Skip to main content Accessibility help

Chitooligosaccharides enhance cold tolerance by repairing photodamaged PS II in rice

  • Jiachun Zhou (a1), Qiao Chen (a1) (a2), Yang Zhang (a1) (a2), Liqiang Fan (a1) (a2), Zhen Qin (a1), Qiming Chen (a1), Yongjun Qiu (a1) (a2), Lihua Jiang (a1) and Liming Zhao (a1) (a2)...


Chitooligosaccharides (COS) are multi-functional foods and nutrients and environmentally friendly biological abiotic-resistance inducing agents for plants. In the current study, the effects and possible mechanisms of COS on improving the cold resistance of rice (II YOU 1259) seedlings were investigated. Compared with the control, a COS pre-soaking treatment enhanced photosynthesis, reduced oxidation damage and led to accumulation of more osmotic regulation substances under chilling treatment. In addition, a novel Deg/HtrA family serine endopeptidase (DegQ) gene, related to COS enhanced rice cold resistance, was identified. Quantitative real-time polymerase chain reaction (qRT-PCR) analysis revealed that transcription of DegQ and psbA (D1 protein encoding gene) were up-regulated in a time-dependent manner by COS treatment under cold stress. With increasing expression of the D1 protein, chlorophyll b content was enhanced correspondingly. The current results suggest that COS could enhance cold stress tolerance of rice by repairing the photodamaged photosystem II, altering osmotic regulation and reducing oxidation damage.


