Reactive oxygen species (ROS) includes a superoxide anion radical a hydroxyl radical, and hydrogen peroxide, and is formed and degraded by an aerobic organism that can cause oxidative damage of all major groups of biomolecules (DNA, protein, lipids, and small cellular molecules) (Halliwell 1999). ROS plays a major role in plant defense against various pathogens (Mittapalli et al. 2007; Torres 2010; Liu et al. 2010). The rapid generation of ROS can lead to a hypersensitive response (HR) that results in a zone of host cell death, which prevents further spread of pathogens (Höglund et al. 2005). In the galls induced by Pseudophacopteron sp. in
Aspidosperma australe, the ROS were more concentrated in the cells of the inner cortex, next to the nymphal chamber (Oliverira and Isaias 2010). Liu et al. (2010) demonstrated that the accumulation of ROS at the gall midge attacked site in three isogenic wheat lines. In our results, methanol extract of galls generally exhibited a relatively higher antioxidant activity than the leaves (Table 5.3) and had significantly higher content of secondary metabolites (Table 5.2). This finding indicates that the gall formation process can cause oxidative damage to plant cells to suppress oxidative damage with a high capacity for free radical scavenging.
In conclusion, polyphenolic and flavonoid contents from galls and their host plant leaves were the most relevant to antioxidant ability.
Scavenging DPPH free radical ability and chelating ferrous ion activity positively correlated to polyphenolic content. Reducing power also related to polyphenolic and flavonic content. Different samples expressed various antioxidant abilities and functions, and the galls generally had better abilities and functions than their host leaves. The organisms possess innate defense mechanisms to scavenge free radicals effectively. The galls demonstrated enhanced levels of antioxidants to balance increased free radicals in their bodies, and may be valuable for use in crude drugs in the
future. This work has several points to investigate in the future; we need to assess the fine effective concentration of anti-oxidant activity in the gall sample, the physiological mechanism of medicine, and understand the role of ROS in gall-inducing insect and plant interaction.
References
Ahn YJ, HS Lee, HS Oh, HT Kim, YH Lee. 2005. Antifungal activity and mode of action of Galla rhois-derived phenolics against phytopathogenic fungi. Pesticide Biochem. Physiol. 81: 105- 112
Al-Laith AAA. 2010. Antioxidant components and antioxidant/antiradical activities of desert truffle(Tirmania nivea) from various Middle Eastern origins. J. Food Composition Analysis 23: 15- 22
Allison SD, JC Schultz. 2005. Biochemical responses of chestnut oak to a galling cynipid. J. Chem. Ecol. 31: 151- 166
Bao J, Y Cai, M Sun, G Wang, H Corke. 2005. Anthocyanins, flavonols, and free radical scavenging activity of chinese bayberry (Myrica rubra) extracts and their color properties and stability. J. Agric. Food Chem.
53: 2327-2332.
Butsat S, S Siriamornpun. 2010. Antioxidant capacities and phenolic compounds of the husk, bran and endosperm of Thai rice. Food Chem.
119: 606–613
Chen TK “Extraction of Cordycepin from Cordyceps militaris using Supercritical Carbon Dioxide” (Master diss. Chaoyang University of Technology. 2004:9-13)
Chirinos R, J Galarza, I Betalleluz-Pallardel, R Pedreschi, D Campos. 2010.
Antioxidant compounds and antioxidant capacity of Peruvian camu camu (Myrciaria dubia (H.B.K.) McVaugh) fruit at different maturity stages. Food Chem. 120: 1019–1024
Chu YH, CL Chang, HF Hsu. 2000. Flavonoid content of several vegetables and their antioxidant activity. J. Sci. Food. Agric. 80:
561-566.
Chung AL. 2001. Studies on the Antioxidative Capacity and Functions of Herb Plants. Master diss., Chinese Culture University, Taiwan.
Dinis TCP, VMC Madeira, LM Almeida. 1994. Action of phenolic derivatives (acetaminophen, salicylate, and 5-amino salicylate) as inhibitors of membrane lipid peroxidation and as peroxyl radical scavengers. Arch. Biochem. Biophys. 315: 161-169.
