'-" 2TS5) 2TRR=+-*+VT1VS 1.0#4'-31*7"#1!0' #"

1#!2'-,TTTTST

3.2.2. P. cinnamomi maintenance and zoospore production

*1-* 2#1 -$ P. cinnamomi 5#0# + ',2 ',#" -, SR$ )?Z % 0 .* 2#1

..#,"'6TTT 1"#1!0' #"',1#!2'-,TTTTYTH--1.-0#15#0#.0-"3!#"

!!-0"',%2- +-"'$'#"+#2&-"-$2& 2-$702 ,"/0 ,2S[Y[ 1"#1!0' #"

.0#4'-31*7 1#!2'-, TTTTSRQ 2&# 1.-0# 131.#,1'-, 5 1 "'*32#" 2- $', *

!-,!#,20 2'-,-$S6SRW1.-0#1+7VS$-00--2',-!3* 2'-,T

3.2.3. Inoculation procedures

(&# 0--21 -$ SRV" 7 -*" 1##"*',%1 -$ Z. mays 5#0# ',-!3* 2#" 5'2&

P. cinnamomi8--1.-0#1 1"#1!0' #"',1#!2'-,TTTTSTTQ2- + 6'+3+-$SRR=7

.#0.* ,2T(�--21-$!-,20-*.* ,215#0#',-!3* 2#"5'2& ,'"#,2'! *4-*3+#

-$1"T>T +','+3+ -$ UR !-,20-* ," UR ',-!3* 2#" .* ,21 5#0# #6 +',#"

.#020# 2+#,2.#02'+#V.-',2 ,"2&0##',"#.#,"#,20#.# 21-$# !.#0'+#,2

5#0#!-,"3!2#"T

89 CHAPTER 3: Identification of cellular defence responses to P. cinnamomi in Z. mays roots

3.2.4. Detection of autofluorescence

)-,20-* ," ',-!3* 2#" 0--21 5#0# #6!'1#" 5'2& 1! *.#* 2 RQ VZQ STR

,"SXZ&.'Q ,"!32',2-2&',20 ,14#01#1#!2'-,1 7& ,"31',% 1',%*##"%#"

* "#T 9.'"#0+ * .##*1-$ !-,20-* ,"',-!3* 2#"0--21 2X ," TV&.'5#0#

2 )#,$0-+2'113#"'0#!2*7 #,# 2&2&#.-',21-$',-!3* 2'-,31',%$',#.-',2#"

$-0!#.1T .#!'+#,1 5#0# 4'#5#" 3,"#0 *'%&2 +'!0-1!-.# 6'-1)-. T +-2

.*31Q H#'11Q /ö22',%#,Q UYRURQ /#0+ ,7 ," .&-2-%0 .&#" 31',% ! +#0

+-3,2#" 2- 2&# +'!0-1!-.# >( .( *'"#0Q ' %,-12'! *,1203+#,21 *,!TT

*+ %#15#0# !/3'0#"31',%>(.(-$25 0#4UTRT 32-$*3-0#1!#,!#5 1

4'13 *'1#" 3,"#0 3*20 4'-*#2 #.'V$*3-0#1!#,!# UXW ,+ #6!'2 2'-,Q VTR ,+

#+'11'-,Q 6'-1)-.QH#'11Q##0!307 0!1-30!#Q>WR* +.T

3.2.5. Aniline blue staining for callose detection

) **-1# 5 1 "#2#!2#" 5'2&', 0--2 2'113# 7 12 ',',% 5'2& ,'*',# *3#

!!-0"',% 2- 2&# +#2&-" -$ -32-, ," #4#0 ** S[[RT ,'*',# *3#

#$$#!2'4#*7/3#,! !)%0-3," 32-$*3-0#1!#,!#! 31#" 72&#.0-"3!2'-,-$

.&#,-*'!.-*7+#01T.--21-$!-,20-* ,"',-!3* 2#".* ,215#0##6!'1#"5'2&

1! *.#* 2 RQ TVQ VZQ YTQ STR ," SXZ &.'Q !32 ',2- 2&', 20 ,14#01# 1#!2'-,1 7

& ," 31',% 1',%*#V#"%# * "# ," !*# 0#" -$ 1-*3 *# .'%+#,21 ', [R$ 4 4

#2& ,-* -4#0 TV &T #!2'-,1 5#0# 2&#, 20 ,1$#00#" ',2- S +7 -$ RTW$ 5 4

,'*',# *3# 3$$#0#"',RTSW#"'.-2 11'3+&7"0-%#,-02&-.&-1.& 2#=T>V

," 131.#,"#" -4#0,'%&2 2 0--+ 2#+.#0 230#.(T (K#0# 0',1#"25'!#

5'2&"T>.0'-02- #',%+-3,2#"-,2-%* 111*'"#1',SWRTRR=*WR$4 4

%*7!#0-*T .#!'+#,1 5#0# 4'#5#" 3,"#0 3*20 4'-*#2 #.'V$*3-0#1!#,!# ,"

90 CHAPTER 3: Identification of cellular defence responses to P. cinnamomi in Z. mays roots

.&-2-%0 .&#" 1.0#4'-31*7"#1!0' #"1#!2'-,UTTTVT(&#.0#1#,!#-$! **-1#

5'2&',0--22'113#5 14'13 *'1#" 1 0'%&27#**-5 5&'2#$*3-0#1!#,!#T

3.2.6. Phloroglucinol-HCl staining for detection of lignin

(&#.0#1#,!#-$*'%,',5'2&',0--22'113#5 1"#2#!2#"31',%2&#

.&*-0-%*3!',-* &7"0-!&*-0'! !'" )* 12 ', # 3!&V# ,' ," *31 0#,)-Q

S[[XT.--21-$!-,20-* ,"',-!3* 2#".* ,215#0##6!'1#"5'2& 1! *.#* 2RQ

TVQVZQYTQSTR ,"SXZ&.'Q!32',2-20 ,14#01#1#!2'-,1 7& ," ,"'++#01#"

',S+7-$S$5 4.&*-0-%*3!',-*',YR$4 4#2& ,-*-4#0,'%&2 2.(',2&#

" 0)T#!2'-,15#0#2&#,.* !#"-,%* 111*'"#1 ," "0-.-$)*5 1 ""#"2-

# !&1#!2'-, ,"*#$22-',!3 2#$-0W+',32#1T !-*-30!& ,%#5 14'1' *#

$2#0SVT+',32#1T96!#11)*5 1"0 ',#"$0-+2*'"#1 ,"2#!2'-,15#0#

2&#,+-3,2#"',WRR71"T>$-0*'%&2+'!0-1!-.7T*'"#15#0#4'#5#"5'2&',

TR+',32#1-$12 ',',% ,"2&#.0#1#,!#-$*'%,',5 14'13 *'1#" 1 0#" .30.*#

12 ',5'2&',!#**1T

3.2.7. 3’3’ diaminobenzidine staining for detection of hydrogen peroxide

7"0-%#, .#0-6'"# T>T5 1"#2#!2#"31',% U_U_"' +',- #,8'"',#

12 ',Q !!-0"',%2-2&#+#2&-"-$(&-0" *V)&0'12#,1#,et al. S[[YT.--21

-$!-,20-* ,"',-!3* 2#".* ,215#0##6!'1#"31',% 1! *.#* 2RQUQXQST ,"TV

&.'Q!32',2-20 ,14#01#1#!2'-,1 7& ," ,"20 ,1$#00#"',2- 1-*32'-,-$S+%

+7VS . UTW ," ',!3 2#" ', 2&# " 0) 2 .( $-0 U &T #!2'-,1 5#0#

20 ,1$#00#"2-%* 111*'"#1 ,"+-3,2#"',WRR71"T>$-0*'%&2+'!0-1!-.7T

T>T5 14'13 *'1#" 1 " 0)0#""'1&V 0-5,.0#!'.'2 2#5'2&',!#**1T

91 CHAPTER 3: Identification of cellular defence responses to P. cinnamomi in Z. mays roots

3.2.8. Quantification of lignin using thioglycolic acid derivitisation

*,-0"#02-$302� 11#112�-*#-$*'%,', !!3+3* 2'-,',Z. mays0--21

', 0#1.-,1# 2- P. cinnamomi ',-!3* 2'-,Q /3 ,2'$'! 2'-, -$ *'%,', 5'2&', 0--2

2'113# 5 1 !-,"3!2#" 31',% *'%,',V2&'-%*7!-*'! !'" 7(/ .0#!'.'2 2'-,

+#2&-" 7## et al. TRRSR #-&0 ," ) &'** TRRST .--2 2'113# -$ !-,20-* ,"

',-!3* 2#" .* ,215 1#6!'1#"5'2& 1! *.#* 2RQTVQVZQ YTQ STR ," SXZ&.'

,".* !#"',2-SR+7-$ 1-*32#+#2& ,-*T'%+#,2 2'-, ,"1-*3 *#.&#,-*'!

+ 2#0' *5 1!*# 0#"-4#0YT&5'2&U!& ,%#1-$ 1-*32#+#2& ,-*T('113#

5 1"0'#"$-0TV&', ,-4#, 2UR5) ,"2&#"075#'%&2"T525 10#!-0"#"T*2

5 1131.#,"#"',WRR7SR$4 42&'-%*7!-*'! !'"',T#&7"0-!&*-0'! !'"

4 4)* ,"&# 2#"$-0V&', 5 2#0 2& 2SRR5)T

(&# 131.#,1'-, !-,2 ',',% 2&# 2'113# 5 1 *#$2 2- !--* $-0 SR +',32#1

#$-0# #',%20 ,1$#00#"2- STW+7!#,20'$3%#23 #9..#,"-0$ ,"

!#,20'$3%#" 2SVRRR6%$-0SW+',32#1T( .#0, 2 ,25 1"'1! 0"#" ,"

2�#13*2 ,2.#**#25 15 1&#"5'2&WRR7-$1"T> ,"0#V!#,20'$3%#" 2SV

RRR6%$-0SR+',32#1T( .#0, 2 ,25 1 % ',"'1! 0"#" ,"2 1&#"

.#**#2131.#,"#"',WRR7S#1-"'3+&7"0-6'"#< > ,"4-02#6#"1#4#0 *

2'+#1-4#0T&2-1-*3 '*'1#2&#(/ "#0'4 2'4#1T(ƒ.#,1'-,5 1',!3 2#"

-4#0,'%&2 2V5) ,"!#,20'$3%#" 2SVRRR6%$-0SR+',32#1T( .#0, 2 ,2

5 1"#! ,2#"',2- STW+7!#,20'$3%#23 # ,"2&#.-*7+#0'!(/ "#0'4 2'4#1

5#0# .0#!'.'2 2#" 5'2& 2&# ""'2'-, -$ TRR 7 -$ !-,!#,20 2#" SR< )* ,"

!--*#"-, ,'!# 2&$-0S&T(�#""'1&V 0-5,.0#!'.'2 2#5 1!#,20'$3%#" 2

SVRRR6%$-0SR+',32#1 ,"2&#.#**#25 10#V"'11-*4#"',RTW#< >T(&#

1-0 ,!# -$2�#13*2 ,2-0 ,%#V!-*-30#"1-*32'-,5 1+# 130#" 2UUW

,+ ,"#6.0#11#" 1 UUW,++%VS"T52T

92 CHAPTER 3: Identification of cellular defence responses to P. cinnamomi in Z. mays roots

3.2.9. Inhibition of callose

) **-1# .0-"3!2'-, 5 1 ',&' '2#" ', 0--21 -$ P. cinnamomi',-!3* 2#"

2'113# 7 .0#V20# 2+#,2 -$ TV"#-67VV%*3!-1# TV/ 1-*32'-, 2

!-,!#,20 2'-,-$TWR=#(-, ,"# 3!&V# ,'TRRVTV&.0'-02-P. cinnamomi

',-!3* 2'-,T *,*-5*'%&2Q2&# 20 715#0#* '"$* 2 ,"2�--21-$ Z. mays

1##"*',%15#0#1.0 7#"5'2& 1-*32'-,-$TV/3,2'*1 230 2#"Q*#$2$-0S&2-

',!3 2# 2- #,130# 2&# 1-*32'-, 5 1 1-0 #" 7 2&# 0--21 #$-0# #',%

0#230,#"2-2#+.#0 230#!-,20-**#"%0-52&! ',#21T (-#,130#3.2 )#-$2&#

',&' '2-0Q 1+ **13 1#!2'-,-$',-!3* 2#".* ,20--215#0#1 +.*#" ,"12 ',#"

5'2& ,'*',# *3# 1"#1!0' #"',UTTTW ,"4'13 *'1#"3,"#0!?V#.'$*3-0#1!#,!#Q

,-! **-1#5 1- 1#04#"', ,7-$1#!2'-,1#6 +',#"Q5�# 1P. cinnamomi

',-!3* 2#" 0--21 ,-2 .0#V20# 2#" 5'2& TV/ #6&' '2#" !& 0 !2#0'12'! ! **-1#

!!3+3* 2'-,T

3.2.10. Inhibition of hydrogen peroxide (H2O2)

(&# .0-"3!2'-, -$ T>T 5 1 ',&' '2#" ', 0--21 -$

P. cinnamomi',-!3* 2#"2'113# 7.0#V20# 2+#,2-$7V 1!-0 '! !'" 2

!-,!#,20 2'-,-$SR+#TV&.0'-02-P. cinnamomi ',-!3* 2'-,Q !!-0"',%2-

+-"'$'#" 4#01'-, -$ 2&# .0-2-!-* 7 (&-0" *V)&0'12#,1#, et al. S[[YT (&#

1-*32'-,5 1 ..*'#"2-2�--22'113# 1 1.0 720# 2+#,2 1"#1!0' #"',

UTTT[T(-!-,$'0+2&##$$#!2'4#,#11-$2&# 1!-0 '! !'"20# 2+#,2Q 13 1#!2'-,

-$ 0--21 5#0# 1 +.*#" ," 12 ',#" 5'2& 1 "#1!0' #" ', UTTTYT ,"

4'13 *'1#"3,"#0*'%&2+'!0-1!-.7T<- V12 ',#"2'113#5 1"#2#!2#"', ,7

-$2&##6 +',#" 1!-0 '! !'"V20# 2#"0--21#!2'-,1T

93 CHAPTER 3: Identification of cellular defence responses to P. cinnamomi in Z. mays roots

3.2.11. Quantification of resistance responses following inhibitor treatment P. cinnamomi ',-!3* 2#" .* ,21 5&'!& & " ##, .0#V20# 2#" 5'2& 2&#

!&#+'! * ',&' '2-01 TV/ ," 7V 1!-0 '! !'" 5#0# 2&#, 11#11#" $-0

17+.2-+1 2 RQ TVQ YTQ STR ," SXZ &.'T .#1'12 ,!# -0 131!#.2' '*'27 5 1

11#11#" 72&# ..*'! 2'-,-$2&#W!0'2#0' -32*',#"',)& .2#0T-$2&'12'1R

5&'!& ',!*3"#" +# 130#+#,2 -$ .* ,2 $0#1& 5#'%&2Q 0--2 *#,%2&Q 0--2 *#1'-,

*#,%2&Q0#* 2'4#!&*-0-.&7**!-,2#,2-$2&#*# 4#1Q ,". 2&-%#,%0-52&',0--2

1#%+#,21.* 2#"-,2-PhytophthoraV1#*#!2'4#+#"' T* ,215#0#2&#, 11'%,#"

0#1'12 ,!# 0 2',% 1#" -, 2&# .0#4'-31*7 "#1!0' #" ,3+#0'! * SVW 1! *#

"#4#*-.#"',)& .2#0TT

3.2.12. Statistical analysis of data

0-0 /3 ,2'$'! 2'-, -$ *'%,',Q 2&# 4#0 %# -$ 2&0## ',"#.#,"#,2

#6.#0'+#,215 1! *!3* 2#" ,"2&#+# ,1$-0# !&2'+#V.-',25#0#!-+. 0#"

31',% 2V2#12T 2 2'12'! * , *71'15 1!-,"3!2#"31',%'%+ *-24#01'-,SS

712 2-$25 0#Q ,-1#Q) Q! T/0-3.12& 25#0#,-2,-0+ **7"'120' 32#"

5#0#13 (#!2#"2- # ,,V3&'2,#7. ,)3+(#12T

3.2.13. Tissue sampling for RNA extractions

.--2 2'113# $0-+ !-,20-* ," ',-!3* 2#" Z. mays 5 1 & 04#12#" 1

.0#4'-31*7"#1!0' #"',1#!2'-,TTTTTU 2RQUQXQ[QSTQTVQVZQ ,"YT&.'T

94 CHAPTER 3: Identification of cellular defence responses to P. cinnamomi in Z. mays roots

3.2.14. RNA extraction from Z. mays root tissue using TRIzol

.< 5 1#620 !2#"$0-+Z. mays0--22'113# 1.0#4'-31*7"#1!0' #"',

1#!2'-,TTTTTWT

3.2.15. Primer design and optimisation

< 1#/3#,!#1-$*'%,', ,"&7"0-%#,.#0-6'"#V0#* 2#"%#,#1',Z. mays

," $3** !< 1 5#0# - 2 ',#" $0-+ 2&# < 2'-, * )#,20# $-0 '-2#!&,-*-%7

*,$-0+ 2'-, <)*T0'+#0 1#21 2 *# UTST 5#0# "#1'%,#" 31',% 2&# -,*',#

.0-%0 + 0'+#0U 4#01'-, RTVTRT &22.S $0-"-T5'T+'2T#"3 .0'+#0UT0'+#01

5#0# 17,2'1#" 7 /#,#3-0)1 27 72" ',"+ 01&Q -32& 3120 *' Q WRRYR

&22.S 555T%#,#5-0)1T!-+T 3T

Primer name ID Oligonucleotide sequence (5’ – 3’)

Cinnamoyl CoA Reductase 1 CCoA1_F GTG GTG TTC ACG TCG TCC AT CCoA1_R GCC CTT CTC CTG GAG GTT CT Cinnamoyl CoA Reductase 2 CCoA2_F CAC CGA TGA CCC TGA GAT GA CCoA2_R CAT GCA TGT GTG GCA CTT GT Glutathione S-transferase1 Zm_GST1_F CAG CTC TGC AGG ATG GTG AC Zm_GST1_R TCC ATT CAC CGA TTG ACA GG

Table 3.1. Primer sets used in this study and their corresponding oligonucleotide sequences

!< 5 117,2'1#"$0-+!-,20-* ,"',-!3* 2#"Z. mays.< 1 +.*#1

1#!2'-,1 UTTTSUVUTTTSV 31',% 3.#01!0'.2 *** .#4#01# (0 ,1!0'.2 1# )'2

*,4'20-%#, !!-0"',% 2- + ,3$ !230#0_1 ',1203!2'-,1T S =% -$ .< 5 1 .0#V

20# 2#"5'2&SRK< 1#*0# !2'-, 3$$#0 ,"< 1#**,4'20-%#,Q) 2 *-%3#3

SZRXZVRSWT9 !&0# !2'-,!-,2 ',#"SS=7.< QRTW=70 ,"-+.0'+#01QS=7

"<(1SR+## !&RTW=79)20# 2#""T>Q2&'15 1&# 2#"$-0W+',32#1

95 CHAPTER 3: Identification of cellular defence responses to P. cinnamomi in Z. mays roots

2XW5)Q ,"13 1#/3#,2*7!&'**#"-,'!#$-0W+',32#1T(&#$-**-5',%5 1

""#"2-# !&23 #V=7WK$'012120 ," 3$$#0QS=7((QS=713.#01!0'.2*** ,"

S=7""T>Q ,"2 #',!3 2#" 2TW5)$-0W+',32#1QWR5)$-0VW+',32#1

,"YR5)$-0SW+',32#1T

0'+#015#0#-.2'+'1#"$-0*',# 00 ,%#31',%).Q31',%3,'4#01 *SZ

',2#0, * 12 ," 0"1 I3 ,2 +.< !,'4#01 * SZ ',2#0, * 12 ," 0"Q + '-,Q

! 2 *-%3# 3SYSZ#Q ,#% 2'4# !-,20-*1 5#0# 03, $-0 # !& ).T 9 !& TR =7

0# !2'-, !-,2 ',#" 2&# $-**-5',%S SR =7 /- ( / < -*7+#0 1# 0-+#% Q

! 2 *-%3#3#URRSQZ=7,3!*# 1#$0##5 2#0<03QS=7!< QS=7SZ

.0'+#0. '0T).5 103,31',%2&#$-**-5',%!-,"'2'-,1S*,'2' *"#, 230#[V5)

$-0U+',32#1Q"#, 230#[V5)$-0UR1#!-,"1Q ,,# *',%VZVXR5) 2UR1#!-,"1Q

#62#,1'-, YT5) $-0 #25##, UR 1#!-,"1 $-0 $0 %+#,21 WRR . ," XR

1#!-,"1$-0$0 %+#,21S) Q12#.1TVW5#0#2&#,0#.# 2#"$-0TZUW!7!*#1

"#.#,"',% -, 2&# 0#13*21 $0-+ 2&# *',# 0 0 ,%# ).T (- -.2'+'1# ,,# *',%

2#+.#0 230#1Q 2#+.#0 230#%0 "'#,2).5 1 *1-03,$-0 **.0'+#01 2VZ

XR5) 2T5)',2#04 *1Q31',% /0 "'#,22�+-!7!*#0#*2'#0(�+ *)7!*#0

()VTRRQ/0 "'#,2)7!*#0T

3.2.16. Northern hybridisation and analysis

**,-02�, , *71'1Q',!*3"',%.< %#*#*#!20-.&-0#1'1Q&7 0'"'1 2'-,Q

,3!*#'! !'"20 ,1$#0Q* #**',% ,"$'*+#6.-130#5#0#!-,"3!2#" 1.0#4'-31*7

"#1!0' #"',1#!2'-,1TTTTTWVTTTTUST

96 CHAPTER 3: Identification of cellular defence responses to P. cinnamomi in Z. mays roots

3.3. Results

3.3.1. Rapid lateral root formation viewed under UV-epifluorescence

7 2#0 * 0--2 $-0+ 2'-, - 1#04#" + !0-1!-.'! **7 ', ',-!3* 2#" 0--2

2'113# 2 VZ &.' 5 1 #4'"#,2 TV &.' ', 1YW$ -$ ** P. cinnamomi',-!3* 2#"

20 ,14#01#0--21#!2'-,15&#,4'#5#"3,"#0!?V#.'$*3-0#1!#,!#$'%30#UTTR,-

* 2#0 * 0--2 $-0+ 2'-, 5 1 - 1#04#" ', !-,20-* 0--21 2 2&# 1 +# 2'+# .-',2T

>,# 2- 25- * 2#0 * 0--2 .0'+-0"' 5#0# - 1#04#" #+#0%',% $0-+ #,"-"#0+ *

2'113#-$P. cinnamomi',-!3* 2#"Z. mays0--21 22&'12'+#V.-',2T

Figure 3.2. Growth of post-embryonic lateral root emerging from the endodermal tissue (arrowhead) in P. cinnamomi inoculated roots of Z. mays 24 hpi. Transverse section, unstained. Scale = 80 m.

3.3.2. Production of autofluorescent material in inoculated root tissue

32-$*3-0#1#,!# 5 1 - 1#04#"Q 1 0'%&2 5&'2# . *# 7#**-5 !-*-30Q

5'2&',2&#-32#0+-12* 7#0-$2&##.'"#0+ *2'113#-$!-,20-*+-!)',-!3* 2#"

5'2& 1"T> Z. mays0--21$'%30#UTU T *25 1 *1-"#2#!2#"5'2&',2&#

#.'"#0+'1-$P. cinnamomi',-!3* 2#"Z. mays0--21#!2'-,1 2X ,"TV&.' 2

97 CHAPTER 3: Identification of cellular defence responses to P. cinnamomi in Z. mays roots

+-0# ',2#,1# *#4#*T 2 X &.'Q !#** 5 ** 2&'!)#,',% 5 1 #4'"#,2 1

32-$*3-0#1!#,!# ', 2&# 5 **1 -$ #.'"#0+ * !#**1 ', P. cinnamomi',-!3* 2#"

Z. mays 0--21 "'0#!2*7 #,# 2& 2&# .-',2 -$ ',-!3* 2'-, $'%30# UTU T '1!0#2#

0# 1 -$ ',2#,1# 32-$*3-0#1!#,!#Q 11-!' 2#" 5'2& 22#+.2#" &7.& *

.#,#20 2'-,Q 1 5#** 1 !#** 5 ** 2&'!)#,',% 5#0# 4'13 *'1#" 2 TV &.' $'%30#

UTU!T

#','+ * !)%0-3," 32-$*3-0#1!#,!#Q-2& ,2& 2 11-!' 2#"5'2&

2&'!)#,#" #.'"#0+'1Q -!!300#" 5'2&', !-,20-* 2'113# !0-11 STR 2'+#V!-301#

$'%30#UTV T7TV&.'Q1-+#!#**5 **2&'!)#,',%5'2&',2 1!3* 02'113#-$

P. cinnamomi',-!3* 2#"Z. mays 0--21 5 1 4'1' *# &-5#4#0 2&'1 5 1 ,-2

11-!' 2#"5'2&"'0#!2&7.& *.#,#20 2'-,$'%30#UTV T 2YT&.'Q',2#,1#

32-$*3-0#1!#,!#Q "'0#!2*7 11-!' 2#" 5'2& 22#+.2#" &7.& * .#,#20 2'-, 5 1

4'1' *#2&0-3%&-322&##,"-"#0+'1 ,"12#*#Q',"'! 2',% &'%&*#4#*1-$.&#,-*'!

+ 2#0' * $'%30# UTV!T 7 STR &.'Q 1-+# &7.& * &# "1 5#0# 4'1' *# ', 2&#

#,"-"#0+ ** 7#0 32 32-$*3-0#1!#,!#5 10#120'!2#"2-.&*-#+.-*#1 ,"!#**

5 **15'2&',2#*#$'%30#UTV"T

98 CHAPTER 3: Identification of cellular defence responses to P. cinnamomi in Z. mays roots

a

b

c

hy

Figure 3.3. Autofluorescence in Z. mays roots (a) Suberised epidermis of control root tissue, transverse section, unstained (b) Cell wall thickening in epidermal tissue 6 hpi, directly beneath the point of inoculation, epidermal peel, unstained (c) Autofluorescence in epidermal cells in response to attempted hyphal penetration (hy) 24 hpi, epidermal peel, unstained. Scale bars: a = 50 m, b = 10 m, c = 100 m.