Corresponding author

Author for correspondence: Liming Zhao and Liqiang Fan, E-mail:,


Hide All
Allakhverdiev, SI, Nishiyama, Y, Miyairi, S, Yamamoto, H, Inagaki, N, Kanesaki, Y and Murata, N (2002) Salt stress inhibits the repair of photodamaged photosystem II by suppressing the transcription and translation of psbA genes in Synechocystis. Plant Physiology 130, 14431453.
Allan, CR and Hadwiger, LA (1979) The fungicidal effect of chitosan on fungi of varying cell wall composition. Experimental Mycology 3, 285287.
Barker, M, de Vries, R, Nield, J, Komenda, J and Nixon, PJ (2006) The deg proteases protect Synechocystis sp. PCC 6803 during heat and light stresses but are not essential for removal of damaged D1 protein during the photosystem two repair cycle. Journal of Biological Chemistry 281, 3034730355.
Bates, LS, Waldren, RP and Teare, ID (1973) Rapid determination of free proline for water-stress studies. Plant and Soil 39, 205207.
Ben Ahmed, C, Ben Rouina, B, Sensoy, S, Boukhriss, M and Ben Abdullah, F (2010) Exogenous proline effects on photosynthetic performance and antioxidant defense system of young olive tree. Journal of Agricultural and Food Chemistry 58, 42164222.
Blum, A (2005) Drought resistance, water-use efficiency, and yield potential – are they compatible, dissonant, or mutually exclusive? Australian Journal of Agricultural Research 56, 11591168.
Boonlertnirun, S, Boonraung, C and Suvanasara, R (2008) Application of chitosan in rice production. Journal of Metals, Materials and Minerals 18, 4752.
Bradford, MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry 72, 248254.
Cao, X, Zhong, C, Zhu, L, Zhang, J, Sajid, H, Wu, L and Jin, Q (2017) Glycine increases cold tolerance in rice via the regulation of N uptake, physiological characteristics and photosynthesis. Plant Physiology and Biochemistry 112, 251260.
Chan, Z and Shi, H (2015) Improved abiotic stress tolerance of bermudagrass by exogenous small molecules. Plant Signaling and Behavior 10, e991577.
Chaoui, A, Mazhoudi, S, Ghorbal, MH and El Ferjani, E (1997) Cadmium and zinc induction of lipid peroxidation and effects on antioxidant enzyme activities in bean (Phaseolus vulgaris L.). Plant Science 127, 139147.
Chen, AS, Taguchi, T, Sakai, K, Kikuchi, K, Wang, MW and Miwa, I (2003) Antioxidant activities of chitobiose and chitotriose. Biological and Pharmaceutical Bulletin 26, 13261330.
Chen, W, Guo, C, Hussain, S, Zhu, B, Deng, F, Xue, Y, Geng, M and Wu, L (2016) Role of xylo-oligosaccharides in protection against salinity-induced adversities in Chinese cabbage. Environmental Science and Pollution Research 23, 12541264.
Collard, BCY and Mackill, DJ (2009) Start Codon Targeted (SCoT) polymorphism: a simple, novel DNA marker technique for generating gene-targeted markers in plants. Plant Molecular Biology Reporter 27, 8693.
Datta, K and Datta, SK (2006) Indica rice (Oryza sativa, BR29 and IR64). Methods in Molecular Biology 343, 201212.
Ding, SJ, Shie, MY, Hoshiba, T, Kawazoe, N, Chen, G and Chang, HC (2010) Osteogenic differentiation and immune response of human bone-marrow derived mesenchymal stem cells on injectable calcium-silicate-based bone grafts. Tissue Engineering Part A 16, 23432354.
Dzung, NA, Khanh, VTP and Dzung, TT (2011) Research on impact of chitosan oligomers on biophysical characteristics, growth, development and drought resistance of coffee. Carbohydrate Polymers 84, 751755.
El Sayed, AI, Rafudeen, MS and Golldack, D (2014) Physiological aspects of raffinose family oligosaccharides in plants: protection against abiotic stress. Plant Biology 16, 18.
Fei, Y, Xue, Y, Du, P, Yang, S and Deng, X (2017) Expression analysis and promoter methylation under osmotic and salinity stress of TaGAPC1 in wheat (Triticum aestivum L). Protoplasma 254, 987996.
