Allelopathic potential of Rhus chinensis on seedling growth of radish,semen cassiae and black soyabean

Allelopathic potential of Rhus chinensis on seedling growth of radish,semen cassiae and black soyabean

Long-yuan Liu?Hong-zhi He?Shi-ming Luo?Hua-shou Li

The allelopathic potential of Rhus chinensis Mill on seedling growth of radish(Raphanus sativus L.), semen cassiae(seed of Cassia obtusifolia L.)and black soyabean[Glycine max(L.)Merr.]was investigated under laboratory conditions.The water extracts of seed,leaf and bark of R.chinensis inhibited the germination and the growth of roots and shoots of radish,semen cassiae,and black soyabean.Significant increasing reductions in the germination and growth of roots and shoots were observed as the concentration of water extract increased in all bioassays.The order of allelopathic potentials of the three parts from R.chinensis plant on seed germination of these receptors was leaf＞seed＞bark.However,the inhibition of water extracts from R.chinensis seed on root activity of semen cassiae was more severe than from the other two parts.The allelopathic potential on the germination of radish was higher than that of black soyabean and semen cassiae.The integrated effect index for allelopathic potentialof water extracts of leaf of R.chinensis on radish,semen cassiae and black soyabean seedling growth were -0.88,-0.93 and-0.98 respectively.The results suggested that it is important to minimize the negative allelopathic potential of R.chinensis in agroforestry systems, and to exploitthis traditional Chinese medicinaltree for the production of natural bio-pesticides.

Allelopathy·Rhus chinensis·Water extracts· Seedling growth

Introduction

The species Rhus chinensis Mill(Anacardiaceae)is an important species of the genus Rhus,which contains over 250 individual species,with six species found in China (Djakpo and Yao 2010).It is an important economic tree notonly widely grown in China,butalso widely distributed in temperate,subtropical,and tropical regions,including Japan,Malaysia and India(Rayne and Mazza 2007;Ren et al.2008).As a traditional Chinese medicinal plant,R. chinensis has long been used to treat diseases,such as inflammation,impaludism,endocrine disorder,rheumatism and peptic ulcer disease(Rayne and Mazza 2007;Djakpo and Yao 2010).R.chinensis can also be used as a hostplant for the production of gallnut which is also used in traditional Chinese medicine and as a dyeing material(Tian et al.2009).R.chinensis has been applied in agroforestry systems by intercropping with other plants.It has been revealed that extracts of R.chinensis possess strong antiviral,antibacterial,anticancer,hepatoprotective,antidiarrhealand antioxidantcapacities(Ahn etal.1998;Kim etal. 2005;Gu et al.2007;Wang et al.2008;Chen et al.2009; Djakpo and Yao 2010).Moreover,the root,leaf,stem and fruit of R.chinensis have been used as pesticides byfarmers in South China.Could R.chinensis impose allelopathic effects on its neighboring species within a plant community?We reporthere on the allelopathic potentialof R.chinensis on seedling growth of radish,semen cassiae and black soyabean.

Fig. 1 a Effects of water extracts of leaf, bark and seed of R.chinensis at different concentrations (200, 100, 50, 33 and 25 mgfresh weight of sample per L distilled water) on the germination trendof radish. Data are the means of three replicates with standard errorsshown by vertical bars. b Effects of water extracts of leaf, bark andseed of R. chinensis at different concentrations (200, 100, 50, 33 and25 mg fresh weight of sample per L distilled water) on the germination of semen cassiae. Data are the means of three replicateswith standard errors shown by vertical bars. c Effects of waterextracts of leaf, bark and seed of R. chinensis at different concentrations(200, 100, 50, 33 and 25 mg fresh weight of sample per Ldistilled water) on the germination of black soyabean. Data are themeans of three replicates with standard errors shown by verticalbars

Fig.1 continued

Materials and methods

Extraction

Rhus chinensis Mill and semen Cassiae(seed of Cassia obtusifolia L.)were obtained from the medicine nursery on the Zhongshan campus of Guangdong Pharmaceutical University.Radish(Raphanus sativus L.)and black soyabean[Glycine max(L.)Merr.]were acquired from a local market.

