Host induced gene silencing of Sclerotinia sclerotiorum effector genes for the control of white mold

Maximiano, Mariana Rocha; Souza, Lucas Santos; Santos, Cristiane dos; Aragão, F. J. L.; Dias, Simoni Campos; Franco, Octávio Luiz; Mehta, Ângela

doi:10.1016/j.bcab.2022.102302

Cited by 5 publications

(3 citation statements)

References 53 publications

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“…infection in three RKN-resistant legume species [ 35 ]. Here, an RNAi-inducing binary vector was constructed using previously described parameters [ 36 ] to downregulate the expression of GmPG in soybean hairy roots. Briefly, different fragments (cutoff of 400 bp) of the GmPG coding sequence were inspected for their specificity using nucleotide BLAST (blast.ncbi.nlm.nih.gov/), and those with no hits against public databanks selected for cloning.…”

Section: Methodsmentioning

confidence: 99%

“…The homology of the selected dsRNA with non-target organisms was predicted by siRNA Target Finder ( https://www.genscript.com/tools/sirna-target-finder ) and dsCheck ( http://dscheck.rnai.jp/ ) tools. The specific 400 bp sequence was then synthesized in sense and antisense orientations, separated by a PDK intron from the pRNAi-psiuK vector [ 36 ], and under the control of the constitutive promoter of CaMV35S and the NOS terminator, using the services of Epoch Life Science Inc. (Missouri City, Texas, USA). The entire cassette was then cloned into the binary vector pPZP-empty (hereafter called pPZP-siGmPG), as described above.…”

Section: Methodsmentioning

confidence: 99%

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A novel soybean hairy root system for gene functional validation

et al. 2023

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Agrobacterium rhizogenes-mediated transformation has long been explored as a versatile and reliable method for gene function validation in many plant species, including soybean (Glycine max). Likewise, detached-leaf assays have been widely used for rapid and mass screening of soybean genotypes for disease resistance. The present study combines these two methods to establish an efficient and practical system to generate transgenic soybean hairy roots from detached leaves and their subsequent culture under ex vitro conditions. We demonstrated that hairy roots derived from leaves of two (tropical and temperate) soybean cultivars could be successfully infected by economically important species of root-knot nematodes (Meloidogyne incognita and M. javanica). The established detached-leaf method was further explored for functional validation of two candidate genes encoding for cell wall modifying proteins (CWMPs) to promote resistance against M. incognita through distinct biotechnological strategies: the overexpression of a wild Arachis α-expansin transgene (AdEXPA24) and the dsRNA-mediated silencing of an endogenous soybean polygalacturonase gene (GmPG). AdEXPA24 overexpression in hairy roots of RKN-susceptible soybean cultivar significantly reduced nematode infection by approximately 47%, whereas GmPG downregulation caused an average decrease of 37%. This novel system of hairy root induction from detached leaves showed to be an efficient, practical, fast, and low-cost method suitable for high throughput in root analysis of candidate genes in soybean.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

A novel soybean hairy root system for gene functional validation

et al. 2023

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“…The following HIGS and SIGS studies are not included in the Gebremichael et al., 2021 review and have been used with Fusarium graminearum sterol 14α-demethylase genes FgCYP51A, FgCYP51B and the virulence factor FgCYP51C, in Arabidopsis and barley ( Koch et al., 2019 ), and the Fg00677, Fg08731 (two essential protein kinases), and CYP51 (cytochrome P450 lanosterol C14-α-demethylase) genes in Brachypodium distachyon ( He et al., 2019 ). HIGS has also been used to target Sclerotinia sclerotiorum genes Ssoah1 (regulatory pivot for oxalic acid production) ( Rana et al., 2022 ), Ss-caF1 (putative Ca2+ binding protein), SspG1d (endopolygalacturonase), and SsiTL (integrin) ( Maximiano et al., 2022 ) and when combatting Phakopsora pachyrhizi , three genes (ATC: acetyl‐CoA acyltransferase , GCS_H: glycine cleavage system H protein , and RP_S16: 40S ribosomal protein S16) of eight genes evaluated, reduced the number of pustules of P. pachyrhizi ( Hu et al., 2020 ).…”

Section: Implications Of Cross-kingdom Rnai For Control Of Plant Path...mentioning

confidence: 99%

Mycovirus-encoded suppressors of RNA silencing: Possible allies or enemies in the use of RNAi to control fungal disease in crops

Coy¹,

Plummer²,

Khalifa³

et al. 2022

Front. Fungal Biol.

