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Author M An and R S Z Hai Hong Bi Li Ming Su Shi Ming Luo Author Address mancsueduau Title Rice allelopathy induced by methyl jasmonate and methyl salicylate Year 2007 Journal Journal of Chemical Ecology Volume 33 Issue 5 Pages 1089-1103 Date May ISSN 0098-0331 DOI httpdxdoiorg101007s10886-007-9286-1 Keywords Methyl Salicylate Methyl Jasmonate Rice Allelopathy Echinochloa crus-galli Phenolic Acid Phenylalanine ammonia-lyase Cinnamate 4-hydroxylase Abstract Plants activate signalling system upon attack by insect herbivores and microbial pathogens Methyl jasmonate (MeJA) and methyl salicylate (MeSA) are important signalling molecules which are able to induce plant defence against insect herbivores and microbial pathogens This study tests the hypothesis that allelopathy is an active inducible defence mechanism of plants and JA and SA signalling pathways may activate the allelochemical release We found that exogenously applied MeJA and MeSA to rice (Oryza sativa L) plants enhanced rice allelopathic potentials and accumulation of phenolics increased enzymatic activities and gene transcription of PAL and C4H which are two key enzymes in the phenylpropanoid pathway Aqueous extracts of the leaves of rice IAC165 a putative allelopathic variety showed 25 21 increase in inhibitory effects on root growth of barnyard grass (Ehinohloa rus-galli L) and 18 23 increase in inhibitory effects on the shoot growth after rice plants were treated with MeSA (5 mM) and MeJA (005 mM) respectively compared with the corresponsive control Leaf aqueous extracts of rice Huajingxian1 a putative non-allelopathic variety treated with MeJA and MeSA showed 24 and 63 higher inhibition to the root length of barnyard grass seedlings respectively The root exudates of rice plants treated with MeJA (005 mM) and MeSA (5 mM) for 48h significantly increased their inhibitory effects on root growth of barnyard grass seedlings 3 4-hydroxybenzoic acid (HBA) vanillic acid (VA) coumaric acid (CMA) and ferulic acid (FA) in the leaves accumulated approximately to 53 313 22 and 17-fold higher levels in response to MeJA exposure and accumulated to 33 131 20 and 22-fold higher levels in response to MeSA At fourth leaf ageMeSA and MeJA treatments enhanced the PAL activity in rice leaves up to 523 and 801 and C4H activity increased by 402 and 67 respectively Gene transcription of PAL and C4H in rice leaves significantly increased after treated with MeJA and MeSA These results indicate that allelopathy is an active defence mechanism of plants and plant signals are potentially valuable in the regulation of allelopathy for competing with other plants Call Number CSU281931

Rice allelopathy induced by methyl jasmonate and methyl salicylate

Hai Hong Bi12 Ren Sen Zeng12 Li Ming Su2 Min An3 Shi Ming Luo12

1Research Center for Chemical Ecology South China Agricultural University 2Institute of

Tropical and Sub-tropical Ecology South China Agricultural University Wushan Guangzhou

510642 PR China

3 Environmental and Analytical Laboratories Charles Sturt University Wagga Wagga New

South Wales Australia

Authors for correspondence Dr Ren Sen Zeng

Tel 86-20-85280211 Fax 86-20-85282693

Email rszengscaueducn

Abbreviations used are MeSA methyl salicylate MeJA methyl jasmonate VA vanillic acid

CA caffeic acid FA ferulic acid CMA coumaric acid HBA 3 4-hydroxybenzoic acid PAL

Phenylalanine ammonia-lyase C4H cinnamate 4-hydroxylase

Abstract Plants activate signaling system upon attack by insect herbivores and microbial

pathogens Methyl jasmonate (MeJA) and methyl salicylate (MeSA) are important signaling

molecules which are able to induce plant defense against insect herbivores and microbial

pathogens This study tests the hypothesis that allelopathy is an active inducible defense

mechanism of plants and JA and SA signaling pathways may activate the allelochemical

release We found that exogenously applied MeJA and MeSA to rice (Oryza sativa L) plants

enhanced rice allelopathic potentials and accumulation of phenolics increased enzymatic

activities and gene transcription of PAL and C4H which are two key enzymes in the

phenylpropanoid pathway Aqueous extracts of the leaves of rice IAC165 a putative

allelopathic variety showed 25 21 increase in inhibitory effects on root growth of

barnyardgrass (Ehinohloa rus-galli L) and 18 23 increase in inhibitory effects on the

shoot growth after rice plants were treated with MeSA (5 mM) and MeJA (005 mM)

respectively compared with the corresponsive control Leaf aqueous extracts of rice

Huajingxian1 a putative non-allelopathic variety treated with MeJA and MeSA showed 24

and 63 higher inhibition to the root length of barnyardgrass seedlings respectively The

root exudates of rice plants treated with MeJA (005 mM) and MeSA (5 mM) for 48h

significantly increased their inhibitory effects on root growth of barnyardgrass seedlings 3

4-hydroxybenzoic acid (HBA) vanillic acid (VA) coumaric acid (CMA) and ferulic acid (FA) in

the leaves accumulated approximately to 53 313 22 and 17-fold higher levels in

response to MeJA exposure and accumulated to 33 131 20 and 22-fold higher levels in

response to MeSA At fourth leaf ageMeSA and MeJA treatments enhanced the PAL

activity in rice leaves up to 523 and 801 and C4H activity increased by 402 and

67 respectively Gene transcription of PAL and C4H in rice leaves significantly increased

after treated with MeJA and MeSA These results indicate that allelopathy is an active

defense mechanism of plants and plant signals are potentially valuable in the regulation of

allelopathy for competing with other plants

Key words Methyl Salicylate Methyl Jasmonate Rice Allelopathy Echinochloa crus-galli

Phenolic Acid Phenylalanine ammonia-lyase Cinnamate 4-hydroxylase

INTRODUCTION

Allelopathy is defined as the direct or indirectly harmful or beneficial effects of one plant on

another through the production of chemical compounds that escape into the environment

(Rice 1984) Allelopathy provides an alternative approach for weed management in

sustainable agriculture Rice (Oryza sativa L) is one of the most important crops in the

world and its allelopathy has attracted a great deal of attention since Dilday et al (1989)

demonstrated that some rice varieties have allelopathic potentials against one or more

paddy weeds Rice allelopathy has been extensively studied with respect to screening

allelopathic rice germplasm (Dilday et al1994 Olofsdotter et al 1995 1999) its

allelochemicals (Mattice et al 1998 Kato-Noguchi 2004 Seal et al 2004a b) and genetic

control (Ebana et al 2001 Jensen et al 2001 Zeng et al 2003 He et al 2004) More than

12000 rice accessions have been evaluated in the United States for allelopathic potential to

weeds in paddy field (Dilday et al 1994 1998) Selection of allelopathic rice germplasm

were also conducted in many other countries (Fujii 1992 Garrity et al 1992 Dilday et al

1994 Olofsdotter et al 1995 Chung et al 1997 2000 2001a b Chou 1999 Ahn and

Chung 2000) Several putative allelochemicals have been identified Many phenolic acids

such as p-hydroxybenzoic acid (Chou and Lin 1976) ferulic acid (Chung et al 2000 2001a)

syringic acid caffeic acid sinapic and o-coumaric acid (Olofsdotter et al 1995) have been

isolated from rice plants soil where allelopathic rice lines have been growing and also soils

containing a decomposing rice residues Recent studies indicate that momilactone A and B

play an important role in rice allelopathy (Kato-Noguchi et al 2002 Kato-Noguchi 2004

Chung et al 2006)

The genetic control of allelopathy in rice is being determined Jensen et al (2001)

identified four main-effect QTL genes located on three chromosomes (2 3 and 8) which

collectively explained 35 of the total phenotypic variation of the allelopathic activity in the

population Ebana et al (2001) also identified QTL genes associated with the allelopathic

effect of rice using RFLP markers One of the QTL on chromosome 6 had the largest effect

on the expression of the allelopathic effect of rice and explained 161 of the phenotypic

variation He et al (2005) employed proteomic method to study the molecular mechanism

of crop allelopathy and identified four proteins peroxidase precursor (POD ) thioredoxinM -

type (Trx-m ) 3-hydroxy-3-methylglutaryl- coenzyme A reductase 3 (HGMR) and

phenylalanine ammonialyase (PAL ) The genes encoding four differential proteins were

located on the chromosome 4 7 8 and 12 in rice

Upon attack by herbivores and pathogens plants use allelochemicals to defense

themselves The chemical defense of plants is ubiquitous inducible and involves a complex

network of plant signaling cascades including jasmonates and salicylate signaling pathway to

trigger defense responses Jasmonates and salicylates function as key signal molecules in

plant chemical defense (Kessler and Baldwin 2002) and modulate plant resistance to insects

and pathogens (Creelman and Mullet 1997) Many defense related genes require jasmonic

acid (JA) and salicylic acid (SA) signaling for activation (Reinbothe et al 1994 Thomma et

al 1998 Turner et al 2002) A large body of evident has demonstrated that signaling

pathways initiate and regulate biosynthesis and production of secondary metabolites in

plants Exogenously applied JA induces the production of momilactone A a major

phytoalexin and allelochemical in rice (Nojiri et al 1996) and increases the resistance of

wild plants to insects in the field (Baldwin 1998) The contents of phenolic acids such as

gallic acid catechinic acid pyrocatechol caffeic acid coumaric acid ferulic acid and benzoic

acid increase sharply in the poplar leaves exogenously treated by methyl salicylate (MeSA) and

methyl jasmonate (MeJA) ( An et al 2006) The sakuranetin a flavonoid phytoalexin was also

induced by amino acid conjugates of JA (Shigeru et al 1997) Two acyclic homoterpenes

48-dimethyl-13E7- dimethylnonatriene (homoterpene I) and 4812-trimethyl-13E7E11-

tridecatetraene (homoterpene II) which are of sesquiterpenoid and diterpenoid origin can

be induced by JA at 01 mM to 10 mM in leaves of Phaseolus lunatus and Zea mays (Joumlrn et

al 1994) MeJA increased contents of u-tropine and tropine in jimsonweed seedlings (Fan