R.chinensis plants were isolated from other trees with at least 1 year of normal growth.Bark of R.chinensis was removed from trunks at heights≥50 cm above the ground. Only half of the bark was scraped from the trunk in order to ensure survival of the trees.Healthy leaves in dark green and seeds with white coats were collected.Dried bark, seeds and leaves were separately cut into 6 cm long pieces and then powdered in a grinder until 98%of the powder could pass through No.1 sieve(10 mesh)and 30%of the powder could pass through No.2 sieve(24 mesh).The powder samples were extracted with five times their weight of distilled water in an 8 L beaker with oscillation of 30 min every 12 h for 48 h in the dark.And then the mixture was filtered through a paper filter.The filtrates were collected and adjusted to five levels:200,100,50,33 and 25 mg fresh weight of sample per L distilled water. The solutions were preserved in brown bottles at 4°C.

Bioassays

For seed germination tests,seeds were soaked with 5.25% (w/v)sodium hypochlorite solution for 15 min,rinsed three times with distilled water and then evenly placed on a twolayer paper filter in sterilized 9 cm Petri dishes.Extracts of differentparts of the donor plant were added to dishes until 2 mm above the seed layer.Each treatment with three replicates and an additional distilled water treatment as control were used.Dishes were put in an incubator at constant temperature of 25°C,85%humidity and 12 h lightper day.The volume of the testsolutions was made up to maintain the original level every day and the numbers of germinated seeds were recorded every day for 7 days.The rootlengths were also measured each day.The rootactivity vigor of semen cassiae was measured by TTC Method(Li 2009)on day 7.

Statistical analyses

Data were analyzed by one-way analysis of variance (ANOVA)using SPSS PASW Statistics v18.0 software (SPSS Statistics,Shanghai,China).Duncan’s new multiple range tests were calculated when treatments were significantly different atα=0.05.

The equations for calculation of relative inhibition index (RI)for receptor plant according to Williamson and Richardson(1988)were as follows:

where C and T represent the response of control and treatment,respectively.When RI≥0,it represents stimulation effects and when RI＜0,it shows inhibitory effects. The absolute value of RI represents the magnitude of the effects.The integrated effects were assessed using integrated effect index(IEI),which is the average value of response indexes(RI).

Results

Effects of donor water extracts on receptors’seed germination

As shown in Fig.1,the extracts from differenttissues of R. chinensis significantly inhibited and delayed seed germination of the three receptors.The intensity of inhibition increased with increasing extract concentration.High concentrations of extracts at 100 and 200 mg·L-1apparently totally inhibited the germination of radish.Radish seed completed its germination at the 4th day in the control treatment,but germination of other treatments was delayed up to the 7th day(Fig.1a).Inhibitory effects were stronger on radish than on black soyabean or semen cassiae.

The extracts of different tissues of R.chinensis had significant inhibitory effects on germination rates of the three receptor species.The increase of water extract concentrations resulted in decreasing germination rates.The order of inhibitory intensity of the same concentration of donor water extracts on germination rates of the receptors was leaf＞seed＞bark.Inhibitory effects were stronger on radish than on black soyabean or semen cassiae(Fig.1).

Allelopathic potential of donor water extracts on receptors

Seeds ofthe three receptorspecies had differentsensitivities to the water extracts of leaf,bark,and seed of R.Chinensis (Table 1).The increase of water extract concentrations resulted in increasing inhibitory effects.The IEI values of water extracts of leaf on radish,semen cassiae,and black soyabean were-0.88,-0.93 and-0.98,respectively.The IEI values of water extracts of bark on these three receptors were-0.36,-0.32 and-0.45,respectively.The IEIvalues ofwaterextractsofseed on these three receptorswere-0.69, -0.65 and-0.95,respectively.

