scholarly journals mCherry-Labeled Verticillium dahliae Could Be Utilized to Investigate Its Pathogenicity Process in Nicotiana benthamiana

Genes ◽  
2018 ◽  
Vol 9 (10) ◽  
pp. 508 ◽  
Author(s):  
Xiaofeng Su ◽  
Guoqing Lu ◽  
Latifur Rehman ◽  
Xiaokang Li ◽  
Lu Sun ◽  
...  
Keyword(s):  
Molecular Mechanism ◽  
Verticillium Dahliae ◽  
Nicotiana Benthamiana ◽  
Phytopathogenic Fungus ◽  
Post Inoculation ◽  
Vascular Wilt ◽  
Wild Type ◽  
Type V ◽  
Lateral Root Primordium ◽  
Root Primordium

Verticillium dahliae is a soil-borne phytopathogenic fungus that causes a destructive vascular wilt, but details of the molecular mechanism behind its pathogenicity are not very clear. Here, we generated a red fluorescent isolate of V. dahliae by protoplast transformation to explore its pathogenicity mechanism, including colonization, invasion, and extension in Nicotiana benthamiana, using confocal microscopy. The nucleotide sequences of mCherry were optimized for fungal expression and cloned into pCT-HM plasmid, which was inserted into V. dahliae protoplasts. The transformant (Vd-m) shows strong red fluorescence and its phenotype, growth rate, and pathogenicity did not differ significantly from the wild type V. dahliae (Vd-wt). Between one and three days post inoculation (dpi), the Vd-m successfully colonized and invaded epidermal cells of the roots. From four to six dpi, hyphae grew on root wounds and lateral root primordium and entered xylem vessels. From seven to nine dpi, hyphae extended along the surface of the cell wall and massively grew in the xylem vessel of roots. At ten dpi, the Vd-m was found in petioles and veins of leaves. Our results distinctly showed the pathway of V. dahliae infection and colonization in N. benthamiana, and the optimized expression can be used to deepen our understanding of the molecular mechanism of pathogenicity.

scholarly journals Transcriptome analysis reveals the mechanism by which the biocontrol fungus Chaetomium globosum CEF-082 controls Verticillium wilt in cotton

2019 ◽  
Author(s):  
Yun Zhang ◽  
Na Yang ◽  
Lihong Zhao ◽  
Heqin Zhu ◽  
Canming Tang
Keyword(s):  
Molecular Mechanism ◽  
Verticillium Wilt ◽  
Verticillium Dahliae ◽  
Substantial Reduction ◽  
Enrichment Analysis ◽  
Metabolic Process ◽  
Defence Response ◽  
Chaetomium Globosum ◽  
Post Inoculation ◽  
Mapk Signalling Pathway

Abstract Background: Verticillium wilt of cotton is a serious soil-borne disease that causes a substantial reduction in cotton yield. A previous study showed that the endophytic fungus Chaetomium globosum CEF-082 could control Verticillium wilt of cotton, but the molecular mechanism by which CEF-082 controls Verticillium wilt is still unknown. Results: To study the mechanism by which CEF-082 controls Verticillium wilt, the transcriptome of cotton seedlings pretreated with CEF-082 was sequenced. The results revealed 5638 DEGs 24 h post-inoculation with CEF-082, and 2921 and 2153 DEGs 12 and 48 h post-inoculation with Verticillium dahliae , respectively. At twenty-four hours post-inoculation with CEF-082, KEGG enrichment analysis indicated that the DEGs were mainly enriched in plant-pathogen interaction, MAPK signalling pathway-plant, flavonoid biosynthesis, and phenylpropanoid biosynthesis. There were 1209 DEGs specifically induced after inoculation with CEF-082 and V. dahliae . GO enrichment indicated that these DEGs were mainly enriched in the terms reactive oxygen species metabolic process, hydrogen peroxide metabolic process, defence response, superoxide dismutase activity, and antioxidant activity. Here, many genes, such as ERF, CNGC, FLS2, MYB, GST and CML, were identified that regulate crucial points in defence-related pathways and that may contribute to V. dahliae resistance in cotton. These results provide a basis for the understanding of the molecular mechanism by which biocontrol fungi control Verticillium wilt. Conclusions: In this study, we found that CEF-082 could regulate multiple metabolic pathways in cotton. After treatment with Verticillium dahliae , the defence response of cotton plants pre-inoculated with CEF-082 was strengthened.