2005) and induced triterpenoid synthesis in both Centella asiatica and Galphimia glauca

plantlets (Mangas et al 2006) And MeJA also induced indole glucosinolates biosynthesis in

Arabidopsis (Brader et al 2001) and oilseed rape (Loivamaki et al 2004) Salicylic acid (SA)

is a well-known inducer of plant systematic acquired resistance (SAR) in plantndashpathogen

interactions characterized by induction of defense related gene expression and synthesis of

phenylpropanoids phytoalexins and pathogenesis-related proteins (PR) which result in

disease resistance to subsequent pathogen infections (Meacutetraux 2001 Durrant and

Dong2004 DeVos et al 2005) SA also induces biosynthesis and production of secondary

metabolites in plants (Taguchi et al 2001)

Allelopathy is an important mechanism for plants interfering with their neighbors by

releasing secondary metabolites namely allelochemicals and thereby enhancing plant

survival and reproduction under stress environments But whether the release of

allelochemicals into the environment is passive or active is largely unknown Hereby we

tested the hypothesis that allelopathy is an active inducible defense mechanism of plants

and JA and SA signaling pathways may activate the allelochemical release We exogenously

applied MeJA and MeSA to rice to study changes in allelopathic potentials of rice exudates

and aqueous extracts the enzymatic activities of phenylalanine ammonia-lyase (PAL)

catalyzing the first step in the biosynthesis of phenylpropanoids and cinnamate 4-

hydroxylase (C4H) catalyzing the conversion of cinnamate into 4-hydroxy-cinnamate a key

reaction of the phenylpropanoid pathway and gene expression of the two enzymes

METHODS AND MATERIALS

Plant and chemical materials

Two rice varieties were used in this study a standard rice cultivar with allelopathic potential

was IAC165 (Kim et al 2005) which was provided by International Rice Research Center

and the non-allelopathic rice cultivar Huajingxian 1 was kindly provided by Prof Zhiqiang

Chen in South China Agricultural University Vanillic acid (VA) caffeic acid (CA) 3 4-

hydroxybenzoic acid (HBA) ferulic acid (FA) coumaric acid (CMA) methyl jasmonate (MeJA)

and methyl salicylate (MeSA) were purchased from Sigma (St Louis MO) The

concentrations of MeJA and MeSA were 005 mM and 5 mM respectively TRIzol reagent

AMV reverse transcriptase Taq polymerase deoxynucleotide triphosphates (dNTPs) were

purchased from TaKaRa （Shuzo Co Ltd Shiga Japan) and MOPS DEPC were purchased

from AMRESCO (Solon OH) All solvents used were analytical or HPLC grades

Bioassays

Rice seeds were surface sterilized with 1 NaClO for 30 min rinsed with distilled water and

germinated in Petri dishes for 3 days Twenty seedlings were transplanted to each plastic

pot (10times15 cm) and were grown in a growth chamber kept at 24ndash26degC with 150 μMdm2s

light and a photoperiod of 12-hr light12-hr dark The seedlings were watered and fertilized

with Hoagland nutrient solution every two days

Rice seedlings were sprayed with 005 mM MeJA and 5 mM MeSA and kept in the

growth chamber for 48 hr The leaves and stems were sampled from rice plants then

aqueous extracts were prepared by extracting 8 g samples with 100 ml distilled water for 24

hr The extracts were filtered through filter paper and stored at 4degC until it was used

Echinochloa crus-galli seeds were placed in a beaker and bioassay of allelopathic

potentials of aqueous extract of rice leaves and stems was the same described by Zeng et al

(2001) Root and shoot lengths of the E crus-galli seedlings were determined at 7 days after

treatment

Root exudates

Seeds of rice (10 seeds per beaker) and barnyardyard (20 seeds per beaker) were

germinated and planted in a 1000 ml beaker with 300 ml of 1 agar culture media the agar

media were divided into two equal compartments using a membrane (mesh) with a pore

size of 35 microm for preventing root contact between the two plants but allowing root

exudates to pass through the membrane The rice seedlings were grown in the growth

chamber described above The seedling leaves were plastered with 005 mM MeJA and 5

mM MeSA respectively three times using brush pen at second fourth and sixth leaf ages

The barnyardgrass seedlings and culture medium were wrapped in aluminium foil to

prevent them to contact to signaling compounds when the compounds were applied The

seedlings were watered and fertilized with Hoagland nutrient solution every two days Root

and shoot lengths of the E crus-galli seedlings were determined at 7 days after the last

treatment with signaling compounds at sixth leaf age

Chemical analysis

Rice plants were treated with MeJA and MeSA at fourth leaf age and kept in the growth

chamber for 48h prior to sampling One gram of rice leaves or stems collected from

different treatments was extracted in 125 ml distilled water for 24 hr The water extracts

were partitioned against equal volume of ethyl acetate three times The ethyl acetate

extracts were combined and concentrated to dry form at 40degC under reduced pressure and

then dissolved with 125 ml methanol The methanol extracts were passed through

sterilized filter paper in a syringe The phenolic compounds in the filtrate were analyzed

using an Agilent Technologies HP1100 series HPLC system equipped with an ODS reverse

phase C18 column (250 times 4 mm 5 microm) and diode array detector (G1315 B) monitoring the

absorbance of the elution at 280 and 268 nm The solvent system was 75 methanol and

25 water adjusting pH to 26 with acetic acid Five micro-liters of extracted sample were

injected flow rate was 10 mlmin and temperature was 30 C The pure compounds were

used as standards and phenolic acids were identified by comparison of retention times and

UV spectrum

Enzymatic Activities

Rice plants were treated with signal compounds at fourth leaf age for C4H assay and at

second fourth and sixth leaf ages for PAL assay Prior to the enzymatic activity analysis the

rice plants were sprayed with MeJA (005mM) and MeSA (5 mM) and kept in different

growth chambers for 48 hr PAL activity was assayed based on the method described by

Ramamoorthy et al (2002b) with slight modification Root samples (1 g) were ground using

liquid nitrogen and homogenized in 1 ml of ice cold 01 M Tris-H2SO4 buffer pH 88

containing 7 mM of 2-mercaptoethanol and 1 mM EDTA-Na7 glycerin The homogenate

was centrifuged at 10000 rpm for 15 minutes The supernatant was used as enzyme

analysis

PAL activity was determined as the rate of the conversion of L-phenylalanine to trans-

cinnamic acid at 290 nm The absorbance (OD1) of the reaction mixture containing 05 ml of

enzyme extract 2 mL of 50 mM Tris- H2SO4 buffer (pH 88) and 1 ml of 20 mM L-

phenylalanine in the same buffer was measured and this reaction mixture was incubated in

hot water for 15 min at 40degC The enzyme activity was stopped by adding 6 M HCl and then

OD2 was measured Phenylalanine ammonia-lyase (PAL) activity was determined

spectrophotometrically as described by Edwards and Kessmann (1992) and was expressed in

Ug hr

The analysis of C4H activity accorded to the method described by Lamb et al (1975) with

slight modification Extraction of C4H from rice fresh leaves (1 g) was accomplished by

homogenization of plant material in 2ml potassium phosphate buffer (200 mM pH 75

containing 2 mM 2-mercaptoethanol) After filtration and centrifugation (15 min at 10000

rpm) the supernatant was diluted to 20 times and used for enzymatic analysis The extract

(02 ml) was added to 2 ml reaction buffer (50 mM phosphate buffer containing 2 mM 2-

mercaptoethanol 2 mM trans-cinnamic acid and 05 mM NADPH) Reaction mixture was

incubated for 1 hr at 37degC Absorbance value was measured with 290 nm after reaction

stopped with 6 M HCl and readjusted to pH 11 with 6 M NaOH The cinnamate 4-

hydroxylase C4H activity was expressed in Ug hr

RT-PCR

Rice plants at fourth leaf age were sprayed with MeJA and MeSA and kept in growth

chambers for 48h prior to sampling The total RNA was extracted and isolated according to

the method described by Kiefer et al (2000) with slight modification

02 g fresh material were grinded with a mortar and pestle under liquid nitrogen and the

powdered tissue were transfered into a 2 ml Eppendorf tube add 1000 microl TRIzol reagent

and intermix them incubate for 8-10 min on ice Add 200 microl chloroform and intermix them

incubate at room temperature for 5min and then centrifuge for 15 min at 4degC and 12000

rpm Transfer the supernatant to a 15 ml Eppendorf tube add 500 microl isoamylalcohol vortex

at room temperature for 10 min and centrifuge for another 10 min at 4degC and 13000 rpm

Discard the supernatant wash the pellet with 1 ml 75 ethanol (vv) and kept them in a

refrigerator (-20 degC) until it was used

The expression patterns of defense-related genes in different treated rice leaves were

analyzed using reverse transcription ndashpolymerase chain reaction The degenerate primers

used for amplification of the putative genes were described in Table 1 Actin was used as a

reference RT-PCR reactions were initiated with first strand cDNA synthesis at 42ordmC for 60

min After denaturation of the RNA-cDNA hybrid at 94ordmC for 2 min 40 PCR amplification

cycles (94ordmC for 1 min 56ordmC for 1 min and 72ordmC for 1 min) were run and followed by a final

extension for 7 min Two microl of concentrated (10times) loading dye were added to each reaction

and 5 microl of each sample were run on 12 agarose gel electrophoresis in 1timesTAE buffer

Statistical analyses

All data were evaluated by one-way analysis of variance (ANOVA) with treatment

differences among means tested at P= 005 with Duncanrsquos multiple range test All data for

the root and shoot growth the content of phenolic acid and the enzymatic activities of PAL

and C4H were means from three replicates

RESULTS

Aqueous extract

After MeSA (5 mM) and MeJA (005 mM) treatments the aqueous extracts of the stem

and leaves of rice IAC165 a putative allelopathic variety had significantly stronger allelopathic

effects on both the root and shoot growth of barnyardgrass compared with non-signaling

treated aqueous extracts of rice (control) (Table 2) After application with MeSA (5 mM) and

