scholarly journals The Conifer Root and Stem Rot Pathogen (Heterobasidion parviporum): Effectome Analysis and Roles in Interspecific Fungal Interactions

2019 ◽  
Vol 7 (12) ◽  
pp. 658
Author(s):  
Zilan Wen ◽  
Zhen Zeng ◽  
Fei Ren ◽  
Fred O. Asiegbu
Keyword(s):  
Expression Patterns ◽  
Physical Contact ◽  
Secreted Proteins ◽  
Stem Rot ◽  
Dual Culture ◽  
In Planta ◽  
Zone Formation ◽  
Fungal Interactions ◽  
Small Secreted Proteins

Heterobasidion parviporum Niemelä & Korhonen is an economically important basidiomycete, causing root and stem rot disease of Norway spruce (Picea abies (L.) Karst) in Northern Europe. The H. parviporum genome encodes numerous small secreted proteins, which might be of importance for interacting with mycorrhiza symbionts, endophytes, and other saprotrophs. We hypothesized that small secreted proteins from H. parviporum (HpSSPs) are involved in interspecific fungal interaction. To identify HpSSP-coding genes potentially involved, we screened the H. parviporum effectome and compared their transcriptomic profiles during fungal development and in planta tree infection. We further conducted phylogenetic analysis, and identified a subset of hypothetical proteins with nonpredicted domain or unknown function as HpSSPs candidates for further characterization. The HpSSPs candidates were selected based on high-quality sequence, cysteine residue frequency, protein size, and in planta expression. We subsequently explored their roles during in vitro interaction in paired cultures of H. parviporum with ectomycorrhizal Cortinarius gentilis, endophytic Phialocephala sphaeroides, saprotrophs (Mycena sp., Phlebiopsis gigantea, and Phanerochaete chrysosporium), respectively. The transcriptomic profile revealed that a large proportion of effector candidates was either barely expressed or highly expressed under all growth conditions. In vitro dual-culture test showed that P. sphaeroides and C. gentilis were overgrown by H. parviporum. The barrage zone formation or no physical contact observed in paired cultures with the saprotrophs suggest they had either combative interaction or antibiosis effect with H. parviporum. Several HpSSPs individuals were up- or downregulated during the nonself interactions. The results of HpSSPs gene expression patterns provide additional insights into the diverse roles of SSPs in tree infection and interspecific fungal interactions.

scholarly journals The Yeast Signal Sequence Trap Identifies Secreted Proteins of the Hemibiotrophic Corn Pathogen Colletotrichum graminicola

2008 ◽  
Vol 21 (10) ◽  
pp. 1325-1336 ◽  
Author(s):  
Jorrit-Jan Krijger ◽  
Ralf Horbach ◽  
Michael Behr ◽  
Patrick Schweizer ◽  
Holger B. Deising ◽  
...  
Keyword(s):  
Cell Walls ◽  
Leaf Extract ◽  
Signal Sequence ◽  
Stem Rot ◽  
Colletotrichum Graminicola ◽  
In Planta ◽  
Chain Reaction ◽  
Genes Encoding ◽  
Polymerase Chain

The hemibiotroph Colletotrichum graminicola is the causal agent of stem rot and leaf anthracnose on Zea mays. Following penetration of epidermal cells, the fungus enters a short biotrophic phase, followed by a destructive necrotrophic phase of pathogenesis. During both phases, secreted fungal proteins are supposed to determine progress and success of the infection. To identify genes encoding such proteins, we constructed a yeast signal sequence trap (YSST) cDNA-library from RNA extracted from mycelium grown in vitro on corn cell walls and leaf extract. Of the 103 identified unigenes, 50 showed significant similarities to genes with a reported function, 25 sequences were similar to genes without a known function, and 28 sequences showed no similarity to entries in the databases. Macroarray hybridization and quantitative reverse-transcriptase polymerase chain reaction confirmed that most genes identified by the YSST screen are expressed in planta. Other than some genes that were constantly expressed, a larger set showed peaks of transcript abundances at specific phases of pathogenesis. Another set exhibited biphasic expression with peaks at the biotrophic and necrotrophic phase. Transcript analyses of in vitro-grown cultures revealed that several of the genes identified by the YSST screen were induced by the addition of corn leaf components, indicating that host-derived factors may have mimicked the host milieu.

scholarly journals Evaluation of in-vitro methods to select effective streptomycetes against toxigenic fusaria