99 CHAPTER 3: Identification of cellular defence responses to P. cinnamomi in Z. mays roots

a a b

c d

Figure 3.4. Autofluorescence within Z. mays roots across 120 hpi (a) Control Z. mays root tissue with minimal background autofluorescence (b) P. cinnamomi inoculated Z. mays root tissue at 24 hpi (c) intense autofluorescence at 72 hpi, associated with hyphal penetration (white arrowheads) (d) 120 hpi, attempted hyphal penetration (white arrowheads) and cell wall thickening surrounding phloem poles (yellow arrowheads). Representative samples from 3 independent experiments, transverse sections, unstained. Scale bar = 100 m

3.3.3. Callose production in inoculated root tissue

) **-1#5 1"#2#!2#" 1 7#**-5V5&'2#$*3-0#1!#,!#',P. cinnamomi

',-!3* 2#"Z. mays0--22'113#12 ',#"5'2& ,'*',# *3#T(&0-3%&-322&#STR&

2'+#V!-301#Q $#',2 $*3-0#1!#,!# 5 1 #4'"#,2 5'2&', 2&# -32#0+-12 !#** * 7#0 -$

2&# #.'"#0+'1 -$ !-,20-* 2'113#Q &-5#4#0 ,- ! **-1# .0-"3!2'-, 5 1 "#2#!2#"

$'%30#UTW T00-+TV&.'Q! **-1#"#.-1'2'-,5 1- 1#04#"5'2&',',-!3* 2#"

0--2 2'113#Q 5'2& 2&# *#4#* -$ "#.-1'2'-, %0 "3 **7 ',!0# 1',% 7 YT &.' ,"

0# !&',% + 6'+ **#4#* 2STR&.'T

100 CHAPTER 3: Identification of cellular defence responses to P. cinnamomi in Z. mays roots

(&# *-! 2'-, -$ "#.-1'2'-, *1- 4 0'#" 2&0-3%&-32 2&# #6 +',#" 2'+#V

!-301#T *,'2' **7 2 TV VZ &.' 2&# + (-0'27-$ ! **-1#5 1 "#.-1'2#" ',2&#

#.'"#0+ * ,"!-02'! *0#%'-,1-$2�--21T00-+YT&.'-,5 0"1Q2&#%0# 2#12

"#.-1'2'-, 5 1 "#2#!2#" 5'2&', 2&# 4 1!3* 0 2'113#Q . 02'!3* 0*7 !0-11 1'#4#

.* 2#15'2&',2'#4#23 ##*#+#,21-$2&#.&*-#+Q ," 3,'$-0+$*3-0#1!#,!#

5 1- 1#04#"5'2&',2&#!-02#6 ,".'2&-$2&##,2'0#0--21#!2'-,$'%30#UTW T

2STR&.'Q+3*2'.*#! **-1#. .'** #1300-3,"#".&*-#+2'113# ,"2 **1-$

#,"-"#0+ *!#**1',0#1.-,1#2-&7.& *.#,#20 2'-,R'25 1 *1- 11-!' 2#"5'2&

, 0# -$*-! *'1#"!#**"# 2&$'%30#UTW!T

3.3.4. Lignin production in inoculated root tissue

7'%,',5 1"#2#!2#"5'2&',P. cinnamomi',-!3* 2#"Z. mays0--22'113#

12 ',#" 5'2& .&*-0-%*3!',-*V)* $0-+ YT &.'Q 5'2& 2&# + (-0'27 -$ "#.-1'2'-,

-!!300',% 5'2&', 2&# 4 1!3* 0 2'113#Q &-5#4#0Q ,- *'%,', !-3*" # "#2#!2#"

5'2&', !-,20-* 0--2 2'113# $'%30# UTX T *1-* 2#" *'%,', .*3%1 5#0# - 1#04#"

5'2&',4#11#*#*#+#,21-$67*#+2'113# 2YT&.'$'%30#UTX T7STR&.'Q2#

* 0%# '1-* 2#" .*3%1 -$ *'%,', !-3*" ,-2 # "#2#!2#" ," 2�#5 1 0#* 2'4#*7

3,'$-0+ "'120' 32'-, -$ 2&# .-*7+#0 2&0-3%&-32 2&# #,"-"#0+ * !#**1Q ,"

1300-3,"',%2'#4#23 #1-$.&*-#+2'113#$'%30#UTX!T

101 CHAPTER 3: Identification of cellular defence responses to P. cinnamomi in Z. mays roots

ab

c

Figure 3.5. Callose deposition in Z. mays root tissue viewed under UV-epifluorescence (a) Control (mock inoculated) root tissue, no callose was observed and autofluorescence has been effectively quenched by aniline blue (b) P. cinnamomi inoculated root tissue 72 hpi with callose plugs (yellow arrowheads) (c) P. cinnamomi inoculated root tissue 120 hpi, callose papillae (yellow arrowheads) surrounding localised cell death (white arrowhead). Representative samples from 3 independent experiments, transverse sections, stained with aniline blue. Scale bars: a & b = 100 m, c = 25 m.

102 CHAPTER 3: Identification of cellular defence responses to P. cinnamomi in Z. mays roots

a

b

c

Figure 3.6. Lignin production in Z. mays roots (a) No lignin deposition was observed in control root tissue. Lignin deposition was observed in P. cinnamomi inoculated Z. mays root tissue (b) Lignin within vessel elements of xylem tissue (square bracket) and a surrounding cell

(arrowhead) at 72 hpi. (c) Uniform lignin deposition surrounding phloem poles (arrowhead) and endodermal cell walls at 120 hpi. Representative of 3 independent experiments, transverse sections, phloroglucinol-HCl stained. Scale bars = 20 m.

103 CHAPTER 3: Identification of cellular defence responses to P. cinnamomi in Z. mays roots

3.3.5. Lignin quantification

I3 ,2'$'! 2'-, -$ *'%,', -4#0 2'+#V!-301# -$ SXZ & 0#4# *#" %0 "3 *

!!3+3* 2'-,-$2&#.-*7+#0-4#0YT&.'5'2&',!-,20-* ,"',-!3* 2#"Z. mays

0--22'113#Q$-**-5#" 7 "#!0# 1#$0-+STRVSXZ&.'$'%30#UTYT*,

P. cinnamomi',-!3* 2#"1 +.*#1Q*#4#*1-$*'%,', !!3+3* 2'-,0# !&#" .# )

2 YT &.'Q $2#0 2& 2 2'+#Q *#4#*1 -$ 2&# .-*7+#0 5#0#0#"3!#" 1'%,'$'! ,2*7T

1'+'* 0 . 22#0, 5 1 - 1#04#" ', !-,20-* 1 +.*#1Q &-5#4#0Q 2 VZ &Q *#4#*1

0#"3!#"1*'%&2*7 #$-0#0# !&',% .# ) 2YT&T7'%,',*#4#*10#!-0"#"1&-5#"

1'+'* 0 20#," 2- 2&# . 22#0, -$ *'%,', "#.-1'2'-, - 1#04#" ', ',-!3* 2#"

.&*-0-%*3!',-*V)* 12 ',#" 2'113# 5&#, 4'13 *'1#" 3,"#0 *'%&2 +'!0-1!-.7T

3&'*#2&#. 22#0,-$ !!3+3* 2'-,5 11'+'* 0', -2&!-,20-* ,"',-!3* 2#"

2'113#Q *#4#*1 5#0# 1'%,'$'! ,2*7 &'%� 0RTRW ', ',-!3* 2#" 2'113# 5&#,

!-+. 0#"2-!-,20-*1T7STR&.'Q*#4#*1-$2&#.-*7+#05#0#*-5#02& ,2&-1#

0#!-0"#" 2TV&.'Q2&'10#$*#!212�#"3!2'-,- 1#04#"3,"#0*'%&2+'!0-1!-.7

',.&*-0-%*3!',-*V)*V12 ',#"',-!3* 2#"2'113# 22 +#2'+#.-',2T

0.18 0.16 * Control 0.14 Inoculated 0.12 0.1 0.08 * * 0.06

A335nm mgA335nm -1 d. wt 0.04 0.02 0 24 48 72 120 168 Hours post inoculation

Figure 3.7. Lignin accumulation in control (sdH2O) and P. cinnamomi inoculated Z. mays root tissue over 168 hours. * denotes means which are statistically different to controls.

104 CHAPTER 3: Identification of cellular defence responses to P. cinnamomi in Z. mays roots

3.3.6. Hydrogen peroxide production in inoculated root tissue

(&# .0-"3!2'-, -$ &7"0-%#, .#0-6'"# T>T 5 1 4'13 *'1#" ',

P. cinnamomi',-!3* 2#"0--22'113#-$Z. mays 2UQXQTV ,"VZ&.'12 ',#"5'2&

T9 0*7.0-"3!2'-,UX&.'5 1!-,$',#"2-2&#',20 !#**3* 00#%'-,1-$

2&#!#**1-$2&##.'"#0+'1 ,"-32#0* 7#01-$2&#!-02#6Q!-,1'12#,25'2& 0# 1-$

22#+.2#" %#0+ 23 # .#,#20 2'-, $'%30# UTZ T 7 TV V VZ &.'Q T>T 5 1

"#2#!2#" 3,"#0 *'%&2 +'!0-1!-.7 5'2&', &'%&*7 *-! *'1#" 0# 1 -$ 2&# 4 1!3* 0

2'113#UTZ !T !!3+3* 2'-,5 1.0'+ 0'*7!-,$',#"2-'1-* 2#" 0# 15'2&',

2&# $'012 * 7#0 -$ !#**1 ',1'"# 2&# #,"-"#0+'1 ," 2- -,# -0 +-0# !#**1

1300-3,"',% .&*-#+ 2'113#Q 5&'!& 0#$*#!21 2&# . 22#0, -$ 22#+.2#"

P. cinnamomi&7.& *.#,#20 2'-,T

3.3.7. Localised cell death in inoculated root tissue at attempted penetration sites

7-! *'1#" !#** "# 2& 5 1 - 1#04#" ', 3,12 ',#" 0--2 1#!2'-,1 -$

P. cinnamomi',-!3* 2#"Z. maysQ YT &.' 5&#, 4'#5#" 3,"#0 !? #.'V

$*3-0#1!#,!#T'1!0#2#!*312#01-$ #25##, UVX"# "!#**15#0# #4'"#,25'2&',

2&# 4 1!3* 0 2'113#Q ," 2&'1 *-! *'1#" !#** "# 2& 5 1 "( !#,2 2- 120-,%*7

32-$*3-0#1!#,22'113#1300-3,"',%2&#.&*-#+.-*#1 15#** 1',2 **1-$

',"'4'"3 *!#**15'2&',2#*#$'%30#UT[ T!,"#0*'%&2+'!0-1!-.7Q 0# 1-$

!#** "# 2& 5#0# *1- 11-!' 2#" 5'2& ',2#0!#**3* 0 &7.& * .#,#20 2'-, $'%30#

UT[ T

105 CHAPTER 3: Identification of cellular defence responses to P. cinnamomi in Z. mays roots

a

b

c

Figure 3.8. Hydrogen peroxide (H2O2) accumulation was visualised as (a) slight staining at 6 hpi in inoculated root tissue (white arrowheads), epidermal peel (b) by 24 hpi (yellow arrowhead) and (c) 48 hpi (blue arrowheads) it was found in the phloem tissue of P. cinnamomi inoculated roots, transverse sections. Pictures representative of 3 independent experiments, all DAB stain ed. Scale bars: a = 10 m, b & c = 20 m.

106 CHAPTER 3: Identification of cellular defence responses to P. cinnamomi in Z. mays roots

a

b

Figure 3.9. (a) Localised cell death (white arrowhead) and intense autofluorescence at phloem poles (black arrowheads) in the vascular tissue of P. cinnamomi inoculated Z. mays roots at 72 hpi. (b) Areas of cell death (white arrowhead) and intercellular hyphal penetration (yellow arrowhead). Representative of 3 independent experiments, transverse sections, a – unstained, b – stained with 0.5% toluidine blue. Scale bar = 10 m.

3.3.8. Assessment of defence responses following inhibitor treatment

)&#+'! * .0#V20# 2+#,2 -$ Z. mays0--2131',%2&#! **-1#',&' '2-0TV

/ ,"2&#T>T ',&' '2-0 7V 1!-0 '! !'" TV & .0'-0 2- ',-!3* 2'-, "'" ,-2

0#"3!# 0#1'12 ,!# 2- P. cinnamomiT <- 1'%,'$'! ,2 "'$$#0#,!# 5 1 0#!-0"#"

#25##, 3,20# 2#" ," ( *# UTTQ !-*3+, T !&#+'! **7 20# 2#" ( *# UTTQ

!-*3+, V ',-!3* 2#" 0--2 2'113# -4#0 2&# #6 +',#" 2'+#V!-301#T 1#" -,

107 CHAPTER 3: Identification of cellular defence responses to P. cinnamomi in Z. mays roots

2# 0#13*21Q ** P. cinnamomi',-!3* 2#"Z. mays .* ,21Q -2& 20# 2#" ,"

3,20# 2#"Q5#0# 11'%,#" 0#1'12 ,!#0 2',%-$WT

Response post Z. mays Resistance Z. mays Resistance inoculation untreated rating treated rating Increased (no Increased (no Root length difference to 1 difference to 1 controls) controls) Lesion length (% of <10% 1 <10% 1 total root length) Increased (no Increased (no Plant fresh weight difference to 1 difference to 1 controls) controls) Growth on Minimal, Minimal, Phytophthora restricted to site 1 restricted to site 1 selective media of inoculation of inoculation 72 hpi Not reduced (no Not reduced (no Relative chlorophyll difference to 1 difference to 1 amount controls) controls) Final resistance Resistant 5 Resistant 5 assessment

Table 3.2. Resistance ratings at 168 hpi in P. cinnamomi inoculated Z. mays roots in untreated plants and plants treated with chemical inhibitors of callose and H2O2 24 h prior to inoculation.

3.3.9. Expression of lignin and H2O2 related genes in P. cinnamomi inoculated Z. mays root tissue

<-!*# 0',"3!2'-,-$2&#"#$#,!#V0#* 2#"%#,#1',Z. mays0--22'113#',

,7-$2&#P. cinnamomi',-!3* 2#"1 +.*#15 1- 1#04#" !0-112&##6 +',#"

2'+#V!-301#-$YT&T0-02&#*'%,',V0#* 2#"%#,#CCoA12�#5 1,-"'$$#0#,!#

#25##,!-,20-* ,"',-!3* 2#"1 +.*#1 2 !0-112&##6 +',#"2'+#!-301#Q$-0

))- TQ -,*7 *-5V*#4#* 0' -1-+ * ,"',% 5 1 .0#1#,2 !0-11 !-,20-* ,"

',-!3* 2#"1 +.*#1$'%30#UTSR T(&#-6'" 2'4# 3012+ 0)#0%#,# gst1

5 1 "#2#!2#" $0-+ R V VZ &.' ', ',-!3* 2#" Z. mays 0--2 2'113#Q &-5#4#0

#6.0#11'-,*#4#*10#+ ',#"0#* 2'4#*7!-,1'12#,25'2&!-,20-*1 +.*#1 !0-11 **

108 CHAPTER 3: Identification of cellular defence responses to P. cinnamomi in Z. mays roots

2'+#V.-',21T (&# %#,# 5 1 ,-2 "#2#!2#" 2 YT &.' ', ',-!3* 2#" 0--2 2'113#

$'%30#UTSR!T

a Control Inoculated 0 3 6 9 12 0 3 6 9 12

CCoA1

Control Inoculated 0 12 24 48 72 0 12 24 48 72

CCoA1

b Control Inoculated 0 3 6 9 12 0 3 6 9 12

CCoA2

Control Inoculated 0 12 24 48 72 0 12 24 48 72

CCoA2

c Control Inoculated 0 3 6 9 12 0 3 6 9 12 GST1

Control Inoculated 0 12 24 48 72 0 12 24 48 72 GST1

rRNA 0-12 h

rRNA 0-72 h

Figure 3.10. Gene expression of lignin and H2O2-related genes in control and P. cinnamomi inoculated Z. mays root tissue across a 72 h time-course (a) lignin-related CCoA1 (b) CCoA2 (c) gst1. rRNA demonstrates equal loading of samples, representative of two repeats.

109 CHAPTER 3: Identification of cellular defence responses to P. cinnamomi in Z. mays roots

3.4. Discussion

(&'1123"7',4#12'% 2#" ,3+ #0-$)#7!#**3* 0!-+.-,#,215'2&',2&#

0--21-$Z. mays$-**-5',%',-!3* 2'-,5'2& P. cinnamomi. (&'1#,!-+. 11#"2&#

',"'4'"3 *0-*#1-$!-+.-3,"1.0#4'-31*7'+.*'! 2#"',"#$#,!#2-2&#. 2&-%#,Q

',!*3"',% ! **-1#Q *'%,', ," 13 #0',Q 1 5#** 1 2&# *#11 5#** !& 0 !2#0'1#"

*-! *'1#"!#**"# 2&Q 32-$*3-0#1!#,!# ,"2&#.0-"3!2'-,-$T>TT

(&# $-0+ 2'-, -$ * 2#0 * 0--21 ', 0#1.-,1# 2- ',-!3* 2'-, 5'2&

P. cinnamomi.0#4'-31*7- 1#04#"+ !0-1!-.'! **7 2VZ&.'',)& .2#0T5 1

4'13 *'1#"+'!0-1!-.'! **7$0-+TV&.'',Z. maysT9+ 07-,'!0--21Q-0'%', 2',%

$0-+2&##,"-"#0+'1Q& " *0# "7#62#,"#"2-2&##.'"#0+'1 72&'12'+#T *,

131!#.2' *# 1.#!'#1Q !#11 2'-, -$ 0--2 %0-52& $-**-5',% . 2&-%#, ',$#!2'-, '1

!!-+. ,'#" 70#"3!#"! . !'272-2 )#3.,320'#,21 ,"5 2#0T(&# '*'272-

0 .'"*7-32%0-52&#. 2&-%#, 7* 7',%"-5,,#5* 2#0 *0--21Q5&'!&! ,2&#,

',!0# 1#2&# 1-0.2'-,-$,320'#,21 ,"5 2#0Q+ 7.* 7 )#70-*#',2&#

.0#4#,2'-,-$"'1# 1#! 31#" 7P. cinnamomiT

3.4.1. Deposition of autofluorescent material in P. cinnamomi inoculated

Z. mays root tissue

,-2�',"'! 2'-,-$ !2'4#"#$#,!#0#1.-,1#12-P. cinnamomi',

Z. mays 0--21 5 1 2&# "#.-1'2'-, -$ 32-$*3-0#1!#,2 + 2#0' * 1300-3,"',%

!-02'! *!#**1 ,"4 1!3* 02'113#T 32-$*3-0#1!#,!#'1-$2#,31#" 1 ,',"'! 2-0

-$4 0'-31.* ,2 "#$#,!#0#1.-,1#1Q',!*3"',%2&#.Q!#** 5 ** $-02'$'! 2'-,4'

*'%,', ,"13 #0',"#.-1'2'-, ,"2&#.0-"3!2'-,-$.&#,-*'!.-*7+#01Q ,"'2'1

*1-$-3,"', 0# 1 "( !#,22-!#**"# 2&?*##1&-35#01et al. TRRRR4 , 0*#, et al.TRRYT0-**-5',%P. cinnamomi',-!3* 2'-,Q 32-$*3-0#1!#,2+ 2#0' *5 1

110 CHAPTER 3: Identification of cellular defence responses to P. cinnamomi in Z. mays roots

"#2#!2#"',#.'"#0+ *2'113#Q"'0#!2*7 #*-52&#.-',2-$',-!3* 2'-,Q 2 -2&X

," TV &.'Q ',"'! 2',% 0 .'" 0#!-%,'2'-, ," 0#1.-,1# 2- 22#+.2#" . 2&-%#,

.#,#20 2'-, 4' !2'4# !#** 5 ** $-02'$'! 2'-,T 7 YT &.'Q + 11'4# +-3,21 -$

32-$*3-0#1!#,2+ 2#0' *5#0#"#.-1'2#"5'2&',2 1!3* 02'113# ,"!#**5 **

2&'!)#,',% 5 1 #4'"#,2 ', 2&# 1'#4# 23 # #*#+#,21 -$ 2&# .&*-#+T (&# 0#13*21

- 2 ',#"',2&'1123"7 0# !-,1'12#,2 5'2&2&-1#',-2�.* ,21.#!'#1 % ',12

-2� ,#!0-20-.&'! ," &#+'V '-20-.&'! . 2&-%#,1T (&# .0-"3!2'-, -$

32-$*3-0#1!#,2 + 2#0' * & 1 ##, 0#.-02#" ', "#$#,!# 2- ,#!0-20-.&'!

. 2&-%#,1 ," ', 0#1'12 ,!# 2- P. infestans = +-3, et al. S[[[T 2 1'2#1 -$

22#+.2#" .#,#20 2'-,Q 7 2&# ,#!0-20-.& Botrytis alliiQ 2&# "#.-1'2'-, -$

.&#,-*'! + 2#0' * 0#13*2#" ', "'12',!2 32-$*3-0#1!#,!#Q 5&#, 4'#5#" 3,"#0

!? #.'$*3-0#1!#,!# +'!0-1!-.7 #!731)7 et al. S[[[T 32-$*3-0#1!#,!# 5 1

*1-- 1#04#"',+#1-.&7** !#**1 "( !#,22- *-! *'1#"!#**"# 2&',-* , !# #

!*-,#15&'!&' '2#" -2&$3** ,". 02' *0#1'12 ,!#2-P. infestans

?*##1&-35#01et al. TRRRT

3.4.2. Hydrogen peroxide (H2O2) accumulation in P. cinnamomi inoculated

Z. mays root tissue

(&# !!3+3* 2'-, -$ &7"0-%#, .#0-6'"# T>T ', Z. mays0--21',

0#1.-,1#2-P. cinnamomi',-!3* 2'-,5 1- 1#04#" 2 -2&U&.' ,"X&.'',

#.'"#0+ * 2'113#Q "'0#!2*7 #,# 2& 2&# .-',2 -$ ',-!3* 2'-,T 7 TV ," VZ &.'Q

'1-* 2#" T>T .0-"3!2'-,5 1 #4'"#,2 5'2&',2 1!3* 02'113#T(-" 2#Q2&#

.0-"3!2'-, -$ T>TQ ', 0#1.-,1# 2- ',$#!2'-, 7 P. cinnamomi& 1,-2 ##,

"#1!0' #" ', 2&# 0--2 2'113# -$ 0#1'12 ,2 .* ,21T (&# !!3+3* 2'-, -$ 0# !2'4#

-67%#,1.#!'#1Q13!& 1T>T'1-$2#, 11-!' 2#"5'2&2&#.T.V*')#!#**"# 2&

111 CHAPTER 3: Identification of cellular defence responses to P. cinnamomi in Z. mays roots

5 1 "#+-,120 2#" 5'2&', 2&# 0--21 -$ 2&# 0#1'12 ,2 3120 *' , , 2'4# *#%3+#

Acacia pulchella ('..#22 ," # * (!83) S[Y[ ," 2�# '1 1-+# #4'"#,!# 2-

13%%#12 '2 -!!300#" ', 2&# 0--21 -$ 0#1'12 ,2 !*-,#1 -$ 2&# 20## 1.#!'#1

C. calophylla) &'** ,"#!)-+ S[[TT-5#4#0Q',P. cinnamomi',-!3* 2#"

0 '"-.1'1Q.V*')#!#**"# 2&Q ,"2&#.0-"3!2'-,-$T>T5 1,-2- 1#04#"',

0--22'113# 32-!!300#"',*# $2'113# .--)#1 et al. TRRZT *, 2&'1 123"7Q 2&#

.0-"3!2'-,-$T>T ..# 012- #!-00#* 2#"5'2&2&#-!!300#,!#-$.V*')#!#**

"# 2&',!#**11300-3,"',%. 2&-%#, 22 !)$'%30#UTZ!Q$'%30#UT[ T

*, %#,#0 * 2&# .0-"3!2'-, -$ 0# !2'4# -67%#, 1.#!'#1 .>Q ',!*3"',%

T>TQ'1 #*'#4#"2-'+.#"#!-*-,'1 2'-,-$.* ,22'113# 7 '-20-.&'!. 2&-%#,1

," 2- $ !'*'2 2# !-*-,'1 2'-, -$ ,#!0-20-.&'! . 2&-%#,1 /* 8# 0--) TRRWR

=*'# #,12#', ," .-5# TRRZT-5#4#0Q 1#4#0 * #6!#.2'-,1 2- 2&'1 03*# & 4#

##,"#+-,120 2#"', -2&&-12 ,",-,V&-120#1'12 ,!#T(&#.0-"3!2'-,-$.>

'10 .'"*7',"3!#"',0#1'12 ,2!3*2'4 01-$.-2 2-Solanum tuberosum$-**-5',%

',$#!2'-, 7 P. infestans ," 0#13*21 ', #$$#!2'4# !-,2 ',+#,2 -$ "'1# 1#

?*##1&-35#01 et al. TRRRT .#!#,2*7Q '2 5 1 1&-5, 2& 2 ', 2&# 1'1!'! !'"V

"#$'!'#,2 2-+ 2- +32 ,2 sitiensQ 0#1'12 ,!# 2- 2&# ,#!0-20-.&'! $3,%31 Botrytis cinerea',4-*4#"0 .'" !!3+3* 2'-,-$T>T',2&##.'"#0+ *!#**5 **-$*# 4#1

11#* #0%& et al. TRRYT(# 123"'#1 "#+-,120 2# 2&# '+.-02 ,!# -$ # 0*7

',"3!2'-,-$T>T',0#1.-,1#2-. 2&-%#, 22 !)Q13%%#12',%2& 22.##"-$

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-6'" 1#Q 0#13*2#" ', ',!0# 1#" 131!#.2' '*'27 11#* #0%& et al. TRRYT *, 2&#

2-+ 2- !3*2'4 0 Lycopersicon esculentum #'**TQ #6-%#,-31 ..*'! 2'-, -$ oV

112 CHAPTER 3: Identification of cellular defence responses to P. cinnamomi in Z. mays roots

7"0-67#2&7*-032',Q5&'!&!&#+'! **7',"3!#"2&#.0-"3!2'-,-$.>Q0#13*2#"

',&'%&*#4#*1-$0#1'12 ,!#2-B. cinerea # *-*#.18 ," !0 ,#) TRRTT

*,2#0#12',%*7 ', 2&'1 123"7Q 5&'*# T>T 5 1 11-!' 2#" 5'2& 22#+.2#"

P. cinnamomi 22 !)Q1! 4#,%',%-$T>T',Z. mays0--2131',% 1!-0 '! !'""'"

,-2 0#13*2 ', ',!0# 1#" *#4#*1 -$ 2&# . 2&-%#,Q -0 13 1#/3#,2 0#"3!2'-, -$

0#1'12 ,!#',2&#.* ,2T2 ',',%-$0--22'113#1#!2'-,15&'!&& " ##,20# 2#"

5'2& 1!-0 '! !'""'",-21&-5.0-"3!2'-,-$T>TQ',"'! 2',%2& 22&#',&' '2-0

5 112'**& 4',% ,#$$#!2" 2 ,-21&-5,T (&'1 5-0) *1- 13%%#121 2& 2Q ',

Z. maysQ*'%,','1,-2 0#/3'0#+#,2$-00#1'12 ,!#2-2&#. 2&-%#, 12&#$', *

12#. ', *'%,', '-17,2'1 '1 ! 2 *71#" 7 .#0-6'" 1#1 5&'!& 0#/3'0# T>T

? ,!#et al.,S[ZRT0302�',4#12'% 2'-,1',2-2�#* 2'-,1&'.-$T>T ,"2&#

.0-"3!2'-, -$ -2� !-+.-3,"1 ',4-*4#" ', !#** 5 ** $-02'$'! 2'-, 5-3*" #

4 *3 *#,#6212#.',2&'10#1# 0!&T

3.4.3. Callose production in P. cinnamomi inoculated Z. mays root tissue

) **-1#5 1"#2#!2#" 2 **#6 +',#"2'+#V.-',21',

P. cinnamomi',-!3* 2#"0--2112 ',#" 5'2& ,'*',# *3#T#.-1'2'-,-$! **-1#

. .'** # 5 1 #4'"#,2 2 .-',21 -$ 22#+.2#" &7.& * .#,#20 2'-, 5'2&', 2&#

4 1!3* 0 2'113# 2 -2& YT ," STR &.'T ,# 0*'#0123"7-$Z. mays 0--21

$-**-5',% P. cinnamomi ',-!3* 2'-, "#+-,120 2#" 2&# $-0+ 2'-, -$ ! **-1#

. .'** #TV&.'',0#1.-,1#2-2&#. 2&-%#,3#2� ##et al. S[ZVR',!&et al.

S[ZWT -5#4#0Q 2&'1 0#13*2 5 1 ,-2 ',4#12'% 2#" $302� -4#0 *-,%#0 2'+#V

!-301#Q,-',&' '2'-,123"'#15#0#!-,"3!2#" ,"2�#/3'0#+#,2$-0! **-1#',

0#1'12 ,!#2-2&#. 2&-%#,0#+ ',#"3,!*# 0T*,.0#4'-31123"'#1', 0 '"-.1'1

*# 4#1Q ! **-1# "#.-1'2'-, 5 1 - 1#04#" $0-+ ST &-301 .-12 ',-!3* 2'-, &.'

