In collaboration with Payame Noor University and Iranian Biotechnology Society

Document Type : Research Paper

Authors

1 Ph.D. Candidate, Biotechnology and Plant Breeding Department, Ferdowsi University of Mashhad, Mashhad, Iran.

2 Assistant Professor, Biotechnology and Plant Breeding Department, Ferdowsi University of Mashhad, Mashhad, Iran.

3 Assistant Professor, Higher Education Complex of Shirvan, University of Bojnord, Shirvan, Iran.

Abstract

Broomrape (Orobanche aegyptica) is a notorious parasitic plant that cause significant production loss. Here we report analysis of publicly available RNA-seq data for broomrape, coupled with experimental verification of part of the results. After quality control of raw illumine reads, qualified reads were mapped against Orobanch transcriptome. Differential gene expression analysis, performed by using DESeq package, identified 391 differentially expressed genes between seed imbibition and haustorium attachment stages. The expression of orthologs of these genes in close relatives of Orobanch, which are parasitic, hemi- or non-parasitic, was investigated. From 391 identified genes, 87 genes showed high levels of expression in parasitic relatives and not in non-parasitic ones. Based on these analyses the 87 genes were considered as candidate genes involved in establishment of parasitic interaction between Orobanch and its host. The expression of nine of these genes were checked experimentally in flower tissues of Orobanch and in tissues sampled from the attachment site on the host root. The expression of Or2094, which a putative serine-carboxy peptidase, was detected only in the attachment site, supporting the role of this gene in establishment of the parasitic interaction. The results of this work will pave the way for future genetic engineering projects to use host-induced gene silencing strategy to enhance resistance to Orobanch in host crop plants.