MeJA (005 mM) the stem aqueous extracts of rice IAC165 increased inhibition by 24 32

to root growth and by 38 27 to shoot growth of tested plants respectively And the

aqueous extracts of the leaves showed 25 21 increase in inhibitory effects on root growth

of barnyardgrass and 18 23 increase in inhibitory effects on the shoot growth after rice

plants were treated by MeSA (5 mM) and MeJA (005 mM) respectively compared with the

corresponsive control (Table 2)

After signaling treatments aqueous extracts of both leaves and stems of rice

Huajingxian1 a putative non-allelopathic variety also showed stronger inhibition on the root

length of barnyardgrass compared with control aqueous extract without signal treatment

(Table 2) The stem aqueous extracts of Huajingxian1 plants treated with MeJA (005 mM) and

MeSA (5 mM) for 48 hr showed 36 and 63 higher inhibition on the root growth of

barnyardgrass compared with the control Similarly leaf aqueous extracts of Huajingxian1

plants treated with MeJA and MeSA were 24 and 63 more inhibition to the root length of

barnyardgrass seedlings respectively (Table 2) Shoot length of barnyardgrass seedlings

treated with aqueous extracts of signaling induced Huajingxian1 plants did not differ from

that on the control (Table 2)

The bioassays result showed that both the allelopathic and non-allelopathic rice plants

enhanced their allelopathic potentials to barnyardgrass plants after treated with signaling

compounds MeSA and MeJA

Root exudates

The root exudates of rice plants treated with MeJA (005 mM) and MeSA (5 mM) for 48h

significantly increased their inhibitory effects on root growth of barnyardgrass seedlings

compared with that of control plants without signal compound induction (Table 3)

Application of MeJA increased inhibitory effects of Huajingxian1 and IAC165 by 24 and 17

respectively and MeSA application increased by 20 and 19 respectively Signal

compounds increased allelopathic potentials of root exudates of both allelopathic and non-

allelopathic rice plants

Phenolic acids

Phenolic acids in the leaves and stems of rice IAC 165 were analyzed using HPLC after

treated with MeSA (5 mM) and MeJA (005 mM) for 48h at the fourth leaf age An

accumulation of HBA VA CMA and FA was observed in the signaling treated leaves of rice

compared with the control (Fig1) CMA and FA accumulated in the treated stems of rice

(Fig2) The HBA VA CMA and FA in the leaves accumulated approximately to 53 313 22

and 17-fold higher levels in response to MeJA exposure and accumulated to 33 131 20

and 22-fold higher levels in response to MeSA (Fig1)

VA CMA and FA were not detected in non-treated rice stems while MeJA induced high

accumulation of VA CMA and FA in the stems and MeSA induced accumulation of CMA

and FA in the stems (Fig2)

PAL and C4H activity

PAL catalyzes the first step in the biosynthesis of phenylpropanoids The activities of

PAL in the rice leaves after treatment with MeSA (5 mM) and MeJA (005 mM) for 48 hr

significantly increased compared with the control The PAL activities in the leaves treated

with MeSA and MeJA at the second leaf age for 48 hr increased by 556 and 319

respectively compared with the control At fourth leaf ageMeSA and MeJA enhanced the

PAL activity up to 523 and 801 The PAL activities increased by 625 and 627

respectively when the leaves were treated with MeSA and MeJA at the sixth leaf age for 48

hr ( Fig3)

Forty-eight hours after treatment with MeSA and MeJA at the fourth leaf age C4H

activity in rice IAC165 leaves increased by 402 and 67 respectively compared to leaves

without signaling treatment (Fig 4)

Induction of PAL and C4H Transcripts

To determine whether MeJA and MeSA enhance the allelopathic potentials and

phenolic acids by inducing transcription of the genes encoding key enzymes PAL and C4H

responsible for biosynthesis of phenylpropanoids the expression patterns of PAL and C4H

genes were analyzed using RT-PCR from the leaves of rice IAC165 treated with MeSA (5 mM)

and MeJA (005 mM) for 48 hr at the fourth leaf age Both MeJA and MeSA induced

accumulation of PAL and C4H transcripts over basal levels present in control leaves (Fig 5)

DISCUSSION

Plants are frequently subjected to insect herbivory pathogen infection and competition

from surrounding plants However plants have evolved a diversity of allelochemicals to

defend themselves against herbivorous insects and microbial pathogens and to produce

allelopathic effects on their neighbors Plant chemical defense against herbivores and

pathogens are inducible and regulated by the jasmonic acid (JA) and salicylic acid signaling

pathways Exogenous addition of JA can increase the resistance of wild plants to insects in

the field (Baldwin 1998) While many studies have shown that signal compounds could

induce plant defense against insect herbivores and microbial pathogens rarely have those

experiments been conducted to demonstrate that signaling pathways are involved in plant

allelopathy

This study found that rice allelopathy could be induced by exogenous application of

signaling compounds MeJA and MeSA The allelopathic potentials of the root exudates of

rice increased after treatment with MeJA and MeSA (Table 3) Aqueous extracts of both

leaves and stems of MeSA and MeJA-treated rice Hajingxian1 and IAC165 also enhanced

their inhibitory effects on barnyardgrass root growth (Table 2)

Further HPLC analysis showed that phenolic acids including VA CA CMA FA and HBA in

rice leaves accumulated after treated by MeJA and MeSA for 48h (Fig1) CA CMA and FA

were detected in the stems after treated by MeJA and MeSA but they were not detected in

the stems of non-treated control plants (Fig2) VA was also detected in the stem after

treated by MeJA This increase in phenolic acids after signaling application may be

responsive for the corresponsive increase in allelopathic potentials in root exudates and

aqueous extracts of induced rice plants Similar changes of several phenolic acids in poplar

plants treated with MeJA were reported by An et al (2006) Phenolics are believed in most

studies to be primary allelochemicals in rice (Chung et al 2000 2001a Olofsdotter et al

1995) Greater amounts of trans-ferulic acid p-hydroxybenzoic acid and caffeic acid were

detected in the exudates of allelopathic cultivars (Seal et al 2004b) Five phenolics

including caffeic p-hydroxybenzoic vanillic syringic and p-coumaric acids from rice

exudates were best correlated with the observed allelopathic effects of rice on arrowhead

(Sagittaria montevidensis) root growth (Seal et al 2004b) MeJA induced multiple

biosynthetic pathways including the shikimic acid pathway (producing methyl salicylate) the

octadecanoid pathway (producing cis-jasmone) as well as the mevalonate-dependent and -

independent terpenoid pathways (producing mono- and sesquiterpenes) (Gundlach et al

1992 Martin et al 2002 Thaler 1996) The induction of these pathways were found in

many agricultural species such as tomato lima bean and corn (Thaler et al 1996 Dicke et

al 1999 Schmelz et al 2003) Martin et al (2003) recently demonstrated that MeJA

application induced the activities of both constitutive and novel terpene syntheses in

Norway spruce

Exogenous application of MeJA (005 mM) and MeSA ( 5 mM) increased the PAL activity

in rice at all three leaf ages (second fourth and sixth) tested (Figure 3) and also increased

the C4H activity at fourth leaf age in rice leaves (Figure 4)

Genes including PAL and C4H from major phenylpropanoid biosynthetic pathways involved

in phenolic acid production are induced by exogenously applied MeJA and MeSA RT-PCR

showed that the PAL and C4H mRNA level in the induced groups were consistently greater

than that in the control (Figure 5)

These results suggest that signaling compounds up-regulate gene transcription of PAL

and C4H and then increase enzymatic activities and accumulation of phenolic acids which

results in enhancement in allelopathic potentials in rice These findings have important

ecological significance in the sense that allelopathy is an active defense mechanism of

plants and JA and SA signaling pathways may control the allelochemical release and

allelopathic potentials That suggests that plants may activate signaling systems to release

allelochemicals into the environments to interfere with neighborrsquos growth in response to

competition stresses etc Wang et al (2005) found that rice plants increased their

allelopathic potentials when they competed with barnyardgrass flatsedge (Cyperus

difformis) ducksalad (Monochoria vaginalis Burmf) Bidens pilosa plants increase

allelopathic potentials in drought season (Zeng and Luo 1995) These findings indicate that

plant chemical defenses against herbivores microbial pathogens and other plants

(allelopathy) have common mechanism in which signaling transduction plays a vital role

Cross resistance may exist in different plant chemical defenses Centaurea maculosa one of

the most destructive invasive plants in North America exudes far higher amounts of (plusmn)-

catechin an allelochemical known to have inhibitory effects on native plants and exhibit

more intense negative effects on natives when attacked by larvae of two different root

boring biocontrol insects and a parasitic fungus (Thelen et al 2005) Plant signals are

potentially valuable in the regulation of allelopathy for competing with other plants

Understanding signaling pathways involved in allelopathy phenomena can provide valuable

insights into whole chemical defense in plants and optimizing chemically based pest

management programs

Acknowledgments This research was supported by Natural Science Foundation of China

(30370246 30670331) and Natural Science Foundation of Guangdong Province (Group Project

039254 04105977) National 973 project of China (2006CB100200) Guangdong Science amp

Technology Plan Program (2004B20501010) and Program for New Century Excellent Talents in

University (NCET-04-0830) to RSZ We thank Dr Arthur Zangerl for technical assistance and

Allen Lawrence and Henan Zeng for help with insect rearing

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CHON S U COUTTS J H and NELSON C J 2000 Effects of light growth media and seedling

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CHUNG I M KIM J T and KIM S H 2006 Evaluation of allelopathic potential and

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CHUNG I M Kim K H Ahn J K JU HJ 1997 Allelopathic potential evaluation of rice

cultivars on Echinochloa crus-galli Korean J Weed Sci17 52ndash58

CREELMAN R A and MULLET J E1997 Biosynthesis and action of jasmonates in plants

Annu Rev Plant Physiol Plant Mol Biol 48 355ndash381

DrsquoCunha G B 2005 Enrichment of phenylalanine ammonia lyase activity of Rhodotorula

yeast Enzyme and Microbial Technology 36 498ndash502

DEVOS M VAN OOSTEN V R POECKE R M VAN PELT J A POZO M J MUELLER M J

BUCHALA A J METRAUX J P VAN LOON L C DICKE M and PIETERSE C M (2005)