Gallegos-Infante JA, NE Rocha-Guzman, RF Gonzalez-Laredo, J Pulido-Alonso. 2010. Effect of processing on the antioxidant properties of extracts from Mexican barley (Hordeum vulgare) cultivar. Food Chem. 119: 903–906
Gordon MH. 1990. The mechanism of antioxidant action in vitro. Pages 1~18 In BJF Hudson eds. Food antioxidants. Elsevier Applied
Science, London, New York.
Gould KS, J McKelvie, KR Markham. 2002. Do anthocyanins function as antioxidants in leaves? Imaging of H2O2 in red and green leaves after mechanical injury. Plant Cell Environ. 25:1261–1269.
Govindarajan R, M Vijayakumar, P Pushpangadan. 2005. Antioxidant approach to disease management and the role of ‘Rasayana’ herbs of Ayurveda. J. Ethnopharmacology 99: 165–178.
Halliwell B. 1999. Establishing the significance and optimal intake of dietary antioxidants: the biomarker concept. Nutrition Rev. 57:
104–113.
Hartley SE. 1998. The chemical composition of plant galls: are levels of nutrients and secondary compounds controlled by gall-former.
Oecologia 113: 492-501.
Höglund S, S Larsson, G Wingsle. 2005. Both hypersensitive and non-hypersensitive response are associated with resistance in Salix viminalis against the gall midge Dasineura marginemtorquens. J. Exp.
Bot.56: 3215- 3222.
Huang DJ, CD Lin, HJ Chen, YH Lin. 2004. Antioxidant and antiproliferative activities of sweet potato constituents. Bot. Bull. Acad.
Sin. 45: 179-186.
Huang WY. 2003. The antioxidant capacity of flavonoids in red indigenous vegetables and their protection role in DNA oxidative damage of Lymphocytes. Master diss., Chinese Culture University, Taiwan.
Huang YC, CC Liaw, SC Chen. 2009. The Research and Development of Natural Products from Ferns in Taiwan. Chem. (The Chinese Chemical Society, Taipei) 67:13-31.
Jia Z, M Tang, J Wu. 1999. The determination of flavonoid content in mulberry and their scavenging effects on superoxide radicals. Food Chem. 64: 555-559.
Kähkönen MP, M Heinonen. 2003. Antioxidant activity of anthocyanins and their aglycons. J. Agric. Food Chem. 51: 628-633.
Karageorgou P, Y Manetas. 2006. The importance of being red when young:
anthocyanins and the protection of young leaves of Quercus coccifera from insect herbivory and excess light. Tree Physiol. 26, 613-621.
Karban R, AA Agrawal. 2002. Herbivore offense. Annu. Rev. Ecol. Syst.
33:641-64.
Kaur G, M Athar, MS Alam. 2008. Quercus infectoria galls possess
antioxidant activity and abrogates oxidative stress-induced functional alterations in murine macrophages. Chem. Biol. Interact. 171: 272–282.
Kim HP, SY Kim, EJ Lee, YC Kim. 1997. Zeaxanthin dipalmitate from Lycium Chinese has hepatoprotective activity. Res. Commun. Mol.
Pathol. Pharmacol. 97: 301-314.
Kraus C, G Spiteller. 1997. Comparison of phenolic compounds from galls and shoots of Picea glauca. Phytochem. 44: 59- 67
Lin KH, PY Chao, CM Yang, WC Cheng, HF LO, TR Chang. 2006. The effects of flooding and drought stress on the antioxidant constituents in sweet potato leaves. Bot. Stud. 47: 417-426.
Liu GQ, WX Liu, MC Wei, H Sun. 2005. Development Situation and Perspectives on Chitin and Chitosan from Insects. J. Northwest Forestry University. 20:143-146.
Liu X, CE Williams, JA Nemacheck, H Wang, S Subramanyam, C Zheng, MS Chen. 2010. Reactive oxygen species are involved in plant defense against a gall midge. Plant Physiol. 152: 985- 999.
Mancinelli AL, CP Huang-Yang, P Lindquist, OR Anderson, I Rabino.
1975. Photochontrol of anthocyanin synthesis, Ⅲ. The action oh streptomycin on the synthesis of chlorophyll and anthocyanin. Plant
Physiol. 55: 251-257.