113 CHAPTER 3: Identification of cellular defence responses to P. cinnamomi in Z. mays roots

5'2&P. cinnamomi8--1.-0#1Q',!0# 1',%2-"#,1#. .'** # 7VZ&.'.- ',1-,

,") &'**TRRUQ.--)#1et al. TRRZT

-5#4#0Q',2&'1123"7Q',&' '2'-,-$! **-1#5'2&TV/"'",-20#13*2',

',!0# 1#" . 2&-%#, 131!#.2' '*'27 ', Z. maysQ 5'2& ** ',-!3* 2#" .* ,21 .0#V

20# 2#" 5'2& 2&# ',&' '2-0 1!-0',% 0#1'12 ,!# 0 2',% -$ WT (# 0#13*21 0#

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,#!0-20-.&'!. 2&-%#,1T *,"3!#"0#1'12 ,!#', 0 '"-.1'12- 2&# ,#!0-20-.&

Alternaria brassicicola 5 1 0#4#01#" $-**-5',% 20# 2+#,2 5'2& 2&# ! **-1#

',&' '2-0 TV"#-67VV%*3!-1# (-, ," # 3!&V# ,' TRRVT .#1'12 ,!# 2- 2&#

,#!0-20-.& Plectosphaerella cucumerina 5 1 *1- *-!)#" ', 2&# ! **-1#

"#$'!'#,2 +32 ,2 pmr4-1 (-, ," # 3!&V# ,' TRRVT -5#4#0Q .0#4'-31

123"7 ', *# $ ," 0--2 2'113# -$ P. cinnamomi ',-!3* 2#" 0 '"-.1'1 .* ,21

1&-5#"2& 2. 2&-%#,',$#!2'-,5 1,-2#,& ,!#"',pmr4.--)#1et al. TRRZT

.#!#,2 5-0) 13%%#121 2& 2 2&# "#.-1'2'-, -$ ! **-1# ', 0#1.-,1# 2- . 2&-%#,

',$#!2'-,'1&'%&*74 0' *#Q ,"! , # $$#!2#" 72&# +-3,2-$T>T5'2&',

.* ,22'113#73, et al.TRSST(&'1 ""1$302#'%&22-2&# 0%3+#,22& 2Q

5&'*#! **-1#+ 71#04# 1 !-+.-,#,2-$2�#1'12 ,!#0#1.-,1#2-

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0#1'12 ,!#2-2&#. 2&-%#,T

3.4.4. Lignin deposition in P. cinnamomi inoculated Z. mays root tissue

3&'*#*'%,',"#.-1'2'-,',!0# 1#"-4#02'+#',P. cinnamomi',-!3* 2#"

0--21Q2�#5 1,-3,'$-0+',!0# 1#-$2&#.-*7+#0',0#1.-,1#2-2&#. 2&-%#,

-4#0 2&'1 2'+# !-301#T *1-* 2#" .*3%1 -$ *'%,',Q '"#,2'$'#" 5'2&', 2&# 4 1!3* 0

2'113# -$ ',-!3* 2#" 0--21Q 5#0# ,-2 11-!' 2#" 5'2& 22#+.2#" &7.& *

114 CHAPTER 3: Identification of cellular defence responses to P. cinnamomi in Z. mays roots

.#,#20 2'-,T0302�',4#12'% 2'-,31',%2(/ '-!&#+'! * 11 72-/3 ,2'$7

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2-2 *"07+ 11-#0( ,et al. TRRUR * #01&#'+et al. TRSST(&'1+ 7 !!-3,2

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#$$#!2'4#*7!-,2 ',#"Q 1- 1#04#"',)& .2#0T1#!2'-,TTUTSR',P. cinnamomi

',-!3* 2#"0--21#!2'-,1T

3.4.5. Expression of defence-related genes in Z. mays root tissue following

P. cinnamomi inoculation

<-!*# 0',"3!2'-,-$2&#*'%,',V0#* 2#"%#,#1))- S-0))- TQ-02&#

-6'" 2'4# 3012V0#* 2#" %#,# gst15 1#4'"#,2',P. cinnamomi',-!3* 2#"

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2'113#',0#1.-,1#2-P. cinnamomi',-!3* 2'-,.--)#1et al.TRRZT({"7Q

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"3# 2- 2&# "'$$#0#,!# ', "#$#,!# 0#1.-,1#1 #25##, #3"'!-21 ," +-,-!-21T

115 CHAPTER 3: Identification of cellular defence responses to P. cinnamomi in Z. mays roots

-5#4#0Q 2# 0#13*21 0# ,-2 130.0'1',% %'4#, 2&# * !) -$ .0#4'-31 0#1# 0!&

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. 3!'27-$',$-0+ 2'-, 4 '* *#T

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2& 2!#**5 **$-02'$'! 2'-,',2&#$-0+-$! **-1#Q*'%,', ,".&#,-*'!!-+.-3,"1

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5&'!&'10#120'!2#"2-!#**11300-3,"',%&7.& *.#,#20 2'-,Q'1 *1- 1'%,'$'! ,2

!-+.-,#,2 -$ 2&# "#$#,!# 0#1.-,1#Q *-,% 5'2& 2&# # 0*7 .0-"3!2'-, -$ T>TT

3&'*# ,- 1',%*# !-+.-3," !-3*" # '"#,2'$'#" ', 2&'1 123"7 1 1.#!'$'! **7

!-,20-**',%2�#1'12 ,!#0#1.-,1#2-P. cinnamomiQ2&'1"-#1,-213%%#122& 2

2#!-+.-3,"1"-,-2.* 7 ,'+.-02 ,20-*#',"#$#,!#2-2&#. 2&-%#,T 2

2&#!#**3* 0*#4#*Q2&'1',"'! 2#12& 20#1'12 ,!#', Z. mays0--21-!!301 1 !-V

-0"', 2#""#$#,!#0#1.-,1#2-2&#. 2&-%#,5&'!&',4-*4#12'+#*7',"3!2'-,

-$ +3*2'.*# !-+.-3,"1T *2 & 1 *1- &'%&*'%&2#" 2&# 0#/3'0#+#,2 $-0 %#,-+#V

5'"# , *71'1',2-"#$#,!#0#1.-,1#12-2&#. 2&-%#,T

116 CHAPTER 4: Gene expression profiling of defence responses to P. cinnamomi

Chapter 4: Gene expression profiling of responses to Phytophthora cinnamomi in Zea mays root tissue

4.1. Introduction

*,)& .2#0TQ2&#Zea mays-Phytophthora cinnamomi . 2&-1712#+ 5 1

-.2'+'1#" ,"0#1'12 ,!#-$2&#+-"#*+-,-!-22-P. cinnamomi5 1#12 *'1&#"

31',% /3 ,2'2 2'4# 1! *#T (&'1 ','2' * #12 *'1&+#,2 -$ Z. mays 1 0#1'12 ,2

+-"#*5 1#11#,2' *2-#,130#2& 20#13*21- 2 ',#"',20 ,1!0'.2'-, *.0-$'*',%

5-3*" #0#.0#1#,2 2'4#-$ 203#0#1'12 ,!#0#1.-,1#2-2&#. 2&-%#,T

1 "'1!311#" ', )& .2#0 SQ --+7!#2# .* ,2 . 2&-%#,1 & 4# 0#!#'4#"

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*#4#* -, 2&# ',2#0 !2'-, #25##, &-12V1.#!'$'! Phytophthora1.#!'#113!& 1

P. infestansQ P. sojae ,"Q +-0# 0#!#,2*7Q P. brassicae (7*#0 TRRYT -5#4#0Q

2�#0#+ ',1 + (-0% .',2,"#012 ,"',%-$+-*#!3* 0.* ,20#1.-,1#12-

%#,#0 *'12 Phytophthora1.#!'#113!& 1P. cinnamomi.0302�+-0#Q5&'*#

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0#+ ', .--0*7 3,"#012--" 4 , 3#12 et al. TRRUR >)3 0 ," 3*'28 TRRWR a 8,'#51) et al. TRSRT (&# "#4#*-.+#,2 -$ 120 2#%'#1 2- .0-4'"# "30 *#

#,& ,!#"0#1'12 ,!#2-P. cinnamomi5'2&',131!#.2' *#1.#!'#1Q#1.#!' **72&-1#

"#1'%, 2#" 1 43*,#0 *# -0 #," ,%#0#"Q '1 "#.#,"#,2 -, , ',!0# 1#"

3,"#012 ,"',% -$ )#7 0#1'12 ,!# +#!& ,'1+1 ', 0#1.-,1# 2- . 2&-%#, 22 !)T

*"#,2'$'! 2'-,-$!-+.-3,"1-0. 2&5 712& 2!-,20-*0#1'12 ,!#!-3*"2&#,*# "

117 CHAPTER 4: Gene expression profiling of defence responses to P. cinnamomi

2- 2&# "#4#*-.+#,2 -$ #$$#!2'4# 20# 2+#,21 2& 2 5'** .0-4'"# *-,%V2#0+

.0-2#!2'-,-$131!#.2' *#1.#!'#1T

3120 *' , , 2'4#%0 11#1 0#%#,#0 **72&#$'012.* ,212- 0#!-*-,'1# ,

0# $-**-5',% P. cinnamomi',!301'-,) &'**et al.TRRZQ&-5#4#0Q%#,-+#

1#/3#,!',% -$ 13!& 1.#!'#1 & 1 ,-2 7#2 ##, 22#+.2#"T *2 5 1 2�#$-0#

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-2&%#,#1 ,"%#,#-0"#05'2&',2&#%0 11#1Q"#1.'2#4 0' 2'-,1',%#,-+#1'8#

," 2&# ,3+ #0 -$ !&0-+-1-+#1 3!)*#0 et al. TRRXR . ',-5'!8 ,"

#,,#28#,TRRXQ) ,"#* ," )#TRRZT0302�+-0#Q.0#*'+', 07123"'#1',

2&# 0--21 -$ 2&# 3120 *' , , 2'4# %0 11 Lomandra filiformis 0#4# *#" 1'+'* 0

+ !0-1!-.'! 0#1.-,1#1 2- P. cinnamomi',-!3* 2'-, 1Z. mays, ',!*3"',% 2&#

0 .'""#4#*-.+#,2-$0#120'!2#"*#1'-,1"'0#!2*7 #*-52&#.-',2-$',-!3* 2'-,

..#,"'6STRT

4.1.1. Z. mays as a model for molecular analysis of responses to P. cinnamomi 1.0#4'-31*7+#,2'-,#"QZ. mays'1 ,#+#0%',%+-"#*%0 111.#!'#1$-0

2&#$'#*"-$%#,-+'!1 ," 2TTX/ .Q'2 '12&#$'012%'% 1#V1'8#".* ,2%#,-+#

2- # 1#/3#,!#" . ',-5'!8 ," #,,#28#, TRRXR ) ,"#* ," )# TRRZT

#/3#,!',%!-++#,!#"',TRRWQ "0 $21#/3#,!#5 1!-+.*#2#"',0# 03 07

TRRZQ ," 2&'1 5 1 '+.0-4#" ', TRR[ ," TRSR 2- XK !-4#0 %# 0#13*2',% ',

&'%&V/3 *'27"0 $2%#,-+#1#/3#,!#!&, *#et al.TRR[R0#3'**#2et al.TRSST

,,-2 2'-,-$2&#%#,-+#'1!-,2',3',%5'2&2#!-," 11#+ *7!-,1'12',%-$

3." 2#" ,,-2 2'-,1 ,"5-0)',%%#,#1#21 &22.S 555T+ '8#1#/3#,!#T-0%T

118 CHAPTER 4: Gene expression profiling of defence responses to P. cinnamomi

(&# $$7+#20'6 /#,#)&'. + '8# %#,-+# 00 7 !300#,2*7 !-,2 ',1 SYQWWW

.0- #1#21 $-0 SVQZWR 20 ,1!0'.21 5&'!& 0#.0#1#,21 SUQUU[ %#,#1 ', 2&# + '8#

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$-0 +-*#!3* 0 "#$#,!# 0#1.-,1#1 -$ %0 11 1.#!'#1 5&'!& '1 0#1'12 ,2 2-

P. cinnamomiT

4.1.2. Genomics for plant-pathogen interactions

(#-$%#,-+'!2#!&,-*-%'#1Q',!*3"',%< +'!0- 00 71Q0#13*21',

,#$$#!2'4# ," !!30 2#+#2&-"-$%#,#0 2',% * 0%# +-3,2-$" 2 ', 1&-02

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*, TRRSQ "'1!*-130# ," !-++3,'! 2'-, 12 ," 0"1 $-0 +'!0- 00 7

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96.#0'+#,2#* #90 8+ et al.TRRSR. 7,#0et al.TRRXT',!#2& 22'+#Q

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2#!&,-*-%7Q. 02'!3* 0*731',%!-++#0!' * 00 71Q& 1 ##,$'0+*7#12 *'1&#" 1

119 CHAPTER 4: Gene expression profiling of defence responses to P. cinnamomi

, !!30 2#Q0#*' *# ,"&'%&*7#$$#!2'4#+-*#!3* 02--*5&'!&"-#1,-20#/3'0#

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120 CHAPTER 4: Gene expression profiling of defence responses to P. cinnamomi

3I + 0 et al. TRRZR ) &'** et al. TRRZT * ,21 !-+.0-+'1#" ',

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121 CHAPTER 4: Gene expression profiling of defence responses to P. cinnamomi

4.2.Materials and methods

4.2.1. Plant growth and maintenance

Zea mays 1##"1 5#0# %0-5, ', 1-'*V$0## .* ,2 %0-52& 1712#+

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4.2.2. Pathogen growth and maintenance

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4.2.3. Inoculation procedures

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122 CHAPTER 4: Gene expression profiling of defence responses to P. cinnamomi

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4.2.4. Experimental design

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123 CHAPTER 4: Gene expression profiling of defence responses to P. cinnamomi

each (n=15) 030309.CEL Af1c2003-15- Replicate 3 Replicate Roots excisedRoots pooled Tissue Z. mays RNA extracted

(n=15 ) (n=15 Labelled Labelled scanned 030309.CEL Hybridised GeneChip Af1c2003-11- Affymetrix Replicate 2 Replicate Quality check Purified RNA: RNA: Purified Inoculated Roots excised pooled Tissue RNA extracted Z. mays (n=15) 030309.CEL Replicate 1 Replicate Af1c2003-7- Roots excised pooled Tissue Z. mays RNA extracted illustrating steps the experiments independent of three 6 h Data normalised (RMA) (RMA) normalised Data Control6h_Inoculated6h.exe (n=15) Information in this flow chart is produced according microarray produced is to chart geneInformation in flow this 030309.CEL Replicate 3 Replicate Af1c2003-13- Roots excised pooled Tissue Z. mays RNA extracted (n=15) scanned Labelled Labelled Control GeneChip 030309.CEL Affymetrix Replicate 2 Replicate Af1c2003-9- Hybridised Roots excised pooled Tissue Quality check Purified RNA: RNA: Purified Z. mays RNA extracted (n=15) Replicate 1 Replicate 030309.CEL Af1c2003-5- Tissue pooled Tissue Roots excised Roots RNA extracted Z. mays Flow outlining chart experimentaldesign for microarray analysis, -point, only the 6 h time-point is shown here. here. shown is h time-point-point, only 6 the expression (MGED) data minimum society and information microarrayabout a experiment (MIAME) guidelines. Figure 4.1. for identical were steps All files. .exe as data normalised of generation the to plants experimental of treatment initial from time

124 CHAPTER 4: Gene expression profiling of defence responses to P. cinnamomi

4.2.5. Extraction and purification of total RNA

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125 CHAPTER 4: Gene expression profiling of defence responses to P. cinnamomi

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126 CHAPTER 4: Gene expression profiling of defence responses to P. cinnamomi

Figure 4.2. NanoDrop spectrophotometer scan showing a typical yield (ng/L) and purity (260/280) achieved for all samples of Z. mays root tissue used in microarray experiments.

Microarray analysis

4.2.7. Array processing – labelling, hybridisation and scanning

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127 CHAPTER 4: Gene expression profiling of defence responses to P. cinnamomi

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128 CHAPTER 4: Gene expression profiling of defence responses to P. cinnamomi

4.2.8. Bioinformatics analysis

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Figure 4.3. Volcano plot with genes arranged according to biological (fold-change) and statistical significance (log10-transformed p-value). Control samples are plotted against inoculated samples with down-regulation of control (¯2 -¯32) indicating up-regulation of inoculated samples. Differentially expressed genes with high biological and statistical significance are circled. Green arrow indicates the gene with the highest biological significance (fold-change) purple arrow indicates gene with the highest statistical significance (P-value).

129 CHAPTER 4: Gene expression profiling of defence responses to P. cinnamomi

4.2.9. Biological pathway analysis

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4.2.10. Data submission to public repository

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130 CHAPTER 4: Gene expression profiling of defence responses to P. cinnamomi

4.3. Results

4.3.1. Differential gene expression in Z. mays roots following inoculation with P. cinnamomi zoospores

(0 ,1!0'.2'-, *.0-$'*',%-$ Z. mays0--21Q ', 0#1.-,1# 2- ',$#!2'-, 5'2&

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131 CHAPTER 4: Gene expression profiling of defence responses to P. cinnamomi

$3,!2'-, **7 ,,-2 2#" %#,#1 2& 2 5#0# "-5,V0#%3* 2#" ', ',-!3* 2#" 0--21 2

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Biotic stress Oxidative burst

6.00% Protein/enzyme function Cell wall changes

23.00%

60.00% 11.00%

Figure 4.4. Functionally annotated differentially expressed genes which were identified in P. cinnamomi inoculated Z. mays roots across both time points (6 hpi and 24 hpi).

4.3.2. Early defence responses to P. cinnamomi initiated in Z. mays roots

(&# +-12 &'%&*7 "'$$#0#,2' **7 #6.0#11#" %#,#1 .0#1#,2 2 X &.' ',

P. cinnamomi ',-!3* 2#" Z. mays 0--2 2'113# 5#0# 2&# .32 2'4# 1#1/3'2#0.#,#

!7!* 1# umi3 H+TSVV[XTST S 2Q )#7 0 ,!& .-',2 #,87+# ', .&72- *#6',

'-17,2'1Q 5&'!& 5 1 3.V0#%3* 2#" SZV$-*"Q ," 2&# ) 30#,# 17,2& 1# an2

H+ $$6TSTTSTS 2Q5&'!&#,!-"#1 !* 11**2#0.#,#17,2& 1#Q5&'!&5 13.V

0#%3* 2#" SRV$-*"T ,3+ #0 -$ )#7 "#$#,!#V0#* 2#" %#,#1 5#0# *1- &'%&*7

"'$$#0#,2' 2#"1W$-*"',Z. mays0--21 2X&.'5'2&P. cinnamomi8--1.-0#1Q

',!*3"',%Q2-3,"V',"3!#"1#0',#.0-2# 1#',&' '2-03*SH+SZXTSTS 2Q

2&# 2& 3+ 2',V*')# . 2&-%#,#1'1V0#* 2#" .0-2#', W .W H+TXXW[TST S 2Q 2&#

%#0+',V*')# .0-2#', /7T H+TWRUTST S 2 ," , !'"'! !* 11 * !&'2', 1#

132 CHAPTER 4: Gene expression profiling of defence responses to P. cinnamomi uiu5(chnH+TZVYTSTS 22 *#VTSTT-2&/7T ,"3*S0#+ ',#"3.V

0#%3* 2#" 2TV&.'Q *2&-3%&2&#'0*#4#*-$#6.0#11'-,5 10#"3!#"2-UTV$-*"

,"UTU$-*"0#1.#!2'4#*72 *#VTTTT

4.3.3. Up-regulation of detoxifying glutathione S-transferase transcripts

>$ 2&# VU $3,!2'-, **7 ,,-2 2#" %#,#1 3.V0#%3* 2#" 2 X &.' ',

P. cinnamomi',-!3* 2#"Z. mays 0--21Q Y #,!-"#" %*32 2&'-,# V20 ,1$#0 1#

/( %#,#1Q 5&'!& 0# ',"3!#" 3,"#0 120#11 !-,"'2'-,1Q ',!*3"',% . 2&-%#,

22 !)Q ,"1#04#2-.0-2#!2.* ,2!#**1 ,"2'113#1$0-+-6'" 2'4#120#11! 31#"

7 2&# %#,#0 2'-, -$ 0# !2'4# -67%#, 1.#!'#1 # 001Q TRRVR /-,% et al. TRRWR

..*#2 *TTRR[T (#%#,#1',!*3"#"%st30 H+ZXVTSTS 25&'!&5 13.V

0#%3* 2#" XTZ $-*"Q gst8 H+TWU[TST S 2 3.V0#%3* 2#" UT[ $-*"Q gst24

H+TXTYTST S 2 3. UTX $-*", gst7 H+TTVXTST S 2, gst25 H+TWVWTSTS 2, gst23 H+TWXSTST S 2, ," gst15 H+TWVWTSTS 2 5&'!& 5#0# 3.V0#%3* 2#"

#25##,STXTTR$-*"2 *#VTST

4.3.4. Differential expression of JA and Et genes

#4#0 *%#,#1"'0#!2*7',4-*4#"',2&# '-17,2'1. 2&5 75#0# *1-

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17,2& 1#aos STX$-*" ,"*'.-67%#, 1#7>)WVTV[WSTW$-*"2 *#VTSTT ,

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2 *# VTS 5�# 1 2&# #2&7*#,#V0#1.-,1'4#V$ !2-0V*')# .0-2#', 9.0S

H+TSSVVSTSTS 25 1"-5,V0#%3* 2#" 7TV&.'VUTS$-*"2 *#VTVT

133 CHAPTER 4: Gene expression profiling of defence responses to P. cinnamomi

4.3.5. Induction of pathogen degrading enzymes, toxins and enzyme inhibitors

/#,#1#,!-"',% ,-2�'+.-02 ,2!* 11-$.0-2#',1Q. 2&-%#,',&' '2-01

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2X&.'Q ,"25-207.1',',&' '2-01Q7>)WVSXZZH+TUYWTSTS 2 2TTY$-*" ,"

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#$$#!21-,. 2&-%#,1 ,". 2&-%#,V"#0'4#".0-2#',1T

134 CHAPTER 4: Gene expression profiling of defence responses to P. cinnamomi

et et al.

(Yin (Yin Z. mays produced inproduced 1994) 2000) et al. et al. (Vick and Zimmerman 2005)

et al. 1997; Shen Annotation pointKey branch enzyme in phytoalexin (terpenoid) biosynthesis pathogen response to elicitation al. synthase terpene II class a Encodes (Harris inhibitorWound-induced serineprotease 2001) Gaut, and (Tiffen (Velazhahan protein Thaumatin-like 1999) and (Bernier resistance Stress/pathogen 2001)Berner Stress responses, including pathogen detoxification of xenobiotic attack, compounds (Marrs 1996) elicitor attack, pathogen response, Stress treatment (Wu JA - converts precursor linolenic to acid acid jasmonic 1986) identity100% sequence to in ethylene step final the Catalyses biosynthesis including responses, stress in Implicated attack, detoxification of pathogen compoundsxenobiotic (Marrs 1996) zeamatin-like protein zeamatin-like

Fold- Change 18.1 10.0 7.7 7.5 7.1 6.8 5.3 5.2 4.2 4.1 3.9

-5 Gene Symbol Umi3 an2 WIP1 PR GLP2 gst30 uiu5(chn) OPR1 LOC542299 ACO31 GST8

-5

-transferase 8 S-transferase 30

-like protein 2-like protein -oxo-phytodienoic reductase acid Gene Title putative sesquiterpene cyclase synthase2kaurene protein induced wound pathogenesis related protein germin glutathione chitinase I class acidic 12 hypothetical LOC542299 acc oxidase glutathione S

Probeset ID Probeset Zm.14496.1.A1_at ZmAffx.12.1.S1_at Zm.186.1.S1_at Zm.6659.1.A1_at Zm.503.1.A1_at Zm.864.1.S1_at Zm.847.1.S1_at Zm.10676.1.S1_at Zm.281.1.S1_s_at Zm.8714.1.A1_at Zm.539.1.A1_at

135 CHAPTER 4: Gene expression profiling of defence responses to P. cinnamomi

et al. 1992) et al. et

2002)

(Lunn 2003; Berger et al. -related protein erine - protease inhibitor inhibitor - protease erine Acidic chitinase Acidic S including responses, stress in Implicated attack, detoxification of pathogen compoundsxenobiotic (Marrs 1996) Defence function Unknown (serine-protease response Stress inhibitor) followingInduced stress stimuli, provides protection against oxidative stress (Considine protein wall Cell Heat shock protein biosynthesis in tryptophan Intermediate pathway production to of secondary (Wright metabolites catalyses stimuli, stress following Induced final pathway in step the of sucrose biosynthesis 2007) Terpenoid/isoprenoid biosynthesis (Kim 2010) including responses, stress in Implicated 3.9 3.7 3.6 3.5 2.9 2.8 2.7 2.4 2.4 2.2 2.2 2.1 2.0

chitinase LOC100037814 LOC541837 inra2(prp) IDP158 IDP370 aox3 LOC542268 hsp18a orp2 LOC542661 Hds1 gst7

18a diphosphate synthasediphosphate

- Birk serine protease inhibitor -

-transferase GST 24 -related protein -phosphatase -phosphatase

PRm 3 Bowmanputative glutathione S defence protein IDP158 serineputative type endopeptidase inhibitor alternative oxidase AOX3 precursor proteincell wall putative (put.); 18 kda heat shock protein orange pericarp2 sucrose 4 hydroxymethylbutenyl glutathione transferase7

Zm.1085.1.A1_a_at Zm.1085.1.A1_a_at Zm.4270.2.A1_a_at Zm.627.1.A1_at Zm.1506.1.A1_at Zm.4213.1.A1_at Zm.3332.1.A1_at Zm.6023.1.S1_at Zm.14951.1.S1_a_at Zm.311.1.A1_at Zm.4340.1.A1_at Zm.8470.1.S1_at Zm.10841.1.A1_at Zm.246.1.A1_at

136 CHAPTER 4: Gene expression profiling of defence responses to P. cinnamomi . 2007)

et al 2007) et al. 2007) 2008) et al. s to unstable s unstable to allene detoxification of et al. R), which inactivates HC secondary metabolites a precursor toa the

stress acclimation (Zdunek- . 2005; Berger Implicated in stress responses, including including responses, stress in Implicated attack, detoxification of pathogen compoundsxenobiotic (Marrs 1996) Catalyses first step in tryptophan biosynthesis, of production (Kriechbaumer enzyme Molybdenum-containing in involved and LipsZastocka 2003) stress Abiotic stress in implicated enzyme Iron-sulfur responses (Weidemann Methionine biosynthesis (Lyi including responses, stress in Implicated attack, pathogen compoundsxenobiotic (Marrs 1996) Stress-related. in Located apoplast, catalyses irreversible hydrolysis of glucose and sucrose (Chourey to fructose et al Encodes NADPH -dependent HC toxin reductase (HCT Briggs (Johal 1992). and toxin JA related. Conversion of lipoxygenase- derived acid fatty epoxides (supply precursors for JA 1.9 1.9 1.9 1.8 1.7 1.6 1.6 1.6 1.6 1.6

-2

LOC541838 TSA zmAO LOC100135422 APRL1 LOC541873 gst23 incw1 hm1 aos

-2 -methyltransferase-1 -methyltransferase-1 -transferase GST 25 -phosphosulfate reductase 1 reductase -phosphosulfate glutathione S synthase alpha tryptophan aldehyde oxidase induced drought proteinearly adenosine 5' homocysteine S glutathione transferase23 invertase1 wall cell Helminthosporium carbonum susceptibility1 synthase oxide allene

Zm.551.1.A1_at Zm.13645.1.S1_at Zm.9562.1.S1_at Zm.1449.2.S1_a_at Zm.12014.1.A1_at Zm.13424.1.A1_at Zm.561.1.A1_at Zm.409.1.S1_at Zm.27.1.S1_at Zm.6107.1.A1_at

137 CHAPTER 4: Gene expression profiling of defence responses to P. cinnamomi

2008) et al. 2002) et al.

activity Implicated in stressImplicated responses, including of detoxification attack, pathogen compounds 1996)xenobiotic (Marrs Terpene/isoprenoid biosynthesis (Carretero-Paulet kinase Protein CatalysesJA-related. the incorporation of molecular oxygen into free fatty (mainlyacids polyunsaturated linoleic linolenic and whose acid) acid areproducts fatty hydroperoxides, form oxylipins to used (Gao protein wall Cell 1.6 1.6 1.6 1.5 1.5 zoospores, arranged in descending order of fold change.