Signal signature and transcriptome changes of Arabidopsis during pathogen and insect

attack Plant Microbe Interact 18 923-937

DICKE M GOLS R LUDEKING D and POSTHUMUS MA1999Jasmonic acid and herbivory

differentially induce carniviore-attracting plant volatiles in lima bean plants J Chem

Ecol251907-1922

DILDAY R H LIN J and YAN W G Identification of allelopathy in the USDA-ARS rice

germplasm collection Aus J Exp Agric 1994 34 901- 910

DILDAY R H YAN W G MOLDENHAUER K A K and GRAVOIS K A 1998 Allelopathic activity

in rice for controlling major aquatic weeds In Olofsdotter M (ed) Allelopathy in

RiceManila IRRI 7-26

DILDAY RH LIN J and YAN W 1994 Identification of allelopathy in the USDA-ARS rice

germplasm collection Aust J Exp Agric34907-910

DURRANT W E and DONG X 2004 Systemic acquired resistance Annu Rev Phytopathol

42 185-209

EBANA K YAN WG DILDAY R H NAMAI H and OKUNO K 2001 Analysis of QTL associated

with the allelopathic effect of rice using water soluble extracts Breeding Sci 5147-51

EDWARDS R and KESSMANN H 1992 Isoflavonoid phytoalexins and their biosynthetic

enzymes In Molecular Plant Pathology A Practical Approach S J Gurr M J

McPherson and D J Bowles eds (Oxford IRL Press) pp 45ndash62

FAN D 2005 Effects of glyphosatechlorsulfuron and methyl jasmonate on growth and

alkaloid biosynthesis of jimsonweed (Datura stramonium L) Pestic Biochem

Physiol8216ndash26

FUJII Y1992The potential for biological control of paddy and aquatic weeds with

allelopathy Allelopathic effect of some rice varieties Proceedings of the International

Symposium on Biological Control and Integrated Management of Paddy and Aquatic

Weeds Tsukuba Japan pp 305ndash320

GARRITY D P MOVILLOn M and MODDY K 1992 Differential weed suppression ability in

upland rice cultivars Agron J 84586ndash591

GRANEacuteLI E and JOHANSSON N 2003 Increase in the production of allelopathic substances by

Prymnesium parvum cells grown under N- or P-deficient conditions Harmful Algae

2135-145

GUNDLACH H MUELLER MJ KUTCHAN TM and ZENK MH (1992) Jasmonic acid is a

signal transducer in elicitor-induced plant cell cultures Proc Natl Acad Sci USA 89

2389ndash2393

HE H Q LIN W X LIANG Y Y SONG B Q KE Y Q GUO Y C Hand LIANG K J 2005

Analyzing the molecular mechanism of crop allelopathy by using differential

proteomics Acta Ecology Sinica 25(12)3141－3146

HE H Q SHEN L H XIONG J JIA X L LIN W X and WU H 2004 Conditional genetic

effect of allelopathy in rice (Oryza sativa L) under different environmental conditions

Plant Growth Regul 44211-218

JENSEN L B COURTOIS B SHIN L LI Z K OLOFSDOTTER M and MAULEON R P 2001

Locating genes controlling allelopathic effects against barnyardgrass in upland rice

Agron J 93 21-26

JOumlRN H JENS D SIEGFRIED B and WILHELM B 1994 Herbivore-induced volatilesThe

emission of acyclic homoterpenes from leaves of Phaseolus lunatus and Zea mays can

be triggered by a -glucosidase and jasmonic acid FEBS Letters 352(2)146-150

KATO-NOGUCHI H 1999 Effect of light irradiation on allelopathic potential of germinating

maize Phytochemistry521023-1027

KATO-NOGUCHI H 2004 Allelopathic substance in rice root exudates Rediscovery of

momilactone B as an allelochemical J Plant Physiol 161271-276

KATO-NOGUCHI H INO T SATA N YAMAMURA S 2002 Isolation and identification of a potent

allelopathic substance in rice root exudates Physiol Plantarum115401-405

KESSLER A and BALDWIN I T 2002 Plant response to insect herbivory the emerging

molecular analysis Annual Review Plant Biology 53299-328

KIEFER E HELLER W and ERNST D 2000 A simple and efficient protocol for isolation of

functional RNA from plant tissues rich in secondary metabolites Plant Molecular

Biology Reporter 18 33ndash39

KIM K U and SHIN D H 1998 Rice allelopathy research in Korea In Olofsdotter M eds

Allelopathy in Rice Manila IRRI 39-44

KIM K U SHIN D H and LEE I J and Kim H Y 2000 Rice allelopathy in Korea pp57-82

in K U Kim and D H Shin (eds) Rice Allelopathy Kyunpook National University Taegu

Korea

KIM S Y MADRID A V PARK S T YANG S J and OLOFSDOTTER M 2005 Evaluation of rice

allelopathy in hydroponics Weed Research 45 74ndash79

LOBON N C GALLEGO J C A DIAZ T S and GARCIA J C E 2002 Allelopathic potential of

Cistus ladanifer chemicals in response to variations of light and temperature

Chemoecology 12139-145

LOIVAMAKI M HOLOPAINEN J K and NERG A M 2004 Chemical changes induced by

methyl jasmonate in oilseed rape grown in the laboratory and in the field J Agric Food

Chem 52 7607 -7613

MANGAS S BONFILL M OSUNA L MOYANO E TORTORIELLO J CUSIDO RM PINOL MT and

PALAZON J 2006 The effect of methyl jasmonate on triterpene and sterol metabolisms

of Centella asiatica Ruscus aculeatus and Galphimia glauca cultured plants

Phytochemistry 672041-2049

MANGAS S BoNFILL M OSUNA L MOYANO E TORTORIELLO J CUSIDO R M PINOL M T

and PALAZƠN J 2006 The effect of methyl jasmonate on triterpene and sterol

metabolisms of Centella asiatica Ruscus aculeatus and Galphimia glauca cultured

plants Phytochemistry 67 2041ndash2049

MARTIN D M GERSHENZON J and BOHLMANN J 2003 Induction of volatile terpene

biosynthesis and diurnal emission by methyl jasmonate in foliage of Norway Spruce

Plant Phsyiol1321586-1599

MARTIN D THROLL D GERSHENZON J and BOHLMANN J 2002 Methyl jasmonate

induces traumatic resin ducts terpenoid resin biosynthesis and terpenoid

accumulation in developing xylem of Norway spruce stems Plant Physiol 129 1003ndash

1018

MATTICE J LAVY T SKULMAN B and DILDAY R H 1998 Searching for allelochemicals in rice

that control ducksalad In Olofsdotter M (ed) Allelopathy in Rice International Rice

Research Institute Manila pp 81ndash98

MEacuteTRAUX J P 2001 Systemic acquired resistance and salicylic acid current state of

knowledge Eur J Plant Pathol 107 13-18

NOJIRI H SUGIMORI M YAMANE H NISHIMURA Y YAMADA A SHIBUYA N KODAMA O

MUROFUSHI N and OMORI T 1996 Involvement of jasmonic acid in elicitor-induced

phytoalexin production in suspension-cultured rice cells Plant Physiol 110387-392

OLOFSDOTTER M NAVAREZ D REBULANAN M and STREIBIG J C 1999 Weed suppressing rice

cultivars ndash does allelopathy play a role Weed Res39441-454

OLOFSDOTTER M 2001 Rice A step toward use of Allelopathy Allelopathy in Natural and

Managed Ecosystems Agron J 93 3-8

OLOFSDOTTER M NAVAREZ D and MOODY K1995 Allelopathic potential in rice (Oryza sativa

L) germplasm Ann Appl Biol127543-560

OLOFSDOTTER M REBULANAN M MADRID A WANG D L NAVAREZ D and OLK D C 2002

Why Phenolic acids are unlikely primary allelochemicals in rice J Chem Ecol

28(1)229-243

PRAMANIK M H R NAGAI M ASAO T and MATSUI Y 2000 Effects of temperature and

photoperiod on phytotoxic root exudates of cucumber (Cucumis sativus) in hydroponic

culture J Chem Ecol 261953-1967

RAMAMOORTHY V T RAGUCHANDER and R SAMIYAPPAN 2002a Enhancing resistance of tomato

and hot pepper to pythium diseases by seed treatment with fluorescent

pseudomonads European Journal of plant pathology 108 429-441

RAMAMOORTHY V T RAGUCHANDER and R SAMIYAPPAN 2002b Induction of defence-related

proteins in tomato roots treated with Pseudomonas fluorescens Pf1 and Fusarium

oxysporum fsp lycopersici Plant and Soil 23955-68

REINBOTHE S Mollenhauer B REINBOTHE C 1994 JIPs and RIPs the regulation of plant gene

expression by jasmonates in response to environmental cues and pathogens The Plant

Cell 61197ndash1209

RICE E L1984 AllelopathyNew YorkAcademic Press

SCHMELZ E A ALBORN H T BANCHIO E and TUMLINSON J H 2003 Quantitative

relationships between induced jasmonic acid levels and volatile emission in Zea mays

during Spodoptera exigua herbivory Planta 216665-673

SEAL A N HAIG T and PRATLEY J E 2004 Evaluation of putative allelochemicals in rice

root exudates for their role in the suppression of arrowhead root growth J Chem Ecol

30 1663-1678

SEAL A N PRATLEY J E HAIG T and AN M 2004 Identification and quantitation of

compounds in a series of allelopathic and non-allelopathic rice root exudates J Chem

Ecol 301647-1662

SEIGLER D S 1996 Chemistry and mechanisms of allelopathic interactions Agron J 88876-