Effects of donor water extracts on receptors seedlings’root length

Comparison of the five longest seedling root lengths by treatment,showed thatwater extracts of leaf,bark and seed of R.chinensis significantly inhibited the root growth of radish,semen cassiae and black soyabean(Fig.2).Increaseof water extract concentration resulted in increasing inhibitory effects on seedling rootgrowth.The average root lengths of semen cassia and black soyabean declined to 50.92 and 62.26%at least.However,the five longest average root length of radish were significantly reduced to 41%of the control even at 25 mg·L-1.The inhibition rates were much higher with higher concentrations.

Table 1 Relative inhibition index(RI)and integrated effect index(IEI)of the three receptor plants by R.chinensis water extracts

Fig.2 Effects of water extracts of leaf,bark and seed of R.chinensis at different concentrations on root length of radish a semen cassiae b black soyabean c seedlings.Bars are standard errors and values sharing a common lowercase letter are not significantly different at p＜0.05

Effects of donor water extracts on semen cassiae root activity

Alltissues from R.chinensis exhibited significantinhibition effects on rootactivity vigor of seedlings of semen cassiae. The greatestinhibition effectwas associated with the seeds of R.chinensis and the leastinhibition effectwas associated with the leaves of R.chinensis.Waterextracts at25 mg·L-1from leaf,bark and seed of R.chinensis reduced rootactivity vigor to 82.43,32.97 and 27.28%,respectively.The root activity vigorof semen cassiae decreased to 22.98,21.68 and 5.74%respectively at50 mg·L-1concentration from leaf, bark,and seed(Table 2).

Discussion

This study demonstrated that R.chinensis displayed allelopathic activity and influenced the emergence and seedling growth of radish,semen cassia and black soyabean.The effects varied depending on the tissue and extract concentration,the target species and the growth parameters measured.With increasing concentration,leaf extracts of R. chinensis caused more significant delay and reduction of seed germination of target species than extracts from barks and seeds extracts.Delayed seed germination by allelopathic extract could be further strengthened by osmotic effects on rate of absorption,delayed initiation of germination and especially cell elongation(Black 1989).At the same concentration,RI on germination of leaf extract treatment was lower than thatof bark or seed extracttreatment.The IEIof water extracts of leaves on black soyabean was-0.98,the lowest of all treatments.Seeds of R.chinensis affected seedling development and growth of the three species,but their effectwas especially strong on radish.

Seed extracts of R.chinensis displayed more consistent effects than did extracts ofleaforbark on targetspecies.Root length was reduced in all species by the water extracts of seed,leaf and bark of R.chinensis.Inhibitory effecton root length,also reported in other experiments(Tongma et al. 2001;Shiraishi et al.2002),may affect the competitive balance between species during the establishmentstage.

Allelopathy is the effect of one species(especially plant) on the growth and development of other species through the release of chemical substances(Kohli et al.1998;Reigosa et al.1999).Allelopathy has been shown to play an important role in forests,influencing the composition of vegetation,and also providing an explanation for the patterns of forest regeneration.Several reports addressed the allelopathic effects of trees on other trees or natural vegetation in the field(Rietveld et al.1983;Klionsky et al. 2011).The allelopathic potential of trees on other trees or naturalvegetation has also been assessed(Chou etal.1989;Souto et al.2001;Fang et al.2009;Zhang et al.2010; Valera-Burgos et al.2012).There are also some research on the allelopathic effect of trees on agricultural crops.A significant allelopathic effect of trees on growth and yield of agricultural crops was noticed in the field,such as Populus deltoides on wheat and winter season crops(Singh et al.1998,2001),and E.tereticornis on crops(Singh and Kohli 1992).Moreover,the allelopathic potentials of trees were also tested using laboratory bioassay with crops,such as Adina cordifolia,A.nepalensis,Celtis australis and Prunus cerasoides on Eleusine coracana,G.max and Hordeum vulgare(Bhatt and Todaria 1990),Picea halepensis on Lactuca sativa and Linum strictum(Fernandez et al.2006),P.halepensis and Quercus coccifera(Alrababah etal.2009),E.grandis on R.sativus and Phaseolus aureus(Zhang et al.2010),Araucaria angustifolia(Braine et al.2012),P.schrenkiana n(Ruan et al.2011).Yu et al. (2003)found that crude root exudates and extracts of cucumber,as well as compounds purified from these sources,increased the activity of SOD and POD in cucumber plants,and decreased stomatal conductance, transpiration and net photosynthesis.Their work also suggested that the presence of superoxide anions also increased in treated roots as the concentrations of root extracts/exudates increased.Root activity can reflect the levelof dehydrogenase activity(Zhao etal.1998),which is an important physiological parameter of root activity.Our results indicated that root activity vigor of semen cassiae was reduced by the water extracts of seeds,leaves and bark of R.chinensis.The root activities of semen cassiae were significantly reduced to 82.43,32.97 and 27.28%of the CK under 25 mg·L-1extracts from leaf,bark and seed of R.chinensis respectively.Seed extracts of R.chinensis displayed more consistent effects than did extracts of leaf and bark on root activity of semen cassiae.