LOC542630 IDP154 ptk1 LOC542495 LOC542268 P. cinnamomi roots at 6 hpi with Z. mays Z. (put.); putative putative (put.);

-transferase GST 15

glutathione S dxr protein kinase1 protein Lipoxygenase protein wall cell

Annotated genes up-regulated inAnnotated Zm.545.1.S1_at Zm.298.1.S1_a_at Zm.168.1.A1_at Zm.445.1.S1_at Zm.14951.1.S1_x_at Table 4.1.

138 CHAPTER 4: Gene expression profiling of defence responses to P. cinnamomi

1999) et al. et

- Protease inhibitor inhibitor - Protease inhibitor - Protease

cidic chitinase Annotation Berner and (Bernier resistance Stress/pathogen 2001) Wound-induced serine inhibitor protease (Tiffen 2001) Gaut, and Serine Defence Serine acquisition Phosphate biotic in implicated inhibitor - protease Serine defence stress/pathogen (Chen A expansion wall Cell

3.4 3.3 2.7 2.3 2.2 2.1 2.0 1.9 1.9 Fold- Change zoospores

P. cinnamomi Gene Symbol Gene Symbol GLP2 WIP1 LOC541688 inra2(prp) sci1 LOC732717 LOC100037814 chitinase xet1

roots at 24 hpi with Z. mays Z.

Birk serine protease inhibitor -

endotransglycosylase homolog1 -chymotrypsin homolog1 inhibitor -related protein -like protein 2-like protein

Gene Title germin protein induced wound trypsin inhibitor defence subtilisin transporterinorganic 3 phosphate Bowmanputative PRm 3 xyloglucan

Annotated genes up-regulated in Annotated Probeset ID ID Probeset Zm.503.1.A1_at Zm.186.1.S1_at Zm.375.1.S1_at Zm.1506.1.A1_at Zm.301.1.A1_a_at Zm.1921.1.S1_at Zm.4270.2.A1_a_at Zm.1085.1.A1_a_at Zm.704.1.S1_at Table 4.2.

139 CHAPTER 4: Gene expression profiling of defence responses to P. cinnamomi

2007) et al. -responsive (Landoni (Landoni -responsive enlargement Annotation Annotation al. 2001) (Wu et loosening wall cell enlargement, Cell wall Cell Gibberellin zoospores zoospores Fold-Change Fold-Change 1.9 1.9 1.7 1.5 1.5 - - - - P. cinnamomi

Gene Symbol Gene Symbol gar1 LOC542341 an1 expB6 roots 6hpi withat Z. mays

Gene Title

-expansin 6-expansin beta responsive1 gibberellin Ribosomal S8 protein anther ear1

Annotated genes down-regulatedAnnotated in Probeset ID ID Probeset Zm.674.1.S1_at Zm.202.1.S1_at Zm.7012.8.S1_a_at Zm.228.1.S1_at Table 4.3.

140 CHAPTER 4: Gene expression profiling of defence responses to P. cinnamomi

, induced in response to reduced . 1996) Annotation respiration Cellular Terpenoid/isoprenoid synthesis respiration Cellular Photosynthesis Photosynthesis Photosynthesis Nitrogen metabolism/regulation Nitrogen metabolism/regulation Nitrogen metabolism Ethylene nitrite a siroheme, of synthesis in Involved reductase (Sakakibara nutrition plant in availability nitrate et al in hypoxic responseInduced to stress (Silva- 2003)al. etCardenas -1.6 -1.6 -1.6 -1.6 -1.7 -1.7 -2.5 -2.6 -2.6 -2.8 -3.1 -3.7 -5.0 Fold- Change zoospores zoospores

Gene

Symbol Symbol P. cinnamomi pgd3 TPS8 adh1 LOC542713 LOC542713 LOC542713 gpm108b nar gln1 ERF1 sum1 LOC541815

roots 24at hpi with methyltransferase1 Z. mays

Gene Title

-NADP reductase precursor -NADP reductase precursor -NADP reductase precursor -responsive protein factor-like 1 reductase (NAD(P)H) (NAD(P)H) reductase phosphogluconate dehydrogenase3 phosphogluconate Terpene synthase 8 alcohol dehydrogenase1 Ferredoxin Ferredoxin ferredoxin nitrite reductase nitrate glutamine synthetase1 ethylene siroheme uroporphyrinogen hemoglobin

_at

a_at Annotated genes down-regulated inAnnotated Probeset ID ID Probeset Zm.507.1.A1_at Zm.4909.1.A1_at Zm.1540.1.S1_a_at Zm.4068.1.S1_ Zm.4068.2.S1_a Zm.4068.2.S1_a_at Zm.13263.1.A1_at Zm.125.1.S1_at Zm.3369.1.A1_at Zm.11441.1.S1_at Zm.131.1.A1_at Zm.485.1.A1_at Table 4.4.

141 CHAPTER 4: Gene expression profiling of defence responses to P. cinnamomi

Gene title Gene symbol p-value Fold-change putative sesquiterpene cyclase Umi3 0.00275091 18.06 kaurene synthase2 an2 9.11E-05 9.97 wound induced protein WIP1 0.00128992 7.73 pathogenesis related protein-5 PR-5 0.00173186 7.51 germin-like protein 2 GLP2 0.00208655 7.12 glutathione transferase30 gst30 0.000208714 6.76 acidic class I chitinase uiu5(chn) 9.72E-05 5.28 12-oxo-phytodienoic acid reductase OPR1 0.0106663 5.15 hypothetical LOC542299 LOC542299 0.00150635 4.22 acc oxidase ACO31 0.00109082 4.10 glutathione S-transferase 8 GST8 0.00164418 3.91 PRm 3 chitinase 2.93E-06 3.86 putative Bowman-Birk serine protease inhibitor LOC100037814 0.000297671 3.68 glutathione S-transferase GST 24 LOC541837 0.000707703 3.63 defence-related protein inra2(prp) 0.001381 3.48 IDP158 protein IDP158 4.88E-06 2.91 putative serine type endopeptidase inhibitor IDP370 0.0190955 2.79 alternative oxidase AOX3 precursor aox3 0.00369429 2.73 cell wall protein (put.); putative LOC542268 0.0110077 2.45 18 kda heat shock protein18a hsp18a 0.00354126 2.39 orange pericarp2 orp2 0.000991058 2.23 sucrose-phosphatase LOC542661 0.00606355 2.18 hydroxymethylbutenyl 4-diphosphate synthase Hds1 0.000103437 2.14 glutathione transferase7 gst7 0.0305949 1.95 glutathione S-transferase GST 25 LOC541838 0.00767372 1.93 tryptophan synthase alpha TSA 0.000462491 1.91 aldehyde oxidase-2 zmAO-2 0.0394588 1.85 early drought induced protein LOC100135422 0.0490061 1.75 adenosine 5'-phosphosulfate reductase 1 APRL1 0.00881366 1.67 homocysteine S-methyltransferase-1 LOC541873 0.0476552 1.63 glutathione transferase23 gst23 0.000321693 1.61 cell wall invertase1 incw1 0.0205222 1.61 Helminthosporium carbonum susceptibility1 hm1 0.0038633 1.61 glutathione S-transferase GST 15 LOC542630 0.000432673 1.58 dxr protein IDP154 0.00252354 1.58 protein kinase1 ptk1 0.000323845 1.58 Lipoxygenase LOC542495 0.0402037 1.51 cell wall protein (put.); putative LOC542268 0.0344627 1.50 anther ear1 an1 0.0496119 -1.51 Ribosomal protein S8 LOC542341 0.0132247 -1.55 gibberellin responsive1 gar1 0.00289269 -1.74 beta-expansin 6 expB6 0.0301653 -1.91

Table 4.5. Functional groups of annotated genes differentially Stress (biotic/abiotic) expressed at 6 hpi. Note: Most of these genes are stress responsive, Oxidative burst however specific functions have been highlighted here as separate JA/Et biosynthesis categories. Protein/Enzyme function Cell Wall changes

142 CHAPTER 4: Gene expression profiling of defence responses to P. cinnamomi

Gene title Gene symbol p-value Fold-change germin-like protein 2 GLP2 0.0242253 3.38 wound induced protein WIP1 0.0217726 3.32 trypsin inhibitor LOC541688 0.023595 2.70

defence-related protein inra2(prp) 0.0118868 2.33 subtilisin-chymotrypsin inhibitor homolog1 sci1 0.0261308 2.24 inorganic phosphate transporter 3 LOC732717 0.0381141 2.07 putative Bowman-Birk serine protease inhibitor LOC100037814 0.0120057 2.00 PRm 3 chitinase 0.000648735 1.88 xyloglucan endotransglycosylase homolog1 xet1 0.0457709 1.86 phosphogluconate dehydrogenase3 pgd3 0.040183 -1.56 terpene synthase 8 TPS8 0.0300799 -1.61 alcohol dehydrogenase1 adh1 0.00906332 -1.64 ferredoxin-NADP reductase precursor LOC542713 0.0195773 -1.70 ferredoxin-NADP reductase precursor LOC542713 0.0125664 -1.71 ferredoxin-NADP reductase precursor LOC542713 0.00931129 -2.46 nitrite reductase gpm108b 0.0294941 -2.55 nitrate reductase (NAD(P)H) nar 0.0372076 -2.56 glutamine synthetase1 gln1 0.00607575 -2.80 ethylene-responsive factor-like protein 1 ERF1 0.0491673 -3.06 siroheme uroporphyrinogen methyltransferase1 sum1 0.0203032 -3.70 hemoglobin LOC541815 0.0135025 -4.96

Table 4.6. Functional groups of annotated genes differentially Stress (biotic/abiotic) expressed at 24 hpi. Primary metabolism Enzymes/Primary metabolism JA/Et related

4.3.6. MAPMAN biological pathway analysis optimised for Z. mays (0 ,1!0'.2'-, * !& ,%#1',!#**3* 0 $3,!2'-,1Q0#1.-,1#1 2- '-2'!120#11

,"&-0+-,# '-17,2'15#0#4'13 *'1#" 7'+.-02',% **-$2&# ,-0+ *'1#"

" 2 Q !-+.0'1',% ** USY "'$$#0#,2' **7 #6.0#11#" %#,#1 ',2- 2&# # # < 2--*

5&'!& 5 1 -.2'+'1#" $-0 Z. maysT # (-0 !#**3* 0 . 2&5 71Q '"#,2'$'#" 7

# # <-,2-*-%7 1 #',%1'%,'$'! ,2*73.V0#%3* 2#"',',-!3* 2#"0--22'113#

2 X &.'Q ',!*3"#" 20 ,1!0'.2'-, * 0#%3* 2'-,Q '-2'! ," '-2'! 120#11Q .<

17,2'1Q .0-2#', +-"'$'! 2'-, ," "#%0 " 2'-,Q 0#"-6 0# !2'-,1 ," #,87+#

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143 CHAPTER 4: Gene expression profiling of defence responses to P. cinnamomi

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92. 2&5 70#%3* 2'-,Q ,"/(V0#* 2#"%#,#1T

144 CHAPTER 4: Gene expression profiling of defence responses to P. cinnamomi

a

b

Figure 4.5. MAPMAN pathway analysis of Z. mays Affymetrix normalised data illustrating an overview of the genes involved in important cellular functions of the interaction of P. cinnamomi with Z. mays at a) 6 hpi and b) 24 hpi. Blue indicates up-regulated genes, red indicates down-regulated genes, deeper colour intensity indicates higher level of expression.

145 CHAPTER 4: Gene expression profiling of defence responses to P. cinnamomi

a

b

Figure 4.6. MAPMAN diagram of Z. mays Affymetrix genechip normalised data representing genes involved in biotic stress in Z. mays roots following inoculation with P. cinnamomi a) At 6 hpi b) 24 hpi. Blue indicates genes that are up-regulated; red indicates genes that are down-regulated.

146 CHAPTER 4: Gene expression profiling of defence responses to P. cinnamomi

4.4. Discussion

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147 CHAPTER 4: Gene expression profiling of defence responses to P. cinnamomi

0#1.-,1#-$Z. mays0--212-',-!3* 2'-,5'2&P. cinnamomiQ5'2&2&#+ (-0'27-$

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4.4.1. Early up-regulation of phytoalexins

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9 0*7 3.V0#%3* 2'-, X &.' ', ',-!3* 2#" Z. mays 0--21 ',"'! 2#1 2& 2

148 CHAPTER 4: Gene expression profiling of defence responses to P. cinnamomi

.&72- *#6', #6.0#11'-, '1 20 ,1!0'.2'-, **7 0#%3* 2#"Q ," 2&'1 + 7 # $302�

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149 CHAPTER 4: Gene expression profiling of defence responses to P. cinnamomi

.0-4'"#"2&#$'012#4'"#,!#2& 2 V0#* 2#"%#,#1 0##6.0#11#"',0--22'113#',

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150 CHAPTER4:GeneexpressionprofilingofdefenceresponsestoP.cinnamomi

9.0SQ5&'!&'1#6.0#11#""-5,120# +-$2&# 92 '-17,2'1 ,"1'%, **',%

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Signals(eg.elicitors)

phospholipids Receptor/s Lipase? Phospholipase? Linolenic acid LOX

13(S)hydroperoxylinolenic acid

AOS 12,13epoxylinolenicacid(unstable) AOC 12oxoPDA Defenceresponses 12oxoPDAreductase Geneexpression Jasmonic acid MeJA conjugates Figure4.7. SchematicrepresentationoftheJAbiosyntheticpathwayincludingthegenesup regulated in this study (blue boxes) at 6 hpi in P. cinnamomi inoculated Z. mays roots. Receptionofpathogenderivedsignals(eg.elicitors)onthemembranetriggerstherelease of linolenic acid via a lipase or phospholipase. Linolenic acid is then oxidised to 13 hydroperoxylinolenicacidviatheenzymelipoxygenase(LOX).Alleneoxidesynthase(AOS) converts 13hydroperoxylinolenic acid to an unstable epoxide 12,13epoxylinolenic acid whichrapidlybecomes12oxophytodienoicacid(12oxoPDA)anddefenceresponsesare initiatedhere.Theenzyme12oxoPDAreductaseisthefinalstepintheconversionof12 oxoPDA to JA which activates gene expression and defence responses. JA may then be furthercatabolisedtoitsmethylester(MeJA).Genesupregulated:LOX1.5fold,AOS1.6 fold, 12oxoPDAreductase 5.2 fold. AOC not differentially expressed (white box). Figure assembledfrom:CreelmanandMullet1997;Turner etal.2002;Wasternack2007

151 CHAPTER 4: Gene expression profiling of defence responses to P. cinnamomi

4.4.3. Rapid induction of glutathione S-transferases

(&#$ +'*7-$#,87+#1),-5, 1%*32 2&'-,#V20 ,1$#0 1#1/(1 0#

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152 CHAPTER 4: Gene expression profiling of defence responses to P. cinnamomi

%#0+',*')#.0-2#',/7T5 13.V0#%3* 2#" 2X&.'',P. cinnamomi

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4.4.4. Pathogenesis-related and thaumatin-like proteins

(5-2& 3+ 2',V*')#.0-2#',1(715#0#3.V0#%3* 2#"',P. cinnamomi

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154 CHAPTER 4: Gene expression profiling of defence responses to P. cinnamomi

S[[TR ) &'** ," #!)-+ S[[TT (&# #,87+# .&#,7* * ,',# ++-,' *7 1#

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Probeset ID Gene Title Fold Change Zm.64.1.A1_at cinnamyl alcohol dehydrogenase 1.05151 Zm.2550.1.S1_at cinnamoyl CoA reductase1 1.05827 Zm.86.1.A1_at cinnamoyl CoA reductase2 1.28396 Zm.15903.1.S1_at phenylalanine ammonia-lyase 1.03313 Table 4.7. Genes in the phenylpropanoid pathway which were not differentially expressed at either of the examined time-points.

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155 CHAPTER 4: Gene expression profiling of defence responses to P. cinnamomi

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156 CHAPTER4:GeneexpressionprofilingofdefenceresponsestoP.cinnamomi

sci1

compounds: WIP1,LOC100037814, gst25,gst23,gst15 ProductionofGSTs: Productionofserine IDP370,LOC541688, – proteaseinhibitors: uiu5(chn),prm3,hm1,GLP2 gst30,gst8,gst24,gst7, roots following inoculation with Synthesisofantimicrobial

Z. mays gulatedgenesareinblueboxes,downregulated

pgd3,adh1,nar roots

Z.mays metabolism: P.cinnamomi LOC542713,

Downregulationofprimary

LOC542495 aos, ,includingindividualgenesineachpathway.Upre (lox),ACO31

Overview of the major molecular pathways identified in Biosynthesisof PR5,LOC542299 ProductionofTLPs: OPR1, JA&Etbiosynthesis: Phytoalexins/terpenoids:

secondarymetabolites opr1,an2,hds1,dxr,TSA,orp2 P.cinnamomi genesareinredboxes. Figure 4.8.

157 CHAPTER 4: Gene expression profiling of defence responses to P. cinnamomi

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158 CHAPTER 5: General Discussion

Chapter 5: General Discussion

5.1. Summary

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159 CHAPTER 5: General Discussion

5.2. Characterisation of Z. mays as a resistant monocot model for examining defence mechanisms to P. cinnamomi under controlled conditions

Z. mays.0-4#" ,#6!#**#,20#1'12 ,2+-,-!-2+-"#*$-0.* ,2V

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5.3. Development and optimisation of an objective method to determine resistance to P. cinnamomi under controlled conditions

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160 CHAPTER 5: General Discussion

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161 CHAPTER 5: General Discussion

131!#.2' *# 1.#!'#1Q 5-3*" 1#04# 1 4 *3 *# + , %#+#,2 2--* $-0 , 2'4#

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5.4. Quantification of P. cinnamomi in Z. mays root tissue demonstrated a reduction in pathogen levels over time

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162 CHAPTER 5: General Discussion

5.5. Identification of individual cellular compounds produced in Z. mays root tissue following P. cinnamomi inoculation revealed a multi-faceted defence response

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163 CHAPTER 5: General Discussion

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164 CHAPTER 5: General Discussion

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165 CHAPTER 5: General Discussion

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5.6.1. A key role for the rapid generation of ROS and GSTs in Z. mays root tissue in defence to P. cinnamomi

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166 CHAPTER 5: General Discussion

(#0#13*21Q!-+ ',#"5'2&2�#13*21',2&'1123"7-$0#1.-,1#1',

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5.7. The important role of root tissue in defence responses to P. cinnamomi

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167 CHAPTER 5: General Discussion

2'113#2-',$#!2'-, 7,#!0-20-.&'!0--2. 2&-%#,1',2�#1'12 ,!#.0-!#11T

/'4#,2�#13*21',Z. mays0--21Q$302�%#,-+#V5'"#',4#12'% 2'-,1',2-2&#

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5.8. An increased understanding of defence mechanisms initiated in a resistant plant in response to P. cinnamomi infection

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168 CHAPTER 5: General Discussion

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169 CHAPTER5:GeneralDiscussion

Pathogeninfection:effectorproteins,germtubepenetration Plantrecognitionofnonselfsignals

0– 6hpi

Defencegeneexpression Localisedcelldeath,H2O2 6hpi

GSTs

Detoxificationofphytotoxicproducts

Defencegeneexpression Cellwallfortification reduced callose,phenolics 24 48hpi

Restrictedlesions lateralrootformation Continuedcellwall fortificationinvasculature Lignin,callose,phenolics

48+ hpi

Continuedplantgrowth,pathogenlevelsreduced

DiseaseResistance

Figure 5.1. Schematic representing the induction and approximate timing of defence responses identified in this research in Z. mays root tissue to P.cinnamomiinoculation.

P.cinnamomi(healthy) P.cinnamomi(dead)

170 CHAPTER 5: General Discussion

5.9. Final conclusions

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172 REFERENCES

References:

#02-,Q#Q3'*1-,Q Q) &'**Q#S[[[(#-$.-2 11'3+.&-1.&-, 2#2- !-,20-*Phytophthora cinnamomi',, 2'4#4#%#2 2'-, 2 ,%*#1# Q?'!2-0' Q Australasian Plant PathologyQTZQTTWVTUV #02-,Q#Q3'*1-,Q Q'**Q ,") &'**Q#TRRS#4#*-.+#,2-$"'1# 1# ! 31#" 7 Phytophthora cinnamomi ', + 230# Xanthorrhoea australisQ Australian Journal of BotanyQ49QTR[VTS[ 3I + 0Q Q )& 'Q #V0Q 73-Q Q -,%Q 0Q ##,%'12#Q ( TRRZ (-+ 2- .0-2#', )', 1#S +#"' 2#11'%, **',%-$.* ,20#1.-,1#12-,#!0-20-.&'!$3,%' ," ',1#!2� '4-07QThe Plant CellQ20QS[XVVS[ZU * Q.Q0#'QHQ 72-,QQ7'3Q1Q#--0#Q7Q# '#QQ)-&,QQ_ 1!#,8-Q#Q /-0"-,Q Q .-1#Q =)Q # 02',Q /Q ( ,)1*#7Q Q -38 7#,Q #Q &,Q ##Q /'-4 ,,-,'Q TRRV 9(1Q !< +'!0- 00 71Q ," %#,# #6.0#11'-, .0-$'*',%S2--*1$-0"'11#!2',%.* ,2.&71'-*-%7 ,""#4#*-.+#,2QThe Plant JournalQ39QX[YVYSV * #01&#'+QQ 04'**Q Q.- #021Q=Q#"#0-$$Q.Q2 #&#*',Q TRSS Plant Cell WallsQ/ 0* ,"!'#,!#Q( 7*-000 ,!'1/0-3.Q77)Q<#51-0)Q! QVUR. **'1-,QQ)3'QKQ %#Q/Q 0'.-30Q#TRRX#'!0- 00 7" 2 , *71'1S$0-+ "'1 00 72-!-,1-*'" 2'-, ," !-,1#,131Q Nature Reviews: GeneticsQ7QWWV XW 1'#% 3Q0>Q= !.08 )Q#Q ,'#*Q/Q-& ,11-,Q#Q2#,*'"QQ# ) Q#S[[[ '-!&#+'! * ',2#0 !2'-,1 -$ !-,'$#0 1##"*',% 0--2 5'2& Fusarium1..TQ Canadian Journal of MicrobiologyQ45Q[TUV[UW ,"#01#,Q . TRRV '-*-%7 ," 1712#+ 2'!1 -$ #0-)-,2 ," & .2-.&72# *% #QAmerican Journal of BotanyQ91QSWRZVSWTT ,"#01-,Q Q (& 2!�Q 70 ," ',%&Q = TRRW * ,2 "#$#,!# 0#1.-,1#1S !-,1#04 2'-, #25##,+-"#*1 ,"!0-.1QFunctional Plant BiologyQ32QTSV UV 11#* #0%&QQ)304#01Q=Q00 ,ç Q)Q 3"#, #02Q=Q?37*12#)#Q#Q? , 0#31#%#+Q0Qö$2#Q#TRRY.#1'12 ,!#2-Botrytis cinerea',sitiensQ , 1!'1'! !'"V"#$'!'#,2 2-+ 2- +32 ,2Q ',4-*4#1 2'+#*7 .0-"3!2'-, -$ &7"0-%#, .#0-6'"# ," !#** 5 ** +-"'$'! 2'-,1 ', 2&# #.'"#0+'1Q Plant PhysiologyQ144QSZXUVSZYY

173 REFERENCES

15 2'V< '0Q .Q ='0 ,Q /Q '4 )3+ 0Q =)Q (&-+ 1Q / TRSR #-*#!3* 0 !& 0 !2#0'1 2'-,-$ ,--+7!#2#V0#1.-,1'4#.0-2#',%#,#$0-+Zingiber zurumbet, Plant Molecular Biology ReporterQ28QSTZVSUW 8#4#"-QQ7',-V<#2-Q(Q( 4 0#1Q.#TRRZ(&# ,#!0-20-.& Botrytis cinerea ',"3!#1 ,-,V&-12 27.# ** 0#1'12 ,!# +#!& ,'1+ ', Pinus pinaster 131.#,1'-,!3*230#"!#**1QPlant Cell PhysiologyQ49QUZXVU[W !)Q =Q #Q Q ='+Q =!Q &',Q S[[Z )*-,',% ," !2#0' * #6.0#11'-, -$ 1#1/3'2#0.#,# !7!* 1#Q )#7 0 ,!& .-',2 #,87+# $-0 2&# 17,2'1 -$ 1#1/3'2#0.#,-'".&72- *#6',! .1'"'-*',!?V!& **#,%#"*# 4#1-$Capsicum annuumQPlant Cell PhysiologyQ39QZ[[V[RV #0%#0QQ',& Q =Q.-'21!&Q(TRRY* ,2.&71'-*-%7+##21.&72-. 2&-*-%7S .* ,2 .0'+ 07 +#2 -*'1+ ," .* ,2V. 2&-%#, ',2#0 !2'-,1Q Journal of Experimental BotanyQ58QVRS[VVRTX #0,'#0Q0 ,"#0, Q TRRS/#0+',1 ,"%#0+',V*')#.0-2#',1S.* ,2"-V ** .0-2#',1T325& 2"-2"-#6 !2*7[Plant Physiology and BiochemistryQ 39QWVWVWWV #00-! *V7- -Q #Q #-*', Q Q -* ,-Q . TRRT )-,12'232'4# #6.0#11'-, -$ Ethylene-Response-Factor1 ', 0 '"-.1'1 !-,$#01 0#1'12 ,!# 2- 1#4#0 * ,#!0-20-.&'!$3,%'QThe Plant JournalQ29QTUVUT *#',Q VQ )-32-1V(&M4#,-2Q Q # 0'-,Q Q -,!Q # TRRT 00-+ #*'!'2',1 2- *'.'" 20 ,1$#0 .0-2#',1S ,#5 ',1'%&2 ', !#** 1'%, **',% ',4-*4#" ', .* ,2 "#$#,!#+#!& ,'1+1QTrends in Plant ScienceQ7,T[UVT[X -#0( ,Q3Q. *.&QQ 3!�Q# TRRU7'%,', '-17,2'1Q Annual Review of Plant BiologyQ54QWS[VWVX 0 8+ Q Q',% +.QQI3 !)#, 31&QQ�*-!)Q/Q.#**+ ,QQ2-#!)& 02Q)Q !&QQ ,1-0%#Q3Q **Q) Q) 312-,Q)et al. TRRS #','+3+ ',$-0+ 2'-, -32 +'!0- 00 7 #6.#0'+#,2 #* #9V2-5 0" 12 ," 0"1 $-0+'!0- 00 7" 2 QNatureQ29QUXWVUYS 0-5,QS[[Y* ,2 2&-*-%7S ,>4#04'#5',Plant Pathogens and Plant DiseasesQ0-5,Q0 ,">%*#Q9"1Q.-!)4 *#3 *'! 2'-,1$-02&#'4'1'-, -$-2 ,7Q!,'4#01'27-$<#59,%* ,"Q<3QWWX.

