Pathogenic fungi represent one of the major biotic stresses for soybean production across the world. Sclerotinia sclerotiorum, the causal agent of Sclerotinia stem rot, is a devastating fungal pathogen that is responsible for significant yield losses in soybean. In this study, the chitinase gene CmCH1, from the mycoparasitic fungus Coniothyrium minitans, which infects a range of ascomycetous sclerotia, including S. sclerotiorum and S. minor, was introduced into soybean. Transgenic plants expressing CmCH1 showed higher resistance to S. sclerotiorum infection, with significantly reduced lesion sizes in both detached stem and leaf assays, compared to the non-transformed control. Increased hydrogen peroxide content and activities of defense-responsive enzymes, such as peroxidase, superoxide dismutase, phenylalanine ammonia lyase, and polyphenoloxidase were also observed at the infection sites in the transgenic plants inoculated with S. sclerotiorum. Consistent with the role of chitinases in inducing downstream defense responses by the release of elicitors, several defense-related genes, such as GmNPR2, GmSGT-1, GmRAR1, GmPR1, GmPR3, GmPR12, GmPAL, GmAOS, GmPPO, were also significantly upregulated in the CmCH1-expressing soybean after inoculation. Collectively, our results demonstrate that overexpression of CmCH1 led to increased accumulation of H2O2 and up-regulation of defense-related genes and enzymes, and thus enhanced resistance to S. sclerotiorum infection while showing no detrimental effects on growth and development of soybean plants.
This is a preview of subscription content, access via your institution.
Buy single article
Instant access to the full article PDF.
Price excludes VAT (USA)
Tax calculation will be finalised during checkout.
Bolton MD, Thomma BP, Nelson BD (2006) Sclerotinia sclerotiorum (Lib.) de Bary: biology and molecular traits of a cosmopolitan pathogen. Mol Plant Pathol 7:1–16
Bolton MD, Thomma BPHJ, Nelson BD (2010) Sclerotinia sclerotiorum (Lib.) de Bary: biology and molecular traits of a cosmopolitan pathogen. Mol Plant Pathol 7:1–16
Cessna SG, Sears VE, Dickman MB, Low PS (2000) Oxalic acid, a pathogenicity factor for sclerotinia sclerotiorum, suppresses the oxidative burst of the host. Plant Cell 12:2191–2200
Cober ER, Rioux S, Rajcan I, Donaldson PA, Simmonds DH (2003) Partial resistance to white mold in a transgenic soybean line. Crop Sci 43:92–95
Collard BC, Mackill DJ (2008) Marker-assisted selection: an approach for precision plant breeding in the twenty-first century. Philos Trans R Soc Lond 363:557–572
Cunha WG, Tinoco MLP, Pancoti HL, Ribeiro RE, Aragão FJL (2010) High resistance to Sclerotinia sclerotiorum in transgenic soybean plants transformed to express an oxalate decarboxylase gene. Plant Pathol 59:654–660
Dana M, Pintor-Toro T, Cubero B (2006) Transgenic tobacco plants overexpressing chitinases of fungal origin show enhanced resistance to biotic and abiotic stress agents. Plant Physiol 142:722–730
Dias BBA, Cunha WG, Morais LS, Vianna GR, Rech EL, Capdeville G, Aragao FJL (2006) Expression of an oxalate decarboxylase gene from Flammulina sp. in transgenic lettuce (Lactuca sativa) plants and resistance to Sclerotinia sclerotiorum. Plant Pathol 55:187–193
Donaldson PA, Anderson T, Lane BG, Davidson AL, Simmonds DH (2001) Soybean plants expressing an active oligomeric oxalate oxidase from the wheat gf-2.8 (germin) gene are resistant to the oxalate-secreting pathogen Sclerotina sclerotiorum. Physiol Mol Plant Pathol 59:297–307
Du Q, Yang XD, Zhang JH, Zhong XF, Kim KS, Yang J, Xing GJ, Li XY, Jiang ZY, Li QY, Dong YS, Pan HY (2018) Over-expression of the Pseudomonas syringae harpin-encoding gene hrpZm confers enhanced tolerance to Phytophthora root and stem rot in transgenic soybean. Transgenic Res 27:277–288