Meir S, J Kanner, B Akiri, S Philosoph-H. 1995. Determination and 31 involvement of aqueous reducing compounds in oxidative defense systems of various sencescing leaves. J. Agric. Food Chem. 43:
1813-1819.
Mittapalli O, JJ Neal, RH Shukle. 2007. Antioxidant defense response in a galling insect. PNAS 104: 1889–1894.
Motta LB, JE Kraus, A Salatino, MLF Salatino. 2005. Distribution of metabolites in galled and non-galled foliar tissues of Tibouchina pulchra. Biochem. System. Ecol. 33: 971- 981.
Nyman T, R Julkunen-Tiitto. 2000. Manipulation of the phenolic chemistry of willows by gall-inducing sawflies. Proc. Natl. Acad. Sci. USA 97:
13184–13187
Oliveira DC, RMS Isaias. 2010. Cytological and histochemical gradients induced by a sucking insect in galls of Aspidosperma australe Arg.
Muell (Apocynaceae). Plant Sci. 178: 350- 358.
Oyaizu M. 1986. Antioxidative activity of browning products of glucosamine fractionated by organic solvent and thin-layer chromatography. Nippon Shokuhin Kogyo Gakkaishi 35: 771-775.
Park EJ, JM Pezzutto. 2002. Botanicals in cancer chemoprevention. Cancer and Metastasis Reviews 21, 231–255.
Piubelli GC, CB Hoffmann-Campo, IC Arruda, JC Franchini, FM Lara.
2003. Flavonoid increase in soybean as a response to Nezara viridula injury and its effect on insect-feeding preference. J. Chem. Ecol. 29(5):
1223-1233.
Pribitkin ED, G Boger. 2001. Herbal therapy: what every facial plastic surgeon must know. Arch. Facial. Plast. Surg. 3:127-132.
Robak J, IR Gryglewski. 1988. Flavonoids are scavengers of superoxide anions. Biochem. Pharmacol. 37: 837-841.
Sato T, Y Isobe, Y Ehara, M Yamaguchi. 1996. Effect of menaquinone-7 purified from the fermented soybean (natto) in the femoral metaphysic from rat femur (in Japanese). Vitamins 70: 317-322.
Scalzo J, A Politi, B Mezzetti, M Battino. 2005. Plant genotype affects total antioxidant capacity and phenolic contents in fruit. Nutrition 21:
207-213.
Shimada K, K Fujikawa, K Yahara, T Nakamura. 1992. Antioxidative properties of xanthan on the autoxidation of soybean oil in cyclodextrin emulsion. J. Agric. Food Chem. 40: 945-948.
Shorthouse JD, D Wool, A Raman. 2005. Gall-inducing insects – Nature’s most sophisticated herbivores. Basic Appl. Ecol. 6: 407- 411.
Singha A, V Thareja, AK Singh. 2007. Application of neem seed kernel extracts result in mouthpart deformities and subsequent mortality in Nezara viridula (L.) (Hem: Pentatomidae). J. Appl. Entomol. 131:
197-201.
Singleton VL, JA Rossi. 1965. Colorimetry of total phenolics with phosphomolybdic-phosphotungstic acid reagents. Am. J. Enol. Vitic. 16:
144-158.
Smith MA, G Perry, PL Richey, LM Sayre, VE Anderson, MF Beal, N Kowal. 1996. Oxidative damage in Alzheimer’s. Nature 382: 120-121.
Surveswaran S, YZ Cai, H Corke, M Sun. 2007. Systematic evaluation of natural phenolic antioxidants from 133 Indian medicinal plants. Food Chem. 102: 938-953.
Torres MA. 2010. ROS in biotic interactions. Physiol. Plant. 138: 414- 429.
Tung GS, PS Yang, MM Yang. 2006. Pattern Analysis of Galling Host-plants in Taiwan. Taiwan Journal of Forest Science 21:205-214.
Viljanen K, R Kivikari, M Heinonen. 2004. Protein-lipid interactions during liposome oxidation with added anthocyanin and other phenolic
compounds. J. Agric. Food Chem. 52: 1104-1111.
Yang CM, MM Yang, JM Hsu, WN Jane. 2003. Herbivorous insect causes deficiency of pigment-protein complexes in an ovoid cecidomyiid gall of Machilus thunbergii leaf. Bot. Bull. Acad. Sin. 44: 315-321.