Table 2 Effect of R.chinensis water extracts on semen cassiae root activity vigor(mg·g-1·h-1)

Most trees were reported in published studies to have inhibitory effects on crops.In the present study,the water extracts of leaf,bark and seed of R.chinensis also showed significant inhibitory effects on seedling growth of radish, semen cassiae,and black soyabean.This work reveals the existence of different sensitivity of plant species to allelopathic chemicals from R.chinensis.Itmakes these results potentially useful to manage R.chinensis communities and agro-forest systems based on R.chinensis.

In most studies of trees,phenolics were identified as potential allelochemical compounds.Chou et al.(1989) identified phytotoxic phenolics as the allelopathic chemicals of Z.formosana and Chinese fir.Singh et al.(2001)concluded thatthe allelopathic interference of P.detoides with crops was effected by release of phytotoxin phenolics from leavesand litterand accumulation ofthese phytotoxins in the soil.Fernandez et al.(2006)documented that the potential allelochemical compounds of P.halepensis were several phenolic acids(e.g.4-hydroxybenzoic acid and p-coumaric acid).Ruan et al.(2011)also isolated a potential allelochemical phenolic compound 3,4-dihydroxyacetophenone (DHAP)from P.schrenkiana.However,Braine etal.(2012) documented thatthe potentialallelochemicalcompounds of A.angustifolia were ent-kaurene and phyllocladene.R. chinensis also contains various chemical substances with biological activity,including terpenes,flavonoids,tannins, resin,starch,and organic acids(Parveen and Khan 1985; Taniguchi et al.2000).These results suggest that,in some cases,an allelochemical may be directly involved in the production of reactive oxygen species(ROS)whereas the increase in oxidizing enzymes is a secondary response to the increase in free radicals.In other cases,the allelochemical might directly inhibit oxidizing enzymes in some way and cause oxidative damage to the plant(Weir etal.2004).This mightalso explain the allelopathy ofdonorwaterextracts on receptor seedling rootlength and rootactivity in our study.

In conclusion,this study is the first to demonstrate the allelopathic potential of R.chinensis.The results of the bioassay in this work may contribute to our understanding of the allelopathic potential of R.chinensis.It is important to ascertain the allelopathic compounds of R.chinensis and to minimize their negative effects on cultural systems.