174 REFERENCES

0-5,Q ," =# ,#Q S[[Y 11#11+#,2 -$ '1# 1# ," 9$$#!21 -, 1'#*" ', Plant Pathogens and Plant DiseasesQ0-5,Q0 ,">%*#Q9"1Q.-!)4 *# 3 *'! 2'-,1$-02&#'4'1'-,-$-2 ,7Q!,'4#01'27-$<#59,%* ,"Q<3Q WWX. 3!)*#0Q9Q/ 32QQ#!#3**#,Q#TRRX#-*#!3* 0 ,"$3,!2'-, *"'4#01'27 -$+ '8#QCurrent Opinion in Plant BiologyQ9QSYTVSYX 3#**Q ). TRR[ - !# # %#,-+#1S %-',% $0-+ 3, 22 ', *# 2- #!-+',% +-"#*!* "#$-0!-+. 0 2'4#%#,-+'!1QPlant PhysiologyQ149QSSSVSSX ü1!Q.Q-**0'!�Q=Q ,1203% Q.Q'+-,1Q/Q3-*2#0Q#Q00'(2#01Q Q4 , #*#,Q.Q4 ,"#07##Q(Q'#0% 0"#QQ/0-#,#,"'()QQ(ö.1!&QQ?-1QQ * +','Q0Q!&3*8#V7#$#02QS[[Y(&# 0*#7mlo%#,#S ,-4#*!-,20-* #*#+#,2-$.* ,2. 2&-%#,0#1'12 ,!#QCellQ88QX[WVYRW 702Q ,"/0 ,2Q.S[Y[-+#!-,"'2'-,1%-4#0,',%8--1.-0#.0-"3!2'-,', 6#,'! !3*230#1 -$ Phytophthora cinnamomi . ,"1Q Australian Journal of BotanyQ27QSRUVSSW 702Q< ,"-** ,"Q S[YZ*,$#!2'-,-$ 6#,'! Eucalyptus1##"*',%15'2& Phytophthora cinnamomi8--1.-0#1QAustralian Journal of BotanyQ26QSX[V SYX ) &'**Q# ,"3#12#Q/S[ZU0-0+ 2'-,-$! **-1#"#.-1'21 1 0#1.-,1#2- ',$#!2'-, 5'2& Phytophthora cinnamomi, Transactions of the British Mycological SocietyQ80QTUVT[ ) &'**Q#Q7#%%#Q

175 REFERENCES

) &'**Q #Q .--)#1Q 9Q 3'*1-,Q Q /' 1-,Q 7Q #!-3% **Q =7 TRRZ Phytophthora cinnamomi ," 3120 *' _1 '-"'4#01'27S'+. !21Q.0#"'!2'-,1 ,".0-%0#112-5 0"1!-,20-*QAustralian Journal of BotanyQ56QTY[VUSR ) ,"#* Q ," )#Q TRRZ (&# 02 ," "#1'%, -$ %#,#2'! 1!0##,1S + '8#Q Nature Reviews GeneticsQ9QS[TVTRU ) 00#2#0-V 3*#2Q7Q &3+ " Q*Q)3,'**#0 Q

176 REFERENCES

,%*Q 7 ," -,#1Q / TRRS * ,2 . 2&-%#,1 ," ',2#%0 2#" "#$#,!# 0#1.-,1#12-',$#!2'-,QNatureQ411QZTXVZUU ,'#*Q.Q3'*1-,Q Q) &'**Q#TRRUPhytophthora cinnamomi ', , 2'4# 4#%#2 2'-, !-++3,'2'#1 -$ 1-32�, ?'!2-0' +-0.&-*-%'! * 4 0' 2'-, ,". 0 %7,7 +-,%'1-* 2#1QAustralasian Plant PathologyQ32QVRUVVR[ "# 07Q SZYX.#1# 0!',2-2&#, 230#-$2&#.-2 2-$3,%31Phytophthora infestans, Journal of the Royal Agricultural Society of EnglandQ12QTU[VTX[ #. 02+#,2 -$ 312 ', '*'27Q 9,4'0-,+#,2Q 3 2#0Q -.3* 2'-, ," )-++3,'2'#1Q&22.S 555T#,4'0-,+#,2T%-4T 3 '6-,QQ7 .2&-0,Q Q9"5 0"1Q.TRRT* ,2%*32 2&'-,#20 ,1$#0 1#1Q Genome BiologyQ3QURRVTSVURRVTSR - 0-5-*1)'Q#Q(-++#03.Q*)Q&# 0#0Q7Q>_0'#,Q TRRU(&0##!*-, * *',# %#1 -$ Phytophthora cinnamomi ', 3120 *' 0#4# *#" 7 +'!0-1 2#**'2#1QPhytopathologyQ93QX[WVYRV -""1Q< ,". 2&(#,QTRSR* ,2'++3,'27S2-5 0"1 ,',2#%0 2#"4'#5 -$.* ,2V. 2&-%#,',2#0 !2'-,1QNature Reviews: GeneticsQ11QWU[VWVZ -#&*#+ ,,Q /Q 3 &*Q .Q -012Q .Q ?-**Q 7#Q !1 "#*Q Q -0##Q 0Q 2'22Q #Q =*&,#+ ,,QQ-,,#5 *"Q!Q= &+ ,,Q.Q==+.#0QTRRZ .#.0-%0 ++',% + '8#.* ,2S20 ,1!0'.2'-, * ,"+#2 -*'!!& ,%#1 ',"3!#" 7 2&# $3,% * '-20-.&1 Ustilago maydis, The Plant JournalQ 56Q SZSVS[W 0#,2& Q3 %#*1/Q+'2&Q#," **Q>_57#0)Q*04',#/Q ,"*05', / TRRX #4#*-.+#,2 -$ < V 1#" +#2&-" $-0 "#2#!2'-, ," '"#,2'$'! 2'-, -$ &72-.&2&-0 1.#!'#1Q Australasian Plant PathologyQ 35Q SVYVSW[ _-38 Q<=Q)-*/3&-3,Q*Q&# 0#0Q7Q 0"7Q/92TRRV(&#.-2#,2' *-$ $'4# 3#12#0, 3120 *' , , 2'4# Acacia 1.#!'#1 $-0 '-*-%'! * !-,20-* -$ Phytophthora cinnamomi, Australian Journal of BotanyQ52QTXYVTY[ _-38 Q<=Q)-*/3&-3,Q*Q&# 0#0Q7Q 0"7Q/92TRRW 11#11',%2&# .-2#,2' * $-0 '-*-%'! * !-,20-* -$ Phytophthora cinnamomi 7 $'$2##, , 2'4# 3#12#0, 3120 *' , ( 00 &V$-0#12 *#%3+# 1.#!'#1Q Australasian Plant PathologyQ34QWUUVWVR

177 REFERENCES

3,! ,Q# ,"=# ,#QS[[X?#%#2 2'-,!& ,%#1 11-!' 2#"5'2& Phytophthora cinnamomi ,"'21"#!*',#3,"#0Xanthorrhoea australis', =',%* )#< 2'-, * 0)Q?'!2-0' QAustralian Journal of BotanyQ44QUWWVUX[ 3,12 ,Q3 Q.3"+ ,Q(Q&# 0#0Q7Q#--0#Q< Q .Q(Q) *4#0Q#)Q 0+'12# "Q.Q- 0-5-*1)'Q#Q#-00'1-,QQ-5 0"Q=Q>_/ 0 Q9Q)0 ,#Q )Q #**Q Q >_0'#,Q Q #!)-+ Q Q 0"7 /92 TRRZ 90 "'! 2'-, -$ Phytophthora cinnamomi$0-+1.-2',$#!2'-,1',, 2'4#.* ,2!-++3,'2'#1 ',3#12#0, 3120 *' ,"( 1+ ,' Q)#,20#$-0&72-.&2&-0 !'#,!# ," # , %#+#,2$-02&# 3120 *' ,/-4#0,+#,2#. 02+#,2-$2&# 9,4'0-,+#,2Q3 2#0Q#0'2 %# ,"2&# 021T 300 ,2Q39 ,"-,%QKTRRV712#+'! !/3'0#"0#1'12 ,!#QAnnual Review of PhytopathologyQ42QSZWVTR[ 9"#,Q # Q '**Q . Q / *.-2&2& %# TRRR , #$$'!'#,2 '2',% 11 7 $-0 /3 ,2'$'! 2'-,-$Phytophthora cinnamomi',1-'*QPlant PathologyQ49QWSWV WTT 9,4'0-,+#,2 3120 *' TRRS(&0# 2 2#+#,2.* ,$-0"'# !)! 31#" 72&# 0--2V0-2 $3,%31 Phytophthora cinnamomi, 9,4'0-,+#,2 3120 *' Q ) , #00 QVR. 90 Q #Q 7#,)Q )Q #%#,& 0"2Q (30*',%1Q () TRR[ (&# 3,"#0#12'+ 2#" 0-*# -$ 0--21',"#$#,!# % ',12*# $ 22 !)#01QTrends in Plant ScienceQ14QXWUVXW[ 905',Q ) S[ZU Phytophthora: its biology, taxonomy, ecology and pathologyQ +#0'! ,&72-. 2&-*-%'! *-!'#27Q2 3*Q#',,#1-2 QU[T. 905',Q ) .' #'0-Q >= S[[X Phytophthora Diseases WorldwideQ (&# +#0'! ,&72-. 2&-*-%'! *-!'#27Q2T 3*Q#',,#1-2 QWXT. 0#3'**#2Q )Q 7# !&Q 9Q .-%#01Q Q !&, *#Q Q 94#01-*#Q = TRSS )0-. %#,-+# 1#/3#,!',%S*#11-,1 ,"0 2'-, *#1QTrends in Plant ScienceQ16QYYVZZ 0*-0Q S[WX (&# !-+.*#+#,2 07 %#,'! 1712#+1 ', $* 6 ," $* 6 0312Q Advances in GeneticsQ8QT[VWV 0*-0QS[YS)300#,212 231-$2&#%#,#V$-0V%#,#!-,!#.2QAnnual Review of PhytopathologyQ9QTYWVT[X

178 REFERENCES

0*-01Q?Q(-,QQ ) Q/Q# 3!&V# ,'QTRRW 1!'1'! !'" ,"! **-1#S2# + .* 7#01',"#$#,!# % ',12. 2&-%#,1[Journal of PhytopathologyQ153QUYYV UZU 00 ,)#Q. ,"!&0#' #0Q7TRRY3 #0', '-.-*7#12#0$-0+',% .-.* 12'! .* ,2',2#0$ !#1QCurrent Opinion in Plant BiologyQ10QTWTVTW[ 00'# *"#,&-4#,Q Q#2#0&ä,1#*Q)Q=302&QQ=0#38 *#0Q0Q7#$#02V!&3*28QS[[X *"#,2'$'! 2'-, -$ %#,#1 0#/3'0#" $-0 2&# $3,!2'-, -$ ,-,V0 !#V1.#!'$'! mlo 0#1'12 ,!#2-.-5"#07+'*"#5', 0*#7QThe Plant CellQ8QWVSV / -QKQ2 00QQ/ #*Q)Q9,%#* #02&QQ0#311,#0Q*Q(3+*',1-,QTRRZ# '8#[V *'.-67%#, 1# H+7-6U !-,20-*1 "#4#*-.+#,2Q 0--2V1.#!'$'! #6.0#11'-, -$ "#$#,!# %#,#1 ," 0#1'12 ,!# 2- 0--2V),-2 ,#+ 2-"#1Q Molecular Plant Microbe InteractionsQ21Q[ZVSR[ / 0) )*'1Q#Q) *4#0Q#)Q3'*1-,Q Q 0"7Q/92TRRVHabitat alteration caused by introduced plant disease: a significant threat to the conservation of Australian forest faunaQ',T73,,#7Q)-,1#04 2'-,-$ 3120 *' _1$-0#12 $ 3, Q .-7 * H--*-%'! * -!'#27 -$ <#5 -32& 3 *#1Q #-1+ ,Q <3Q ..T Z[[V[SU / 3,2Q .9 S[[W (&# 0#* 2'-,1&'. #25##, .* ,2 "'1# 1# 1#4#0'27 ," 7'#*"Q Annual Review of PhytopathologyQ33QSS[VSVV /#0 21QQ ))#0Q #Q4 ,7--,Q7)TRRT92&7*#,#',1#,1'2'4'27'+. '01 0#1'12 ,!# 2- 1-'* -0,# . 2&-%#,1 ', 2- !!- ," Arabidopsis thalian Q Molecular Plant Microbe InteractionsQ15QSRYZVSRZW /#0 21 Q ))#0Q #Q 7 50#,!#Q )Q !&3-Q 9 Q ö$2#Q #Q 4 , 7--,Q 7) TRRU 92&7*#,#V',1#,1'2'4# 2- !!- 1&-51 "'$$#0#,2' **7 *2#0#" 131!#.2' '*'272-"'$$#0#,2. 2&-%#,1QPhytopathologyQ93QZSUVZTS /* 8# 0--)QTRRW)-,20 12',%+#!& ,'1+1-$"#$#,1# % ',12 '-20-.&'! ," ,#!0-20-.&'!. 2&-%#,1QAnnual Review of PhytopathologyQ43QTRWVTTY /-3(-,Q(Q' -32Q.Q93"#1Q Q# != 7QQ-3 ,',Q7TRRU/#,#1',4-*4#"',2&# '-17,2'1-$*'%,',.0#!301-01',Arabidopsis thaliana, Plant Physiology and BiochemistryQ41QXYYVXZY /0*,5 *"Q <Q /-11Q 9#Q 0#11Q )# TRRZ 2&-%#, 0-$'*#S Phytophthora ramorumS . 2&-%#,5'2& 0#+ 0) *75'"#&-120 ,%#! 31',%13""#,

179 REFERENCES

- )"# 2&-,- )1 ,"0 +-03+ *'%&2-,5--"7-0, +#,2 *1QMolecular Plant PathologyQ9QYT[VYVR /3'** 3+'#QQ ,V)*#+#,2#QQ#15 02#Q)Q# 02',#8Q1Q7 .'#00#Q)Q#30'%,#36Q Q 00'L0#Q '!&-,Q #Q /-$$,#0Q TRRY # *H93 777T 2 1# ," "#4#*-.+#,2 * %#,# #6.0#11'-, .0-$'*',% -$ !#** 5 ** '-17,2'1 ," 11#+ *7',+ '8#QPlant PhysiologyQ143QUU[VUXU /3,,',%Q(= ,") &'**Q#TRR[ 1-'*V$0##.* ,2%0-52&1712#+2-$ !'*'2 2# , *71'1-$.* ,2V. 2&-%#,',2#0 !2'-,1',0--21QJournal of PhytopathologyQ 157QV[YVWRS 1QQ= +-3,QQH-"7Q#)Q' ,%Q.1Q ,"1 )#0Q.9Q) ,-Q7#Q /0 �Q#Q=-"'0 Q)Q. $$ #*#QQ(-02-V * *' -Q(#2 *TTRR[/#,-+# 1#/3#,!# ," , *71'1-$2&#*0'1&.-2 2- $ +',#. 2&-%#, Phytophthora infestans, NatureQ461QU[UVU[Z *1 **Q#S[ZT $-0#121-'*13..0#11'4#2-Phytophthora cinnamomi ," !-,"3!'4# 2- Phytophthora cryptogeaT*T304'4 *Q%#0+', 2'-, ," ',$#!2'4'27-$+7!#*'3+ ,"!&* +7"-1.-0#1QAustralian Journal of BotanyQ 30QSSVTW ++#01!&+'"2Q.S[[[&72- *#6',1S3& 2& 4#5#*# 0,#" $2#0XR7# 01Q Annual Review of PhytopathologyQ37QTZWVURX 0"& +Q . TRRW 2&-%#, 0-$'*#S Phytophthora cinnamomiQ Molecular Plant PathologyQ6QWZ[VXRV 0"& +Q . TRRY )#** '-*-%7 -$ .* ,2V--+7!#2# ',2#0 !2'-,1Q Cellular MicrobiologyQ9QUSVU[ 0"& +Q . ," * !)+ ,Q 7# TRSR #-*#!3* 0 !72-*-%7 -$ PhytophthoraV .* ,2',2#0 !2'-,1QAustralasian Plant PathologyQ39QT[VUW 0"7Q/2Q 00#22QQ&# 0#0Q7TRRS(&#$3230#-$.&-1.&'2# 1 $3,%'!'"#2-!-,20-*2-'* -0,#.* ,2. 2&-%#,Phytophthora cinnamomi ',, 230 *#!-1712#+1QAustralasian Plant PathologyQ30QSUUVSU[ 0+1Q=Q. +'0#8Q*Q# )-02M1QS[[Z*,&' '2'-,-$5-3,"V',"3!#" !!3+3* 2'-,-$ **#,#-6'"#17,2& 1#20 ,1!0'.21',$* 6*# 4#1 7 1.'0', ,"1 *'!7*'! !'"QPlant PhysiologyQ118QSRWYVSRXW

180 REFERENCES

00'1Q 7Q . 0,-Q Q -&,12-,Q Q 0'1'!Q Q K3Q #Q ** 0"Q Q = 2&'0#1 ,Q Q >3#**#2Q(Q#2#0Q.TRRW(&#+ '8#an2 %#,# '1 ',"3!#" 7 Fusarium 22 !) ,"#,!-"#1 ,entV!-. *7*"'.&-1.& 2#17,2& 1#QPlant Molecular BiologyQ59QZZSVZ[V 023,%Q= ,"'#.&-QVTRRY 0#-0"', *0 2',%1! *#1 #22#02& ,.#0!#,2 0 2',%1[ 12 2'12'! * ,"b.17!&-*-%'! *c4'#5QEuphyticaQ155QSWVTX # 2&Q #) TRRR <-,&-12 0#1'12 ,!# ," ,-,1.#!'$'! .* ,2 "#$#,1#1Q Current Opinion in Plant BiologyQ3QUSWVUS[ M+ 27Q =Q )�)Q )Q -+#04'**#Q TRR[ -12V. 2&-%#, 5 0$ 0# 2 2&# .* ,2 !#**5 **QCurrent Opinion in Plant BiologyQ12QVRXVVSU ',!&Q ,"3#12#Q/S[Y[#& 4'-30-$Phytophthora cinnamomi8--1.-0#1 -,0--21-$ 3120 *' ,$-0#121.#!'#1QAustralian Journal of BotanyQ27QXY[V X[S ',!&Q# ,")* 0)#Q 9S[ZT) **-1#$-0+ 2'-,',Zea mays 1 0#1.-,1#2- ',$#!2'-,5'2&Phytophthora cinnamomi, Physiological Plant PathologyQ21Q SSUVSTV ',!&Q#Q3#2� ##Q.Q# **#22Q9Q)* 0)#Q 9S[ZW.#1.-,1#-$Zea mays 0--21 2- ',$#!2'-, 5'2& Phytophthora cinnamomiQ ProtoplasmaQ 126Q SYZV SZY -!)&-*"',%#0Q0Q3-**Q=Q 3#0Q#Q#+ ',1)7QTRRV/#,#2'!"'11#!2'-,-$ 0--2 $-0+ 2'-, ', + '8# Zea mays 0#4# *1 0--2V27.# 1.#!'$'! "#4#*-.+#,2 *.0-%0 ++#1QAnnals of BotanyQ93QUW[VUXZ -!)&-*"',%#0Q 0 ," H'++#0+ ,,Q . TRRZ )-,1#04#" ," "'4#01# +#!& ,'1+1',0--2"#4#*-.+#,2QCurrent Opinion in Plant BiologyQ11QYRV YV -02 Q#Q-31 Q

181 REFERENCES

* #0*'QQ(-++#03.Q*)Q- 0-5-*1)'Q#Q) *4#0Q#)Q 0"7Q/92TRRS &#,-27.'!4 0' 2'-,', !*-, **',# %#-$25-Phytophthora cinnamomi .-.3* 2'-,1 $0-+ 3#12#0, 3120 *' Q Mycological ResearchQ 105Q SRWUV SRXV * #0*'Q Q (-++#03.Q *)Q )-*/3&-3,Q *Q 0"7Q /9 2 TRRT 94 *3 2'-, -$ 0#1'12 ,!# 2- Phytophthora cinnamomi ', 1##"V%0-5, 20##1 ," !*-, * *',#1 -$ Eucalyptus marginata ',-!3* 2#" ', * 2#0 * 0 ,! ," 0--21Q Plant PathologyQ51QVUWVVVT ü!)*#&-4#,Q.Q#!Q)Q=-%#*Q=VTRRS<-,V&-120#1'12 ,!#-$ 0*#7'1 11-!' 2#" 5'2& &7"0-%#, .#0-6'"# 3012 2 1'2#1 -$ 22#+.2#" .#,#20 2'-, 75&# 2.-5"#07+'*"#5$3,%31QMolecular Plant PathologyQ 2QS[[VTRW *!)*#&-4#,Q . TRRY )#** 5 **V 11-!' 2#" +#!& ,'1+1 -$ "'1# 1# 0#1'12 ,!# ,"131!#.2' '*'27QAnnual Review of PhytopathologyQ45QSRSVSTY 3'2#+ Q9Q?*##1&-35#01Q?/ Q00 ,!'1Q#Q= +-3,QTRRU !2'4#"#$#,!# 0#1.-,1#1 11-!' 2#"5'2&,-,V&-120#1'12 ,!#-$ Arabidopsis thaliana2- 2&#--+7!#2#. 2&-%#,Phytophthora infestans, Molecular Plant Pathology, 4,VZYVWRR *0'8 007Q . Q - 1Q Q )-**',Q 0Q # 8#0V 0!* 7Q 1Q ,2-,#**'1Q =Q !�$Q !Q .##"Q(TRRU96.*-0 2'-,Q,-0+ *'1 2'-,Q ,"13++ 0'#1-$&'%& "#,1'27-*'%-,3!*#-2'"# 00 7.0- #*#4#*" 2 QBiostatistics, 4,TV[VTXV *05',Q /Q ) &'**Q # ," 0#,2&Q S[[W Phytophthora', 3120 *' Q Australian Journal of Agricultural ResearchQ46QSUSSVSUUY #$$#01Q < ," # 02',Q S[ZX )-+. 0'1-, -$ 25- +#"' 1#*#!2'4# $-0 Phytophthora ,"Pythium1.#!'#1QPlant DiseaseQ70QSRUZVSRVU -& *Q/ ,"0'%%1QS[[T.#"3!2 1# !2'4'27#,!-"#" 72&#HM1"'1# 1# 0#1'12 ,!#%#,#',+ '8#QScienceQ258Q[ZWV[ZY -,#1Q/ ," ,%*Q7TRRX(&#.* ,2'++3,#1712#+QNatureQ444QUTUVUT[ = !&0--Q ," = !&0--Q TRRY *'!7*'! !'"Q ( 1+-,'! !'" ," #2&7*#,#V +#"' 2#" 0#%3* 2'-, -$ .* ,2 "#$#,1# 1'%, **',%Q Genetic engineeringQ 28Q WWVZU

182 REFERENCES

= +-3,QQ3'2#+ Q9Q?*##1&-35#01Q?/ S[[[.#1'12 ,!#2---+7!#2#1S %#,#0 *0-*#$-02&#&7.#01#,1'2'4#0#1.-,1#[Trends in Plant ScienceQ 4Q S[XVTRR = +-3,Q TRRS <-,&-12 0#1'12 ,!# 2- PhytophthoraS ,-4#* .0-1.#!21 $-0 !* 11'! *.0- *#+QCurrent Opinion in Plant BiologyQ4QT[WVURR = +-3,QTRRU#-*#!3* 0%#,#2'!1-$. 2&-%#,'!--+7!#2#1QEukaryotic CellQ 2QS[SVS[[ = ,%Q 7Q 7'Q Q H& -Q (Q K' -Q 0Q ( ,%Q KQ (&'*+-,7Q .Q #Q Q H&-3Q V# TRRU *,2#0.* 7-$2&#Arabidopsis,-,&-120#1'12 ,!#%#,#<>S5'2& !2#0' * 4'03*#,!#QProceedings of the National Academy of Sciences USAQ100QUWS[V UWTV =#** +Q#= ,")-$$#7Q#S[ZWI3 ,2'2 2'4#!-+. 0'1-,-$2�#1'12 ,!#2- Phytophthora 0--2 0-2 ', 2&0## 4-! "- 0--212-!)1Q Phytopathology, 75, TURVTUV ='+Q V# ," ='+Q V! TRSR )& 0 !2#0'1 2'-, -$ SV&7"0-67VTV9V 32#,7*VVV "'.&-1.& 2# 17,2& 1# %#,# $0-+ Ginko biloba, Molecular Biology ReporterQ37Q[YUV[Y[ =*'# #,12#',Q ,".-5#Q)TRRZ9!-*-%'! *!-121-$ '-20-.&'!4#0131 ,#!0-20-.&'!. 2&-%#,0#1'12 ,!#Q2&#&7.#01#,1'2'4#0#1.-,1# ,"1'%, * 20 ,1"3!2'-,QPlant ScienceQ174QWWSVWWX =--0,##$Q ," '#2#01#Q )# TRRZ )0-11 2 *) ', "#$#,1# 1'%, **',%Q Plant PhysiologyQ146QZU[VZVV =-2!&-,'Q > ," / !&-+-Q 93 TRRX (&# 0# !2'4# -67%#, 1.#!'#1 ,#25-0) . 2&5 71S ,#11#,2' *.0#0#/3'1'2#$-0.#0!#.2'-,-$. 2&-%#, 22 !) ," 2&# !/3'0#""'1# 1#0#1'12 ,!#',.* ,21QJournal of BiosciencesQ31QUZ[V VRV =0# 1Q 7 ," 3'*1-,Q # TRRT .#1'12 ,!# 2- Phytophthora 0--2 0-2 ', !-,2#+.-0 07.&-"-"#,"0-,!3*2'4 01QHortScienceQ37QY[RVY[T =0'#!& 3+#0Q ?Q 3#'% ,%Q 7Q 0'#ß#*+ ,,Q Q 7#28#*Q (Q 00#7Q #Q /'#0*Q Q /* 5'1!&,'%Q9TRRZ)& 0 !2#0'1 2'-,-$2Ï.2-.& ,17,2& 1# *.& 13 3,'2',+ '8#QBMC Plant BiologyQ8Q"-'SSRTSSZX SVYSVTTT[VZVVV

183 REFERENCES

=3) 4') QQ?3G','EQQ?3*#2'EQ#TRRW3.#0-6'"#"'1+32 1#Q.#0-6'"'1#Q ," %#0+',V*')#.0-2#', !2'4'27',.* 1+ +#+ 0 ,#1 ," .-.* 12-$+ '8# 0--21QProtoplasmaQ226QS[SVS[Y =3,)#*Q< ,"0--)1Q#TRRT)0-112 *) #25##,1'%, **',%. 2&5 71', . 2&-%#,"#$#,!#QCurrent Opinion in Plant BiologyQ5QUTWVUUS 7 '"* 5Q3 ,"3'*1-,Q TRRU0*-0'12'! ,"1203!230 *!& 0 !2#0'12'!1-$ !- 12 * &# 2&* ," #6&' '2',% 17+.2-+1 -$ Phytophthora cinnamomi ',$#12 2'-,',2&## 12#0,>25 7. ,%#1Q?'!2-0' QAustralian Journal of BotanyQ51QTZUVT[U 7 + Q) ,"'6-,Q. S[[Y(&#-6'" 2'4# 3012',.* ,2"'1# 1#0#1'12 ,!#Q Annual Review of Plant Physiology and Plant Molecular BiologyQ 48Q TWSV TYW 7 ,"-,'Q #Q ** ?#!!&' Q 0Q / 4 88'Q /Q /'3*','Q Q 7 .-!! Q