885

SHIGERU T RANDEEP R and OSAMU K 1997 Phytoalexin production by amino acid

conjugates of jasmonic cid through induction of aringenin-7-O- methyltransferase a

key enzyme on phytoalexin biosynthesis in rice (Oryza sativa L) FEBS Letters 401239-

242

SHIN D H KIM K U SOHN D S KANG S G KIM H Y LEE I J AND KIM M Y 2000

Regulation of gene expression related to allelopathy pp109-124 in K U Kim and D H

Shin (eds) Rice Allelopathy Kyunpook National University Taegu Korea

TAGUCHI G YAZAWA T HAYASHIDA N and OKAZAKI M 2001 Molecular cloning and

heterologous expression of novel glucosyltransferases from tobacco cultured cells that

have broad substrate specificity and are induced by salicylic acid and auxin Eur J

Biochem 68406-4094

THALER J S STOUT M J KARBAN R and DUFFEY S S 1996 Exogenous jasmonates simulate

insect wounding in tomato plants (Lycopersicon esculentum) in the laboratory and field

J Chem Ecol 221767-1781

Thelen G C Vivanco J M Newingham B Good W Bais H P Landres P Caesar A

and Callaway RM 2005 Insect herbivory stimulates allelopathic exudation by an

invasive plant and the suppression of natives Ecology Letters 8 209-217

THOMMA B P H J EGGERMONT K PENNINCKX I A M A MAUCH-MANI B VOGELSANG R

CAMMUE B P A BROEKART W F 1998 Separate jasmonate-dependent and salicylate-

dependent defense-response pathways in Arabidopsis are essential for resistance to

distinct microbial pathogens Proc Natl Acad Sci USA 9515107-15111

TURNER J G ELLIS C and EVOTO A 2002The jasmonate signal pathway Plant Cell

14S153ndashS164

WANG Y P TANG L H ZHANG H S and FANG X W 2005 Induction effect of some

weeds on the allelopathy of rice varieties Ecology and Environment 14250-252

ZENG D L QIAN Q TENG S Dong G Fujimoto H Yasufumi K Zhu L 2003Genetic

analyses on rice allelopathy Chin Sci Bull 48(1)70-73

ZENG R S and LUO S M1995 Relationship between allelopathic effects of Bidens pilosa

aqueous extracts and rainfall J South China Agri Univ 16(4)69-72

ZENG R S LUO S M SHI M B SHI Y H ZENG Q and TAN H F 2001 Allelopathy of

Aspergillus japonicus on Crops Agron J93 60-64

ZHOU Z H1999 Method of allelopathy bioassay and the affecting factors Ecol Sci 18 35-

38

Figure legends

FIG 1 Enzymatic activity of the phenylalanine ammonia-lyase in the leaves of rice IAC 165

after treatment with MeSA (5 mM) and MeJA (005 mM) for 48 hr at the second fourth and

sixth leaf ages Values are means plusmn standard errors from three replicates Significant

difference (P lt 005) among treatments in each group are indicated by different letters

above bars

FIG 2 Enzymatic activity of the Cinnamate 4-hydroxylase in the leaves of the IAC165 after

treatment with MeSA (5 mM) and MeJA (005 mM) for 48 hr at the second fourth and sixth

leaf ages Values are means plusmn standard errors from three replicates Significant difference (P

lt 005) among treatments in each group are indicated by different letters above bars

FIG 3 Concentrations of phenolic acids in aqueous leaf extracts of rice IAC 165 after

treatment with MeSA (5 mM) and MeJA (005 mM) for 48 hr at the fourth leaf age Values

are means plusmn standard errors from three replicates Significant difference (P lt 005) among

treatments in each group are indicated by different letters above bars VA vanillic acid CA

caffeic acid FA ferulic acid CMA Coumaric acid HBA 3 4-hydroxybenzoic acid

FIG 4 Concentrations of phenolic acids in aqueous stem extracts of rice IAC 165 after





FIG 5 Gene expression of phenylalanine ammonia-lyase (PAL) and cinnamate 4-hydroxylase

(C4H) in the leaves of rice IAC165 in response to treatment with MeSA (5 mM) and MeJA

(005 mM) for 48 hr at the fourth leaf age Actin was used as internal standard of RT-PCR

Table 1 Specific primers for RT-PCR

Gene Accession

number Gene function Specific primer

OsPAL X16099 catalyzing the first step in the

biosynthesis of phenylpropanoids F5-CACAAGCTGAAGCACCACCC-3

R5-GAGTTCACGTCCTGGTTGTG-3

OsC4H OJ1342 a cytochrome P450 catalyzing the

conversion of cinnamate into 4-

hydroxy-cinnamate a key

reaction of the phenylpropanoid

pathway

F5rsquo-CTGGCACCGACGGTCATGTT-3rsquo

R5rsquo-CTGGATGGTGCTTGAGCTTG-3rsquo

Actin X15865 Constitutive actin gene and

internal standard of RT-PCR

F5-ACTGTCCCCATCTATGAAGGA-3rsquo

R5-CTGCTGGAATGTGCTGAGAGA-3

C4H Cinnamate 4-hydroxylase PAL Phenylalanine ammonia-lyase Actin Internal standard

of RT-PCR

TABLE 2 Influences of external application of MeJA (005 mM) and MeSA (5 mM) on effects of

the aqueous extract of rice Huajingxian1 and IAC165 on root length and shoot length of

barnyardgrass

Extraction

organ Treatments

Huajingxian1 IAC165

Root length

(cm)

Shoot length

(cm)

Root length

(cm)

Shoot length

(cm)

Stems

Control 70plusmn049a 60plusmn018a 63plusmn027a 60plusmn049a

MeJA 26plusmn021c 59plusmn042a 42plusmn047b 44plusmn042b

MeSA 45plusmn051b 56plusmn023a 48plusmn04b 37plusmn039b

Leaves




Rice plants were treated at fourth leaf age for 48 hr Values are mean standard error significant



the root exudates of rice Huajingxian1 and IAC165 in agar on root length and shoot length of

barnyardgrass

Rice cultivars Treatments Root length (cm) Shoot length (cm)

Huajingxian 1 Control 109plusmn044a 223plusmn055a

MeJA 84plusmn044b 214plusmn076a

MeSA 88plusmn036b 212plusmn077a

IAC165 Control 104plusmn074a 206plusmn054a





0

150

300

450

600

HBA VA CA CMA FA

Phenolic acid

Conce

ntr

atio

ns

(μgg

FW

)

control

MeJA

MeSAa

c

b

ab

a

a

b

a

b

c

c a aa






0

250

500

750

1000

1250

HBA VA CA CMA FA

Phenolic acid

control

MeJA

MeSA

bb

a

cb

a

b

a

c

b

a

ca aa Co

nce

ntr

atio

ns

(μg

g F

W)





caffeic acid FA ferulic acid CMA coumaric acid HBA 3 4-hydroxybenzoic acid

0

1000

2000

3000

4000

2 4 6

Signal treated leaf age

control

MeJA

MeSA

b

aa

b

aa

a

b

c

PA

L a

ctiv

ity

(U

gh

)





above bars

0

50

100

150

200

control MeJA MeSA

Signal compound

b

c

a

C4H

act

ivity (

Ug

h)


treatment with MeSA (5 mM) and MeJA (005 mM) for 48 hr at the fourth leaf ages Values


treatments are indicated by different letters above bars

control MeJA MeSA control MeJA MeSA

PA

L

C4H




Actin Actin

Rice allelopathy induced by methyl jasmonate and methyl salicylate

Hai Hong Bi12 Ren Sen Zeng12 Li Ming Su2 Min An3 Shi Ming Luo12

1Research Center for Chemical Ecology South China Agricultural University 2Institute of

Tropical and Sub-tropical Ecology South China Agricultural University Wushan Guangzhou

510642 PR China

3 Environmental and Analytical Laboratories Charles Sturt University Wagga Wagga New

South Wales Australia

Authors for correspondence Dr Ren Sen Zeng

Tel 86-20-85280211 Fax 86-20-85282693

Email rszengscaueducn

Abbreviations used are MeSA methyl salicylate MeJA methyl jasmonate VA vanillic acid

CA caffeic acid FA ferulic acid CMA coumaric acid HBA 3 4-hydroxybenzoic acid PAL

Phenylalanine ammonia-lyase C4H cinnamate 4-hydroxylase





























INTRODUCTION





















Chung et al 2006)
































































Bioassays













treatment

Root exudates













Chemical analysis














UV spectrum










analysis








Ug hr











RT-PCR

























RESULTS

Aqueous extract
























Root exudates








Phenolic acids



















hr ( Fig3)











DISCUSSION










allelopathy



























Norway spruce





























management programs







References


















20921ndash928



Crop Sci 4544-49
















Ecol251907-1922









42 185-209








Physiol8216ndash26









2135-145



2389ndash2393









Agron J 93 21-26



















Korea








Chem 52 7607 -7613















1018

















28(1)229-243












Cell 61197ndash1209







30 1663-1678



Ecol 301647-1662


885




242







Biochem 68406-4094



J Chem Ecol 221767-1781









14S153ndashS164










38

Figure legends





above bars



















Gene Accession









pathway








of RT-PCR



barnyardgrass

Extraction

organ Treatments

Huajingxian1 IAC165

Root length

(cm)

Shoot length

(cm)

Root length

(cm)

Shoot length

(cm)

Stems




Leaves








barnyardgrass










0

150

300

450

600

HBA VA CA CMA FA

Phenolic acid

Conce

ntr

atio

ns

(μgg

FW

)

control

MeJA

MeSAa

c

b

ab

a

a

b

a

b

c

c a aa






0

250

500

750

1000

1250

HBA VA CA CMA FA

Phenolic acid

control

MeJA

MeSA

bb

a

cb

a

b

a

c

b

a

ca aa Co

nce

ntr

atio

ns

(μg

g F

W)






0

1000

2000

3000

4000

2 4 6


control

MeJA

MeSA

b

aa

b

aa

a

b

c

PA

L a

ctiv

ity

(U

gh

)





above bars

0

50

100

150

200

control MeJA MeSA

Signal compound

b

c

a

C4H

act

ivity (

Ug

h)