Differential symptom development and viral RNA loads in ten Nicotiana benthamiana accessions infected with the tobamovirus yellow tailflower mild mottle virus

Plant Disease ◽  
2021 ◽  
Author(s):  
Weinan Xu ◽  
Yuxia Guo ◽  
Hua Li ◽  
Krishnapillai Sivasithamparam ◽  
Michael G.K. Jones ◽  
...  
Keyword(s):  
Nicotiana Benthamiana ◽  
Geographical Location ◽  
Viral Rna ◽  
Mottle Virus ◽  
Wild Plants ◽  
Post Inoculation ◽  
Wild Type ◽  
Reverse Transcription Pcr ◽  
Systemic Hypersensitivity ◽  
Rna Polymerase 1

Yellow tailflower mild mottle virus (YTMMV, genus Tobamovirus) was identified from wild plants of solanaceous species in Australia. Nicotiana benthamiana is a species indigenous to the arid north of Australia. N. benthamiana accession RA-4 or the ‘lab’ type, which has a mutant, functionally-defective, RNA-dependent RNA polymerase 1 (Rdr1) gene (Nb-Rdr1m), has played a significant role in plant virology, but little study has been done on responses to virus infection by other accessions of N. benthamiana. All wild-collected N. benthamiana accessions used in this study harboured wild-type Rdr1 genes (Nb-Rdr1). We compared symptoms of YTMMV infection and viral RNA load on RA-4 and nine wild-collected accessions of N. benthamiana from mainland Western Australia, an island, and the Northern Territory. After inoculation with YTMMV, RA-4 plants responded with systemic hypersensitivity and all individuals were dead 35 days post-inoculation (dpi). Plants of wild-collected accessions exhibited a range of symptoms, from mild to severe, and some, but not all, individuals died in the same period. Quantitative reverse transcription PCR revealed that the Rdr1 mutation was not a predictor of viral RNA load or symptom severity. For example, wild-collected A019412 plants carried over twice the viral RNA load of RA-4 plants, but symptom expression was moderate. For plants of most accessions, viral RNA load did not increase after 10 dpi. The exception was plants of accession Barrow-1, where viral RNA load was low until 15 dpi, after which it increased over 29-fold. This study revealed differential responses by N. benthamiana accessions to infection by an isolate of YTMMV. The Rdr1 gene, whether mutant or wild-type, did not appear to influence viral RNA load or disease expression. Genetic diversity of the ten N. benthamiana lines in some cases reflected geographical location, but in other lines this was not so.

scholarly journals Transcriptome analysis reveals the defense mechanism of cotton against Verticillium dahliae in the presence of the biocontrol fungus Chaetomium globosum CEF-082

2019 ◽  
Author(s):  
YUN ZHANG ◽  
Na Yang ◽  
Lihong Zhao ◽  
Heqin Zhu ◽  
Canming Tang(New Corresponding Author)
Keyword(s):  
Molecular Mechanism ◽  
Verticillium Wilt ◽  
Verticillium Dahliae ◽  
Defense Response ◽  
Defense Mechanism ◽  
Substantial Reduction ◽  
Metabolic Process ◽  
Chaetomium Globosum ◽  
Post Inoculation ◽  
Cotton Plants

Abstract Background: Verticillium wilt of cotton is a serious soil-borne disease that causes a substantial reduction in cotton yields. A previous study showed that the endophytic fungus Chaetomium globosum CEF-082 could control Verticillium wilt of cotton, and induce a defense response in cotton plants. However, the comprehensive molecular mechanism governing this response is not yet clear. Results: To study the signalling mechanism induced by CEF-082, the transcriptome of cotton seedlings pretreated with CEF-082 was sequenced. The results revealed 5638 DEGs at 24 h post inoculation with CEF-082, and 2921 and 2153 DEGs at 12 and 48 h post inoculation with Verticillium dahliae, respectively. At 24 h post inoculation with CEF-082, KEGG enrichment analysis indicated that the DEGs were enriched mainly in the plant-pathogen interaction, MAPK signalling pathway-plant, flavonoid biosynthesis, and phenylpropanoid biosynthesis pathways. There were 1209 DEGs specifically induced only in cotton plants inoculated with V. dahliae in the presence of the biocontrol fungus CEF-082, and not when cotton plants were only inoculated with V. dahliae. GO analysis revealed that these DEGs were enriched mainly in the following terms: ROS metabolic process, H2O2 metabolic process, defense response, superoxide dismutase activity, and antioxidant activity. Moreover, many genes, such as ERF, CNGC, FLS2, MYB, GST and CML, that regulate crucial points in defense-related pathways were identified and may contribute to V. dahliae resistance in cotton. These results provide a basis for understanding the molecular mechanism by which the biocontrol fungus CEF-082 increases the resistance of cotton to Verticillium wilt. Conclusions: The results of this study showed that CEF-082 could regulate multiple metabolic pathways in cotton. After treatment with V. dahliae, the defense response of cotton plants preinoculated with CEF-082 was strengthened.