184 REFERENCES

7-0#,8-Q>Q'/3#0 1Q.Q ,!V#00 ,-QQ-* ,-Q.TRRU92&7*#,#0#1.-,1# $ !2-0S',2#%0 2#11'%, *1$0-+#2&7*#,# ,"( 1+-, 2#. 2&5 71',.* ,2 "#$#,!#QThe Plant CellQ15QSXWVSYZ 73Q #Q ( ,%Q KQ H&-3Q V# TRRS 0 '"-.1'1 NHO1 '1 0#/3'0#" $-0 %#,#0 * 0#1'12 ,!# % ',121#3"-+-, 1 !2#0' QThe Plant CellQ13QVUYVVVY 73, Q 9Q 12-0Q ?Q .- #02Q Q 0*-01Q ?Q # 3!&V# ,'Q Q (-,Q TRSS ) **-1# "#.-1'2'-,S +3*2'V$ !#2#" .* ,2 0#1.-,1#Q Molecular Plant Microbe InteractionsQ24QSZUVS[U 77'Q#QH&-3QKQ=-!&' ,Q7?Q7'Q7TRRY'-!&#+'! * ,"+-*#!3* 0 !& 0 !2#0'1 2'-, -$ 2&# &-+-!712#',# V+#2&7*20 ,1$#0 1# $0-+ 0-!!-*' Brassica oleracea4 0TitalicaQPhytochemistryQ68QSSSTVSSS[ # ," *Q ," #'20 Q TRRY .#',$-0!#+#,2 -$ !#** 5 ** ', 0--21 -$ Lycopersicon esculentum 2&0-3%& ',"3!2'-, -$ .&#,-*'! !-+.-3,"1 ," *'%,', 7 #*'!'2-01Q Physiological and Molecular Plant PathologyQ 71Q TRSV TR[ # *-*#.18 Q ! ," !0 ,#)Q TRRT oV7"0-67#2&7*-032',V+#"' 2#" #,& ,!#+#,2-$2-+ 2-0#1'12 ,!#2-Botrytis cinerea"#.#,"1-, 3012 -$0# !2'4#-67%#,1.#!'#1QJournal of PhytopathologyQ150QXSXVXTV # 0)1Q/) ,"= 11 7Q01S[YV#2#!2'-,-$Phytophthora cinnamomi',1-'*Q Australian ForestryQ63QS[ZVTRU # 001Q= S[[X(&#$3,!2'-,1 ,"0#%3* 2'-,-$%*32 2&'-,#1V20 ,$#0 1#1', .* ,21QAnnual Review of Plant Physiology, Plant Molecular BiologyQ47QSTYV SWZ # 3!&V# ,'Q ,"*31 0#,)-Q S[[X0-"3!2'-,-$1 *'!7*'! !'".0#!301-01 '1 + (-0$3,!2'-,-$.&#,7* * ,',# ++-,' V*7 1#',2�#1'12 ,!#-$ 0 '"-.1'12-Peronospora parasiticaQThe Plant CellQ8QTRUVTST #!) 00#,Q =7Q #!)-+ Q Q &# 0#0Q 7Q 0"7Q /2 TRRW (&# 0-*# -$ !&* +7"-1.-0#1-$Phytophthora cinnamomi 0#4'#5QAustralasian Plant PathologyQ34QUUUVUUZ #!731)7Q.Q#,,#22Q#Q# *#Q#Q7#5'1Q#Q/ 1)',Q ,"# ,1$'#*"Q3 S[[[ )#** 5 ** *2#0 2'-,1 ," *-! *'8#" !!3+3* 2'-, -$ $#03*-7*VU_V +#2&-67270 +',#',-,'-,#.'"#0+'1 21'2#1-$ 22#+.2#".#,#20 2'-, 7

185 REFERENCES

Botrytis allii 0# 11-!' 2#"5'2& !2',.-* 0'1 2'-,Q.#0-6'"'1# !2'4'27 ," 13..0#11'-,-$$* 4-,-'" '-17,2'1QThe Plant JournalQ17QWTUVWUV ##'#0Q ,"/#&0',%Q)TRRZ %3'"#2-2&#',2#%0 2#" ..*'! 2'-,-$-,V*',# " 2 +',',%2--*1$-02&#',$#0#,!#-$%#,#$3,!2'-,1 22ڰ#+1*#4#*Q Journal of BiotechnologyQ3QSUYWVSUZY ##+ 0MQ0% ,'1 2'-, ," 4 0' '*'27 -$ 2&# + '8# %#,-+#QCurrent Opinion in Plant BiologyQ9QSWYVSXU #'**#2Q 1 Q ,, Q )Q )* 7Q <=Q -,%,3 ,Q 3Q '+-,Q #Q 3#0!)V.#'!& 02Q Q 31 #*Q 0# TRSR *,, 2# '++3,# 0#1.-,1#1 !2'4 2#" ', 0 '"-.1'1 0--21 7 +'!0- #V 11-!' 2#" +-*#!3* 0 . 22#0,1Q The Plant CellQ 22Q [YUV [[R #-&0Q/ ,") &'**Q#TRRS.#* 2'4#0-*#1-$%*7!#-**',Q*'%,', ,"2&# &7.#01#,1'2'4# 0#1.-,1# ," 2&# ',$*3#,!# -$ ', !-+. 2' *# ," ',!-+. 2' *# ',2#0 !2'-,1 -$ 1-7 # ,1 5'2& Phytophthora sojaeQ Physiological and Molecular Plant PathologyQ58QUSVVS #71-0#= ,".73)V#QTRRV <-,&-120#1'12 ,!#S &-5+3!&"- 5# ),-5[ Trends in Plant ScienceQ9,[YVSRV < /4'QQ0 00#Q/Q , &3( Q/Q) .#**Q(QH&3Q)Q)&0'12-3QTRSR3&#,+-0#'1 #22#0S+3*2'%#,##,%',##0',%',.* ,21QTrends in Plant ScienceQ15QVZVWX <#+!&#,)-Q Q=3,8#QQ0#311,#0Q*Q=-*-+'#21Q#TRRX3.*'! 2#+ '8#SUV *'.-67%#, 1#%#,#1 0#"'$$#0#,2' **70#%3* 2#" 7!'0! "' ,0&72&+Q!-*" 120#11Q5-3,"',%Q. 2&-%#,',$#!2'-, ,"&-0+-, *20# 2+#,21QJournal of Experimental BotanyQ57QUYXYVUYY[ <#22*#2-,QTRRX "'1!311'-,-$12 2'12'! *+#2&-"1$-0"#1'%, ," , *71'1-$ +'!0- 00 7#6.#0'+#,21$-0.* ,21!'#,2'121QThe Plant CellQ18QTSSTVTSTS <#5&--)Q 0 ," -"%#0Q 0 S[YT (&# 0-*# -$ Phytophthora cinnamomi', 3120 *' , ,"<#5H# * ,"$-0#121QAnnual Review of PhytopathologyQ10Q T[[VUTX

186 REFERENCES

<'"#0+ ,Q(Q/#,#2#2Q*Q037#0#Q(Q/##1Q.Q2',28'Q Q7#%0 ,"Q#Q00'2'%QQ #-1',%#0Q 9 S[[W 2&-%#,#1'1V0#* 2#" .VS .0-2#',1 0# ,2'$3,% *S *1-* 2'-, ," !& 0 !2#0'1 2'-, -$ 2&0## SVV)'*-" *2-, .0-2#',1 -$ 2-+ 2- ," -$ 1'! .VS -$ 2- !!- 5'2& ',&' '2-07 !2'4'27 2-5 0"1 Phytophthora infestansQPlant PhysiologyQ108QSYVTY <'*11-,Q V9 S[[W .#+-2# 1#,1',% ," '+ %# , *71'1 ', .* ,2 . 2&-*-%7Q Annual Review of PhytopathologyQ15QVZ[VWTY <'2 Q #Q 9**'1Q # ," # ""#,Q 7? TRRU .#*' '*'27 ," !!30 !7 -$ 4'13 * #12'+ 2'-, -$ &-+-.1'1 7# $ *'%&2 -$ 120 5 #007Q PhytopathologyQ 93Q [[WVSRRW <*0, #0%#0Q( ,"7'.) Q?TRRW<-,V&-120#1'12 ,!#',.* ,21S,#5',1'%&21 ',2- ,-*".&#,-+#,-,QMolecular Plant PathologyQ6QUUWVUVW >_0'#,Q. ,"4 ,03%%#,Q )S[[T !!30 !7Q.0#!'1'-, ,"!-00#* 2'-, 2-7'#*"*-11-$"'1# 1#1#4#0'271! *#1$-0!-0)70--2-$*#223!#Q PhytopathologyQ82Q[SV[X >_0'#,Q Q3'**' +1Q_)-,,#**Q. ," ,1203% Q.TRRX(ê2#à2ê2#',1'"# .* ,2!#**S#12 *'1&',% !-+. 2' '*'27 #25##, .* ,21 ," '-20-.&'! $3,%' ," --+7!#2#1Q New PhytologistQ171QX[[VYSZ >)3 0 Q ," 3*'28Q () TRRW .--2 "#$#,1# 0#1.-,1#1 2- $3,% * . 2&-%#,1S +-*#!3* 0.#01.#!2'4#QPlant and SoilQ274QTSWVTTX >1 -30,Q 9 TRRS * ,2 +#!& ,'1+1 2& 2 %'4# "#$#,!# % ',12 1-'* -0,# "'1# 1#1QAustralasian Plant PathologyQ30Q[[VSRT 35#*1Q7Q*,8MQQ/--11#,1Q TRR[ 1+-, 2#V',"3!' *#%#,#S5& 2"-#1'2 +# ,[Trends in Plant ScienceQ14QZYV[S &'**'.1Q ,"3#12#Q/S[ZV0'#*"0#1'12 ,!#',2&0##, 2'4#+-,-!-27*#"-, 1.#!'#1 2& 2 !-*-,'8# ',"'%#,-31 1!*#0-.&7** $-0#12 $2#0 ',4 1'-, 7 Phytophthora cinnamomi, Australian Journal of BotanyQ32QUU[VUWT '* # +Q . Q )-*/3&-3,Q *Q &# 0#0Q Q 0"7Q /2 TRRR &-1.&'2# !-,!#,20 2'-,S'21#$$#!2-,.&72-2-6'!'2717+.2-+1 ,"!-*-,'1 2'-, 7

187 REFERENCES

Phytophthora cinnamomi ', 2&0## 3,"#012-0#7 1.#!'#1 -$ Eucalyptus marginata$-0#12QAustralasian Plant PathologyQ29,ZXV[W -"%#0Q 0Q -#.#*Q .0Q H#,2+#7#0Q / S[XW 11-!' 2'-, -$ Phytophthora cinnamomi 5'2& "'1# 1#-$Eucalyptus marginata$-0#12',3#12#0, 3120 *' QPlant Disease ReporterQ49Q[VUV[VY -"%#0Q0 ," 1&2-,QS[YRPhytophthora cinnamomi',"7',%4#%#2 2'-, -,2&#0'1 ,#. ,%#1Q?'!2-0' QAustralian Forest ResearchQ4QUUVUX -"%#0Q0 ,"0-5,Q#S[Z[?#%#2 2'-," + %#! 31#" 7Phytophthora cinnamomi -, "'1230 #" 1'2#1 ', 2#+.#0 2# 0 ',$-0#121 ', 5#12#0, ( 1+ ,' QAustralian Journal of BotanyQ37QVVUVVZR -* ,"Q Q =302'V *',2Q Q 3'11#0Q .Q 0 22Q .)Q <#*1-,Q . TRRZ & "#1 -$ %0 7S2-0*"-$/3 ,2'2 2'4#"'1# 1#0#1'12 ,!#QTrends in Plant ScienceQ 14QTSVT[ -** 0"Q#Q#'11-,Q0Q7'Q1Q>&*0-%%#QTRRZ3'*"',%*'.'" 00'#01S '-17,2'1-$!32', ,"13 #0',QTrends in Plant ScienceQ13QTUXVTVX 07!#QQ9"5 0"1Q3Q/ "#)QTRRT'120' 32'-,-$Phytophthora cinnamomi 2"'$$#0#,21. 2' *1! *#1S3&#,! , ,#% 2'4#0#13*2 #!-,1'"#0#" .-1'2'4#*7[Austral EcologyQ27QVW[VVXT . ',-5'!8Q ," #,,#28#,Q 7 TRRX (&# + '8# %#,-+# 1 +-"#* $-0 #$$'!'#,21#/3#,!# , *71'1-$* 0%#.* ,2%#,-+#1QCurrent Opinion in Plant BiologyQ9QSV[VSWX . + +--02&7Q?Q. %3!& ,"#0Q(Q +'7 .. ,Q.TRRT*,"3!2'-,-$"#$#,!#V 0#* 2#".0-2#',1',2-+ 2-0--2120# 2#"5'2&Pseudomonas fluorescens$S ,"Fusarium oxysporum$T1.TlycopersiciQPlant and SoilQ239QWWVXZ . ,"1Q . S[TT 20##.) ,)#0 4 , = ,##*Q 4#0--08 )2 "--0 Phytophthora cinnamomi ,T 1.T 20'.# ! ,)#0 -$ !',, +-, ! 31#" 7 Phytophthora cinnamomi ,T1.TMededeelingen van het Instituut voor PlantenziektenQ54Q SVWU . 7,#0Q (0Q .-!! V#00 Q Q .#**+ ,Q (Q ) 312-,Q )Q 0 0,#Q Q -**-5 7Q 9Q *0#8 007Q. Q7'3QQ# '#0QQ#'**#0Q#et al.TRRX 1'+.*#1.0# "1&##2V 1#" #* #9V13..-02'4# $-0+ 2 $-0 +'!0- 00 7 " 2 S # /9V( Q BMC BioinformaticsQ7Q"-'SSRTSSZX SVYSVTSRWVYVVZ[

188 REFERENCES

.#'2#0Q

189 REFERENCES

!&#,)Q #Q )&--Q Q 3-,%Q )7 TRR[ #'!0- 00 7 , *71#1 2- 123"7 .* ,2 "#$#,!# ," 0&'8-1.�#V+'!0- # ',2#0 !2'-,1Q CAB Reviews, Perspectives in Agriculture, Veterinary Science, Nutrition and Natural ResourcesQ4QSVSV !&'+-*#0V>_.-30)#Q .Q .'!& 0"1-,Q #Q #*'20#,,')-$$Q ) TRRS H# + 2', ',&' '21 207.1', ," IV +7* 1# !2'4'2'#1Q Applied and Environmental MicrobiologyQ67QTUXWVTUXX !&, *#QQ3 0#QQ03*2-,Q.Q2#',Q)Q3#'Q0Q 12#0, )QQ7' ,%Q)QH& ,%Q Q03*2-,Q7Q/0 4#1Q( Qet al.TRR[(&#YU+ '8#%#,-+#S!-+.*#6'27Q "'4#01'27Q ,""7, +'!1QScienceQ326QSSSTVSSSW !&-0, !)Q Q 3'2#+ Q 9Q ) ,-Q 7#Q -8)302Q (>Q >*'4 Q .Q ? , ++#Q #Q !&5'8#0QQ. $$ #*#QQ)& .. 0-V/ 0!' Q Q#%0#2',Q#9Q-1QQ 1QQ H-"7Q#)Q<31 3+Q)Q3',QQ(&',#1Q#Q= +-3,QTRR[(#,2&',%12- ),-5 -32--+7!#2##$$#!2-01QMolecular Plant PathologyQ10QY[WVZRU !&0#' #0Q 7Q 00 ,)#Q .Q 7#11'0#Q . TRRW '-!&#+'! * !& 0 !2#0'8 2'-, -$ #*-,% 1# !2'4'27',!-0,Zea mays7T0--21QPhytochemistryQ66QSUSVSUZ !&3*8#V7#$#02Q ," ,1203% Q . TRSS +-*#!3* 0 #4-*32'-, 07 !-,!#.2 !-,,#!2',% ,-,&-12 0#1'12 ,!#Q . 2&-%#, &-12 0 ,%#Q ," . 2&-%#, 1.#!' 2'-,QTrends in Plant ScienceQSXQSSYVSTW ##+Q.)S[ZV'1# 1#',!'"#,!# ,"1#4#0'270#* 2'-,1&'.1QAnnual Review of PhytopathologyQ22QSUUVSWR &# 0#0Q7Q)0 ,#Q)9Q)-!&0 ,#Q TRRVI3 ,2'$'! 2'-,-$2ƒ!#.2' '*'27-$ 2&#, 2'4#$*-0 -$2&#-32&3#12-2 ,'! *0-4',!#Q3#12#0, 3120 *' Q 2-Phytophthora cinnamomi, Australian Journal of BotanyQ52QVUWVVVU &# 0#0Q7Q'**-,Q#Q=', *QQ3#&0'%Q.#TRSR(#+.-0 * ,"1. 2' *1-'* ',-!3*3+1 "7, +'!1 $-**-5',% Phytophthora cinnamomi',4 1'-,-$ Banksia woodland ,"Eucalyptus marginata $-0#12 '-+#1 -$ 1-32&V 5#12#0, 3120 *' QAustralasian Plant PathologyQ39QT[UVUSS &#,QQH&#,%QHQ--,#0Q=TRRR + '8#1#1/3'2#0.#,#!7!* 1#%#,# ',"3!#" 7 ',1#!2 � '4-07 ," 4-*'!'2',S )& 0 !2#0'1 2'-, -$ 5'*"V27.# ,"+32 ,2 **#*#1QPNASQ97QSVZRYVSVZST '*4 V) 0"#, 1Q.*Q.'! 0"QQ %*'-QQ'**Q.TRRU#+-%*- ', ,"&7.-6'! !!*'+ 2'-,',+ '8#0--22'.1QRussian Journal of Plant PhysiologyQ50QZTSV ZTX

190 REFERENCES

'++-,"1Q S[XX Host index of plant diseases in QueenslandQ I3##,1* ," #. 02+#,2-$0'+ 07*,"3120'#1QSSS. ',"&3Q Q)&',2 + , ,'QQ0 ,"2Q QH ,'1Q#Q!-$'#*"Q.Q-& *Q/TRRZ %3 0"' ,-$%0 11#1S1.#!'$'!-0'%', ,"!-,1#04 2'-,-$ 3,'/3#"'1# 1#V 0#1'12 ,!#%#,#',2&#%0 11*',# %#QProceedings of the National Academy of SciencesQ105QSYXTVSYXY *31 0#,)-Q ,"!&* '!&Q<7TRRU-5,7+'*"#5-$Arabidopsis thaliana ! 31#" 7Hyaloperonospora parasitica$-0+#0*7Peronospora parasitica Molecular Plant PathologyQ4QSW[VSYR -32-,Q/ ,"#4#0 **QS[[R'12-!&#+'! * ,"!&#+'! *#4'"#,!#$-0 *'%,', !!3+3* 2'-,"30',%2&##6.0#11'-,-$0#1'12 ,!#2-*# $0312$3,%'', 5&# 2QPhysiological and Molecular Plant PathologyQ36QVZUVV[V 2 &*Q9 ,"'1&-.Q/TRRR* ,2V. 2&-%#, 0+10 !#1 22&#+-*#!3* 0 *#4#*QCurrent Opinion in Plant BiologyQ3QT[[VURV 2-,#Q ," )* 0)#Q 9 S[[T Chemistry and Biology of (1-3)--glucansQ 7 (0- #!,'4#01'270#11Q?'!2-0' Q 3120 *' ( )#+-2-QQ 0"& +Q .Q-,#1Q TRRW'$$#0#,!#1',!#**"# 2&',"3!2'-, 7 Phytophthora #*'!'2',1 0# "#2#0+',#" 7 1'%, * !-+.-,#,21 "-5,120# +-$# )', 1#)', 1#',"'$$#0#,21.#!'#1-$<'!-2' , ," !3*2'4 01 -$ Brassica rapa ,"Raphanus sativusQ Plant PhysiologyQ 138Q SV[SVSWRV (#,& )#,Q.Q7#4',#Q Q0'11-,Q70Q'6-,Q. Q7 + Q)S[[W03,!2'-,-$2&# -6'" 2'4# 3012 ', &7.#01#,1'2'4# "'1# 1# 0#1'12 ,!#Q Proceedings of the National Academy of SciencesQ92QVSWZVVSXU (& 2!�Q70Q ,"#01-,QQ',%&Q=TRRW* ,2"#$#,!#0#1.-,1#1S5& 2 & 4#5#*# 0,2$0-+ 0 '"-.1'1QFunctional Plant BiologyQ32QSVS[ (&'++Q>Q*=1',%Q>Q/' -,Q1Q< %#*Q Q##7#0QQ=0*%#0QQ#* '%QQ#***#0Q7 Q .&##Q1Q2'22Q#TRRV# #

~~191 REFERENCES~~

~~!-+.-1'2'-,Q ,"0#* 2'-,1&'.2-. 02' *0#1'12 ,!# 2- Phytophthora sojae, Plant PhysiologyQ144QT[[VUSS (&-0" *V)&0'12#,1#,Q Q H& ,%Q HQ 3#'Q 1Q )-**',%#Q S[[Y 3 !#**3* 0~~

*-! *'1 2'-, -$ T>T ',.* ,21TT>T !!3+3* 2'-,',. .'** # ," &7.#01#,1'2'4#0#1.-,1#"30',%2&# 0*#7V.-5"#07+'*"#5',2#0 !2'-,Q The Plant JournalQ11QSSZYVSS[V (&-0" *V)&0'12#,1#,Q TRRU 00#1& ',1'%&21 ',2- .0-!#11#1 -$ ,-,&-12 0#1'12 ,!#QCurrent Opinion in Plant BiologyQ6QUWSVUWY ('#0#,1Q =0#VQ (&-++ Q Q . (#,"0 Q Q / 0+'#0Q #Q 9%%#0+-,2Q =Q 0-35#0Q #Q #,,',!)6Q * # Q 0-#) #02Q 30Q ) ++3#Q TRRT Esa1Q , 0 '"-.1'1 +32 ,2 5'2& #,& ,!#" 131!#.2' '*'27 2- 0 ,%# -$ ,#!0-20-.&'! $3,% * . 2&-%#,1Q 1&-51 "'12-02#" ',"3!2'-, -$ "#$#,1# 0#1.-,1#1 7 0# !2'4# -67%#, %#,#0 2',% !-+.-3,"1Q The Plant JournalQ29QSUSVSVR ('$$#,Q ,"/ 32QTRRS#-*#!3* 0#4-*32'-,-$2-3,"V',"3!#"1#0',# .0-2# 1#',&' '2-0wip1 ',Zea ,"0#* 2#"%#,#0 QMolecular Biology and EvolutionQ18QTR[TVTSRS ('..#22Q ,"# * (!83)Q<S[Y[*,2#0 !2'-,-$Phytophthora cinnamomi ," 0#1'12 ,2&-12QAcacia pulchellaQPhytopathologyQ69QYXVVYYT (-,Q ," # 3!&V# ,'Q TRRV JV +',-V 3270'! !'"V',"3!#" 0#1'12 ,!# % ',12 ,#!0-20-.&'! . 2&-%#,1 '1 1#" -, V"#.#,"#,2 .0'+',% $-0 ! **-1#QThe Plant JournalQ38QSS[VSUR (-,QQ_ **#11 ,"0-Q#Q-30"'#Q?Q ) Q/Q= 0*#,QQ#*"Q#Q# 3!&V# ,'QQ (30*',%1Q(TRRX0'+',% 7 '0 -0,#1'%, *1 --121"'0#!2 ,"',"'0#!2 0#1'12 ,!#',+ '8#QThe Plant JournalQ49QSXVTX (03('**-Q#Q=-%#*Q=VQü!)#*&-4#,Q . TRRV 3.#0-6'"# ," &7"0-%#, .#0-6'"#.* 7"'$$#0#,20-*#1',2&#,-,&-12',2#0 !2'-,-$ 0*#7 ," 5&# 2 5'2& ', ..0-.0' 2# $-0+ # 1.#!' *#1 -$ Blumeria graminis, Molecular PlantMicrobe InteractionsQ17QURVVUST (031-4Q1Q#5#* +Q

~~192 REFERENCES~~

(30,#0Q/Q9**'1Q)Q#4-2-Q TRRT(&#( 1+-, 2#1'%, *. 2&5 7Q The Plant CellQSWUVSXV (7*#0 #Q (0'. 2&7 , H& ,%QKQ#& *QQ' ,%Q.1Q #021Q Q 00#"-,"-Q0Q 62#0Q 7Q #,1 11-,Q Q #7,-,Q 7Q et al. TRRX Phytophthora %#,-+# 1#/3#,!#13,!-4#0#4-*32'-, 07-0'%',1 ,"+#!& ,'1+1-$. 2&-%#,#1'1T Science313QSTXSVXT (7*#0Q#TRRYPhytophthora sojaeS0--20-2. 2&-%#,-$1-7 # , ,"+-"#* --+7!#2#QMolecular Plant PathologyQ8QSVZ !.. * . 2'Q.Q# 0#)Q#Q 7##QV=Q< ) 1&'+ QQ( ,%Q 1Q*#"%#Q#=Q'6-,Q . Q#71-0#Q=TRR[/*- *%#,##6.0#11'-,.0-$'*',%"30',%Medicago truncatula-Phymatotrichopsis omnivora',2#0 !2'-,0#4# *1 0-*#$-0 ( 1+-,'! !'"Q #2&7*#,# ," 2&# $* 4-,-'" . 2&5 7 ', "'1# 1# "#4#*-.+#,2QMolecular Plant Microbe InteractionsQ22QYVSY !1 "#*Q Q -0##Q 0Q < %#*Q Q 7-&1#Q #Q )8#"')V971#, #0%Q Q 2'22Q # TRR[ %3'"#2-31',%# .# ,2-4'13 *'1# ,"!-+. 0#>+'!1" 2 ',.* ,21S ! 1#123"7',2&#!0-.1.#!'#1Q# '8#QPlant Cell and EnvironmentQ 32Q STSSVSTT[ ? , 0*#,Q Q 2 21Q #Q ? , = ,Q 7 TRRV *,"3!2'-, -$ .0-%0 ++#" !#** "# 2& ', *'*7 7 2&# $3,% * . 2&-%#, Botrytis elliptica, Molecular Plant PathologyQ5QWW[VWYV ? , 0*#,QQ3-*2#0',%Q9Q2 21Q#Q? ,= ,Q 7TRRY'12-!&#+'! * ," %#,#2'! , *71'1 -$ &-12 ," ,-,V&-12 ',2#0 !2'-,1 -$ 0 '"-.1'1 5'2& 2&0##Botrytis1.#!'#1S ,'+.-02 ,20-*#$-0!#**"# 2&!-,20-*QMolecular Plant PathologyQ8QVSVWV ? ,!#Q)Q='0)Q(= ,"--"Q.(S[ZR7'%,'$'! 2'-, 1 +#!& ,'1+-$ "'1# 1#0#1'12 ,!#QAnnual Review of PhytopathologyQ18QTW[VTZZ ? ,"#0'#8#,Q9 ,"-,#1Q/S[[Z* ,2"'1# 1#V0#1'12 ,!#.0-2#',1 ," 2&#%#,#V$-0V%#,#!-,!#.2QTrends in Biochemical SciencesQ23QVWVVVWX ? ,"#0--0,Q. ,"= +-3,QTRRZ00-+%3 0"2-"#!-7S ,#5+-"#*$-0 .#0!#.2'-,-$.* ,2. 2&-%#,#$$#!2-01QPlant CellQ20QTRR[VTRSY ? ,= ,Q 7TRRX7'!#,!#"2-)'**S2&#*'$#127*#-$ ,#!0-20-.&'!.* ,2 . 2&-%#,QTrends in Plant ScienceQ11QTVYVTWU

~~193 REFERENCES~~

? , 7--,Q 7)Q .#.Q #Q '#2#01#Q )# TRRX '%,'$'! ,!# -$ ',"3!' *# "#$#,!#V 0#* 2#" .0-2#',1 ', ',$#!2#" .* ,21Q Annual Review of PhytopathologyQ 44Q SUWVSXT ? ,3#12QQ ..' &Q Q/-5Q< .TRRU "4 ,!#1',0#1# 0!&-,--+7!#2# 0--2. 2&-%#,1QPhysiological and Molecular Plant PathologyQ62Q[[VSSU ?#* 8& & ,Q.Q 22 Q=Q#32&3)0'1&, ,QS[[[(&#.VW$ +'*7S2& 3+ 2',V *')# .0-2#',1Q ', Pathogenesis-related proteins in plantsQ 22 ," #32&3)0'1&, ,#"1).)0#11Q0*-0'" Q! QT[S. ?'!)Q ," H'++#0+ ,Q)S[ZX )& 0 !2#0'1 2'-,-$STV-6-V.&72-"'#,-'! !'"0#"3!2 1#',!-0,QPlant PhysiologyQ80QTRTVTRW ?'( 7 ,Q Q &-!)#7Q Q 7é4#1/3#Q ) Q )--)Q Q 0-51#Q S[[Z 0-*# $-0 ( 1+-, 2#',. 2&-%#,"#$#,!#-$ 0 '"-.1'1QProceedings of the National Academy of Sciences USAQ95QYTR[VYTSV ?*##1&-35#01Q ?/ Q 4 , --'(#5##02Q 3Q /-4#01Q 0Q = +-3,Q Q )-*-,Q 7( TRRR(&#&7.#01#,1'2'4#0#1.-,1#'1 11-!' 2#"5'2&&-12 ,",-,&-12 0#1'12 ,!#2-Phytophthora infestans, PlantaQ210QZWUVZXV ?-%2Q(TRSR&#,7*.0-. ,-'" '-17,2'1QMolecular PlantQ3QTVTR 3 ,%Q = 7V)Q 7'Q Q 9!)#0Q . TRRT 92&7*#,# '-17,2'1 ," 1'%, **',% ,#25-0)1QThe Plant CellQ14QSUSVSWS 3 12#0, !)Q ) TRRY 1+-, 2#1S , 3." 2# -, '-17,2'1Q 1'%, * 20 ,1"3!2'-, ," !2'-,',.* ,2120#110#1.-,1#Q%0-52& ," "#4#*-.+#,2QAnnals of BotanyQ100QXZSVX[Y 3'#"#+ ,,Q/Q=-.0'4-4 Q Q!&,#'"#0Q#Q#01!& !&Q)Q.#1)'Q.Q=-.0'4 Q TRRY (&# 0-*# -$ 2&# ,-4#* "#,-1',# W_V.&-1.&-13*$ 2# 0#"3!2 1# ', 0#%3* 2'-, -$ 13*.& 2# 11'+'* 2'-, -$ Physcomitrella patensQ Plant Molecular BiologyQ65QXXYVXYX 3#12#Q/ ,"( 7*-0QS[YS(&#',4 1'-,-$, 2'4#$-0#12 7Phytophthora cinnamomi, Australian Journal of BotanyQ19QTZSVT[V 3#12#Q/ ,"?'2& , %#Q=S[YZ9$$#!2-$Phytophthora cinnamomi-, +'!0- ' *.-.3* 2'-,1 11-!' 2#"5'2&2�--21-$$-0#12$*-0 QAustralian Journal of BotanyQ26QSWUVSXY