PA

L

C4H




Actin Actin





























INTRODUCTION





















Chung et al 2006)
































































Bioassays













treatment

Root exudates













Chemical analysis














UV spectrum










analysis








Ug hr











RT-PCR

























RESULTS

Aqueous extract
























Root exudates








Phenolic acids



















hr ( Fig3)











DISCUSSION










allelopathy



























Norway spruce





























management programs







References


















20921ndash928



Crop Sci 4544-49
















Ecol251907-1922









42 185-209








Physiol8216ndash26









2135-145



2389ndash2393









Agron J 93 21-26



















Korea








Chem 52 7607 -7613















1018

















28(1)229-243












Cell 61197ndash1209







30 1663-1678



Ecol 301647-1662


885




242







Biochem 68406-4094



J Chem Ecol 221767-1781









14S153ndashS164










38

Figure legends





above bars



















Gene Accession









pathway








of RT-PCR



barnyardgrass

Extraction

organ Treatments

Huajingxian1 IAC165

Root length

(cm)

Shoot length

(cm)

Root length

(cm)

Shoot length

(cm)

Stems




Leaves








barnyardgrass










0

150

300

450

600

HBA VA CA CMA FA

Phenolic acid

Conce

ntr

atio

ns

(μgg

FW

)

control

MeJA

MeSAa

c

b

ab

a

a

b

a

b

c

c a aa






0

250

500

750

1000

1250

HBA VA CA CMA FA

Phenolic acid

control

MeJA

MeSA

bb

a

cb

a

b

a

c

b

a

ca aa Co

nce

ntr

atio

ns

(μg

g F

W)






0

1000

2000

3000

4000

2 4 6


control

MeJA

MeSA

b

aa

b

aa

a

b

c

PA

L a

ctiv

ity

(U

gh

)





above bars

0

50

100

150

200

control MeJA MeSA

Signal compound

b

c

a

C4H

act

ivity (

Ug

h)






PA

L

C4H




Actin Actin
















Chung et al 2006)
































































Bioassays













treatment

Root exudates













Chemical analysis














UV spectrum










analysis








Ug hr











RT-PCR

























RESULTS

Aqueous extract
























Root exudates








Phenolic acids



















hr ( Fig3)











DISCUSSION










allelopathy



























Norway spruce





























management programs







References


















20921ndash928



Crop Sci 4544-49
















Ecol251907-1922









42 185-209








Physiol8216ndash26









2135-145



2389ndash2393









Agron J 93 21-26



















Korea








Chem 52 7607 -7613















1018

















28(1)229-243












Cell 61197ndash1209







30 1663-1678



Ecol 301647-1662


885




242







Biochem 68406-4094



J Chem Ecol 221767-1781









14S153ndashS164










38

Figure legends





above bars



















Gene Accession









pathway








of RT-PCR



barnyardgrass

Extraction

organ Treatments

Huajingxian1 IAC165

Root length

(cm)

Shoot length

(cm)

Root length

(cm)

Shoot length

(cm)

Stems




Leaves








barnyardgrass










0

150

300

450

600

HBA VA CA CMA FA

Phenolic acid

Conce

ntr

atio

ns

(μgg

FW

)

control

MeJA

MeSAa

c

b

ab

a

a

b

a

b

c

c a aa






0

250

500

750

1000

1250

HBA VA CA CMA FA

Phenolic acid

control

MeJA

MeSA

bb

a

cb

a

b

a

c

b

a

ca aa Co

nce

ntr

atio

ns

(μg

g F

W)






0

1000

2000

3000

4000

2 4 6


control

MeJA

MeSA

b

aa

b

aa

a

b

c

PA

L a

ctiv

ity

(U

gh

)





above bars

0

50

100

150

200

control MeJA MeSA

Signal compound

b

c

a

C4H

act

ivity (

Ug

h)






PA

L

C4H




Actin Actin








































Bioassays













treatment

Root exudates













Chemical analysis














UV spectrum










analysis








Ug hr











RT-PCR

























RESULTS

Aqueous extract
























Root exudates








Phenolic acids



















hr ( Fig3)











DISCUSSION










allelopathy



























Norway spruce





























management programs







References


















20921ndash928



Crop Sci 4544-49
















Ecol251907-1922









42 185-209








Physiol8216ndash26









2135-145



2389ndash2393









Agron J 93 21-26



















Korea








Chem 52 7607 -7613















1018

















28(1)229-243












Cell 61197ndash1209







30 1663-1678



Ecol 301647-1662


885




242







Biochem 68406-4094



J Chem Ecol 221767-1781









14S153ndashS164










38

Figure legends





above bars



















Gene Accession









pathway








of RT-PCR



barnyardgrass

Extraction

organ Treatments

Huajingxian1 IAC165

Root length

(cm)

Shoot length

(cm)

Root length

(cm)

Shoot length

(cm)

Stems




Leaves








barnyardgrass










0

150

300

450

600

HBA VA CA CMA FA

Phenolic acid

Conce

ntr

atio

ns

(μgg

FW

)

control

MeJA

MeSAa

c

b

ab

a

a

b

a

b

c

c a aa






0

250

500

750

1000

1250

HBA VA CA CMA FA

Phenolic acid

control

MeJA

MeSA

bb

a

cb

a

b

a

c

b

a

ca aa Co

nce

ntr

atio

ns

(μg

g F

W)






0

1000

2000

3000

4000

2 4 6


control

MeJA

MeSA

b

aa

b

aa

a

b

c

PA

L a

ctiv

ity

(U

gh

)





above bars

0

50

100

150

200

control MeJA MeSA

Signal compound

b

c

a

C4H

act

ivity (

Ug

h)






PA

L

C4H




Actin Actin














UV spectrum










analysis








Ug hr











RT-PCR

























RESULTS

Aqueous extract
























Root exudates








Phenolic acids



















hr ( Fig3)











DISCUSSION










allelopathy



























Norway spruce





























management programs







References


















20921ndash928



Crop Sci 4544-49
















Ecol251907-1922









42 185-209








Physiol8216ndash26









2135-145



2389ndash2393









Agron J 93 21-26



















Korea








Chem 52 7607 -7613















1018

















28(1)229-243












Cell 61197ndash1209







30 1663-1678



Ecol 301647-1662


885




242







Biochem 68406-4094



J Chem Ecol 221767-1781









14S153ndashS164










38

Figure legends





above bars



















Gene Accession









pathway








of RT-PCR



barnyardgrass

Extraction

organ Treatments

Huajingxian1 IAC165

Root length

(cm)

Shoot length

(cm)

Root length

(cm)

Shoot length

(cm)

Stems




Leaves








barnyardgrass










0

150

300

450

600

HBA VA CA CMA FA

Phenolic acid

Conce

ntr

atio

ns

(μgg

FW

)

control

MeJA

MeSAa

c

b

ab

a

a

b

a

b

c

c a aa






0

250

500

750

1000

1250

HBA VA CA CMA FA

Phenolic acid

control

MeJA

MeSA

bb

a

cb

a

b

a

c

b

a

ca aa Co

nce

ntr

atio

ns

(μg

g F

W)






0

1000

2000

3000

4000

2 4 6


control

MeJA

MeSA

b

aa

b

aa

a

b

c

PA

L a

ctiv

ity

(U

gh

)





above bars

0

50

100

150

200

control MeJA MeSA

Signal compound

b

c

a

C4H

act

ivity (

Ug

h)






PA

L

C4H




Actin Actin






RT-PCR

























RESULTS

Aqueous extract
























Root exudates








Phenolic acids



















hr ( Fig3)











DISCUSSION










allelopathy



























Norway spruce





























management programs







References


















20921ndash928



Crop Sci 4544-49
















Ecol251907-1922









42 185-209








Physiol8216ndash26









2135-145



2389ndash2393









Agron J 93 21-26



















Korea








Chem 52 7607 -7613















1018

















28(1)229-243












Cell 61197ndash1209







30 1663-1678



Ecol 301647-1662


885




242







Biochem 68406-4094



J Chem Ecol 221767-1781









14S153ndashS164










38

Figure legends





above bars



















Gene Accession









pathway








of RT-PCR



barnyardgrass

Extraction

organ Treatments

Huajingxian1 IAC165

Root length

(cm)

Shoot length

(cm)

Root length

(cm)

Shoot length

(cm)

Stems




Leaves








barnyardgrass










0

150

300

450

600

HBA VA CA CMA FA

Phenolic acid

Conce

ntr

atio

ns

(μgg

FW

)

control

MeJA

MeSAa

c

b

ab

a

a

b

a

b

c

c a aa






0

250

500

750

1000

1250

HBA VA CA CMA FA

Phenolic acid

control

MeJA

MeSA

bb

a

cb

a

b

a

c

b

a

ca aa Co

nce

ntr

atio

ns

(μg

g F

W)






0

1000

2000

3000

4000

2 4 6


control

MeJA

MeSA

b

aa

b

aa

a

b

c

PA

L a

ctiv

ity

(U

gh

)





above bars

0

50

100

150

200

control MeJA MeSA

Signal compound

b

c

a

C4H

act

ivity (

Ug

h)






PA

L

C4H




Actin Actin

RESULTS

Aqueous extract
























Root exudates








Phenolic acids



















hr ( Fig3)











DISCUSSION










allelopathy



























Norway spruce





























management programs







References


















20921ndash928



Crop Sci 4544-49
















Ecol251907-1922









42 185-209








Physiol8216ndash26









2135-145



2389ndash2393









Agron J 93 21-26



















Korea








Chem 52 7607 -7613















1018

















28(1)229-243












Cell 61197ndash1209







30 1663-1678



Ecol 301647-1662


885




242







Biochem 68406-4094



J Chem Ecol 221767-1781









14S153ndashS164










38

Figure legends





above bars



















Gene Accession









pathway








of RT-PCR



barnyardgrass

Extraction

organ Treatments

Huajingxian1 IAC165

Root length

(cm)

Shoot length

(cm)

Root length

(cm)