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Plants, fungi, and many other eukaryotes have evolved an RNA interference (RNAi) mechanism that is key for regulating gene expression and the control of pathogens. RNAi inhibits gene expression, in a sequence-specific manner, by recognizing and deploying cognate double-stranded RNA (dsRNA) either from endogenous sources (e.g. pre-micro RNAs) or exogenous origin (e.g. viruses, dsRNA, or small interfering RNAs, siRNAs). Recent studies have demonstrated that fungal pathogens can transfer siRNAs into plant cells to suppress host immunity and aid infection, in a mechanism termed cross-kingdom RNAi. New technologies, based on RNAi are being developed for crop protection against insect pests, viruses, and more recently against fungal pathogens. One example, is host-induced gene silencing (HIGS), which is a mechanism whereby transgenic plants are modified to produce siRNAs or dsRNAs targeting key transcripts of plants, or their pathogens or pests. An alternative gene regulation strategy that also co-opts the silencing machinery is spray-induced gene silencing (SIGS), in which dsRNAs or single-stranded RNAs (ssRNAs) are applied to target genes within a pathogen or pest. Fungi also use their RNA silencing machinery against mycoviruses (fungal viruses) and mycoviruses can deploy virus-encoded suppressors of RNAi (myco-VSRs) as a counter-defence. We propose that myco-VSRs may impact new dsRNA-based management methods, resulting in unintended outcomes, including suppression of management by HIGS or SIGS. Despite a large diversity of mycoviruses being discovered using high throughput sequencing, their biology is poorly understood. In particular, the prevalence of mycoviruses and the cellular effect of their encoded VSRs are under-appreciated when considering the deployment of HIGS and SIGS strategies. This review focuses on mycoviruses, their VSR activities in fungi, and the implications for control of pathogenic fungi using RNAi.

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Synthetic modulation of ROS scavenging during host—Sclerotinia sclerotiorum interaction: a new strategy for the development of highly resistant plants

Ding,

Yan,

Zhao

et al. 2024

Phytopathol Res

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Sclerotinia sclerotiorum is a widespread fungal pathogen responsible for significant crop losses across the globe. The challenge of breeding resistant varieties is exacerbated by the fungus's sophisticated pathogenic mechanisms. A pivotal factor in the host-pathogen interaction is the regulation of reactive oxygen species (ROS) within both the fungi and the host plants. However, there is currently no efficient strategy to leverage this interaction mechanism for developing disease-resistant crop varieties. Here, we introduce an engineered ROS scavenging system designated as syn-ROS for impairing ROS neutralization in S. sclerotiorum while concurrently fortifying it in the host. The syn-ROS system comprises gene silencing constructs targeting the S. sclerotiorum Cu/Zn superoxide dismutase (SsSOD) and its copper chaperone (SsCCS), alongside overexpression constructs for the Arabidopsis thaliana AtSOD1 and AtCCS. Transgenic plants carrying the syn-ROS system demonstrated a marked enhancement in resistance to S. sclerotiorum. Upon infection, the expression of SsSOD and SsCCS was reduced, while the expression of AtSOD1 and AtCCS was enhanced in syn-ROS transgenic plants. Moreover, the infected syn-ROS plants showed decreased Cu/Zn SOD enzyme activity and elevated ROS concentrations within the fungal cells. In contrast, the cells of A. thaliana manifested increased Cu/Zn SOD enzyme activity and lowered ROS levels. Collectively, these findings suggest a novel and promising approach for contriving plants with robust resistance by synthetically manipulating ROS scavenging activities in the interaction between the host and S. sclerotiorum.

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Host induced gene silencing of Sclerotinia sclerotiorum effector genes for the control of white mold

Cited by 5 publications

References 53 publications

A novel soybean hairy root system for gene functional validation

A novel soybean hairy root system for gene functional validation

Mycovirus-encoded suppressors of RNA silencing: Possible allies or enemies in the use of RNAi to control fungal disease in crops

Synthetic modulation of ROS scavenging during host—Sclerotinia sclerotiorum interaction: a new strategy for the development of highly resistant plants

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