~~194 REFERENCES~~

3#12#Q/ ,"# 0)1Q/)S[ZY(&# '-*-%7-$Phytophthora cinnamomi', 3120 * 1' ,$-0#121QAnnual Review of PhytopathologyQ25QTRYVTT[ 3#12#Q / ," =#,,#"7Q S[[Y .#%#,#0 2'-, -$ 131!#.2' *# , 2'4# 1.#!'#1 $-**-5',% "#!*',#-$Phytophthora cinnamomi-4#0 .#0'-"-$TR7# 01 -,"#$',#".*-21',2&#/0 +.' ,1Q3#12#0,?'!2-0' QAustralian Journal of BotanyQ45QSXYVS[R 3#12#Q/TRRT*,2#0 !2'-, #25##,Phytophthora cinnamomi ,"?'!2-0' , , 2'4#.* ,21.#!'#1%0-5',%',2'*"QAustralian MycologistQ21QXVVYT 3#12#Q /Q 0-5,Q =Q =#,,#"7Q Q 3 *1&#Q ( TRRT Phytophthora cinnamomi ',$#12 2'-, TVV7# 0 123"7 -$ 4#%#2 2'-, !& ,%# ', $-0#121 ," 5--"* ,"1 -$ 2&# /0 +.' ,1Q 3#12#0, ?'!2-0' Q Australian Journal of BotanyQ50QTVYVTYV 3#12#Q / TRRU (&# "'# !) !7!*# ', ?'!2-0' , $-0#121S URV7# 0 123"7 -$ !& ,%#1 ! 31#" 7 &72-.&2&-0 !',, +-+' ', ?'!2-0' , -.#, $-0#121Q 5--"* ,"1 ,"&# 2&* ,"1QAustralasian Plant PathologyQ32QTVYVTWX 3#2� ##Q.Q',!&QQ-,'%Q*Q)* 0)#Q S[ZV.#1.-,1#-$Zea mays0--212- ',$#!2'-,5'2&P. cinnamomi**S2&#!-02#6 ,"12#*#QProtoplasmaQ126QSZZV S[Y 3&'2$'#*"Q0Q&# Q.Q/'**#,Q=Q& 5Q=S[ZS?-* 2'*#!-+.-,#,21$0-+2&# 0--21-$Acacia pulchella.T0 ,"2&#'0#$$#!2-,Phytophthora cinnamomi . ,"1QAustralian Journal of BotanyQ29QS[WVTRZ 3'*1-,Q Q #02-,QQ) &'**Q#TRRR.#* 2'-,1&'.1 #25##,1'2#$ !2-01 ," "'120' 32'-, -$ Phytophthora cinnamomi ', 2&# 9 12#0, >25 7 . ,%#1Q ?'!2-0' QAustralian Journal of BotanyQ48QTVYVTXR 3'1#Q.Q#-1!-3Q#Q-%" ,-4#Q Q3&'2& +Q TRRY(0 ,1!0'.2.0-$'*',% ',&-12V. 2&-%#,',2#0 !2'-,1QAnnual Review of PhytopathologyQ45QUT[V UX[ 30'%&2Q Q #-#&*#,) +.Q ) Q #00-2Q /Q <#3$$#0Q #/Q )-,#Q =) S[[T (&# + '8# 36-20-.&'!+32 ,2orange pericarp'1"#$#!2'4#',"3.*'! 2#%#,#1 $-0207.2-.& ,17,2& 1#JQThe Plant CellQ4QYSSVYS[

~~195 REFERENCES~~

33Q Q =0'8Q 7Q 3'"&-*+Q # S[[V #-*#!3* 0 , *71'1 -$ 25- !< !*-,#1 #,!-"',% !'"'! !* 11 * !&'2', 1# ', + '8#Q Plant PhysiologyQ 105Q SR[YV SSRW 33Q 1Q ###*#7Q .Q )-1%0-4#Q TRRS , *71'1 ," #6.0#11'-, -$ 2&# IV #6. ,1', ," JV#6. ,1', %#,# $ +'*'#1 ', + '8#Q Plant PhysiologyQ 126Q TTTVTUT 1',QQ##'Q7Q<#5+ ,QQ !)Q=Q)& ..#**QS[[Y.#%3* 2'-,-$1#1/3'2#0.#,# !7!* 1#%#,##6.0#11'-,QPlant PhysiologyQ115QVUYVVWS H#**#0&-$$Q

~~196 Appendix~~

~~Appendix 1.0: .#120'!2#"*#1'-,$-0+ 2'-,',2&# 3120 *' ,, 2'4#%0 11 Lomandra filiformis.~~

a

b

Figure A1. The Australian native grass Lomandra filiformis roots 72 hpi with P. cinnamomi zoospores, showing similar restricted lesions to those in Z. mays roots. (a) L. filiformis (b) root section indicating lesion formation directly beneath the point of inoculation. Scale = 1.5 cm and 0. 25 cm respectively.

~~197~~

~~Appendix~~

~~Appendix 1.1: Zea mays online genomic resources:~~

~~&22.S 555T+ '8#1#/3#,!#T-0%~~

~~&22.S 555T+ '8#%" T-0%~~

~~&22.S 555T+ '8#+ .T-0%~~

~~&22.S %0 +#,#T-0%~~

~~&22.S 555T.&72-8-+#T,#2 ',"#6T.&.~~

~~198~~

~~Appendix~~

Appendix 2.0. Plant growth trials for Z. mays (5- 1-'*V%0-5, +#2&-"1 5#0# 20' **#" ', 2&'1 123"7S !-,4#,2'-, * .-21 $'%30# TT ," .30.-1# 3'*2 0--2 - 1#04 2'-, -6#1 $'%30# TT T )-,4#,2'-, *.-21.0-4'"#"#6!#**#,2%0-52&!-,"'2'-,1 325#0#3,13'2 *#$-0 "'0#!20--2',-!3* 2'-,1T*, ,#$$-022-!-+ 22&'1.0- *#+Q.30.-1#V 3'*20--2 - 1#04 2'-, -6#1 5#0# "#1'%,#" ," !-,1203!2#" 90 * 12'!1Q /##*-,%Q ?'!2-0' 2- **-5 $-0 "'0#!2 0--2 ',-!3* 2'-, ," - 1#04 2'-, -$ "'1# 1# .0-%0#11'-,T(&#&',%#"Q!*# 0#01.#6$0-,2!-3*" ## 1'*7-.#,#"5'2&-32 "'1230 ,!#2-2&#.* ,20--21T

~~Figure A.2. Comparison of soil grown plant methods (a) conventional pots (b) Purpose-built root observation boxes. Scale bar = 10 cm.~~

~~199~~

~~Appendix~~

~~Appendix 2.1. Isolation of P. cinnamomi from field collected soil samples~~

a

b

~~c d~~

Figure A3. The process of P. cinnamomi isolation from field collected soil samples (a) soil is collected from the base of X. australis showing early signs of chlorosis (arrowhead) (b) individual soil samples are labelled and mixed with water to form a slurry (c) isolation of P. cinnamomi (black arrow) on Phytophthora selective media from within the roots of E. sieberi d) plug of P. cinnamomi transferred to CV8 media following 72 h incubation.

~~200~~

~~Appendix~~

~~Appendix 2.2: Media and solutions for P. cinnamomi growth and maintenance~~

~~10% CV8 agar medium Reagent Quantity~~

~~V8 Vegetable Juice 150 mL~~

~~Calcium carbonate 1.5 g~~

~~Bacteriological agar 15 g~~

) )>U 5 1 !-+ ',#" 5'2& ?Z (3'!# ) +. #**_1 -3.1 3120 *' Q 7#+,-1Q ?'!2-0' ," 12'00#" $-0 STW &-301T (&# +'6230# 5 1 2&#, !* 0'$'#" 7 !#,20'$3% 2'-, 2SWRR6%$-0W+',32#1TSRR+7-$13.#0, 2 ,25 1!-+ ',#"

5'2& [RR +7 1"T> ," $'*2#0#" 5'2&', $3,,#* 2&0-3%&3& 2+ , <3+ #0 T $'*2#0. .#0U##3& 2+ ,.0'-02-. "(312+#,22-XTWT !2#0'-*-%'! * % 0 5 1 ""#" ,"2-*32'-, 32-!* 4#" 2STS5)$-0TR+',32#1 ,".-30#"',2- [!+.#20'.* 2#1T

~~5% CV8 broth Reagent Quantity~~

~~V8 Vegetable Juice 150 mL~~

~~Calcium carbonate 1.5 g~~

) )>U 5 1 !-+ ',#" 5'2& ?Z (3'!# ) +. #**_1 -3.1 3120 *' Q 7#+,-1Q ?'!2-0' ," 12'00#" $-0 STW &-301T (&# +'6230# 5 1 2&#, !* 0'$'#" 7 !#,20'$3% 2'-, 2SWRR6%$-0W+',32#1TSRR+7-$13.#0, 2 ,25 1!-+ ',#"

~~5'2& [RR +7 1"T> ,"$'*2#0#".0'-02-. "(312+#,22-XTWT0-2&5 1 12#0'*'1#" 7 32-!* 4',% ,"!--*#"2-.(.0'-02-31#T~~

~~Mineral salts solution Reagent Quantity~~

~~Calcium nitrate tetrahydrate Ca(NO3) 4H2O 3.08 g~~

~~Magnesium sulphate heptahydrate MgSO4 7H2O 1.49 g~~

~~Potassium nitrate KNO3 0.51 g~~

~~(&# -4#5#0#+ "#3.2-S75'2&1"T> ,"2-*32'-,5 1%#,2*7&# 2#" $-0 UR +',32#1 .0'-0 2- #',% 32-!* 4#"T (&# 1-*32'-, 5 1 !--*#" 2- 0--+ 2#+.#0 230# #$-0#2&# ""'2'-,-$S+712#0'*'1#"!&#* 2#"'0-,1-*32'-,T~~

~~201~~

~~Appendix~~

~~Chelated iron solution Reagent Quantity~~

~~EDTA Disodium salt 0.625 g~~

~~Iron Sulphate heptahydrate FeSO4 7H2O 1.245 g~~

~~Potassium hydroxide KOH 0.375 g~~

9( ,"0#>VYT>5#0#"'11-*4#"',WR+71"T>5'2&%#,2*#&# 2T>,!# !-+.*#2#*7!--*#"QRTUYW%=>5 1 ""#" ,"2-*32'-,12#0'*'1#"2&0-3%& RTVW=+QTW++170',%#$'*2#02(3$$07,+#+ 0 ,#Q !0-"'1!Q **Q?'!2-0' Q 3120 *' T.#+ ',',%1-*32'-,5 112-0#" 2V5)$-03.2-S7# 0T

~~Phytophthora selective media (PARPH) Reagent Quantity~~

~~Pimaricin 0.8 mL of 2.5%~~

~~Ampicillin 0.13 g~~

~~Rifampicin 0.01 g~~

~~Pentachloronitrobenzene (PCNB) 0.15 g~~

~~Hymexazol 0.07 g~~

~~Corn meal agar 17 g~~

~~Ethanol 5 mL~~

~~)-0, +# * % 0 5 1 "'11-*4#" ', S 7 1"T> ," 32-!* 4#" 2 STS5) $-0 TR +',32#1T'+ 0'!',Q +.'!'**',Q)< ,"&7+#6 8-*5#0#2&#,"'11-*4#"',VR~~

+7 1"T>T.'$ +.'!',5 1"'11-*4#"',#2& ,-*Q+'6#" 74-02#6',%$-0XR 1#!-,"1Q ," ""#"2-2&#"'11-*4#" ,2' '-2'!1T(&#+'6230#5 112#0'*'1#" 2&0-3%& RTVW=+170',%#$'*2#0 ," ""#"2-2&#!-0,+# *+#"' T(&# . 5 1 2&#, 12'00#" $-0 SR +',32#1 #$-0# .-30',% ',2- .#20' .* 2#1 =#** + ," )-$$#7S[ZWT

~~Preparation and sterilisation of Miracloth #'0 !*-2&) * '-!&#+Q7 -** Q) 5 1!32',2-Z!+"' +#2#0!'0!*#1 ,"2&# "'1!15#0#03,3,"#02 .5 2#0$-0TR+',32#1T(K#0#2&#,20 ,1$#00#"2-~~

-'*',% 5 2#0 $-0 SR +',32#1 ," 2&#, 0',1#" ', 1"T> $-0 SR +',32#1T (&'1 -'*',% ,"0',1',%.0-!#115 10#.# 2#"2&0##2'+#12-#,130#2&#.-0#1-$2&# !*-2&& "$3**7#6. ,"#"T(&##'0 !*-2&5 12&#,.* !#"5'2&',%* 11.#20'

~~.* 2#1 + 6'+3+ SZ !'0!*#1 .#0 .* 2# ," !-4#0#" 5'2& TR +7 1"T>T (&# .* 2#15#0#50 ..#"',25-* 7#01-$ *3+','3+$-'* ,"12#0'*'1#" 7 32-!* 4#T~~

~~202~~

~~Appendix~~

~~Appendix 2.3. Phytophthora DNA identification~~

~~Table A1: Phytophthora – specific PCR~~

~~PCR Reagents x1 reaction (L) x6 reaction (L) sdH2O 15.3 91.8~~

~~10x PCR Buffer 2.0 12.0~~

~~MgCl2 (25 mM) 1.0 6.0 A2_Forward primer 0.5 3.0 5’ ACTTTCCACGTGAACCGTTTCAA 3’ I2_Reverse primer 0.5 3.0 5’ GATATCAGGTCCAATTGAGATGC 3’ dNTPs (2.5M each, total 10 M) 0.5 3.0~~

~~Taq (1 unit) 0.2 1.2~~

~~+ gDNA~~

~~Total Volume 20.0 120~~

~~Table A2: P. cinnamomi restriction digest~~

~~MspI (L) RsaI (L) TaqI (L) dH2O 2.5 dH2O 2.5 dH2O 1.5~~

~~NE Buffer 2 1.0 NE Buffer 2 1.0 NE Buffer 3 1.0~~

~~BSA 1.0~~

~~MspI 0.5 RsaI 0.5 TaqI 0.5~~

~~PCR mix 6.0 PCR mix 6.0 PCR mix 6.0~~

~~Incubate at 37°C Incubate at 37°C Incubate at 65°C~~

~~203~~

~~Appendix~~

~~Appendix 2.4. The Application of Koch’s Postulates to Plant Pathology 0-5,S[[Y~~

ST (&# -0% ,'1+ +312 # !-,1'12#,2*7 11-!' 2#" 5'2& 2&# "'1# 1# 17+.2-+1T TT (&# -0% ,'1+ 11-!' 2#" 5'2& 2&# "'1# 1# +312 # '1-* 2#" ', .30# 6#,'!!3*230#T UT # *2&7.* ,21-$2 +#1.#!'#1 ,"!3*2'4 01&-3*" #',-!3* 2#" 131.#!2#" . 2&-%#,T *$ 2&# -0% ,'1+ '1 2&# . 2&-%#,Q '2 5'** ',$#!2 2&# .* ,2 ,"0#.0-"3!#17+.2-+11'+'* 02-2&-1#-,2&#.* ,2$0-+5&'!& '25 1'1-* 2#"T VT (&# . 2&-%#,1&-3*" 2&#, #0#'1-* 2#"$0-+ 2&# 02'$'!' **7',-!3* 2#" "'1# 1#"2'113#T*$2�#'1-* 2#"-0% ,'1+'12 +# 12&#-,#'1-* 2#" $0-+2&#-0'%', *"'1# 1#".* ,22&#,=-!&_1.-123* 2#1& 4# ##, 1 2'1$'#" ," 2&# -0% ,'1+ & 1 ##, .0-4#" 2- # 2&# ! 31# -$ 2&# "'1# 1#T

~~204~~

~~Appendix~~

~~Appendix 2.5. Solutions and wash conditions for Northern Hybridisation~~

~~1M Sodium Phosphate Buffer pH 7.2~~

~~XZTV+7S#< T>V"'7"0 2#SVST[W% 7~~

~~USTX+7S#< T>V"'7"0 2#SWX% 7~~

~~Church Buffer SR+7S#-"'3+&-1.& 2#3$$#0 SR+7SR$5 4~~

~~Membrane washes:~~

(x',%#,!7-$+#+ 0 ,#5 1'1"#.#,"#,2-, !)%0-3," ,"1'%, * *#4#*1"#2#!2#"31',% /#'%#0!-3,2#0Q2&#&'%'%, *Q2&#+-0# 120',%#,22 1 #/3'0#"T*,%#,#0 *Q&'% *2!-,2#,25'2&',2 1& 1-*32'-,10#13*21', +-0#120',%#,25 1&T

~~2 ," 0"5 1&!-,"'2'-,131#"',2&'1123"7$-0+#+ 0 ,#15'2& !-3,2-$0SW !.1 0#*'12#"S~~

~~TK)QRTS$ SW+',0--+2#+. TK)QRTS$ SW+',WW5)0-!)',% RTTK)QRTS$ SW+',WW5)0-!)',%~~

~~Wash solutions 2X SSC / 0.1% SDS (1 L) TRK) SRR+7 U#< )*QURR+#20'1-"'3+!'20 2#.Y5'2&)* SR$ SR+7~~

~~"T> [RR+7~~

~~205~~

~~Appendix~~

~~Appendix 3.0. “Pineapple” RNA Extraction Procedure ) 88-,#**' et al. S[[ZT~~

('113#5 1&-+-%#,'1#"3,"#0*'/3'" ,'20-%#,', .0#V!--*#"+-02 0Q 2&#&-+-%#, 2#5 12&#,20 ,1$#00#"',2-SW+7!#,20'$3%#23 #!-,2 ',',%V +7-$#620 !2'-, 3$$#0 ,"4-02#6#"$-0T+',32#1TS+7-$SRR$#2& ,-*5 1 ""#" ," 2&# 1-*32'-, 4-02#6#" $-0 $302�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�#13*2 ,2.#**#25 15 1&#" 5'2&W+7 ZR$ #2& ,-*Q 2&# 23 #',4#02#"-,!# #$-0# #',%!#,20'$3%#" 2URRR6%$-0W+', 2V5)T(&# 13.#0, 2 ,2 5 1 0#+-4#" ," 2&# 23 # ',4#02#" ," **-5#" 2- '0 "07T (&#

.#**#2 5 1 2&#,0#131.#,"#"', U+7 9)V20# 2#""T>QS+7-$ Z#*'2&'3+ !&*-0'"# 5 1 2&#, ""#" ," 2&# 131.#,1'-, ',!3 2#" -4#0,'%&2 2 VTR5)T*2 5 12&#,!#,20'$3%#" 2URRR6%$-0SW+',T(&#.#**#25 1 5 1&#"',Y+7 ZR$#2& ,-*Q',4#02#"2-+'6 ,"!#,20'$3%#" 2URRR6%$-0W+',32#1T(&# #2& ,-* 5 1 0#+-4#" ," 2&# 23 # 5 1 0'#$*7 1.3,Q #$-0# 2&# .#**#2 5 1

0#131.#,"#"',URR=7-$9)V20# 2#""T>TUR=7-$U#1-"'3+ !#2 2# ," XRR =7 -$ SRR$ #2& ,-* 5 1 ""#" ," 2&# 23 # ',!3 2#" 2 VZR5) $-0 UR +',32#1T( #5 12&#,!#,20'$3%#"$-0UR+',32#1 2 V5)Q2&#.#**#2 5 1 5 1&#" ', ZR$ #2& ,-* ," "0'#" 3,"#0 4 !33+ *',#T *2 5 1 2&#,

~~0#131.#,"#"',WR=79)V20# 2#""T>T~~

~~206~~

~~Appendix~~

Reagents required for Pineapple extraction method: U#-"'3+ !#2 2# V[TT%',TRR+7 W#-2 11'3+ !#2 2# Z#7'2&'3+!&*-0'"# UUT[%',SRR+7 Extraction Buffer (100 mL) SW+7-$S#(0'1.YTW5'2&1-*'" -0'! !'" TR+7-$SR$ SR+7-$RTW#9(

~~207~~

~~Appendix~~

Appendix 4.0. Microarray data: Differentially expressed genes: ** %#,#1 5&'!& 5#0# '"#,2'$'#" 1 #',% "'$$#0#,2' **7 #6.0#11#" ', P. cinnamomi',-!3* 2#"Z. mays0--22'113# 0#1&-5,',2&# $-**-5',%2 *#1 ," ',!*3"# 2&-1# %#,#1 5&'!& 5#0#,-2$3,!2'-, **7 ,,-2 2#"T (# 2 *#1 5#0#%#,#0 2#"$-**-5',%,-0+ *'1 2'-, ,"4'13 *'1 2'-,-$" 2 -32.32Q 15#** 112 2'12'! * , *71'1T

~~208~~

~~Appendix~~

UniGene ID Archival Un Zm.9845 Zm.9845 Zm.15827 Zm.15827 Zm.14496 Zm.87786 ---Zm.8610Zm.9297 Zm.8610 --- Zm.19364 Zm.9297 Zm.186Zm.10304 Zm.10304 Zm.186Zm.6659 --- Zm.503 Zm.6659 Zm.8726 Zm.66876Zm.9731 Zm.8726 Zm.9093 Zm.9731 Zm.864 Zm.9093 --- Zm.864Zm.7654--- Zm.7654 ------Zm.9644Zm.847 Zm.97592 --- Zm.10676 --- Zm.93771 Zm.33576 Zm.9932 Zm.9932 ---Zm.2227 Zm.2227 Zm.9106 Zm.9106 Zm.10167 Zm.95010 Zm.94188 Zm.14663 Zm.75213 Zm.5882Zm.1450 Zm.94767 Zm.1450 Zm.281Zm.2301 Zm.281 Zm.2301 Zm.16272 Zm.16272 Zm.8714 Zm.8714 6.921 7.5103 6.8414 5.5021 5.0415 4.1642 4.0999 4.07955 18.0563 9.96864 9.33135 9.02707 7.96169 7.72908 7.71519 7.11864 6.95224 6.76019 5.97588 5.53548 5.32392 5.29398 5.28386 5.15265 5.00964 4.97005 4.89232 4.85222 4.80964 4.67314 4.52037 4.22114 4.13803 4.08572 00275091 cyclase Umi3 0. Probeset ID Gene Title Symbol p-value Fold-Change Zm.15827.1.A1_at ------0.000922634 Zm.14496.1.A1_at putative sesquiterpene ZmAffx.12.1.S1_at kaurene synthase2 an2 9.11E-05 Zm.8610.1.A1_atZm.9297.1.A1_atZm.2136.1.S1_at ---Zm.186.1.S1_at ---Zm.10304.1.S1_at ---Zm.6659.1.A1_at induced wound Zm.503.1.A1_at protein ---Zm.8726.1.A1_at pathogenesis related protein-5Zm.9731.1.A1_at germin-likeZm.9093.1.A1_at protein 2 ---Zm.864.1.S1_at --- PR-5Zm.7909.1.A1_at --- WIP1Zm.7654.1.A1_at glutathioneZm.9486.1.A1_at transferase30 ---Zm.10586.1.A1_at 0.00173186 ---Zm.9644.1.A1_at 0.00128992 ------GLP2Zm.847.1.S1_at ------Zm.10676.1.S1_at ------gst30Zm.9932.1.A1_at acidicZmAffx.1198.1.A1_at --- 0.00208655 class 12-oxo-phytodienoic chitinase I acid reductaseZm.2227.1.A1_at ------OPR1 5.95E-05Zm.9106.1.A1_at 0.000208714 5.73E-05Zm.10167.1.A1_at ------1.07E-05Zm.14663.1.S1_s_at ------Zm.5882.1.A1_at ------0.0106663 --- 1.56E-05 uiu5(chn)Zm.1450.1.S1_atZm.281.1.S1_s_at ------Zm.2301.1.A1_at --- 0.0268917 ---Zm.16272.1.A1_at 9.72E-05 hypothetical LOC542299 --- 0.0424054 Zm.8714.1.A1_at --- 0.00278029 ---Zm.9845.1.S1_at ------acc oxidase 4.57E-05 --- 2.79E-05 --- LOC542299 --- 2.28E-06 4.00E-07 0.00150635 0.000285129 ------0.000124044 0.00525066 --- 0.000342651 --- ACO31 0.000635517 0.0206015 --- 7.13E-05 --- 0.00109082 0.000240203 1.01E-05 --- 0.000332024 0.0001669 0.000307602

~~209~~

~~Appendix~~

Zm.9554Zm.1871 Zm.67119 Zm.539 Zm.1871 --- Zm.539 Zm.1085Zm.16538 Zm.95692 Zm.16538 Zm.12612 --- Zm.12612 Zm.3332 Zm.67113 ---Zm.1595Zm.11924 Zm.92797 Zm.11924 Zm.14677 --- Zm.14677 Zm.6023--- Zm.97935 --- Zm.4270 Zm.92782 Zm.627--- Zm.627 Zm.8822Zm.1506 Zm.96646 --- Zm.38616 --- Zm.11838 Zm.11838 Zm.7687--- Zm.7687 --- Zm.11191 Zm.11191 Zm.17181 Zm.17181 ------Zm.10022 Zm.10022 Zm.10175 Zm.10175 Zm.7385 --- Zm.13232 Zm.97414 Zm.98532 Zm.4213Zm.8525 --- Zm.18148 Zm.82751 Zm.84292 Zm.17110 Zm.17110 ------4.00336 3.99237 3.90878 3.89262 3.85752 3.76774 3.74985 2.79094 2.78488 2.76185 2.74021 2.73939 2.73321 2.72485 3.68179 3.62593 3.54451 3.52271 3.47912 3.41329 3.35335 3.34417 3.21226 3.21064 3.18423 3.15552 3.06119 3.05272 3.04739 2.96468 2.90528 2.88906 2.81048 2.80949 2.80512 0.000297671 LOC1000378 IDP370 0.0190955 putative Bowman-Birk serine protease putative serine type endopeptidase inh Zm.9554.1.A1_at ------8.75E-05 Zm.1871.1.A1_atZm.539.1.A1_atZm.18344.1.A1_at ---Zm.1085.1.A1_a_at glutathioneZm.16538.1.S1_at S-transferase --- 8 PRm 3Zm.12612.1.A1_atZm.4270.2.A1_a_at ------GST8 0.00164418 --- chitinase ------2.93E-06 0.000177555 --- 2.69E-05 0.00014183 0.00684432 Zm.12518.1.A1_at ------0.0012928 Zm.1595.1.S1_atZm.11924.1.A1_atZm.14677.1.S1_at ---Zm.6023.1.S1_at ---Zm.17271.7.A1_x_at ------alternative oxidase precursor AOX3 aox3 0.00369429 ------0.0291738 2.02E-06 1.86E-05 0.00439605 Zm.627.1.A1_at glutathione S-transferase 24 GST LOC541837 0.000707703 Zm.14226.1.A1_atZm.8822.2.A1_atZm.1506.1.A1_at ---Zm.17271.6.S1_x_at ---Zm.11838.1.A1_at --- defence-related proteinZm.7687.1.A1_atZm.10238.1.A1_at ---Zm.11191.1.A1_at ---Zm.17181.1.S1_at ---Zm.7734.1.A1_at ---Zm.10022.1.A1_at --- inra2(prp)Zm.10175.1.A1_at ---Zm.7385.1.A1_at --- 0.001381Zm.13232.1.S1_at ---Zm.4213.1.A1_at ------Zm.8525.1.A1_at ---Zm.18148.1.A1_at --- IDP158 protein ---Zm.17110.1.S1_at ---Zm.9615.1.A1_at ------Zm.3332.1.A1_at --- 3.35E-05 ------0.000254451 0.0169599 ------0.000180089 --- 0.00014232 --- IDP158 0.0112502 --- 0.000136075 2.37E-05 ------4.88E-06 0.00138689 4.90E-07 --- 0.00137536 --- 0.000425648 --- 0.00196578 --- 0.000290231 1.21E-05 0.00449402 0.000897051