Shoot length

(cm)

Stems




Leaves








barnyardgrass










0

150

300

450

600

HBA VA CA CMA FA

Phenolic acid

Conce

ntr

atio

ns

(μgg

FW

)

control

MeJA

MeSAa

c

b

ab

a

a

b

a

b

c

c a aa






0

250

500

750

1000

1250

HBA VA CA CMA FA

Phenolic acid

control

MeJA

MeSA

bb

a

cb

a

b

a

c

b

a

ca aa Co

nce

ntr

atio

ns

(μg

g F

W)






0

1000

2000

3000

4000

2 4 6


control

MeJA

MeSA

b

aa

b

aa

a

b

c

PA

L a

ctiv

ity

(U

gh

)





above bars

0

50

100

150

200

control MeJA MeSA

Signal compound

b

c

a

C4H

act

ivity (

Ug

h)






PA

L

C4H




Actin Actin



Phenolic acids



















hr ( Fig3)











DISCUSSION










allelopathy



























Norway spruce





























management programs







References


















20921ndash928



Crop Sci 4544-49
















Ecol251907-1922









42 185-209








Physiol8216ndash26









2135-145



2389ndash2393









Agron J 93 21-26



















Korea








Chem 52 7607 -7613















1018

















28(1)229-243












Cell 61197ndash1209







30 1663-1678



Ecol 301647-1662


885




242







Biochem 68406-4094



J Chem Ecol 221767-1781









14S153ndashS164










38

Figure legends





above bars



















Gene Accession









pathway








of RT-PCR



barnyardgrass

Extraction

organ Treatments

Huajingxian1 IAC165

Root length

(cm)

Shoot length

(cm)

Root length

(cm)

Shoot length

(cm)

Stems




Leaves








barnyardgrass










0

150

300

450

600

HBA VA CA CMA FA

Phenolic acid

Conce

ntr

atio

ns

(μgg

FW

)

control

MeJA

MeSAa

c

b

ab

a

a

b

a

b

c

c a aa






0

250

500

750

1000

1250

HBA VA CA CMA FA

Phenolic acid

control

MeJA

MeSA

bb

a

cb

a

b

a

c

b

a

ca aa Co

nce

ntr

atio

ns

(μg

g F

W)






0

1000

2000

3000

4000

2 4 6


control

MeJA

MeSA

b

aa

b

aa

a

b

c

PA

L a

ctiv

ity

(U

gh

)





above bars

0

50

100

150

200

control MeJA MeSA

Signal compound

b

c

a

C4H

act

ivity (

Ug

h)






PA

L

C4H




Actin Actin

DISCUSSION










allelopathy



























Norway spruce





























management programs







References


















20921ndash928



Crop Sci 4544-49
















Ecol251907-1922









42 185-209








Physiol8216ndash26









2135-145



2389ndash2393









Agron J 93 21-26



















Korea








Chem 52 7607 -7613















1018

















28(1)229-243












Cell 61197ndash1209







30 1663-1678



Ecol 301647-1662


885




242







Biochem 68406-4094



J Chem Ecol 221767-1781









14S153ndashS164










38

Figure legends





above bars



















Gene Accession









pathway








of RT-PCR



barnyardgrass

Extraction

organ Treatments

Huajingxian1 IAC165

Root length

(cm)

Shoot length

(cm)

Root length

(cm)

Shoot length

(cm)

Stems




Leaves








barnyardgrass










0

150

300

450

600

HBA VA CA CMA FA

Phenolic acid

Conce

ntr

atio

ns

(μgg

FW

)

control

MeJA

MeSAa

c

b

ab

a

a

b

a

b

c

c a aa






0

250

500

750

1000

1250

HBA VA CA CMA FA

Phenolic acid

control

MeJA

MeSA

bb

a

cb

a

b

a

c

b

a

ca aa Co

nce

ntr

atio

ns

(μg

g F

W)






0

1000

2000

3000

4000

2 4 6


control

MeJA

MeSA

b

aa

b

aa

a

b

c

PA

L a

ctiv

ity

(U

gh

)





above bars

0

50

100

150

200

control MeJA MeSA

Signal compound

b

c

a

C4H

act

ivity (

Ug

h)






PA

L

C4H




Actin Actin





























management programs







References


















20921ndash928



Crop Sci 4544-49
















Ecol251907-1922









42 185-209








Physiol8216ndash26









2135-145



2389ndash2393









Agron J 93 21-26



















Korea








Chem 52 7607 -7613















1018

















28(1)229-243












Cell 61197ndash1209







30 1663-1678



Ecol 301647-1662


885




242







Biochem 68406-4094



J Chem Ecol 221767-1781









14S153ndashS164










38

Figure legends





above bars



















Gene Accession









pathway








of RT-PCR



barnyardgrass

Extraction

organ Treatments

Huajingxian1 IAC165

Root length

(cm)

Shoot length

(cm)

Root length

(cm)

Shoot length

(cm)

Stems




Leaves








barnyardgrass










0

150

300

450

600

HBA VA CA CMA FA

Phenolic acid

Conce

ntr

atio

ns

(μgg

FW

)

control

MeJA

MeSAa

c

b

ab

a

a

b

a

b

c

c a aa






0

250

500

750

1000

1250

HBA VA CA CMA FA

Phenolic acid

control

MeJA

MeSA

bb

a

cb

a

b

a

c

b

a

ca aa Co

nce

ntr

atio

ns

(μg

g F

W)






0

1000

2000

3000

4000

2 4 6


control

MeJA

MeSA

b

aa

b

aa

a

b

c

PA

L a

ctiv

ity

(U

gh

)





above bars

0

50

100

150

200

control MeJA MeSA

Signal compound

b

c

a

C4H

act

ivity (

Ug

h)






PA

L

C4H




Actin Actin

References


















20921ndash928



Crop Sci 4544-49
















Ecol251907-1922









42 185-209








Physiol8216ndash26









2135-145



2389ndash2393









Agron J 93 21-26



















Korea








Chem 52 7607 -7613















1018

















28(1)229-243












Cell 61197ndash1209







30 1663-1678



Ecol 301647-1662


885




242







Biochem 68406-4094



J Chem Ecol 221767-1781









14S153ndashS164










38

Figure legends





above bars



















Gene Accession









pathway








of RT-PCR



barnyardgrass

Extraction

organ Treatments

Huajingxian1 IAC165

Root length

(cm)

Shoot length

(cm)

Root length

(cm)

Shoot length

(cm)

Stems




Leaves








barnyardgrass










0

150

300

450

600

HBA VA CA CMA FA

Phenolic acid

Conce

ntr

atio

ns

(μgg

FW

)

control

MeJA

MeSAa

c

b

ab

a

a

b

a

b

c

c a aa






0

250

500

750

1000

1250

HBA VA CA CMA FA

Phenolic acid

control

MeJA

MeSA

bb

a

cb

a

b

a

c

b

a

ca aa Co

nce

ntr

atio

ns

(μg

g F

W)






0

1000

2000

3000

4000

2 4 6


control

MeJA

MeSA

b

aa

b

aa

a

b

c

PA

L a

ctiv

ity

(U

gh

)





above bars

0

50

100

150

200

control MeJA MeSA

Signal compound

b

c

a

C4H

act

ivity (

Ug

h)






PA

L

C4H




Actin Actin





Ecol251907-1922









42 185-209








Physiol8216ndash26









2135-145



2389ndash2393









Agron J 93 21-26



















Korea








Chem 52 7607 -7613















1018

















28(1)229-243












Cell 61197ndash1209







30 1663-1678



Ecol 301647-1662


885




242







Biochem 68406-4094



J Chem Ecol 221767-1781









14S153ndashS164










38

Figure legends





above bars



















Gene Accession









pathway








of RT-PCR



barnyardgrass

Extraction

organ Treatments

Huajingxian1 IAC165

Root length

(cm)

Shoot length

(cm)

Root length

(cm)

Shoot length

(cm)

Stems




Leaves








barnyardgrass










0

150

300

450

600

HBA VA CA CMA FA

Phenolic acid

Conce

ntr

atio

ns

(μgg

FW

)

control

MeJA

MeSAa

c

b

ab

a

a

b

a

b

c

c a aa






0

250

500

750

1000

1250

HBA VA CA CMA FA

Phenolic acid

control

MeJA

MeSA

bb

a

cb

a

b

a

c

b

a

ca aa Co

nce

ntr

atio

ns

(μg

g F

W)






0

1000

2000

3000

4000

2 4 6


control

MeJA

MeSA

b

aa

b

aa

a

b

c

PA

L a

ctiv

ity

(U

gh

)





above bars

0

50

100

150

200

control MeJA MeSA

Signal compound

b

c

a

C4H

act

ivity (

Ug

h)






PA

L

C4H




Actin Actin









Agron J 93 21-26



















Korea








Chem 52 7607 -7613















1018

















28(1)229-243












Cell 61197ndash1209







30 1663-1678



Ecol 301647-1662


885




242







Biochem 68406-4094



J Chem Ecol 221767-1781









14S153ndashS164










38

Figure legends





above bars



















Gene Accession









pathway








of RT-PCR



barnyardgrass

Extraction

organ Treatments

Huajingxian1 IAC165

Root length

(cm)

Shoot length

(cm)

Root length

(cm)

Shoot length

(cm)

Stems




Leaves








barnyardgrass










0

150

300

450

600

HBA VA CA CMA FA

Phenolic acid

Conce

ntr

atio

ns

(μgg

FW

)

control

MeJA

MeSAa

c

b

ab

a

a

b

a

b

c

c a aa






0

250

500

750

1000

1250

HBA VA CA CMA FA

Phenolic acid

control

MeJA

MeSA

bb

a

cb

a

b

a

c

b

a

ca aa Co

nce

ntr

atio

ns

(μg

g F

W)






0

1000

2000

3000

4000

2 4 6


control

MeJA

MeSA

b

aa

b

aa

a

b

c

PA

L a

ctiv

ity

(U

gh

)





above bars

0

50

100

150

200

control MeJA MeSA

Signal compound

b

c

a

C4H

act

ivity (

Ug

h)