Foyer, CH and Noctor, G (2005) Oxidant and antioxidant signalling in plants: a re-evaluation of the concept of oxidative stress in a physiological context. Plant, Cell and Environment 28, 10561071.
Freguson, IB, Watkins, CB and Harman, JE (1983) Inhibition by calcium of senescence of detached cucumber cotyledons. Plant Physiology 71, 182186.
Fu, J, Sun, Y, Chu, X, Xu, Y and Hu, T (2014) Exogenous 5-aminolevulenic acid promotes seed germination in Elymus nutans against oxidative damage induced by cold stress. PLoS ONE 9, e107152.
Hamed, I, Özogul, F and Regenstein, JM (2016) Industrial applications of crustacean by-products (chitin, chitosan, and chitooligosaccharides): a review. Trends in Food Science and Technology 48, 4050.
Hart, KN, Kwon, IK, Lohakara, JD, Heo, S and Chae, BJ (2007) Chito-oligosaccharides as an alternative to antimicrobials in improving performance, digestibility and microbial ecology of the gut in weanling pigs. Asian Australasian Journal of Animal Sciences 20, 556562.
Haußühl, K, Andersson, B and Adamska, I (2001) A chloroplast DegP2 protease performs the primary cleavage of the photodamaged D1 protein in plant photosystem II. EMBO Journal 20, 713722.
Havaux, M and Tardy, F (1997) Thermostability and photostability of photosystem II in leaves of the Chlorina-f2 barley mutant deficient in light-harvesting chlorophyll a/b protein complexes. Plant Physiology 113, 913923.
Hirano, S and Nagao, N (1989) Effects of chitosan, pectic acid, lysozyme and chitinase on the growth of several phytopathogens. Agricultural and Biological Chemistry 53, 30653066.
Hu, M, Shi, Z, Zhang, Z, Zhang, Y and Li, H (2012) Effects of exogenous glucose on seed germination and antioxidant capacity in wheat seedlings under salt stress. Plant Growth Regulation 68, 177188.
Huesgen, PF, Schuhmann, H and Adamska, I (2009) Deg/HtrA proteases as components of a network for photosystem II quality control in chloroplasts and cyanobacteria. Research in Microbiology 160, 726732.
Jiang, FY, Li, Y and Weng, BQ (2002) Review on physiology of chilling stress and chilling resistance of plants. Fujian Journal of Agricultural Sciences 17, 190195.
Jing, P, Zou, J, Kong, L, Hu, S, Wang, B, Yang, J and Xie, G (2016) OsCCD1, a novel small calcium-binding protein with one EF-hand motif, positively regulates osmotic and salt tolerance in rice. Plant Science 247, 104114.
Jumaa, M, Furkert, FH and Müller, BW (2002) A new lipid emulsion formulation with high antimicrobial efficacy using chitosan. European Journal of Pharmaceutics and Biopharmaceutics 53, 115123.
Khan, MIR, Fatma, M, Per, TS, Anjum, NA and Khan, NA (2015 a) Salicylic acid-induced abiotic stress tolerance and underlying mechanisms in plants. Frontiers in Plant Science 6, 462.
Khan, MS, Ahmad, D and Khan, MA (2015 b) Utilization of genes encoding osmoprotectants in transgenic plants for enhanced abiotic stress tolerance. Electronic Journal of Biotechnology 18, 257266.
Kiani, SP, Grieu, P, Maury, P, Hewezi, T, Gentzbittel, L and Sarrafi, A (2007) Genetic variability for physiological traits under drought conditions and differential expression of water stress-associated genes in sunflower (Helianthus annuus L.). Theoretical and Applied Genetics 114, 193207.
Kieselbach, T and Funk, C (2003) The family of Deg/HtrA proteases: from Escherichia coli to Arabidopsis. Physiologia Plantarum 119, 337346.
Korkmaz, A, Korkmaz, Y and Demirkıran, AR (2010) Enhancing chilling stress tolerance of pepper seedlings by exogenous application of 5-aminolevulinic acid. Environmental and Experimental Botany 67, 495501.