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Aly, R., Dubey, N. K., Yahyaa, M., Abu-Nassar, J., & Ibdah, M. (2014). Gene silencing of CCD7 and CCD8 in Phelipanche aegyptiaca by tobacco rattle virus system retarded the parasite development on the host. Plant Signaling & Behavior, 9(8), e29376. Anders, S., & Huber, W. (2010). Differential expression analysis for sequence count data. Nature Precedings, 1-1. Andrews, S. (2017). FastQC: a quality control tool for high throughput sequence data. 2010. Bolger, A. M., Lohse, M., & Usadel, B. (2014). Trimmomatic: a flexible trimmer for Illumina sequence data. Bioinformatics, 30(15), 2114-2120. Deng, Q., Wang, X., Zhang, D., Wang, X., Feng, C., & Xu, S. (2017). BRS1 function in facilitating lateral root emergence in Arabidopsis. International Journal of Molecular Sciences, 18(7), 1549. El-Maarouf-Bouteau, H., Moreau, E., Errakhi, R., & Sallé, G. (2008). A diffusible signal from germinating Orobanche ramosa elicits early defense responses in suspension-cultured Arabidopsis thaliana. Plant Signaling & Behavior, 3(3), 189-193. Govindarajulu, M., Epstein, L., Wroblewski, T., & Michelmore, R. W. (2015). Host‐induced gene silencing inhibits the biotrophic pathogen causing downy mildew of lettuce. Plant Biotechnology Journal, 13(7), 875-883. Hegenauer, V., Fürst, U., Kaiser, B., Smoker, M., Zipfel, C., Felix, G.,.. . Albert, M. (2016). Detection of the plant parasite Cuscuta reflexa by a tomato cell surface receptor. Science, 353(6298), 478-481. Heide-Jørgensen, H. (2008). Parasitic flowering plants: Brill. Hogenhout, S. A., Van der Hoorn, R. A., Terauchi, R., & Kamoun, S. (2009). Emerging concepts in effector biology of plant-associated organisms. Molecular Plant-Microbe Interactions, 22(2), 115-122. Jones, J. D., & Dangl, J. L. (2006). The plant immune system. Nature, 444(7117), 323. Koch, A., Kumar, N., Weber, L., Keller, H., Imani, J., & Kogel, K.-H. (2013). Host-induced gene silencing of cytochrome P450 lanosterol C14α-demethylase–encoding genes confers strong resistance to Fusarium species. Proceedings of the National Academy of Sciences, 110(48), 19324-19329. Li, H., & Durbin, R. (2009). Fast and accurate short read alignment with Burrows–Wheeler transform. Bioinformatics, 25(14), 1754-1760. Li, J., Lease, K. A., Tax, F. E., & Walker, J. C. (2001). BRS1, a serine carboxypeptidase, regulates BRI1 signaling in Arabidopsis thaliana. Proceedings of the National Academy of Sciences, 98(10), 5916-5921. Li, J., & Timko, M. P. (2009). Gene-for-gene resistance in Striga-cowpea associations. Science, 325(5944), 1094-1094. Liu, J., Blaylock, L. A., Endre, G., Cho, J., Town, C. D., VandenBosch, K. A., & Harrison, M. J. (2003). Transcript profiling coupled with spatial expression analyses reveals genes involved in distinct developmental stages of an arbuscular mycorrhizal symbiosis. The Plant Cell, 15(9), 2106-2123. Mower, J.P., Stefanović, S., Hao, W., Gummow, J. S., Jain, K., Ahmed, D., & Palmer, J.D. (2010). Horizontal acquisition of multiple mitochondrial genes from a parasitic plant followed by gene conversion with host mitochondrial genes. BMC Biology, 8(1), 1-16. Musselman, L. J. (1980). The biology of Striga, Orobanche, and other root-parasitic weeds. Annual Review of Phytopathology, 18(1), 463-489. Petersen, T. N., Brunak, S., Von Heijne, G., & Nielsen, H. (2011). SignalP 4.0: discriminating signal peptides from transmembrane regions. Nature Methods, 8(10), 785-786. Roney, J.K., Khatibi, P.A., & Westwood, J.H. (2007). Cross-species translocation of mRNA from host plants into the parasitic plant dodder. Plant Physiology, 143(2), 1037-1043. Song, Y., & Thomma, B. P. (2018). Host‐induced gene silencing compromises Verticillium wilt in tomato and Arabidopsis. Molecular Plant Pathology, 19(1), 77-89. Warnes, G. R., Bolker, B., Bonebakker, L., Gentleman, R., Liaw, W. H. A., Lumley, T.,.. . Schwartz, M. (2015). gplots: Various R programming tools for plotting data. Win, J., Chaparro-Garcia, A., Belhaj, K., Saunders, D., Yoshida, K., Dong, S.,.. . Hogenhout, S. (2012). Effector biology of plant-associated organisms: concepts and perspectives. Paper presented at the Cold Spring Harbor Symposia on Quantitative Biology. Xi, Z., Wang, Y., Bradley, R. K., Sugumaran, M., Marx, C. J., Rest, J. S., & Davis, C. C. (2013). Massive mitochondrial gene transfer in a parasitic flowering plant clade. PLoS Genetics, 9(2), e1003265. Yang, Z., Wafula, E. K., Honaas, L. A., Zhang, H., Das, M., Fernandez-Aparicio, M.,.. . Der, J. P. (2015). Comparative transcriptome analyses reveal core parasitism genes and suggest gene duplication and repurposing as sources of structural novelty. Molecular Biology and Evolution, 32(3), 767-790. Zhang, D., Qi, J., Yue, J., Huang, J., Sun, T., Li, S.,.. . Wang, L. (2014). Root parasitic plant Orobanche aegyptiaca and shoot parasitic plant Cuscuta australis obtained Brassicaceae-specific strictosidine synthase-like genes by horizontal gene transfer. BMC Plant Biology, 14(1), 1-14. Zhang, T., Jin, Y., Zhao, J.-H., Gao, F., Zhou, B.-J., Fang, Y.-Y., & Guo, H.-S. (2016). Host-induced gene silencing of the target gene in fungal cells confers effective resistance to the cotton wilt disease pathogen Verticillium dahliae. Molecular Plant, 9(6), 939-942.