PA

L

C4H




Actin Actin



Chem 52 7607 -7613















1018

















28(1)229-243












Cell 61197ndash1209







30 1663-1678



Ecol 301647-1662


885




242







Biochem 68406-4094



J Chem Ecol 221767-1781









14S153ndashS164










38

Figure legends





above bars



















Gene Accession









pathway








of RT-PCR



barnyardgrass

Extraction

organ Treatments

Huajingxian1 IAC165

Root length

(cm)

Shoot length

(cm)

Root length

(cm)

Shoot length

(cm)

Stems




Leaves








barnyardgrass










0

150

300

450

600

HBA VA CA CMA FA

Phenolic acid

Conce

ntr

atio

ns

(μgg

FW

)

control

MeJA

MeSAa

c

b

ab

a

a

b

a

b

c

c a aa






0

250

500

750

1000

1250

HBA VA CA CMA FA

Phenolic acid

control

MeJA

MeSA

bb

a

cb

a

b

a

c

b

a

ca aa Co

nce

ntr

atio

ns

(μg

g F

W)






0

1000

2000

3000

4000

2 4 6


control

MeJA

MeSA

b

aa

b

aa

a

b

c

PA

L a

ctiv

ity

(U

gh

)





above bars

0

50

100

150

200

control MeJA MeSA

Signal compound

b

c

a

C4H

act

ivity (

Ug

h)






PA

L

C4H




Actin Actin












Cell 61197ndash1209







30 1663-1678



Ecol 301647-1662


885




242







Biochem 68406-4094



J Chem Ecol 221767-1781









14S153ndashS164










38

Figure legends





above bars



















Gene Accession









pathway








of RT-PCR



barnyardgrass

Extraction

organ Treatments

Huajingxian1 IAC165

Root length

(cm)

Shoot length

(cm)

Root length

(cm)

Shoot length

(cm)

Stems




Leaves








barnyardgrass










0

150

300

450

600

HBA VA CA CMA FA

Phenolic acid

Conce

ntr

atio

ns

(μgg

FW

)

control

MeJA

MeSAa

c

b

ab

a

a

b

a

b

c

c a aa






0

250

500

750

1000

1250

HBA VA CA CMA FA

Phenolic acid

control

MeJA

MeSA

bb

a

cb

a

b

a

c

b

a

ca aa Co

nce

ntr

atio

ns

(μg

g F

W)






0

1000

2000

3000

4000

2 4 6


control

MeJA

MeSA

b

aa

b

aa

a

b

c

PA

L a

ctiv

ity

(U

gh

)





above bars

0

50

100

150

200

control MeJA MeSA

Signal compound

b

c

a

C4H

act

ivity (

Ug

h)






PA

L

C4H




Actin Actin



J Chem Ecol 221767-1781









14S153ndashS164










38

Figure legends





above bars



















Gene Accession









pathway








of RT-PCR



barnyardgrass

Extraction

organ Treatments

Huajingxian1 IAC165

Root length

(cm)

Shoot length

(cm)

Root length

(cm)

Shoot length

(cm)

Stems




Leaves








barnyardgrass










0

150

300

450

600

HBA VA CA CMA FA

Phenolic acid

Conce

ntr

atio

ns

(μgg

FW

)

control

MeJA

MeSAa

c

b

ab

a

a

b

a

b

c

c a aa






0

250

500

750

1000

1250

HBA VA CA CMA FA

Phenolic acid

control

MeJA

MeSA

bb

a

cb

a

b

a

c

b

a

ca aa Co

nce

ntr

atio

ns

(μg

g F

W)






0

1000

2000

3000

4000

2 4 6


control

MeJA

MeSA

b

aa

b

aa

a

b

c

PA

L a

ctiv

ity

(U

gh

)





above bars

0

50

100

150

200

control MeJA MeSA

Signal compound

b

c

a

C4H

act

ivity (

Ug

h)






PA

L

C4H




Actin Actin

Figure legends





above bars



















Gene Accession









pathway








of RT-PCR



barnyardgrass

Extraction

organ Treatments

Huajingxian1 IAC165

Root length

(cm)

Shoot length

(cm)

Root length

(cm)

Shoot length

(cm)

Stems




Leaves








barnyardgrass










0

150

300

450

600

HBA VA CA CMA FA

Phenolic acid

Conce

ntr

atio

ns

(μgg

FW

)

control

MeJA

MeSAa

c

b

ab

a

a

b

a

b

c

c a aa






0

250

500

750

1000

1250

HBA VA CA CMA FA

Phenolic acid

control

MeJA

MeSA

bb

a

cb

a

b

a

c

b

a

ca aa Co

nce

ntr

atio

ns

(μg

g F

W)






0

1000

2000

3000

4000

2 4 6


control

MeJA

MeSA

b

aa

b

aa

a

b

c

PA

L a

ctiv

ity

(U

gh

)





above bars

0

50

100

150

200

control MeJA MeSA

Signal compound

b

c

a

C4H

act

ivity (

Ug

h)






PA

L

C4H




Actin Actin


Gene Accession









pathway








of RT-PCR



barnyardgrass

Extraction

organ Treatments

Huajingxian1 IAC165

Root length

(cm)

Shoot length

(cm)

Root length

(cm)

Shoot length

(cm)

Stems




Leaves








barnyardgrass










0

150

300

450

600

HBA VA CA CMA FA

Phenolic acid

Conce

ntr

atio

ns

(μgg

FW

)

control

MeJA

MeSAa

c

b

ab

a

a

b

a

b

c

c a aa






0

250

500

750

1000

1250

HBA VA CA CMA FA

Phenolic acid

control

MeJA

MeSA

bb

a

cb

a

b

a

c

b

a

ca aa Co

nce

ntr

atio

ns

(μg

g F

W)






0

1000

2000

3000

4000

2 4 6


control

MeJA

MeSA

b

aa

b

aa

a

b

c

PA

L a

ctiv

ity

(U

gh

)





above bars

0

50

100

150

200

control MeJA MeSA

Signal compound

b

c

a

C4H

act

ivity (

Ug

h)






PA

L

C4H




Actin Actin



barnyardgrass

Extraction

organ Treatments

Huajingxian1 IAC165

Root length

(cm)

Shoot length

(cm)

Root length

(cm)

Shoot length

(cm)

Stems




Leaves








barnyardgrass










0

150

300

450

600

HBA VA CA CMA FA

Phenolic acid

Conce

ntr

atio

ns

(μgg

FW

)

control

MeJA

MeSAa

c

b

ab

a

a

b

a

b

c

c a aa






0

250

500

750

1000

1250

HBA VA CA CMA FA

Phenolic acid

control

MeJA

MeSA

bb

a

cb

a

b

a

c

b

a

ca aa Co

nce

ntr

atio

ns

(μg

g F

W)






0

1000

2000

3000

4000

2 4 6


control

MeJA

MeSA

b

aa

b

aa

a

b

c

PA

L a

ctiv

ity

(U

gh

)





above bars

0

50

100

150

200

control MeJA MeSA

Signal compound

b

c

a

C4H

act

ivity (

Ug

h)






PA

L

C4H




Actin Actin



barnyardgrass










0

150

300

450

600

HBA VA CA CMA FA

Phenolic acid

Conce

ntr

atio

ns

(μgg

FW

)

control

MeJA

MeSAa

c

b

ab

a

a

b

a

b

c

c a aa






0

250

500

750

1000

1250

HBA VA CA CMA FA

Phenolic acid

control

MeJA

MeSA

bb

a

cb

a

b

a

c

b

a

ca aa Co

nce

ntr

atio

ns

(μg

g F

W)






0

1000

2000

3000

4000

2 4 6


control

MeJA

MeSA

b

aa

b

aa

a

b

c

PA

L a

ctiv

ity

(U

gh

)





above bars

0

50

100

150

200

control MeJA MeSA

Signal compound

b

c

a

C4H

act

ivity (

Ug

h)






PA

L

C4H




Actin Actin

0

150

300

450

600

HBA VA CA CMA FA

Phenolic acid

Conce

ntr

atio

ns

(μgg

FW

)

control

MeJA

MeSAa

c

b

ab

a

a

b

a

b

c

c a aa






0

250

500

750

1000

1250

HBA VA CA CMA FA

Phenolic acid

control

MeJA

MeSA

bb

a

cb

a

b

a

c

b

a

ca aa Co

nce

ntr

atio

ns

(μg

g F

W)






0

1000

2000

3000

4000

2 4 6


control

MeJA

MeSA

b

aa

b

aa

a

b

c

PA

L a

ctiv

ity

(U

gh

)





above bars

0

50

100

150

200

control MeJA MeSA

Signal compound

b

c

a

C4H

act

ivity (

Ug

h)






PA

L

C4H




Actin Actin

0

250

500

750

1000

1250

HBA VA CA CMA FA

Phenolic acid

control

MeJA

MeSA

bb

a

cb

a

b

a

c

b

a

ca aa Co

nce

ntr

atio

ns

(μg

g F

W)






0

1000

2000

3000

4000

2 4 6


control

MeJA

MeSA

b

aa

b

aa

a

b

c

PA

L a

ctiv

ity

(U

gh

)





above bars

0

50

100

150

200

control MeJA MeSA

Signal compound

b

c

a

C4H

act

ivity (

Ug

h)






PA

L

C4H




Actin Actin

0

1000

2000

3000

4000

2 4 6


control

MeJA

MeSA

b

aa

b

aa

a

b

c

PA

L a

ctiv

ity

(U

gh

)





above bars

0

50

100

150

200

control MeJA MeSA

Signal compound

b

c

a

C4H

act

ivity (

Ug

h)






PA

L

C4H




Actin Actin

0

50

100

150

200

control MeJA MeSA

Signal compound

b

c

a

C4H

act

ivity (

Ug

h)






PA

L

C4H




Actin Actin


PA

L

C4H




Actin Actin

researchoutput.csu.edu 2719.pdf · phenylpropanoid pathway. aqueous extracts of the leaves of rice...

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