Dutton MV, Evans CS (1996) Oxalate production by fungi: its role in pathogenicity and ecology in the soil environment. Can J Microbiol 42:881–895
Godoy G, Steadman JR, Dickman MB, Dam R (1990) Use of mutants to demonstrate the role of oxalic acid in pathogenicity of Sclerotinia sclerotiorum on Phaseolus vulgaris. Physiol Mol Plant Pathol 37:179–191
Guo X, Wang D, Gordon SG, Helliwell E, Smith T, Berry SA, St. Martin SK, Dorrance AE (2008) Genetic mapping of QTLs underlying partial resistance to Sclerotinia sclerotiorum in soybean PI 391589A and PI 391589B. Crop Sci 48:1129–1139
Hu X, Bidney DL, Yalpani N, Duvick JP, Crasta O, Folkerts O, Lu GH (2003) Overexpression of a gene encoding hydrogen peroxide-generating oxalate oxidase evokes defense responses in sunflower. Plant Physiol 133:170–181
Kabbage M, Yarden O, Dickman MB (2015) Pathogenic attributes of Sclerotinia sclerotiorum: switching from a biotrophic to necrotrophic lifestyle. Plant Sci 233:53–60
Karmakar S, Molla KA, Chanda PK, Sarkar SN, Datta SK, Datta K (2016) Green tissuespecific co-expression of chitinase and oxalate oxidase 4 genes in rice for enhanced resistance against sheath blight. Planta 243:115–130
Kim KS, Min JY, Dickman MB (2008) Oxalic acid is an elicitor of plant programmed cell death during Sclerotinia sclerotiorum disease development. Mol Plant Microbe Interact 21:605–612
Kumar V, Parkhi V, Kenerley CM, Rathore KS (2009) Defense-related gene expression and enzyme activities in transgenic cotton plants expressing an endochitinase gene from Trichoderma virens in response to interaction with Rhizoctonia solani. Planta 230:277–291
Kumar V, Chattopadhyay A, Ghosh S, Irfan M, Chakraborty N, Chakraborty S, Datta A (2016) Improving nutritional quality and fungal tolerance in soya bean and grass pea by expressing an oxalate decarboxylase. Plant Biotechnol J 14:1394–1405
Livingstone DM, Hampton JL, Phipps PM, Grabau EA (2005) Enhancing resistance to Sclerotinia minor in peanut by expressing a barley oxalate oxidase gene. Plant Physiol 137:1354–1362
Lou Y, Han Y, Yang L, Wu M, Zhang J, Cheng J, Wang M, Jiang D, Chen W, Li G (2016) CmpacC regulates mycoparasitism, oxalate degradation and antifungal activity in the mycoparasitic fungus Coniothyrium minitans. Environ Microbiol 17:4711–4729
Mccaghey M, Willbur J, Ranjan A, Grau CR, Chapman S, Diers B, Groves C, Kabbage M, Smith DL (2017) Development and evaluation of Glycine max germplasm lines with quantitative resistance to Sclerotinia sclerotiorum. Front Plant Sci 8:1495. doi:https://doi.org/10.3389/fpls.2017.01495
Pedras MSC, Ahiahonu PWK (2004) Phytotoxin production and phytoalexin elicitation by the phytopathogenic fungus Sclerotinia sclerotiorum. J Chem Ecol 30:2163–2179
Peltier AJ, Bradley CA, Chilvers MI, Malvick DK, Mueller DS, Wise KA, Esker PD (2012) Biology, yield loss and control of Sclerotinia stem rot of soybean. J Integr Pest Manag 3:1–7
Ranjan A, Jayaraman D, Grau C, Hill JH, Whitham SA, Ané JM, Smith DL, Kabbage M (2017) The pathogenic development of Sclerotinia sclerotiorum in soybean requires specific host NADPH oxidases. Mol Plant Pathol 19:700–714
Ranjan A, Westrick NM, Jain S, Piotrowski JS, Ranjan M, Kessens R, Stiegman L, Grau CR, Conley SP, Smith DL, Kabbage M (2019) Resistance against Sclerotinia sclerotiorum in soybean involves a reprogramming of the phenylpropanoid pathway and up-regulation of antifungal activity targeting ergosterol biosynthesis. Plant Biotechnol J 17:1567–1581
Ren L, Li GQ, Han YC, Jiang DH, Huang HC (2007) Degradation of oxalic acid by Coniothyrium minitans and its effects on production and activity of β-1,3-glucanase of this mycoparasite. Biol Control 43:1–11