scholarly journals Transcriptome analysis reveals the defense mechanism of cotton against Verticillium dahliae in the presence of the biocontrol fungus Chaetomium globosum CEF-082

2019 ◽  
Author(s):  
YUN ZHANG ◽  
Na Yang ◽  
Lihong Zhao ◽  
Heqin Zhu ◽  
Canming Tang
Keyword(s):  
Molecular Mechanism ◽  
Verticillium Wilt ◽  
Verticillium Dahliae ◽  
Defense Response ◽  
Defense Mechanism ◽  
Substantial Reduction ◽  
Metabolic Process ◽  
Chaetomium Globosum ◽  
Post Inoculation ◽  
Cotton Plants

Abstract Background: Verticillium wilt of cotton is a serious soil-borne disease that causes a substantial reduction in cotton yields. A previous study showed that the endophytic fungus Chaetomium globosum CEF-082 could control Verticillium wilt of cotton, and induce a defense response in cotton plants. However, the comprehensive molecular mechanism governing this response is not yet clear. Results: To study the signalling mechanism induced by CEF-082, the transcriptome of cotton seedlings pretreated with CEF-082 was sequenced. The results revealed 5638 DEGs at 24 h post inoculation with CEF-082, and 2921 and 2153 DEGs at 12 and 48 h post inoculation with Verticillium dahliae , respectively. At 24 h post inoculation with CEF-082, KEGG enrichment analysis indicated that the DEGs were enriched mainly in the plant-pathogen interaction, MAPK signalling pathway-plant, flavonoid biosynthesis, and phenylpropanoid biosynthesis pathways. There were 1209 DEGs specifically induced only in cotton plants inoculated with V. dahliae in the presence of the biocontrol fungus CEF-082, and not when cotton plants were only inoculated with V. dahliae . GO analysis revealed that these DEGs were enriched mainly in the following terms: ROS metabolic process, H 2 O 2 metabolic process, defense response, superoxide dismutase activity, and antioxidant activity. Moreover, many genes, such as ERF , CNGC , FLS2 , MYB , GST and CML , that regulate crucial points in defense-related pathways were identified and may contribute to V. dahliae resistance in cotton. These results provide a basis for understanding the molecular mechanism by which the biocontrol fungus CEF-082 increases the resistance of cotton to Verticillium wilt. Conclusions: The results of this study showed that CEF-082 could regulate multiple metabolic pathways in cotton. After treatment with V. dahliae , the defense response of cotton plants preinoculated with CEF-082 was strengthened.

scholarly journals Transcriptome analysis reveals the cotton defense mechanism in response to Verticillium dahliae in the presence of the biocontrol fungus Chaetomium globosum CEF-082

2019 ◽  
Author(s):  
Yun Zhang ◽  
Na Yang ◽  
Lihong Zhao ◽  
Heqin Zhu ◽  
Canming Tang
Keyword(s):  
Molecular Mechanism ◽  
Verticillium Wilt ◽  
Verticillium Dahliae ◽  
Defense Response ◽  
Substantial Reduction ◽  
Enrichment Analysis ◽  
Metabolic Process ◽  
Chaetomium Globosum ◽  
Post Inoculation ◽  
Cotton Plants

