Draft Genome Sequence Resource of CBPPT1, a ‘<i>Candidatus</i> Phytoplasma trifolii’-Related Strain Associated with Potato Purple Top Disease in the Columbia Basin, U.S.A.

Wei, Wei; Shao, Jonathan; Bottner-Parker, Kristi D.; Zhao, Yan

doi:10.1094/pdis-08-22-1788-a

Cited by 3 publications

(1 citation statement)

References 27 publications

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“…To obtain the full-length gene-encoding PPT-IDP, genomic DNA was extracted from PPT-infected tomato plants using a previously established protocol (see above). PCR amplification was then performed using specific primer pairs PPT-IDP-F and PPT-IDP-R ( Supplementary Table S1 ), designed based on the PPT-IDP gene sequence (Accession number JANHJP000000000) [ 28 ]. The pDONR221 entry vector (Invitrogen, Waltham, MA, USA) in the Gateway cloning system was utilized to create a recombinant entry clone by recombining the PPT-IDP PCR product with the vector using BP Clonase II enzyme (Thermo Fisher Scientific, Waltham, MA, USA).…”

Section: Methodsmentioning

confidence: 99%

The Endoplasmic Reticulum Is a Key Battleground between Phytoplasma Aggression and Host Plant Defense

Inaba,

Kim,

Zhao

et al. 2023

Cells

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Phytoplasmas are intracellular plant pathogens that heavily rely on host cell nutrients for survival and propagation due to their limited ability to synthesize essential substrates. The endoplasmic reticulum (ER), which plays a vital role in various cellular processes, including lipid and protein biosynthesis, is an attractive target for numerous intracellular pathogens to exploit. This study investigated the impact of potato purple top (PPT) phytoplasma infection on the ER in tomato plants. Abnormal accumulation of ER-resident proteins, disrupted ER network structures, and formation of protein aggregates in the phloem were observed using confocal microscopy and transmission electron microscopy, indicating a phytoplasma-infection-induced disturbance in ER homeostasis. The colocalization of phytoplasmas with the accumulated ER-resident proteins suggests an association between ER stress, unfolded protein response (UPR) induction, and phytoplasma infection and colonization, with the ER stress response likely contributing to the host plant’s defense mechanisms. Quantitative real-time PCR revealed a negative correlation between ER stress/UPR activation and PPT phytoplasma titer, implying the involvement of UPR in curbing phytoplasma proliferation. Inducing ER stress and activating the UPR pathway effectively decreased phytoplasma titer, while suppressing the ER-resident protein, binding immunoglobulin protein (BiP) increased phytoplasma titer. These results highlight the ER as an intracellular battleground where phytoplasmas exploit host components for survival and multiplication, while host plants deploy defense mechanisms to counteract the invasion. Understanding the intricate interactions between phytoplasmas and plant hosts at the subcellular level, particularly within the ER, provides valuable insights for developing new strategies to control phytoplasma diseases.

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Section: Methodsmentioning

confidence: 99%

The Endoplasmic Reticulum Is a Key Battleground between Phytoplasma Aggression and Host Plant Defense

Inaba,

Kim,

Zhao

et al. 2023

Cells

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Phytoplasma DNA Enrichment from Sugarcane White Leaves for Shotgun Sequencing Improvement

Lohmaneeratana,

Gutiérrez,

Thamchaipenet

et al. 2024

Plants

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Sugarcane white leaf (SCWL) disease, caused by Candidatus Phytoplasma sacchari, poses a significant threat to sugarcane cultivation. An obligate parasite, phytoplasma is difficult to culture in laboratory conditions, making the isolation of its DNA from the massive amount of plant host DNA extremely challenging. Yet, the appropriate amount and quality of plant microbiome-derived DNA are key for high-quality DNA sequencing data. Here, a simple, cost-effective, alternative method for DNA isolation was applied using a guanidine-HCl-hydroxylated silica (GuHCl-Silica)-based method and microbiome DNA enrichment based on size-selective low-molecular-weight (LMW) DNA by PEG/NaCl precipitation. qPCR analysis revealed a significant enrichment of phytoplasma DNA in the LMW fraction. Additionally, the NEBNext Microbiome DNA enrichment kit was utilized to further enrich microbial DNA, demonstrating a remarkable increase in the relative abundance of phytoplasma DNA to host DNA. Shotgun sequencing of the isolated DNA gave high-quality data on the metagenome assembly genome (MAG) of Ca. Phytoplasma sacchari SCWL with completeness at 95.85 and 215× coverage. The results indicate that this combined approach of PEG/NaCl size selection and microbiome enrichment is effective for obtaining high-quality genomic data from phytoplasma, surpassing previous methods in efficiency and resource utilization. This low-cost method not only enhances the recovery of microbiome DNA from plant hosts but also provides a robust framework for studying plant pathogens in complex plant models.

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Recent advances of the causal agent of yellow leaf disease (YLD) on areca palm (<i>Areca catechu</i> L.)

Khan,

Zhao,

Wang

et al. 2023

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Draft Genome Sequence Resource of CBPPT1, a ‘Candidatus Phytoplasma trifolii’-Related Strain Associated with Potato Purple Top Disease in the Columbia Basin, U.S.A.

Cited by 3 publications

References 27 publications

The Endoplasmic Reticulum Is a Key Battleground between Phytoplasma Aggression and Host Plant Defense

The Endoplasmic Reticulum Is a Key Battleground between Phytoplasma Aggression and Host Plant Defense

Phytoplasma DNA Enrichment from Sugarcane White Leaves for Shotgun Sequencing Improvement

Recent advances of the causal agent of yellow leaf disease (YLD) on areca palm (<i>Areca catechu</i> L.)

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