Identification and expression of genes in response to cassava bacterial blight infection
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Cassava bacterial blight (CBB) caused by Xanthomonas axonopodis pv. manihotis (or XAM) is a serious disease of cassava (Manihot esculenta Crantz). In this study, quantitative trait loci (QTL) associated with CBB infection were identified in the F1 progenies of a cross between the “Huay Bong 60” and “Hanatee” cassava cultivars. The phenotype of disease severity was observed at 7, 10, and 12 days after inoculation (DAI). A total of 12 QTL were identified, of which 5, 6, and 1 were detected in 7, 10, and 12 DAI samples, respectively. Among all identified QTL, CBB14_10dai_1, CBB14_10dai_2, and CBB14_12dai showed the most significant (P < 0.0001) associations with CBB infection, and explained 21.3, 13.8, and 26.5% of phenotypic variation, respectively. Genes underlying the QTL were identified and their expression was investigated in resistant and susceptible cassava plants by real-time quantitative RT-PCR. The results identified candidate genes that showed significant differences in expression between resistant and susceptible lines, including brassinosteroid insensitive 1-associated receptor kinase 1-related (Manes.04G059100), cyclic nucleotide-gated ion channel 2 (Manes.02G051100), and autophagy-related protein 8a-related (Manes.17G026600) at 7 DAI, and regulator of nonsense transcripts 1 homolog (Manes.17G021900) at both 7 and 12 DAI. The expression pattern of all genes showed higher levels in resistant (B82, B32, B20, and B70) as compared to susceptible (HB60, B100, B95, and B47) plants. Overall, this study has identified QTL and markers linked to CBB infection trait, and identified candidate genes involved in CBB resistance. This information will be of use for better understanding defense mechanisms in cassava to bacterial blight disease.
KeywordsCassava Cassava bacterial blight Xanthomonas axonopodis pv. manihotis QTL SSR markers Quantitative real-time PCR
We thank Mr. Rungsi Charaensataporn, Department of Agriculture, for providing the XAM stock and Miss Saowaree Tangkasakul, Field Crops Experiment Station Ladbuakao, Sikhio, Nakhon Ratchasima, Thailand for providing plant materials.
PT, SS, NS, and KT conceived, designed, and performed the experiments. PT, SS, NS, and KT analyzed the data. PT, DRS, and KT wrote the manuscript.
This research is partially supported by the Center of Excellence on Agricultural Biotechnology (AG-BIO/PERDO-CHE), Thailand (AG-BIO/60-005-003), and Mahidol University. . PT was supported for Ph.D. study by the AG-BIO/PERDO-CHE.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
Human and animal rights and informed consent
This article does not contain any studies with human participants or animals performed by any of the authors.
- Belhassen BB, Abbassian A, Alesiani A et al. (2016) Food outlook. Biannual Report on Global Food Markets Food and Agriculture Organization of the United Nations (FAO), RomeGoogle Scholar
- Bio-Rad (2006) Real-time PCR applications guide. Bio-Rad Laboratories, Inc.Google Scholar
- Chin K, DeFalco TA, Moeder W, Yoshioka K (2013) The Arabidopsis cyclic nucleotide-gated ion channels AtCNGC2 and AtCNGC4 work in the same signaling pathway to regulate pathogen defense and floral transition. Plant Physiol 163:611–624. https://doi.org/10.1104/pp.113.225680 CrossRefPubMedPubMedCentralGoogle Scholar
- Delannoy E et al. (2005) Resistance of cotton towards Xanthomonas campestris pv. malvacearum. Annu Rev Phytopathol 43:63–82. doi: https://doi.org/10.1146/annurev.phyto.43.040204.140251 CrossRefPubMedGoogle Scholar