Ahn YJ,Lee CO,Kweon JH,Ahn JW,Park JH(1998)Growth inhibitory effects of Galla Rhois-derived tannins on intestinal bacteria.J Appl Microbiol 84:439–443

Alrababah MA,Tadros MJ,Samarah NH,Ghosheh H(2009) Allelopathic effects of Pinus halepensis and Quercus coccifera on the germination of Mediterranean crop seeds.New Forest 38:261–272

Bhatt BP,Todaria NP(1990)Studies on the allelopathic effects of some agroforestry tree crops of Garhwal Himalaya.Agrofor Syst 12:251–255

Black M(1989)Seed research-past,present and future.In:Taylorson RB(ed)Recentadvances in the developmentand germination of seeds.Plenum,New York,pp 1–6

Braine JW,Curcio GR,Wachowicz CM,Hansel FA(2012)Allelopathic effects of Araucaria angustifolia needle extracts in the growth of Lactuca sativa seeds.J For Res 17:440–445

Chen JC,Ho TY,Chang YS,Wu SL,Li CC,Hsiang CY(2009) Identification of Escherichia coli enterotoxin inhibitors from traditional medicinal herbs by in silico,in vitro,and in vivo analyses.J Ethnopharmacol 121:372–378

Chou CH,Chang SJ,Cheng CM,Wang YC,Hsu FH,Den WH(1989) The selective allelopathic interaction of a pasture-forest intercropping in Taiwan II.Interaction between kikuyu grass and three hardwood plants.Plant Soil 116:207–215

Djakpo O,Yao WR(2010)Rhus Chinensis and Galla Chinensis–folklore to modern evidence:review.Phytother Res24:1739–1747

Fang B,Yu S,Wang Y,Qiu X,Cai C,Liu S(2009)Allelopathic effects of Eucalyptus urophylla on ten tree species in south China.Agrofor Syst 76:401–408

Fernandez C,Lelong B,Vila B,Me′vy JP,Robles C,Greff S, Dupouyet S,Bousquet-Me′lou A(2006)Potential allelopathic effect of Pinus halepensis in the secondary succession:an experimental approach.Chemoecology 16:97–105

Gu Q,Wang RR,Zhang XM,Wang YH,Zheng YT,Zhou J,Chen JJ (2007)A new benzofuranone and anti-HIV constituents from the stems of Rhus chinensis.Planta Med 73:279–282

Kim SH,Park HH,Lee S,Jun CD,Choi BJ,Kim SY,Kim SH,Kim DK,Park JS,Chae BS,Shin TY(2005)The anti-anaphylactic effect of the gallof Rhus javanica is mediated through inhibition of histamine release and inflammatory cytokine secretion.Int Immunopharmacol 15:1820–1829

Klionsky SM,Amatangelo KL,Waller DM(2011)Above-and belowground impacts of European buckthorn(Rhamnus cathartica)on four native forbs.Restor Ecol 19:728–737

Kohli RK,Batish D,Singh HP(1998)Allelopathy and its implications in agroecosystems.J Crop Prod 1:169–202

Kordali S,Cakir A,Ozer H,Cakmakci R,Kesdek M,Mete E(2008) Antifungal,phytotoxic and insecticidalproperties of essentialoil isolated from Turkish Origanum acutidens and its three components,carvacrol,thymol and p-cymene.Bioresour Technol 99:8788–8795

Lee IS,OH SR,Ahn KS(2001)Semialactone,isofouquierone peroxide and fouquierone,three new dammarane triterpenes from Rhus javanica.Chem Pharm Bull 49:1024

Li L(2009)Plant physiology module experiment guidance.Science Press,Beijing,pp 23–26

Parveen N,Khan NU(1985)Phenolic constituents of Rhus semialata leaves.Planta Med 5:467

Rayne S,Mazza G(2007)Biologicalactivities of extracts from sumac (Rhus spp.):a review.Plant Food Hum Nutr 62:165–175

Reigosa MJ,Sanchez-Moreiras A,Gonzalez L(1999)Ecophysiological approach in allelopathy.Crit Rev Plant Sci18:577–608

Ren Z,Zhu B,Wang D,Ma E,Su D,Zhong Y(2008)Comparative population structure of Chinese sumac aphid Schlechtendalia chinensis and its primary host-plant Rhus chinensis.Genetica 132:103–112

Rietveld WJ,Schlesinger RC,Kessler KJ(1983)Allelopathic effects of black walnut on European black alder coplanted as a nurse species.J Chem Ecol 9:1119–1133