Kuang, YJ, Ye, GP and Qin, CQ (2009) Influence of oligochitosan on cold resistance of rice seedling. Hubei Agricultural Sciences 48, 15681571.
Kumar, V and Yadav, SK (2009) Proline and betaine provide protection to antioxidant and methylglyoxal detoxification systems during cold stress in Camellia sinensis (L.) O. Kuntze. Acta Physiologiae Plantarum 31, 261269.
Kumar, S, Kaur, G and Nayyar, H (2008) Exogenous application of abscisic acid improves cold tolerance in chickpea (Cicer arietinum L.). Journal of Agronomy and Crop Science 194, 449456.
Kumar, SG, Rahman, MA, Lee, SH, Hwang, HS, Kim, HA and Yun, JW (2009) Plasma proteome analysis for anti-obesity and anti-diabetic potentials of chitosan oligosaccharides in ob/ob mice. Protemics 9, 21492162.
Kurepin, LV, Ivanov, AG, Zaman, M, Pharis, RP, Hurry, V and Hüner, NPA (2017) Interaction of glycine betaine and plant hormones: protection of the photosynthetic apparatus during abiotic stress. In Hou, H, Najafpour, M, Moore, G and Allakhverdiev, S (eds), Photosynthesis: Structures, Mechanisms and Applications. Dordrecht, the Netherlands: Springer, Cham, pp. 185202.
Leborgne, N, Teulieres, C, Travert, S, Rols, MP, Teissie, J and Boudet, AM (1995) Introduction of specific carbohydrates into Eucalyptus gunnii cells increases their freezing tolerance. European Journal of Biochemistry 229, 710717.
Lei, P, Xu, Z, Liang, J, Luo, XH, Zhang, YX, Feng, XH and Xu, H (2016) Poly(γ-glutamic acid) enhanced tolerance to salt stress by promoting proline accumulation in Brassica napus L. Plant Growth Regulation 78, 233241.
Li, Y, Yin, H, Wang, Q, Zhao, XM, Du, YG and Li, FL (2009) Oligochitosan induced Brassica napus L. production of NO and H2O2 and their physiological function. Carbohydrate Polymers 75, 612617.
Lichtenthaler, HK and Buschmann, C (2001) Chlorophylls and carotenoids measurement and characterization by UV-VIS. In Wrolstad, RE, Acree, TE, An, H, Decker, EA, Penner, MH, Reid, DS, Schwartz, SJ, Shoemaker, CF and Sporns, P (eds), Current Protocols in Food Analytical Chemistry (CPFA). New York, USA: John Wiley, pp. F4.3.1F4.3.8.
Lin, ZF, Peng, CL, Lin, GZ, Ou, ZY, Yang, CW and Zhang, JL (2003) Photosynthetic characteristics of two new chlorophyll b-less rice mutants. Photosynthetica 41, 6167.
Lin, WL, Hu, XY, Zhang, WQ, Rogers, WJ and Cai, WM (2005) Hydrogen peroxide mediates defence responses induced by chitosans of different molecular weights in rice. Journal of Plant Physiology 162, 937944.
Liu, H, Zhang, YH, Yin, H, Wang, WX, Zhao, XM and Du, YG (2013) Alginate oligosaccharides enhanced Triticum aestivum l. tolerance to drought stress. Plant Physiology and Biochemistry 62, 3340.
Luo, C, He, XH, Chen, H, Ou, SJ and Gao, MP (2010) Analysis of diversity and relationships among mango cultivars using Start Codon Targeted (SCoT) markers. Biochemical Systematics and Ecology 38, 11761184.
Marco, F, Bitrián, M, Carrasco, P, Rajam, MV, Alcázar, R and Tiburcio, AF (2015) Genetic engineering strategies for abiotic stress tolerance in plants. In Bahadur, B, Venkat Rajam, M, Sahijram, L and Krishnamurthy, K (eds), Plant Biology and Biotechnology. New Delhi, India: Springer, pp. 579609.
Marta, B, Szafrańska, K and Posmyk, MM (2016) Exogenous melatonin improves antioxidant defense in cucumber seeds (Cucumis sativus L.) germinated under chilling stress. Frontiers in Plant Science 7, 575,.
Mendis, E, Kim, MM, Rajapakse, N and Kim, SK (2007) An in vitro cellular analysis of the radical scavenging efficacy of chitooligosaccharides. Life Sciences 80, 21182127.
Minorsky, PV (2003) Raffinose oligosaccharides. Plant Physiology 131, 11591160.
Mir, BA, Mir, SA, Khazir, J, Tonfack, LB, Cowan, DA, Vyas, D and Koul, S (2015) Cold stress affects antioxidative response and accumulation of medicinally important with anolides in Withania somnifera (L.) Dunal. Industrial Crops and Products 74, 10081016.