- Doyle JJ, Doyle JL (1987) A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochem Bull 19:11–15Google Scholar
- Lagrimini LM, Vaughn J, Erb WA, Miller SA (1993) Peroxidase overproduction in tomato: wound-induced polyphenol deposition and disease resistance. Hortscience 28:218–221Google Scholar
- Lehmann E (1975) Nonparametrics: statistical methods based on ranks. McGrew-Hill, San FranciscoGoogle Scholar
- Liu P, Wang L, Wong S-M, Yue GH (2016) Fine mapping QTL for resistance to VNN disease using a high-density linkage map in Asian seabass. Sci Rep 6:32122. https://doi.org/10.1038/srep32122 https://www.nature.com/articles/srep32122#supplementary-information
- Liu X-H et al. (2017) Autophagy-related protein MoAtg14 is involved in differentiation, development and pathogenicity in the rice blast fungus Magnaporthe oryzae. Sci Rep 7:1415–1425. https://doi.org/10.1038/srep40018 https://www.nature.com/articles/srep40018#supplementary-information
- Lopez C, Soto M, Restrepo S et al. (2005) Gene expression profile in response to Xanthomonas axonopodis pv. manihotis infection in cassava using a cDNA microarray. Plant Mol Biol 57:393–410. https://doi.org/10.1007/s11103-004-7819-3
- Muengula-Manyi M, Nkongolo KK, Bragard C, Tshilenge-Djim P, Winter S, Kalonji-Mbuyi A (2012) Incidence, severity and gravity of cassava mosaic disease in savannah agro-ecological region of DR-Congo: analysis of agro-environmental factors. Am J Plant Sci 03:512–519. https://doi.org/10.4236/ajps.2012.34061 CrossRefGoogle Scholar
- Ooijen JW, Voorrips RE (2001) JoinMap 3.0 Software for the calculation of genetic linkage maps. Plant Research International, Wageningen, the NetherlandsGoogle Scholar
- Prince DC, Drurey C, Zipfel C, Hogenhout SA (2014) The leucine-rich repeat receptor-like kinase brassinosteroid insensitive1-associated kinase1 and the cytochrome p450 phytoalexin deficient3 contribute to innate immunity to aphids in arabidopsis. Plant Physiol 164:2207–2219. https://doi.org/10.1104/pp.114.235598 CrossRefPubMedPubMedCentralGoogle Scholar
- SPSS Inc (2010) IBM SPSS Statistics 19.0 core system user’s guide. New YorkGoogle Scholar
- Sukhuman W, Supajit S, Ratchadaporn T, Opas B, Duncan RS, Kanokporn T (2015) Validation of a reference gene for transcript analysis in cassava (Manihot esculenta Crantz) and its application in analysis of linamarase and -hydroxynitrile lyase expression at different growth stages. Afr J Biotechnol 14:745–751. https://doi.org/10.5897/ajb2014.14316 CrossRefGoogle Scholar
- Van Ooijen JW, Boer MP, Jansen RC, Maliepaard C (2002) MapQTL 4.0. Software for the calculation of QTL positions on genetic maps. Plant Research International, Wageningen, the Netherlands,Google Scholar
- Whankaew S, Poopear S, Kanjanawattanawong S, Tangphatsornruang S, Boonseng O, Lightfoot DA, Triwitayakorn K (2011) A genome scan for quantitative trait loci affecting cyanogenic potential of cassava root in an outbred population. BMC Genomics 12:266. https://doi.org/10.1186/1471-2164-12-266 CrossRefPubMedPubMedCentralGoogle Scholar
- Wydra K, Zinsou V, Jorge V, Verdier V (2004) Identification of pathotypes of Xanthomonas axonopodis pv. manihotis in Africa and detection of quantitative trait loci and markers for resistance to bacterial blight of cassava. Phytopathology 94:1084–1093. https://doi.org/10.1094/PHYTO.2004.94.10.1084 CrossRefPubMedGoogle Scholar
- Zhang J, Liu T, Feng R et al. (2015) Genetic Map Construction and Quantitative Trait Locus (QTL) Detection of Six Economic Traits Using an F2 Population of the Hybrid from Saccharina longissima and Saccharina japonica. PLoS ONE 10(5):e0128588Google Scholar