Allelopathic Effects and Identification of Allelochemicals in Grape Root Exudates

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1 Acta Horticulturae Sinica 1 1,3,* Vitis vinifera L. Red Globe V. riparia V. labrusca Beta LC-MS g ml -1 / PAL SOD MDA PAL SOD 69.16% 10.35% MDA LC-MS 1 β V. amurensis V. riparia Shanhe 2 S A X Allelopathic Effects and Identification of Allelochemicals in Grape Root Exudates GUO Xiu-wu 1 LI Kun 1,3,* SUN Ying-ni 1 ZHANG Li-heng 1 HU Xi-xi 2 and XIE Hong-gang 3 1 College of Horticulture Shenyang Agricultural University Shenyang China 2 The Daqing Branch of Heilongjiang Academy of Agricultural Sciences Daqing Heilongjiang China 3 Liaoning Agricultural Vocation-Technical College Yingkou Liaoning China Abstract: In order to explore the allelopathy of grape root exudates tissue cultured plantlets of Red Globe Vitis vinifera L. in laboratory and cutting seedlings of Beta V. riparia V. labrusca by pot experiment were used to examine the effects of grape exudates on plant growth. Laboratory tests showed g ml -1 root dry weight/root exudates volume grape root exudates significantly inhibited growth of grape plantlets the inhibition being stronger at higher root exudate concentrations. Pot experiment indicated that low-concentration root exudates promoted plant growth and increased soluble sugar content starch content and protein content of leaves. The difference of PAL and SOD activities between nycytx-30-zp * Author for correspondence xhgbox@163.com

2 the treatments and the control were not significant and MDA content decreased under low concentration. High-concentration root exudates inhibited plant growth significantly and PAL and SOD activity were higher than the control by 69.16% and 10.35% respectively and MDA content increased. p-hydroxybenzoic acid and salicylic acid were determined by LC-MS in grape root exudates which had significant allelopathy effect on tissue culture plantlets. Other compounds in root exudates of grape were inferred as palmitic acid 1-heneicosanol citric acid aconitic acid β-sitosterol phthalic acid and gallic acid. Key words grape root exudates allelopathy allelochemicals Allelopathy Rice 1974 Wittacker & Feeny 1971 Singh et al Vitis riparia V. labrusca Beta 1 L 5 30 min 15 min 5 h 4 1 L ml 0.4 g ml -1 / g ml -1 3g ml Vitis vinifera L. Red Globe 39 ml MS 1 ml 0.22 µm 0.01 g ml -1 / ) 0.10 g ml g ml -1 1 ml L Hoagland 2 30

3 6 863 min 15 min 2 d ml E ml ml E200 ml ml E ml 5 2 kg d 300 ml CIRAS-1 HANSATECH Oxy-Lab G SOD NBT MDA 1994 PAL 0.5 g r min min 40 µl 0.6 ml ph ml 0.01 mol L -1 4 ml min 290 nm OD LC-MS ml XAD ml Sigma 98.0% 99.0% >95.0% C mm 0.1% 0 ~ 15 min 35% ~ 50% 15 ~ 20 min 50% ~ 95% 5 min 10 min 1.0 ml min nm 20 µl psi 8.0 L min ~ 400 m/z V. amurensis V. riparia Shanhe 2 39 ml 1 ml 0.22 µm mol L DPS Duncan s g ml -1 1

4 / g ml -1 Treatment /cm Plant height 1 Table 1 Allelopathic effects of grape root exudates on tissue culture seedlings /g /cm Fresh weight of shoots Max root length Root number b b 2.10 a 6 b b c c 0.50 b 2 c c Control 4.10 a a 2.75 a 8 a a - P<0.05 /g Fresh weight of roots Note: - means non determination. Significant differences between treatments are indicated by different letters P< g ml g ml g ml -1 4 Fig. 1 Effects of grape root exudates on tissue culture plantlets growth 1 Root exudates 0.01 g ml -1 2 Root exudates 0.10 g ml -1 3 Root exudates 0.50 g ml -1 4 Control. 2.2 E % 14.58% 28.08% 19.11% E100 E200 E % %

5 Table 2 Allelopathic effects of grape root exudates on pot seedlings Treatment /cm Plant height /cm Stem diameter /g /g / Fresh weight of shoots Fresh weight of roots µmol m -2 s -1 Net photosynthetic rate / µmol min -1 g -1 Rate of root respiration Control c a c 5.60 b 11.3 b bc E a a a 8.21 a 12.4 a a E b a b 7.83 a 12.0 a ab E d b d 4.53 c 8.2 c c P<0.05 Note Significant differences between treatments are indicated by different letters P< Table 3 Effects of grape root exudates on physiological indices of pot seedlings Treatment /% / PAL / SOD / /% Soluble mg g -1 unit mg -1 min -1 FWunit g -1 FW Starch content sugar content Protein content PAL activity SOD activity MDA / nmol g -1 MDA content Control c ab c 321 b 599 b b E a a a 279 b 597 b b E b a b 283 b 591 b 9.60 c E d b d 543 a 661 a a P<0.05 Note Significant differences between treatments are indicated by different letters P<0.05. PAL E100 E200 E % SOD MDA E300 > > E100 > E200 E300 SOD 10.35% LC-MS 4.06 µg ml -1 4 Allelochemicals 4 LC-MS Table 4 LC-MS detection results of allelochemicals of grape root exudates m/z /min Time of apex p-hydroxybenzoic acid Salicylic acid Palmitic acid Heneicosanol Citric acid Aconitic acid β β-sitosterol Phthalic acid Gallic acid /(µg ml -1 ) Content - P<0.05 Note - means non determination. Significant differences between treatments are indicated by different letters P<0.05.

6 LC-MS β mol L mol L % 86.06% 81.54% 98.17% 5 Table 5 Inhibition effect of allelochemical on grape tissue culture seedlings / mol L -1 /cm /g /g Treatment Concentration Plant height Fresh weight of shoots Root number Fresh weight of roots Control 4.2a a 11a a p-hydroxybenzoic a b 12a b a c 7b c b d 6b d Control 4.2b a 11b a Salicylic acid a a 14a b ab b 12ab c c c 3c d P<0.05 Note Significant differences between treatments are indicated bydifferent letters P <0.05) Pramanik 2001 et al SOD PAL Fren 1975 Ralph E100 E200

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