PeerJ ◽  
2019 ◽  
Vol 7 ◽  
pp. e6905 ◽  
Author(s):  
Elena Maria Colombo ◽  
Cristina Pizzatti ◽  
Andrea Kunova ◽  
Claudio Gardana ◽  
Marco Saracchi ◽  
...  
Keyword(s):  
Plant Pathogens ◽  
Culture Media ◽  
Screening Methods ◽  
Dual Culture ◽  
In Planta ◽  
In Vitro Method ◽  
In Vitro Methods ◽  
Strain Diversity ◽  
Fungal Plant Pathogens

Biocontrol microorganisms are emerging as an effective alternative to pesticides. Ideally, biocontrol agents (BCAs) for the control of fungal plant pathogens should be selected by an in vitro method that is high-throughput and is predictive of in planta efficacy, possibly considering environmental factors, and the natural diversity of the pathogen. The purpose of our study was (1) to assess the effects ofFusariumstrain diversity (N= 5) and culture media (N= 6) on the identification of biological control activity ofStreptomycesstrains (N= 20) againstFusariumpathogens of wheat in vitro and (2) to verify the ability of our in vitro screening methods to simulate the activity in planta. Our results indicate that culture media,Fusariumstrain diversity, and their interactions affect the results of an in vitro selection by dual culture assay. The results obtained on the wheat-based culture media resulted in the highest correlation score (r= 0.5) with the in planta root rot (RR) inhibition, suggesting that this in vitro method was the best predictor of in planta performance of streptomycetes against Fusarium RR of wheat assessed as extension of the necrosis on the root. Contrarily, none of the in vitro plate assays using the media tested could appropriately predict the activity of the streptomycetes against Fusarium foot rot symptoms estimated as the necrosis at the crown level. Considering overall data of correlation, the activity in planta cannot be effectively predicted by dual culture plate studies, therefore improved in vitro methods are needed to better mimic the activity of biocontrol strains in natural conditions. This work contributes to setting up laboratory standards for preliminary screening assays ofStreptomycesBCAs against fungal pathogens.

scholarly journals Inhibitory Effect of Trichoderma asperellum Isolate against Ralstonia solanacearum Causing Brinjal Wilt

2021 ◽  
Vol 24 (2) ◽  
pp. 107-120
Author(s):  
SMN Islam ◽  
SS Siddique ◽  
MZH Chowdhury ◽  
NJ Mishu
Keyword(s):  
Ralstonia Solanacearum ◽  
Disease Incidence ◽  
Its Region ◽  
Inhibitory Effect ◽  
Dual Culture ◽  
Trichoderma Asperellum ◽  
In Planta ◽  
Quantitative Test ◽  
Trichoderma Isolate

A native Trichoderma isolate was collected from the agricultural soil of Gazipur. This isolate was identified as a Trichoderma asperellum through morphology and analysis of internal transcribed spacer (ITS) region of ribosomal RNA gene sequence and reconstruction of the phylogenetic tree. The antagonistic effects of the newly identified T. asperellum isolate were assessed against brinjal bacterial wilt caused by Ralstonia solanacearum both in vitro and in planta. Both qualitative and quantitative bioassays were conducted in vitro. For qualitative tests, dual culture and antibacterial activity were carried out, and pathogen growth was observed visually. The antagonism of T. asperellum cell free culture filtrate on the growth of R. solanacearum was conducted in a quantitative test. Successful antagonism was recorded after both in vitro qualitative tests. In addition, the lowest colony forming unit was recorded in 100% of CFC (2.4±0.51 ×103 cfu/ml) in quantitative test. The T. asperellum inoculated plant showed low disease incidence (13.33%) when seedlings were challenged with R. solanacearum in planta experiment. Disease incidence was 100% for seedlings when treated with only R. solanacearum. The results showed that the isolated and identified T. asperellum isolate suppressed R. solanacearum growth in vitro and protected the seedling from wilting in planta. Therefore, this isolate could be considered as a potential isolate. Ann. Bangladesh Agric. (2020) 24(2) : 107-120

scholarly journals MEKANISME PARASITISME TRICHODERMA HARZIANUM TERHADAP FUSARIUM OXYSPORUM PADA SEMAI ACACIA MANGIUM

2015 ◽  
Vol 15 (1) ◽  
pp. 72
Author(s):  
Susanti Tasik ◽  
Siti Muslimah Widyastuti ◽  
Harjono .
Keyword(s):  
Fusarium Oxysporum ◽  
Trichoderma Harzianum ◽  
Acacia Mangium ◽  
Culture Method ◽  
Inhibition Mechanism ◽  
Dual Culture ◽  
In Planta ◽  
Know How ◽  
Soil Borne Fungi