~~210~~

~~Appendix~~

Zm.10841 Zm.10841 Zm.17971 Zm.17971 Zm.8697Zm.17344 Zm.93793 Zm.17344 Zm.1419 Zm.67083 ---Zm.10101 Zm.83288 Zm.11900 Zm.11900 Zm.13778 --- Zm.82174 Zm.10134 Zm.10134 Zm.15084 Zm.30910 Zm.15532 Zm.98798 Zm.6105--- Zm.6105 Zm.9646Zm.8743 Zm.9646 Zm.14951 Zm.8743 --- Zm.97193 Zm.1663Zm.311 Zm.1663 Zm.12227 Zm.94463 Zm.94787 ---Zm.3225Zm.16973 Zm.81275 Zm.16973 Zm.9851 --- Zm.17644 Zm.9851 Zm.17644 Zm.11645 Zm.11645 Zm.11950 Zm.41285 Zm.11809 Zm.84239 Zm.4340--- Zm.82337 Zm.10103 Zm.10103 --- Zm.8470Zm.14951 Zm.8470 Zm.94358 Zm.9687 Zm.9687 Zm.8357 Zm.97498 2.5409 2.4853 2.4808 2.2577 2.1849 2.1458 2.14203 2.13767 2.13553 2.13268 2.13036 2.68023 2.67666 2.65945 2.62684 2.47153 2.46715 2.45956 2.45223 2.44928 2.39649 2.38842 2.38415 2.36866 2.36056 2.31995 2.28675 2.28538 2.28359 2.25754 2.23402 2.21567 2.18541 2.16767 2.14715 Hds1 0.000103437 hydroxymethylbutenyl 4-diphosphate s Zm.17971.1.A1_atZm.8697.1.A1_atZm.17344.1.S1_at ---Zm.1419.1.S1_at ------0.000331455 0.0156986 0.0113104 0.00245133 Zm.1601.1.S1_at ------1.80E-05 Zm.10101.1.A1_atZm.11900.1.A1_atZm.13778.1.A1_at ---Zm.10134.1.A1_at ------0.000273056 0.00147448 0.00715255 0.000282685 Zm.15084.1.A1_at ------0.0206251 Zm.15532.1.S1_at ------0.000254608 Zm.6105.1.S1_atZm.8478.1.A1_a_atZm.9646.1.A1_at ------Zm.8743.1.A1_atZm.14951.1.S1_a_at ---Zm.1663.1.A1_at cell wall protein (put.); --- putativeZm.311.1.A1_atZm.12227.1.S1_at ---Zm.5748.1.A1_at LOC542268 18 kda heat shockZm.3225.1.S1_at --- protein18a 0.0110077 Zm.16973.1.S1_at ---Zm.9851.1.A1_at ---Zm.17644.1.A1_at --- hsp18aZm.11645.1.A1_at ---Zm.11950.1.A1_at ------0.00354126 ------0.00187236 0.0391875 --- 0.00182882 --- 0.000288103 ------0.00419915 ------0.00105413 --- 0.00248257 --- 0.000296424 --- 0.000163213 0.0294903 0.0254645 6.74E-06 1.03E-05 Zm.11809.1.A1_atZm.4340.1.A1_atZm.11803.1.A1_at ---Zm.10103.1.A1_at pericarp2 orange Zm.8470.1.S1_at ------sucrose-phosphatase orp2 LOC542661 0.00606355 0.000991058 ------0.0270305 0.00229056 0.0269331 Zm.14951.2.S1_x_atZm.9687.1.A1_at ---Zm.8357.1.A1_at ------0.00840631 0.024307 0.041856 Zm.10841.1.A1_at

~~211~~

~~Appendix~~

Zm.12391 Zm.95681 Zm.8365 Zm.8365 ---Zm.1995Zm.2707 Zm.85422 Zm.15533 Zm.98559 --- Zm.15533 Zm.11214 Zm.78825 Zm.7515Zm.17613 Zm.7515 Zm.17613 Zm.16992 Zm.12316 ---Zm.1956 Zm.1956 ------Zm.13228 Zm.13228 Zm.17809 Zm.28212 Zm.246 --- Zm.9101 Zm.68178 Zm.551 Zm.9101 Zm.10830 Zm.551 Zm.10830 Zm.13645 Zm.66926 ---Zm.17589 Zm.98093 Zm.9699Zm.4051 Zm.9699 --- Zm.9783 Zm.4051 Zm.9783 Zm.15278 Zm.97740 Zm.10136 Zm.10136 Zm.9562Zm.11657 Zm.96728 Zm.79170 Zm.11679 Zm.11679 Zm.18269 Zm.21324 Zm.7275--- Zm.97181 Zm.9765 Zm.9765 ------1.872 1.809 2.0997 2.0047 1.8066 1.79588 2.12867 2.12699 2.12462 2.10018 2.07884 2.07876 2.05926 2.04329 2.02251 1.99009 1.96863 1.95267 1.94047 1.93477 1.93373 1.91407 1.91161 1.90021 1.89822 1.88081 1.87417 1.86884 1.85345 1.85217 1.84903 1.84157 1.82792 1.81139 1.79958 Zm.8365.1.A1_at ------0.0196983 Zm.19319.1.A1_at ------0.000186191 Zm.1995.1.S1_atZm.2707.1.S1_atZm.15533.1.S1_at ---Zm.11214.1.S1_at ------0.00182109 0.00475303 0.00770767 0.00102758 Zm.7515.1.A1_atZm.17613.1.A1_atZm.16992.1.A1_at ---Zm.8478.2.A1_x_at ---Zm.1956.1.S1_at ------Zm.10172.1.A1_at ------0.0031704 0.00196259 9.17E-05 0.0028828 0.0012968 0.00289541 Zm.13228.1.S1_atZm.17809.1.A1_atZm.246.1.A1_at ---Zm.9101.1.A1_at ---Zm.551.1.A1_at glutathione transferase7Zm.10830.1.S1_at ---Zm.13645.1.S1_at glutathione S-transferaseZm.17263.1.A1_at --- 25 GST Zm.17589.1.A1_at tryptophan synthase alpha LOC541838Zm.9699.1.A1_at ---Zm.4051.1.A1_at --- gst7 0.00767372 Zm.9783.1.A1_at ---Zm.15278.1.S1_at --- TSA ------0.0305949 ------0.000462491 --- 0.00270576 --- 0.0227401 --- 0.00426532 ------0.0398208 ------0.00265462 --- 0.00149591 0.00513705 0.0384784 0.00357059 0.00406683 Zm.10136.1.A1_atZm.9562.1.S1_atZm.11657.1.A1_at ---Zm.11679.1.A1_at aldehyde oxidase-2Zm.18269.1.S1_at ---Zm.7275.1.A1_at ---Zm.9675.1.A1_at ---Zm.9765.1.A1_at ---Zm.15170.1.A1_at ------zmAO-2 0.0394588 ------0.000603759 ------0.00916223 --- 0.00429384 --- 0.00740585 0.00060184 0.000348052 0.00363255 0.00124954 Zm.12391.1.S1_at ------0.00156207

~~212~~

~~Appendix~~

Zm.1449Zm.10135 Zm.103764 Zm.10135 Zm.18453 Zm.76257 Zm.13494 Zm.13494 ---Zm.2139Zm.3216 Zm.84867 Zm.9842 Zm.73783 ------Zm.9842 Zm.4089Zm.3278 Zm.4089 Zm.13583 Zm.83895 --- Zm.95781 Zm.4190--- Zm.4190 Zm.9811 Zm.9811 --- Zm.12014 Zm.12014 Zm.10827 Zm.99318 Zm.10003 Zm.10003 Zm.9049Zm.9205 Zm.37546 Zm.9205 ------Zm.10623 Zm.75749 Zm.9654 --- Zm.12216 Zm.9654 Zm.25820 Zm.67220 ---Zm.12129 Zm.12129 Zm.1079 Zm.93673 --- Zm.11612 Zm.11612 Zm.14674 Zm.14674 Zm.9833Zm.807 Zm.9833 Zm.807 Zm.3371Zm.4407 Zm.17702 Zm.13424 Zm.4407 Zm.13424 1.743 1.6553 1.6909 1.6308 1.75445 1.75038 1.74709 1.74236 1.73482 1.72345 1.71971 1.70933 1.70545 1.70255 1.69093 1.68463 1.68414 1.67355 1.67345 1.66455 1.66383 1.65681 1.65277 1.64763 1.64347 1.64047 1.63514 1.63487 1.63252 1.78475 1.77807 1.77219 1.77125 1.62655 1.62602 0.0490061 LOC1001354 APRL1 0.00881366 cysteine S-methyltransferase-1 LOC541873cysteine 0.0476552 adenosine 5'-phosphosulfate reductase Zm.10135.1.A1_atZm.18453.1.A1_atZm.13494.1.S1_at ---Zm.17954.1.A1_at ---Zm.2139.1.S1_at ---Zm.3216.1.S1_at ---Zm.9842.1.A1_at ---Zm.1327.1.A1_at ---Zm.4089.1.S1_at ---Zm.3278.1.S1_at ---Zm.13583.1.S1_at ---Zm.4190.1.A1_at ---Zm.16860.1.A1_at ---Zm.9811.1.A1_at ---Zm.12014.1.A1_at ------0.00165232 --- 0.0147278 --- 0.000152949 0.040208 ------0.0131035 --- 0.00250433 --- 0.000910208 --- 0.00933311 --- 0.00603142 --- 0.000215393 0.00802737 0.00122103 0.0076197 0.000804853 Zm.10827.1.S1_atZm.10003.1.A1_atZm.9049.1.A1_x_at ---Zm.9205.1.A1_at ------Zm.11758.1.A1_at ------0.00381191 0.00669933 0.0121194 0.0232843 0.0115807 ZmAffx.71.1.A1_atZm.10623.1.S1_atZm.9654.1.A1_at ---Zm.12216.1.S1_at ---Zm.18470.1.A1_at ---Zm.12129.1.S1_at ---Zm.1079.3.A1_at ---Zm.3371.1.A1_at ------0.0476503 --- 2.52E-05 --- 0.000502483 0.0011801 --- 0.00642912 0.00557447 0.00267611 0.0183668 Zm.11612.1.A1_at ------0.000542754 Zm.14674.1.S1_atZm.9833.1.A1_atZm.807.1.A1_at ---Zm.1449.2.S1_a_at --- early drought induced protein ------0.00484583 0.0192287 0.0042614 Zm.4407.1.A1_atZm.13424.1.A1_at --- homo --- 0.0228351

~~213~~

~~Appendix~~

Zm.27Zm.9556Zm.6107 Zm.9556 Zm.27 Zm.545 Zm.93878 Zm.298 Zm.97225 Zm.13229 Zm.298 Zm.13229 Zm.168Zm.12139 Zm.168 Zm.85848 Zm.14906 Zm.80294 Zm.9368Zm.1461 Zm.9368 --- Zm.1461 Zm.13534 Zm.85488 Zm.10850 Zm.10850 --- Zm.6108Zm.1323 Zm.94314 Zm.2538 Zm.1498 Zm.1498 Zm.561Zm.9749 Zm.561Zm.409 Zm.9749 Zm.409 ---Zm.6685Zm.8326 Zm.79179 Zm.12293 Zm.8326 --- Zm.12293 ---Zm.1315--- Zm.37546 Zm.1579 --- Zm.8307 Zm.67898 Zm.12208 Zm.8307 --- Zm.85863 Zm.18116 Zm.18116 Zm.445Zm.14951 Zm.94716 Zm.97193 Zm.3443Zm.11196 Zm.3443 Zm.11196 1.5756 1.5641 1.6127 1.5447 1.5092 1.60675 1.59553 1.58511 1.58093 1.57883 1.57594 1.56949 1.56908 1.62362 1.62175 1.62139 1.61948 1.61858 1.56135 1.55285 1.61224 1.61074 1.54096 1.53934 1.53818 1.53686 1.53173 1.52638 1.52277 1.52041 1.51565 1.50898 1.50425 1.50344 1.50221 hm1 0.0038633 Helminthosporium carbonum susceptib Zm.9556.1.A1_atZm.6107.1.A1_atZm.545.1.S1_at ---Zm.298.1.S1_a_at allene oxide synthaseZm.13229.1.S1_at glutathione S-transferaseZm.168.1.A1_at dxr protein 15 GST --- LOC542630 0.000432673 protein kinase1 aos 0.000155218 IDP154 --- ptk1 0.00252354 --- 0.00454327 0.000323845 0.00345993 Zm.12139.1.A1_atZm.14906.1.A1_s_atZm.6108.2.S1_at ------0.000678533 0.0119234 0.00881752 Zm.9368.1.A1_at ------0.0234297 Zm.1461.1.A1_atZm.8617.1.A1_atZm.13534.1.A1_at ---Zm.10850.1.S1_at ---Zm.561.1.A1_at ------glutathione transferase23 gst23 0.000321693 ------0.000362609 8.34E-05 5.46E-05 0.0296482 Zm.1323.1.S1_x_atZm.2538.1.A1_at ---Zm.10590.1.S1_at ------0.0386111 0.000547302 0.0155613 Zm.9749.1.S1_a_atZm.409.1.S1_at ---Zm.27.1.S1_at cell wall invertase1 incw1 --- 0.0205222 0.0457608 Zm.6685.1.A1_atZm.8326.1.A1_atZm.12293.1.S1_at ---Zm.9880.1.S1_at ---Zm.1315.1.A1_at ---Zm.19027.1.A1_at ---Zm.1579.1.S1_at ---Zm.8307.2.A1_a_at ---Zm.12208.1.A1_at ------Zm.18116.1.S1_at ------0.0020722 0.000622376 ------0.0041293 --- 0.0324171 --- 0.0387305 0.00185456 0.000358291 0.0460041 0.041789 0.0162769 Zm.445.1.S1_atZm.14951.1.S1_x_atZm.3443.1.A1_at cell wall protein (put.); putative LipoxygenaseZm.11196.1.A1_at ------LOC542268 0.0344627 LOC542495 0.0402037 ------0.045068 0.0268729

~~214~~

~~Appendix~~

root tissue 6 at change) Blue indicates --- Zm.23815 Zm.10740 Zm.66923 Zm.11650 Zm.84700 Zm.14805 Zm.76943 Zm.7882 Zm.7882 Zm.228Zm.16206 Zm.94938 Zm.228Zm.17735 Zm.17735 Zm.8944Zm.759 Zm.8944 Zm.9563 Zm.117536 Zm.18090 Zm.9563 Zm.96533 Zm.7012 Zm.72311 Zm.9879 Zm.18280 Zm.17208 Zm.17208 ------Zm.13239 Zm.77645 Zm.3473 Zm.94529 ------Zm.202 Zm.202 Zm.17781 Zm.17781 Zm.674 Zm.95898 Zm.5608 Zm.66854 Zm.8136 Zm.8136 - Z. mays -1.5569 -1.5634 1.50128 1.50045 1.50018 -1.50514 -1.50522 -1.51077 -1.52359 -1.53304 -1.53883 -1.53896 -1.54933 -1.56173 -1.57177 -1.58639 -1.63822 -1.67825 -1.70468 -1.71635 -1.74011 -1.77725 -1.90719 -1.97882 -2.09909 llin responsive1llin gar1 0.00289269 zoospores, after being subjected to statistical analysis. Includes genes which are not genes statistical which are to analysis. being subjected Includes after zoospores,

All genes identified in microarray analysis as being differentially expressed in in expressed differentially being as analysis microarray in identified genes All P. cinnamomi P. -regulated, red indicates down-regulated genes. hpi with hpi with genes up functionally annotated, unigene assignments arefunctionally shown annotated, in columns 6 & 5 (fold 7. Column Table A3: Zm.9879.2.S1_at ------0.0122071 Zm.17208.1.S1_at ------0.0247891 Zm.11650.1.A1_at ------0.00145674 Zm.14805.1.A1_atZm.7882.1.A1_atZm.228.1.S1_at ------anther ear1 an1 ------0.0496119 0.0027204 0.0231977 Zm.16206.1.S1_atZm.17735.1.S1_atZm.8944.1.S1_at ---Zm.759.1.A1_at ---Zm.9563.1.A1_at ---Zm.18090.1.A1_at ---Zm.7012.8.S1_a_at ---ZmAffx.86.1.S1_at --- Ribosomal protein S8 --- LOC542341 --- 0.0132247 ------0.0102925 --- 0.0115442 --- 0.0349333 0.0270835 0.0341845 0.00322682 4.85E-05 Zm.10740.1.S1_a_at ---Zm.7818.1.A1_at ------0.00379987 0.0332336 Zm.4667.1.A1_at ------0.0196532 Zm.13239.3.S1_at ------0.0306603 Zm.3473.1.S1_at ------0.00883885 ZmAffx.1465.1.S1_at ------0.00344642 Zm.202.1.S1_at gibbere Zm.17781.1.A1_at ------0.0256939 Zm.674.1.S1_at beta-expansin 6 expB6 0.0301653 Zm.5608.1.S1_a_at ------0.0168658 Zm.8136.1.A1_at ------0.019065

~~215~~

~~Appendix~~

UniGene ID Zm.503Zm.186Zm.1450--- Zm.66876 Zm.4524 Zm.186Zm.375 Zm.1450 Zm.9845Zm.1506 Zm.79619 --- Zm.93761 Zm.9845 Zm.38616 Archival UniGen Zm.7566Zm.13494---Zm.2227 Zm.7566 Zm.13494 Zm.16805Zm.1085Zm.2707 Zm.2227 Zm.67336 Zm.704 --- Zm.2549 Zm.95692 Zm.9953 Zm.98559 --- Zm.94357 Zm.898 Zm.2549 Zm.10175 Zm.33549 Zm.8557--- Zm.33267 Zm.10175 Zm.13583 --- Zm.2551 Zm.8557 Zm.95781 Zm.2551 --- Zm.4270 Zm.92782 Zm.16805Zm.16805Zm.301 Zm.67336 Zm.7654 Zm.67336 Zm.2301Zm.10103 Zm.72638 Zm.1921 Zm.7654 --- Zm.2301 Zm.10103 Zm.95951 --- 2.001 3.3785 3.32251 3.14378 2.98573 2.92918 2.70306 2.40141 2.32565 1.96946 1.94516 1.94413 1.92963 1.89744 1.88236 1.86618 1.85549 1.81315 1.77592 1.76921 1.75285 1.74653 1.74296 1.69853 1.69376 1.67616 2.31941 2.25614 2.23941 2.23908 2.18306 2.13059 2.06586 2.04797 Fold-Change LOC100037814 0.012006 ------putative Bowman-Birk serine protease inhibito ------0.020316 0.045816 Zm.186.1.S1_atZm.1450.1.S1_at induced wound proteinZm.9486.1.A1_at ---Zm.4524.1.A1_at ---Zm.375.1.S1_at ---Zm.9845.1.S1_at trypsin inhibitorZm.1506.1.A1_at --- defence-related protein WIP1 0.021773 inra2(prp) LOC541688 ------0.011887 0.023595 ------0.001424 6.10E-05 0.005172 0.005524 Zm.503.1.A1_at germin-like protein 2 GLP2 0.024225 Zm.13494.1.S1_at ---Zm.2136.1.S1_atZm.2227.1.A1_at ---Zm.16805.3.S1_at ------Zm.1085.1.A1_a_at PRm 3Zm.2707.1.S1_atZm.704.1.S1_at ---Zm.2549.1.A1_at xyloglucan endotransglycosylaseZm.9953.1.S1_at homolog1 --- xet1Zm.7909.1.A1_at ---Zm.898.1.A1_a_at ------Zm.10175.1.A1_at ---Zm.8557.1.A1_atZm.8327.1.A1_at ---Zm.13583.1.S1_at 0.045771 ------Zm.2551.1.A1_at ------chitinase ------0.000649 0.019036 0.014563 ------0.04066 0.026134 ------0.012565 --- 0.030435 ------0.002023 0.034892 --- 0.020512 0.043674 0.036889 0.004741 0.007897 0.013171 Zm.7566.1.A1_at ------0.01636 Zm.301.1.A1_a_at homolog1 inhibitor subtilisin-chymotrypsin Zm.7654.1.A1_at sci1Zm.2301.1.A1_at ---Zm.10103.1.A1_at ------Zm.1921.1.S1_atZm.12518.1.A1_at inorganic phosphate transporter 3 ---Zm.4270.2.A1_a_at 0.026131 LOC732717 0.038114 ------0.0039 0.006784 0.030864 0.009563 Probeset IDProbeset Gene TitleZm.16805.3.S1_x_at Zm.16805.3.S1_a_at Symbol p-value

~~216~~

~~Appendix~~

Zm.10209Zm.9101Zm.11698 Zm.15730 ---Zm.11732 Zm.9101 Zm.11698 Zm.6681Zm.13248 Zm.11732 Zm.2477Zm.507 Zm.82526 --- Zm.13248 Zm.2477 Zm.507 ---Zm.3742Zm.4909Zm.1306Zm.1540 Zm.85052 Zm.4068 Zm.4909 --- Zm.4068 Zm.95653 Zm.16890 Zm.1540 Zm.4068 Zm.4068 Zm.93688 Zm.8996Zm.3633 Zm.8996 Zm.92562 Zm.5424Zm.3561Zm.6806Zm.6756 Zm.5424 Zm.13168 Zm.24529 --- Zm.81479 Zm.4672 Zm.95227 Zm.85554 Zm.10097 Zm.7574 Zm.10097 --- Zm.11838Zm.16941 Zm.11838 Zm.16941 Zm.7882 Zm.7882 ---Zm.11214---Zm.15278 Zm.78825 Zm.3225 --- Zm.97740 Zm.81275 Zm.77072 1.524 -1.506 1.5573 -2.3651 1.52184 1.51596 1.51179 1.55481 1.54088 1.66822 1.66575 1.63341 1.60666 1.56945 -1.51504 -1.54775 -1.55756 -1.56373 -1.57023 -1.60573 -1.61193 -1.61218 -1.63398 -1.64012 -1.69671 -1.71463 -1.72254 -1.75548 -1.57366 -1.80492 -1.83186 -1.86942 -2.11027 -2.24535 -2.28599 ------0.041419 Zm.9101.1.A1_atZm.11698.1.A1_at ------Zm.15895.1.A1_at ---Zm.11732.1.A1_at ---Zm.6681.1.A1_atZm.13248.1.S1_at ------Zm.2477.1.A1_atZm.507.1.A1_at ---Zm.8996.1.A1_at phosphogluconate dehydrogenase3 --- pgd3 ------0.040183 ------0.037002 --- 0.007809 0.049537 --- 0.02661 0.019844 0.032399 0.007195 0.013403 Zm.3633.4.A1_at ------0.049658 Zm.3742.1.A1_atZm.4909.1.A1_at ---Zm.1306.1.S1_at synthase Terpene 8Zm.1540.1.S1_a_at --- alcohol dehydrogenase1Zm.4068.1.S1_atZm.4068.2.S1_x_at Ferredoxin-NADP reductase Ferredoxin-NADP precursor reductase precursorZm.16890.1.A1_at ---Zm.5424.1.S1_at LOC542713 LOC542713 --- 0.019577 adh1 0.012566 TPS8 0.009063 --- 0.03008 ------0.001386 --- 0.026118 9.14E-05 0.009517 Zm.3561.1.A1_atZm.6806.1.A1_at ------0.030463 0.046416 Zm.6756.1.S1_s_at ---Zm.13168.1.A1_at ---ZmAffx.442.1.A1_at ---Zm.4672.1.S1_atZm.10097.1.A1_at ------0.034209 0.00498 0.002912 0.039566 0.009284 Zm.16941.1.S1_at ---Zm.10209.1.A1_at ------0.038638 0.037325 Zm.11838.1.A1_at ------0.044124 Zm.11214.1.S1_at ---ZmAffx.685.1.S1_at ---Zm.15278.1.S1_at ---Zm.3225.1.S1_atZm.7882.1.A1_at ------0.008658 0.048272 0.016837 0.012807 0.01563 Zm.8478.1.A1_a_at ------ZmAffx.1462.1.S1_at 0.032672

~~217~~

~~Appendix~~

- root tissue root at 24 hpi Zm.131 Zm.131 Zm.15899 Zm.94770 ------Zm.4068 Zm.4068 Zm.13263Zm.125 Zm.13263 Zm.42640 Zm.3369 Zm.3369 Zm.11441 Zm.11441 Zm.12285 Zm.12285 Zm.13128 Zm.13128 Zm.15899Zm.15899Zm.15899 Zm.94770 Zm.94775 Zm.94775 Zm.485 Zm.485 Z. mays -2.5609 -3.69507 -4.08469 -2.42186 -2.46316 -2.55259 -2.79552 -3.05993 -3.17188 -3.46712 -4.12121 -5.84439 -6.55112 -4.95875 -change) Blue indicates genes up sum1 0.020303 -regulated genes. zoospores, after being subjected statisticalzoospores,to analysis. being subjected after not functionally which Includes are genes

siroheme uroporphyrinogen methyltransferase ------0.02913 0.017366 0.011824 All genes identified in microarray analysis as being differentially expressed in in expressed differentially being as analysis microarray in identified genes All P. cinnamomi regulated, red indicatesregulated, down annotated, unigene assignments are shown in columns 6 & 7. Column 5 (fold 5 7. Column 6 & in columns shown unigene are assignments annotated, Table A4: with Zm.15899.3.A1_at ------0.037849 Zm.10238.1.A1_at ---Zm.15899.3.A1_x_at Zm.15899.1.A1_a_at Zm.15899.4.A1_x_at --- 0.037914 Zm.4068.2.S1_a_at ferredoxin-NADP reductase precursor LOC542713 0.009311 Zm.13263.1.A1_at nitrite reductase gpm108b 0.029494 Zm.125.1.S1_atZm.3369.1.A1_at nitrate reductase (NAD(P)H) glutamine synthetase1 nar gln1 0.037208 0.006076 Zm.11441.1.S1_at ethylene-responsive factor-like protein 1 ERF1 0.049167 Zm.12285.1.A1_at ------0.033713 Zm.13128.1.A1_at ------0.047223 Zm.131.1.A1_at Zm.485.1.A1_at hemoglobin LOC541815 0.013503

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~~Appendix~~

~~Appendix 4.1: Visualisation tools for pre-processing microarray data ?'13 *'1 2'-,-$+'!0- 00 7" 2 5 1!-,"3!2#"31',% 02#)/#,-+'!1 3'2#5&'!&%#,#0 2#"&# 2+ .1$'%30# V ,"4-*! ,-.*-21T~~

~~1 2 3 4 5 6~~

Figure A.4. Heat map representing relative expression values of control (columns 1-3) and P. cinnamomi inoculated (columns 4-6) samples at 6 hpi. Each column represents a single technical replicate, comprising 15 biological replicate roots.

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