Abstract Background: Verticillium wilt of cotton is a serious soil-borne disease that causes a substantial reduction in cotton yields. A previous study showed that the endophytic fungus Chaetomium globosum CEF-082 could control Verticillium wilt of cotton, and induce a defense response in cotton plants. However, the comprehensive molecular mechanism governing this response is not clear. Results: To study the signalling mechanism induced by CEF-082, the transcriptome of cotton seedlings pretreated with CEF-082 was sequenced. The results revealed 5638 DEGs at 24 h post inoculation with CEF-082, and 2921 and 2153 DEGs at 12 and 48 h post inoculation with Verticillium dahliae, respectively. At 24 h post inoculation with CEF-082, KEGG enrichment analysis indicated that the DEGs were enriched mainly in the plant-pathogen interaction, MAPK signalling pathway-plant, flavonoid biosynthesis, and phenylpropanoid biosynthesis pathways. There were 1209 DEGs specifically induced only in cotton plants inoculated with V. dahliae in the presence of the biocontrol fungus CEF-082, and not when cotton plants were only inoculated with V. dahliae. GO enrichment analysis revealed that these DEGs were enriched mainly in the following terms: ROS metabolic process, H2O2 metabolic process, defense response, superoxide dismutase activity, and antioxidant activity. Moreover, many genes, such as ERF, CNGC, FLS2, MYB, GST and CML genes, were identified that regulate crucial points in defence-related pathways and that may contribute to V. dahliae resistance in cotton. These results provide a basis for the understanding of the molecular mechanism by which biocontrol fungus CEF-082 increased the resistance of cotton to Verticillium wilt. Conclusions: The results of this study showed that CEF-082 could regulate multiple metabolic pathways in cotton. After treatment with V. dahliae, the defense response of cotton plants preinoculated with CEF-082 was strengthened.

scholarly journals Host-Dependent Requirement for the Potato leafroll virus 17-kDa Protein in Virus Movement

2002 ◽  
Vol 15 (10) ◽  
pp. 1086-1094 ◽  
Author(s):  
Lawrence Lee ◽  
Peter Palukaitis ◽  
Stewart M. Gray
Keyword(s):  
Amino Acids ◽  
Solanum Tuberosum ◽  
Nicotiana Benthamiana ◽  
Systemic Infection ◽  
Potato Leafroll Virus ◽  
Virus Movement ◽  
Wild Type ◽  
Mature Leaves ◽  
Leafroll Virus ◽  
Virus Distribution

The requirement for the 17-kDa protein (P17) of Potato leafroll virus (PLRV) in virus movement was investigated in four plant species: potato (Solanum tuberosum), Physalis floridana, Nicotiana benthamiana, and N. clevelandii. Two PLRV P17 mutants were characterized, one that does not translate the P17 and another that expresses a P17 missing the first four amino acids. The P17 mutants were able to replicate and accumulate in agroinoculated leaves of potato and P. floridana, but they were unable to move into vascular tissues and initiate a systemic infection in these plants. In contrast, the P17 mutants were able to spread systemically from inoculated leaves in both Nicotiana spp., although the efficiency of infection was reduced relative to wild-type PLRV. Examination of virus distribution in N. benthamiana plants using tissue immunoblotting techniques revealed that the wild-type PLRV and P17 mutants followed a similar movement pathway out of the inoculated leaves. Virus first moved upward to the apical tissues and then downward. The P17 mutants, however, infected fewer phloem-associated cells, were slower than wild-type PLRV in moving out of the inoculated tissue and into apical tissues, and were unable to infect any mature leaves present on the plant at the time of inoculation.

scholarly journals The Auxin-Regulated AP2/EREBP Gene PUCHI Is Required for Morphogenesis in the Early Lateral Root Primordium of Arabidopsis

2007 ◽  
Vol 19 (7) ◽  
pp. 2156-2168 ◽  
Author(s):  
Atsuko Hirota ◽  
Takehide Kato ◽  
Hidehiro Fukaki ◽  
Mitsuhiro Aida ◽  
Masao Tasaka
Keyword(s):  
Lateral Root ◽  
Lateral Root Primordium ◽  
Root Primordium

scholarly journals The ham-2 Locus, Encoding a Putative Transmembrane Protein, Is Required for Hyphal Fusion in Neurospora crassa

Genetics ◽  
2002 ◽  
Vol 160 (1) ◽  
pp. 169-180
Author(s):  
Qijun Xiang ◽  
Carolyn Rasmussen ◽  
N Louise Glass
Keyword(s):  
Neurospora Crassa ◽  
Somatic Cell ◽  
Molecular Mechanism ◽  
Cell Fusion ◽  
Deletion Mutant ◽  
Transmembrane Protein ◽  
Wild Type ◽  
Vegetative Incompatibility ◽  
Hyphal Fusion ◽  
Aerial Hyphae