Ruan X,Li ZH,Wang Q,Pan CD,Jiang DA,Wang GG(2011) Autotoxicity and allelopathy of 3,4-dihydroxyacetophenone isolated from Picea schrenkiana needles.Molecules 16:8874–8893

Shiraishi S,Watanabe I,Kuno K,Fujii Y(2002)Allelopathic activity of leaching from dry leaves and exudates from roots of ground cover plants assayed on agar.Weed Biol Manag 2:133

Singh D,Kohli RK(1992)Impact of Eucalyptus tereticornis Sm. shelterbelts on crops.Agrofor Syst 20:253–266

Singh HP,Batish DR,Kohli RK(1998)Effect of Poplar(Populus deltoids)shelterbelton the growth and yield ofwheatin Punjab, India.Agrofor Syst 40:207–213

Singh HP,Kohli RK,Batish DR(2001)Allelopathic interference of Populus deltoides with some winter season crops.Agronomie 21:139–146

Souto XC,Bolan?o JC,Gonza′lez L,Reigosa MJ(2001)Allelopathic effects of tree species on some soil microbial populations and herbaceous plants.Biol Plant 44:269–275

Taniguchi S,Yazaki K,Yabu UR(2000)Galloylglucoses and riccionidin A in Rhus javanica adventitious root cultures. Phytochemistry 53:357

Tian F,Li B,Ji B,Yang J,Zhang G,Chen Y,Luo Y(2009) Antioxidant and antimicrobial activities of consecutive extracts from Galla chinensis:the polarity affects the bioactivities.Food Chem 113:173–179

Tongma S,Kobayashi K,Usui K(2001)Allelopathic activity of Mexican sunflower[Tithonia diversifolia(Hemsl.)A.Gray]in soil under natural field conditions and different moisture conditions.Weed Biol Manag 1:115–294

Valera-Burgos J,D?′az-Barradas MC,ZunzuneguiM(2012)Effects of Pinus pinea litter on seed germination and seedling performance of three Mediterranean shrub species.Plant Growth Regul 66:285–292

Wang RR,Gu Q,Wang YH,Zhang XM,Yang LM,Zhou J,Chen JJ, Zheng YT(2008)Anti-HIV-1 activities of compounds isolated from the medicinal plant Rhus chinensis.J Ethnopharmacol 117:249–256

Weir TL,Park SW,Vivanco JM(2004)Biochemical and physiological mechanisms mediated by allelochemicals.Curr Opin Plant Biol 7(4):472–479

Williamson GB,Richardson D(1988)Biossays for allelopathy: measuring treatment responses with independent controls. J Chem Ecol 1:181–187

Yu JQ,Ye SF,Zhang MF,Hu WH(2003)Effects of root exudates and aqueous root extracts of cucumber(Cucumis sativus),and allelochemicals on photosynthesis and antioxidant enzymes in cucumber.Biochem Syst Ecol 31:129–139

Zhang D,Zhang J,Yang W,Wu F(2010)Potentialallelopathic effect of Eucalyptus grandis across a range of plantation ages.Ecol Res 25:13–23

Zhao SJ,Liu HS,Dong XC(1998)Experimental guide of plant physiology.Science Press,Beijing

23 September 2013/Accepted:14 January 2014/Published online:27 January 2015

?Northeast Forestry University and Springer-Verlag Berlin Heidelberg 2015

Project funding:The work was financially supported by the Major State Basic Research Development Program(973)of China(No. 2011CB100400)and Sci-technology Program of Guangdong Province (No.0203-E12147).

The online version is available at http://www.link.springer.com

Corresponding editor:Hu Yanbo

L.Liu

School of Chemistry and Chemical Engineering,Guangdong Pharmaceutical University,Zhongshan 528458,China

L.Liu·H.He·S.Luo·H.Li(?)

Key Laboratory of Tropical Agricultural Environment in South China,the Ministry of Agriculture,People’s Republic of China, South China Agricultural University,Guangzhou 510642,China e-mail:lihuashou@scau.edu.cn