Yang MM, GS Tung. 1998. The diversity of insect-induced galls on vascular plants in Taiwan: a preliminary report. USDA For. Serv. North Central For. Expt. Sta. Gen. Tech. Rep. NC-199, pp.44-53
Table 5.1 The gall-inducing insects and their host plants from galls
Gall-inducing insects Host plants Location
Ovoid galls
Diptera：Cecidomyiidae Lauraceae
Leaf Daphnephila taiwanensis Machilus thunbergii
Obovate gall Diptera：Cecidomyiidae Lauraceae
Leaf Daphnephila sueyenae Machilus thunbergii
Nutgalls
Hemiptera：Pemphigidae Anacardiaceae
Leaf Schlechtendalia chinensis Rhus javanica var. roxbur-ghiana
Table 5.2 The content of various antioxidative substances from galls and their host plants
Polyphenols Flavonoids Anthocyanins
Sample (mg gallic acid g-1) (µg g-1) (µmol g-1) Daphnephila taiwanensis 177.53 ±12.70b 55.10 ±6.74a 1.37 ±0.06a Daphnephila sueyenae 89.88 ±12.34c 20.33 ±0.78b 0.39 ±0.03b Machilus thunbergii (leaf) 23.83 ±0.72d 49.33 ±1.53a 0.44 ±0.06b Schlechtendalia chinensis 821.13 ±3.81a 5.55 ±0.09c 0.10 ±0.01c Means within a column with different superscripts are significantly different.
Table 5.3 The ability of various antioxidative functions from galls and their host plants
DPPH Fe2+ ion Superoxide anion
Sample (5mg mL-1) scavenging activity (%)
scavenging activity (%)
scavenging
activity (%) Reducing power Daphnephila taiwanensis 87.41 ± 0.42b 40.25 ±1.05c 47.68 ±0.77c 2.19 ±0.01b Daphnephila sueyenae 85.46 ± 1.73b 60.68 ±7.05b 57.66 ±0.29a 2.18 ±0.01b Machilus thunbergii (leaf) 83.50 ± 0.68b 19.59 ±0.87d 27.80 ±2.12d 2.18 ±0.01b Schlechtendalia chinensis 91.68 ± 0.22a 82.98 ±2.47a 53.73 ±1.06b 2.32 ±0.01a Means within a column with different superscripts are significantly different.
Chapter 6:
Conclusion
There are four main topics that be made a description of this dissertation: (1) In contrast to the host plant leaves, herbivorous insects alter both chlorophyll biosynthetic and degradation pathways of galls on host plants, and insect-induced galls may utilize Chl→ Chlide→ Pchlide and Chlide→ Phe→ Pchlide as the respective major and minor degradative routes. (2) Furthermore, the stomata were not found on the surface of galls. Photosynthetic performance was also evaluated by measuring chlorophyll fluorescence and gas exchanges. When compared photosynthesis in infested leaves and scale-free leaves, insect infestation didn’t alter photosynthetic efficiency. In addition, the carbon and nitrogen contents of gall tissues dropped, but the C/N ratio of galls was higher than that of leaves. The glucose and fructose contents of gall tissues were also higher than that of leaves.
The results suggest that the leaf-derived cecidomyiid galls are novel sinks in Machilus thunbergii leaves. (3) Overally the fluorescence and reflectance analyses suggested that insect infestations reduce the photosystem II efficiency of gall tissues. In addition, there were highly significant and positive relationships of PRI and CHB with Fv/Fm, indicating that gall infections induced physiological changes that could also be detected using reflectance spectra. (4) The results show that most of the samples expressed various antioxidant abilities and functions. The galls had higher abilities and functions than their host leaves to scavenge free radicals effectively for balancing the increases free radicals in their bodies.
The cecidomyiid galls and their hosts offer an excellent model system for studying insect-plant interaction. However, it is clear that many interesting issues remained outside the scope of this thesis, and that further studies are needed, such as (1) What are the mechanisms for galling insects to induce the change of chlorophyll biosynthesis and degradation? (2) What are the factors that affect the growth of gall tissues? (3) Can the spectral reflectance be utilized as a tool to study the ecophysiology for galls of other types? (4) What is the mechanism for galling insects to increase antioxidant ability in gall?