Abstract Somatic cell fusion is common during organogenesis in multicellular eukaryotes, although the molecular mechanism of cell fusion is poorly understood. In filamentous fungi, somatic cell fusion occurs during vegetative growth. Filamentous fungi grow as multinucleate hyphal tubes that undergo frequent hyphal fusion (anastomosis) during colony expansion, resulting in the formation of a hyphal network. The molecular mechanism of the hyphal fusion process and the role of networked hyphae in the growth and development of these organisms are unexplored questions. We use the filamentous fungus Neurospora crassa as a model to study the molecular mechanism of hyphal fusion. In this study, we identified a deletion mutant that was restricted in its ability to undergo both self-hyphal fusion and fusion with a different individual to form a heterokaryon. This deletion mutant displayed pleiotropic defects, including shortened aerial hyphae, altered conidiation pattern, female sterility, slow growth rate, lack of hyphal fusion, and suppression of vegetative incompatibility. Complementation with a single open reading frame (ORF) within the deletion region in this mutant restored near wild-type growth rates, female fertility, aerial hyphae formation, and hyphal fusion, but not vegetative incompatibility and wild-type conidiation pattern. This ORF, which we named ham-2 (for hyphal anastomosis), encodes a putative transmembrane protein that is highly conserved, but of unknown function among eukaryotes.

scholarly journals The Valine Anticodon and Valylatability of Peanut Clump Virus RNAs Are Not Essential but Provide a Modest Competitive Advantage in Plants

2000 ◽  
Vol 74 (18) ◽  
pp. 8720-8725 ◽  
Author(s):  
Daiki Matsuda ◽  
Patrice Dunoyer ◽  
Odile Hemmer ◽  
Christiane Fritsch ◽  
Theo W. Dreher
Keyword(s):  
Sequence Analysis ◽  
Nicotiana Benthamiana ◽  
General Characteristic ◽  
Wild Type ◽  
Genomic Rnas ◽  
Viral Rnas ◽  
Fold Reduction ◽  
Direct Competition

ABSTRACT The role of valine aminoacylation of the two genomic RNAs ofPeanut clump virus (PCV) was studied by comparing the amplification in vivo of RNAs with GAC, GΔC, or CCA anticodons in the tRNA-like structure (TLS) present at the 3′ end of each viral RNA. The PCV RNA1 TLS of isolate PCV2 possesses a GAC anticodon and is capable of highly efficient valylation, whereas the RNA2 TLS has a GΔC anticodon that does not support valylation. The presence in RNA1 of GΔC or CCA anticodons that conferred nonvalylatability resulted in about 2- to 4-fold and a 14- to 24-fold reduction, respectively, in RNA accumulations in tobacco BY-2 protoplasts inoculated with the RNA1 variants together with wild-type RNA2(GΔC). No differences in RNA levels were observed among protoplasts inoculated with the three variant RNA2s in the presence of wild-type RNA1(GAC). All combinations of valylatable and nonvalylatable RNAs 1 and 2 were similarly infectious in Nicotiana benthamiana plants, and viral RNAs accumulated to similar levels; all input TLS sequences were present unchanged in apical leaves. In direct competition experiments in N. benthamiana plants, however, both RNA1 and RNA2 with GAC valylatable anticodons outcompeted the nonvalylatable variants. We conclude that valylation provides a small but significant replicational advantage to both PCV RNAs. Sequence analysis of the TLS from RNA2 of a second PCV isolate, PO2A, revealed the presence of an intact GAC valine anticodon, suggesting that the differential valylation of the genomic RNAs of isolate PCV2 is not a general characteristic of PCV.

scholarly journals THE MECHANISM OF HETEROKARYOTIC GROWTH IN VERTICILLIUM DAHLIAE

Genetics ◽  
1974 ◽  
Vol 76 (3) ◽  
pp. 411-422
Author(s):  
John E Puhalla ◽  
John E Mayfield
Keyword(s):  
High Temperature ◽  
Verticillium Dahliae ◽  
Nuclear Migration ◽  
Sharp Contrast ◽  
Wild Type ◽  
Continue Growth ◽  
Hyphal Anastomosis

ABSTRACT Heterokaryons of Verticillium dahliae, forced between complementary auxotrophs, were stable at 21° and resembled the wild type morphologically. In such heterokaryons the hyphal cells were predominantly uninucleate, and no nuclear migration from cell to cell was observed. Heterokaryosis was apparently confined to binucleate, interhyphal, anastomosed cells that arose 1-2 mm behind the colony front. Such anastomosed cells thereby fed and maintained large homokaryotic areas including the colony edge. This stable mosaic colony is in sharp contrast to the heterokaryon of Neurospora.—Heterokaryons of V. dahliae cannot continue growth at 30° because the high temperature prevents hyphal anastomosis. Heterozygous diploids sector out from heterokaryons after 8-12 days at 30°. Interhyphal anastomosed cells are apparently the site of karyogamy.

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