2010 37 6861 868 Acta Horticulturae Sinica 1 1,3,* 1 1 2 3 1 110866 2 163316 3 115009 Vitis vinifera L. Red Globe V. riparia V. labrusca Beta LC-MS 0.01 0.10 0.50 g ml -1 / PAL SOD MDA PAL SOD 69.16% 10.35% MDA LC-MS 1 β V. amurensis V. riparia Shanhe 2 S 663.1 A 0513-353X 2010 06-0861-08 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 110866 China 2 The Daqing Branch of Heilongjiang Academy of Agricultural Sciences Daqing Heilongjiang 163316 China 3 Liaoning Agricultural Vocation-Technical College Yingkou Liaoning 115009 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 0.01 0.10 0.50 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 2009 12 01 2010 05 04 nycytx-30-zp-08 2008204003 * Author for correspondence E-mail xhgbox@163.com
862 7 3 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. 1999 2001 2005 2003 2006 1999 2004 2006 1 1.1 20 1 Vitis riparia V. labrusca Beta 1 L 5 30 min 15 min 5 h 4 1 L 10 10 40 10 ml 0.4 g ml -1 / 4.0-1 20.0 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 -1 0.50 g ml -1 1 ml 2009 6 21 6 3 4 1.2 2009 5 7 20 2 1 L Hoagland 2 30
6 863 min 15 min 2 d 1 300 ml E100 100 ml + 200 ml E200 ml + 100 ml E300300 ml 5 2 kg 6 1 2 d 300 ml 10 7 3 CIRAS-1 HANSATECH Oxy-Lab 2004 4 5 G-250 3 4 SOD NBT MDA 1994 PAL 0.5 g 4 10 000 r min -1 10 min 40 µl 0.6 ml ph 8.80.1 ml 0.01 mol L -1 4 ml 40 15 min 290 nm OD 0.01 3 1.3 LC-MS 40 50 ml XAD-4 5 30 1 ml Sigma 98.0% 99.0% >95.0% C18 250 4.6 mm 0.1% 0 ~ 15 min 35% ~ 50% 15 ~ 20 min 50% ~ 95% 5 min 10 min 1.0 ml min -1 3 7 0.3 280 nm 20 µl 35 25 psi 8.0 L min -1 325 50 ~ 400 m/z 1.4 2 V. amurensis V. riparia Shanhe 2 39 ml 1 ml 0.22 µm 10-4 10-3 10-2 mol L -1 6 3 6 DPS Duncan s 2 2.1 1 0.50 g ml -1 1
864 37 / 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 0.01 3.83 b 0.2096 b 2.10 a 6 b 0.0262 b 0.10 2.91 c 0.1262 c 0.50 b 2 c 0.0143 c 0.50 - - - - - Control 4.10 a 0.3036 a 2.75 a 8 a 0.0830 a - P<0.05 /g Fresh weight of roots Note: - means non determination. Significant differences between treatments are indicated by different letters P<0.05. 1 1 0.01 g ml -1 2 0.10 g ml -1 3 0.50 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 E300 25.82% 14.58% 28.08% 19.11% 2 2.3 3 E100 E200 E300 10.52% 10.67 33.93%
6 865 2 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 24.59 c 0.3538 a 14.03 c 5.60 b 11.3 b 0.487 bc E100 33.80 a 0.3649 a 24.06 a 8.21 a 12.4 a 0.508 a E200 30.19 b 0.3558 a 21.73 b 7.83 a 12.0 a 0.494 ab E300 18.24 d 0.3022 b 10.09 d 4.53 c 8.2 c 0.473 c P<0.05 Note Significant differences between treatments are indicated by different letters P<0.05. 3 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 0.0580 c 0.0150 ab 0.9027 c 321 b 599 b 10.48 b E100 0.0660 a 0.0160 a 1.4888 a 279 b 597 b 10.16 b E200 0.0624 b 0.0151 a 1.2630 b 283 b 591 b 9.60 c E300 0.0519 d 0.0134 b 0.5964 d 543 a 661 a 13.79 a P<0.05 Note Significant differences between treatments are indicated by different letters P<0.05. PAL E100 E200 E300 69.16% SOD MDA E300 > > E100 > E200 E300 SOD 10.35% 3 2.4 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 137 6.0 1.43 Salicylic acid 137 16.8 5.49 Palmitic acid 255 19.6-1 1-Heneicosanol 311 21.5 - Citric acid 191 21.7 - Aconitic acid 173 22.3 - β β-sitosterol 413 23.1 - Phthalic acid 165 24.0 - Gallic acid 169 24.5 - /(µg ml -1 ) Content - P<0.05 Note - means non determination. Significant differences between treatments are indicated by different letters P<0.05.
866 7 3 LC-MS β 1 2.5 10-5 mol L -1 5 10-3 mol L -1 87.43% 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 1.2733a 11a 0.1636a p-hydroxybenzoic 10-5 4.4a 0.5008b 12a 0.1048b 10-4 4.0a 0.4334c 7b 0.0719c 10-3 3.2b 0.1601d 6b 0.0228d Control 4.2b 1.2733a 11b 0.1636a Salicylic acid 10-5 4.9a 1.1088a 14a 0.1027b 10-4 4.7ab 0.7333b 12ab 0.0534c 10-3 3.5c 0.2351c 3c 0.0030d P<0.05 Note Significant differences between treatments are indicated bydifferent letters P <0.05). 3 2005 Pramanik 2001 et al. 2008 2008 2005 SOD PAL Fren 1975 Ralph 1992 2007 E100 E200
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