Mutlu, S, Atıcı, Ö, Nalbantoğlu, B and Mete, E (2016) Exogenous salicylic acid alleviates cold damage by regulating antioxidative system in two barley (Hordeum vulgare L.) cultivars. Frontiers in Life Science 9, 99109.
Nahar, K, Hasanuzzaman, M, Alam, MM and Fujita, M (2015) Exogenous glutathione confers high temperature stress tolerance in mung bean (Vigna radiata L.) by modulating antioxidant defense and methylglyoxal detoxification system. Environmental and Experimental Botany 112, 4454.
Nazar, R, Umar, S and Khan, NA (2015) Exogenous salicylic acid improves photosynthesis and growth through increase in ascorbate-glutathione metabolism and S assimilation in mustard under salt stress. Plant Signaling and Behavior 10, e1003751.
Nishiyama, Y, Allakhverdiev, SI, Yamamoto, H, Hayashi, H and Murata, N (2004) Singlet oxygen inhibits the repair of photosystem II by suppressing translation elongation of the D1 protein in Synechocystis sp. PCC 6803. Biochemistry 43, 1132111330.
Pallen, MJ and Wren, BW (1997) The HtrA family of serine proteases. Molecular Microbiology 26, 209221.
Park, EJ, Jeknic, Z and Chen, TH (2006) Exogenous application of glycinebetaine increases chilling tolerance in tomato plants. Plant and Cell Physiology 47, 706714.
Prasil, O, Adir, N and Ohad, I (1992) Dynamics of photosystem II: mechanism of photoinhibition and recovery in vivo. In Barber, J (ed.), Topics in Photosynthesis, Vol. 11, Amsterdam, the Netherlands: Elsevier Biomedical Press, pp. 295348.
Qian, H, Li, J, Sun, L, Chen, W, Sheng, GD, Liu, W and Fu, Z (2009) Combined effect of copper and cadmium on Chlorella vulgaris growth and photosynthesis-related gene transcription. Aquatic Toxicology 94, 5661.
Qiang, XJ, Yu, GH, Jiang, LL, Sun, LL, Zhang, SH, Wei, LI and Cheng, XG (2015) Thellungiella halophila ThPIP1 gene enhances the tolerance of the transgenic rice to salt stress. Journal of Integrative Agriculture 14, 19111922.
Rathinasabapathi, B (2000) Metabolic engineering for stress tolerance: installing osmoprotectant synthesis pathways. Annals of Botany 86, 709716.
Sakamoto, A and Murata, N (2002) The role of glycine betaine in the protection of plants from stress: clues from transgenic plants. Plant, Cell and Environment 25, 163171.
Sami, F, Yusuf, M, Faizan, M, Faraz, A and Hayat, S (2016) Role of sugars under abiotic stress. Plant Physiology and Biochemistry 109, 5461.
Sen, G, Eryilmaz, IE and Ozakca, D (2014) The effect of aluminium-stress and exogenous spermidine on chlorophyll degradation, glutathione reductase activity and the photosystem II D1 protein gene (psbA) transcript level in lichen Xanthoria parietina. Phytochemistry 98, 5459.
Shen, KT, Chen, MH, Chan, HY, Jeng, JH and Wang, YJ (2009) Inhibitory effects of chitooligosaccharides on tumor growth and metastasis. Food and Chemical Toxicology 47, 18641871.
Soliman, MH, Alayafi, AA, El Kelish, AA and Abu-Elsaoud, AM (2018) Acetylsalicylic acid enhance tolerance of Phaseolus vulgaris L. to chilling stress, improving photosynthesis, antioxidants and expression of cold stress responsive genes. Botanical Studies 59, 6.
Sun, Z, Qi, X, Wang, Z, Li, P, Wu, C, Zhang, H and Zhao, Y (2013) Overexpression of TsGOLS2, a galactinol synthase, in Arabidopsis thaliana enhances tolerance to high salinity and osmotic stresses. Plant Physiology and Biochemistry 69, 8289.
Thomashow, MF (1999) Plant cold acclimation: freezing tolerance genes and regulatory mechanisms. Annual Review of Plant Physiology and Plant Molecular Biology 50, 571599.
