Plant hairy roots enable high throughput identification of antimicrobials against Candidatus Liberibacter spp.

Irigoyen, Sonia; Ramasamy, Manikandan; Pant, Shankar R.; Niraula, Prakash M.; Bedre, Renesh; Gurung, Meena; D, Rossi; Laughlin, Corinne; Gorman, Zachary; Achor, Diann; Levy, Amit; Kolomiets, Michael V.; Sétamou, Mamoudou; Badillo-Vargas, Ismael E.; Avila, Carlos A.; Irey, Mike; Mandadi, Kranthi K.

doi:10.1038/s41467-020-19631-x

Cited by 44 publications

(28 citation statements)

References 99 publications

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“…Thus, it remains unclear whether A4 and A4T are the same or separate strains (T in A4T might merely indicate that A4 is a type strain), and a comparative analysis of the genome and plasmid sequences of A4, A4T, A136, and R1000 may be required to resolve the uncertainty with this nomenclature. R1000 is also known under its strain collection name ATCC43056 [ 63 ] ( Figure 1 ), and in one study as 43056 [ 64 ]. The R1000 genome was published by ATCC in April 2021 [ 63 ], with the results showing a 96.62% average nucleotide identity (ANI) between the genomes of R1000 (ATCC43056) and C58 (ATCC33970) and 98.48% ANI between the genomes of R1000 and A136 (ATCC51350) [ 65 ].…”

Section: Agrobacterium Rhizogenes : a Historical View Of The Widely Used Strains And Their Nomenclaturementioning

confidence: 99%

Hairy CRISPR: Genome Editing in Plants Using Hairy Root Transformation

Kiryushkin

Ilina

Guseva

et al. 2021

Plants

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CRISPR/Cas-mediated genome editing is a powerful tool of plant functional genomics. Hairy root transformation is a rapid and convenient approach for obtaining transgenic roots. When combined, these techniques represent a fast and effective means of studying gene function. In this review, we outline the current state of the art reached by the combination of these approaches over seven years. Additionally, we discuss the origins of different Agrobacterium rhizogenes strains that are widely used for hairy root transformation; the components of CRISPR/Cas vectors, such as the promoters that drive Cas or gRNA expression, the types of Cas nuclease, and selectable and screenable markers; and the application of CRISPR/Cas genome editing in hairy roots. The modification of the already known vector pKSE401 with the addition of the rice translational enhancer OsMac3 and the gene encoding the fluorescent protein DsRed1 is also described.

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Section: Agrobacterium Rhizogenes : a Historical View Of The Widely Used Strains And Their Nomenclaturementioning

confidence: 99%

Hairy CRISPR: Genome Editing in Plants Using Hairy Root Transformation

Kiryushkin

Ilina

Guseva

et al. 2021

Plants

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“…Despite the substantial global economic impact of Liberibacter pathogens, our understanding of the molecular mechanisms involved in host infection and disease progression remains superficial. Although novel methods for screening antimicrobial compounds have been developed [28,29], increased understanding of the virulent mechanism can reveal precise targets in the pathosystems for creating new disease control strategies. More than 200 potential effectors have been reported from Liberibacter pathogens, but only about a dozen have been functionally characterized.…”

Section: Discussionmentioning

confidence: 99%

HPE1, an Effector from Zebra Chip Pathogen Interacts with Tomato Proteins and Perturbs Ubiquitinated Protein Accumulation

Kan

Mendoza-Herrera

Lévy

et al. 2021

IJMS

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The gram-negative bacterial genus Liberibacter includes economically important pathogens, such as ‘Candidatus Liberibacter asiaticus’ that cause citrus greening disease (or Huanglongbing, HLB) and ‘Ca. Liberibacter solanacearum’ (Lso) that cause zebra chip disease in potato. Liberibacter pathogens are fastidious bacteria transmitted by psyllids. Pathogen manipulation of the host’ and vector’s immune system for successful colonization is hypothesized to be achieved by Sec translocon-dependent effectors (SDE). In previous work, we identified hypothetical protein effector 1 (HPE1), an SDE from Lso, that acts as a suppressor of the plant’s effector-triggered immunity (ETI)-like response. In this study, using a yeast two-hybrid system, we identify binding interactions between tomato RAD23 proteins and HPE1. We further show that HPE1 interacts with RAD23 in both nuclear and cytoplasmic compartments in planta. Immunoblot assays show that HPE1 is not ubiquitinated in the plant cell, but rather the expression of HPE1 induced the accumulation of other ubiquitinated proteins. A similar accumulation of ubiquitinated proteins is also observed in Lso infected tomato plants. Finally, earlier colonization and symptom development following Lso haplotype B infection are observed in HPE1 overexpressing plants compared to wild-type plants. Overall, our results suggest that HPE1 plays a role in virulence in Lso pathogenesis, possibly by perturbing the ubiquitin-proteasome system via direct interaction with the ubiquitin-like domain of RAD23 proteins.

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“…The growth of Ishi-1 was determined based on the identification of C Las population and increase of C Las DNA amounts; however, there has been no direct evidence to support the phenomenon of Ishi-1 growth [ 34 ]. C Lso and C Las were able to be maintained in vivo in hairy root explants for 28 and 120 days, respectively [ 35 ]. Similarly, the leaf discs with supplemented glucose show an increase in C Las titer [ 36 ].…”

Section: Transient Culture Of Ca Liberibactermentioning

confidence: 99%

Molecular signatures between citrus and Candidatus Liberibacter asiaticus

Rao

Deng

et al. 2021

PLoS Pathog

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Citrus Huanglongbing (HLB), also known as citrus greening, is one of the most devastating citrus diseases worldwide. Candidatus Liberibacter asiaticus (CLas) is the most prevalent strain associated with HLB, which is yet to be cultured in vitro. None of the commercial citrus cultivars are resistant to HLB. The pathosystem of Ca. Liberibacter is complex and remains a mystery. In this review, we focus on the recent progress in genomic research on the pathogen, the interaction of host and CLas, and the influence of CLas infection on the transcripts, proteins, and metabolism of the host. We have also focused on the identification of candidate genes for CLas pathogenicity or the improvements of HLB tolerance in citrus. In the end, we propose potentially promising areas for mechanistic studies of CLas pathogenicity, defense regulators, and genetic improvement for HLB tolerance/resistance in the future.

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Plant hairy roots enable high throughput identification of antimicrobials against Candidatus Liberibacter spp.

Cited by 44 publications

References 99 publications

Hairy CRISPR: Genome Editing in Plants Using Hairy Root Transformation

Hairy CRISPR: Genome Editing in Plants Using Hairy Root Transformation

HPE1, an Effector from Zebra Chip Pathogen Interacts with Tomato Proteins and Perturbs Ubiquitinated Protein Accumulation

Molecular signatures between citrus and Candidatus Liberibacter asiaticus

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