Rollins JA (2003) The Sclerotinia sclerotiorum pac1 gene is required for sclerotial development and virulence. Mol Plant Microbe Interact 16:785–795
Roslan HA, Anji SB (2011) Characterization of inflorescence-predominant chitinase gene in Metroxylon sagu via differential display. Biotech 1:27–33
Sun XP, Zhao Y, Jia JC, Xie JT, Cheng JS, Liu HQ, Jiang DH, Fu YP (2017) Uninterrupted expression of cmsit1 in a sclerotial parasite coniothyrium minitans leads to reduced growth and enhanced antifungal ability. Front Microbiol 8:2208. https://doi.org/10.3389/fmicb.2017.02208
Telzur N, Abbo S, Myslabodski D, Mizrahi Y (1999) Modified CTAB procedure for DNA isolation from Epiphytic Cacti of the Genera Hylocereus and Selenicereus (Cactaceae). Plant Mol Biol Report 17:249–254
Vuong TD, Diers BW, Hartman GL (2008) Identification of QTL for resistance to Sclerotinia stem rot in soybean plant introduction 194639. Crop Sci 48:2209–2214
Walz A, Zingen-Sell I, Loeffler M, Sauer M (2010) Expression of an oxalate oxidase gene in tomato and severity of disease caused by Botrytis cinerea and Sclerotinia sclerotiorum. Plant Pathol 57:453–458
Wan XQ, Tan JL, Lu SY, Lin CY, Hu YH, Guo ZF (2009) Increased tolerance to oxidative stress in transgenic tobacco expressing a wheat oxalate oxidase gene via induction of antioxidant enzymes is mediated by H2O2. Physiol Plant 136:30–44
Williams B, Kabbage M, Kim HJ, Britt R, Dickman MB (2011) Tipping the balance: Sclerotinia sclerotiorum secreted oxalic acid suppresses host defenses by manipulating the host redox environment. PLoS Pathog 7(6):e1002107
Workneh F, Yang XB (2000) Prevalence of Sclerotinia stem rot of soybeans in the north-central United States in relation to tillage, climate, and latitudinal positions. Phytopathology 90:1375–1382
Yang X, Yang J, Wang Y, He H, Niu L, Guo D, Xing G, Zhao Q, Zhong X, Sui L, Li Q, Dong Y (2019) Enhanced resistance to sclerotinia stem rot in transgenic soybean that overexpresses a wheat oxalate oxidase. Transgenic Res 28(1):103–114
Zhang FL, Ruan XL, Wang X, Liu ZH, Hu LZ, Li CW (2016) Overexpression of a chitinase gene from Trichoderma asperellum increases disease resistance in transgenic soybean. Appl Biochem Biotechnol 180:1–17
Zhao X, Han YP, Li YH, Liu DY, Sun MM, Zhao Y, Lv CM, Li DM, Yang ZJ, Huang L, Teng WL, Qiu LJ, Zheng HK, Li WB (2015) Loci and candidate gene identification for resistance to Sclerotinia sclerotiorum in soybean (Glycine max L. Merr.) via association and linkage maps. Plant J Cell Mol Biol 82:245–255
Zhu WJ, Wei W, Fu YP, Cheng JS, Xie JT, Li GQ, Yi XH, Kang ZS, Dickman MB, Jiang DH (2013) A secretory protein of necrotrophic fungus Sclerotinia sclerotiorum that suppresses host resistance. PLoS ONE 8(1):e53901
This work was supported by grants from the Jilin Provincial Agricultural Science & Technology Innovation Project (CXGC2017JQ013) and China National Novel Transgenic Organisms Breeding Project (2016ZX08004-004). We thank Prof. Daohong Jiang of Huazhong Agricultural University for providing the CmCHI1 gene. We also thank Editage (www.editage.cn) for English language editing.
Conflict of interest
The authors declare that they have no conflict of interest.
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Xiangdong Yang and Jing Yang have contributed equally to this work.
Electronic supplementary material
About this article
Cite this article
Yang, X., Yang, J., Li, H. et al. Overexpression of the chitinase gene CmCH1 from Coniothyrium minitans renders enhanced resistance to Sclerotinia sclerotiorum in soybean. Transgenic Res 29, 187–198 (2020). https://doi.org/10.1007/s11248-020-00190-2