Walsh, NP, Alba, BM, Bose, B, Gross, CA and Sauer, RT (2003) OMP peptide signals initiate the envelope-stress response by activating DegS protease via relief of inhibition mediated by its PDZ domain. Cell 113, 6171.
Wang, M, Chen, Y, Zhang, R, Wang, W, Zhao, X, Du, Y and Yin, H (2015) Effects of chitosan oligosaccharides on the yield components and production quality of different wheat cultivars (Triticum aestivum L.) in Northwest China. Field Crops Research 172, 1120.
Wang, F, Liu, J, Chen, M, Zhou, L, Li, Z, Zhao, Q, Pan, G, Zaidi, SHR and Cheng, F (2016) Involvement of abscisic acid in PSII photodamage and D1 protein turnover for light-induced premature senescence of rice flag leaves. PLoS ONE 11, e0161203.
Wilken, C, Kitzing, K, Kurzbauer, R, Ehrmann, M and Clausen, T (2004) Crystal structure of the DegS stress sensor: how a PDZ domain recognizes misfolded protein and activates a protease. Cell 117, 483494.
Wright, GC and Smith, RCG (1983) Differences between two grain sorghum genotypes in adaptation to drought stress. II. Root water uptake and water use. Australian Journal of Agricultural Research 34, 627636.
Yin, H, Li, Y, Zhang, HY, Wang, WX, Lu, H, Grevsen, K, Zhao, XM and Du, YG (2013) Chitosan oligosaccharides-triggered innate immunity contributes to oilseed rape resistance against Sclerotinia sclerotiorum. International Journal of Plant Sciences 174, 722732.
Yin, H, Du, Y and Dong, Z (2016) Chitin oligosaccharide and chitosan oligosaccharide: two similar but different plant elicitors. Frontiers in Plant Science 7, 522.
Yoon, HJ, Moon, ME, Park, HS, Kim, HW, Im, SY, Lee, JH and Kim, YH (2008) Effects of chitosan oligosaccharide (COS) on the glycerol-induced acute renal failure in vitro and in vivo. Food and Chemical Toxicology 46, 710716.
Zhang, HY, Zhao, XM, Yang, JL, Yin, H, Wang, WX, Lu, H and Du, YG (2011) Nitric oxide production and its functional link with OIPK in tobacco defense response elicited by chitooligosaccharide. Plant Cell Reports 30, 11531162.
Zhang, X, Li, K, Liu, S, Zou, P, Xing, R, Yu, H, Chen, X, Qin, Y and Li, P (2017) Relationship between the degree of polymerization of chitooligomers and their activity affecting the growth of wheat seedlings under salt stress. Journal of Agricultural and Food Chemistry 65, 501509.
Zhao, XM, She, XP, Du, YG and Liang, XM (2007) Induction of antiviral resistance and stimulary effect by oligochitosan in tobacco. Pesticide Biochemistry and Physiology 87, 7884.
Zheng, K, Zhao, J, Lin, D, Chen, J, Xu, J, Zhou, H, Teng, S and Dong, Y (2016) The rice TCM5 gene encoding a novel Deg protease protein is essential for chloroplast development under high temperatures. Rice 9, 13.
Zong, H, Li, K, Liu, S, Song, L, Xing, R, Chen, X and Li, P (2017) Improvement in cadmium tolerance of edible rape (Brassica rapa L.) with exogenous application of chitooligosaccharide. Chemosphere 181, 92100.
Zou, P, Li, K, Liu, S, Xing, R, Qin, Y, Yu, H, Zhou, M and Li, P (2015) Effect of chitooligosaccharides with different degrees of acetylation on wheat seedlings under salt stress. Carbohydrate Polymers 126, 6269.
Zou, P, Li, K, Liu, S, He, X, Zhang, X, Xing, R and Li, P (2016) Effect of sulfated chitooligosaccharides on wheat seedlings (Triticum aestivum L.) under salt stress. Journal of Agricultural and Food Chemistry 64, 28152821.
Zou, P, Tian, X, Dong, B and Zhang, C (2017) Size effects of chitooligomers with certain degrees of polymerization on the chilling tolerance of wheat seedlings. Carbohydrate Polymers 160, 194202.



Full text views

Total number of HTML views: 0
Total number of PDF views: 0 *
Loading metrics...

Abstract views

Total abstract views: 0 *
Loading metrics...

* Views captured on Cambridge Core between <date>. This data will be updated every 24 hours.

Usage data cannot currently be displayed