Mechanism of parasitism of Trichoderma harzianum on Fusarium oxysporum on Acacia mangium seedlings. Fusarium oxysporum is one of the most important soil-borne fungi the causal agent of damping-off disease. Detailed information it needed to know how the pathogen can be inhibited by Trichoderma harzianum. The objective of this research was to investigate the inhibition mechanism of T. harzianum on F. oxysporum in vitro and in planta. Green Flourescent Protein (GFP) T. harzianum was used as biocontrol agent of F. oxysporum. An in vitro inhibition test of T. harzianum was performed using dual culture method. In the in planta inhibition tests, seedlings of A. mangium were applied with GFP T. harzianum two days before inoculation of F. oxysporum; GFP T. harzianum was simultaneously applied with F. oxysporum and GFP T. harzianum was applied two days after inoculation of F. oxysporum. The inhibition effect of T. harzianum GFP was observed at seven days incubation, indicated by attachment of T. harzianum to F. oxysporum hyphae. GFP T. harzianum hyphae covered the colonies of F. oxysporum at 12 days after incubation. The highest life percentage of A. mangium seedlings was found on the treatment of GFP T. harzianum two days before inoculation of F. oxysporum (82.22%), whereas the lowest life percentage was found on seedling applied with GFP T. harzianum two days after inoculation of F. oxysporum (64.44%).

scholarly journals Novel Antifungal Activity of Lolium-Associated Epichloë Endophytes

2020 ◽  
Vol 8 (6) ◽  
pp. 955 ◽  
Author(s):  
Krishni Fernando ◽  
Priyanka Reddy ◽  
Inoka K. Hettiarachchige ◽  
German C. Spangenberg ◽  
Simone J. Rochfort ◽  
...  
Keyword(s):  
Antifungal Activity ◽  
Insect Pests ◽  
Fungal Endophytes ◽  
Dual Culture ◽  
The Novel ◽  
In Planta ◽  
Pasture Production ◽  
Fusarium Sp ◽  
Invertebrate Pests

Asexual Epichloë spp. fungal endophytes have been extensively studied for their functional secondary metabolite production. Historically, research mostly focused on understanding toxicity of endophyte-derived compounds on grazing livestock. However, endophyte-derived compounds also provide protection against invertebrate pests, disease, and other environmental stresses, which is important for ensuring yield and persistence of pastures. A preliminary screen of 30 strains using an in vitro dual culture bioassay identified 18 endophyte strains with antifungal activity. The novel strains NEA12, NEA21, and NEA23 were selected for further investigation as they are also known to produce alkaloids associated with protection against insect pests. Antifungal activity of selected endophyte strains was confirmed against three grass pathogens, Ceratobasidium sp., Dreschlera sp., and Fusarium sp., using independent isolates in an in vitro bioassay. NEA21 and NEA23 showed potent activity against Ceratobasidium sp. and NEA12 showed moderate inhibition against all three pathogens. Crude extracts from liquid cultures of NEA12 and NEA23 also inhibited growth of the phytopathogens Ceratobasidium sp. and Fusarium sp. and provided evidence that the compounds of interest are stable, constitutively expressed, and secreted. Comparative analysis of the in vitro and in planta metabolome of NEA12 and NEA23 using LCMS profile data revealed individual metabolites unique to each strain that are present in vitro and in planta. These compounds are the best candidates for the differential bioactivity observed for each strain. Novel endophyte strains show promise for endophyte-mediated control of phytopathogens impacting Lolium spp. pasture production and animal welfare.

scholarly journals Penapisan isolat rizobakteri indigenos untuk pengendalian (Ganoderma boninense) di pre nursery kelapa sawit (Elaeis guineensis Jacq.)

Jurnal Agro ◽  
10.15575/4665 ◽  
2019 ◽  
Vol 6 (2) ◽  
pp. 110-122
Author(s):  
Yulmira Yanti ◽  
Imam Rifai ◽  
Yogie Aditya Pratama ◽  
Muhammad Ihsan Harahap
Keyword(s):  
Plant Growth ◽  
Oil Palm ◽  
Control Plant ◽  
Stem Rot ◽  
In Planta ◽  
Basal Stem Rot ◽  
Plant Growth Promoting ◽  
Growth Promoting ◽  
And Control

Rizobakteri merupakan kelompok bakteri yang aktif mengkolonisasi akar tanaman, meningkatkan pertumbuhan dan mengendalikan patogen tanaman. Penelitian ini bertujuan untuk memperoleh isolat rizobakteri indigenous  terbaik dalam meningkatkan pertumbuhan kelapa sawit dan mengendalikan penyakit busuk pangkal batang di pre-nursery kelapa sawit secara in planta serta karakterisasi kemampuan antagonisnya secara in vitro. Penelitian bersifat eksperimental terdiri atas 3 tahap dengan menggunakan Rancangan Acak Lengkap (RAL): (1) Isolasi dan karakterisasi isolat rizobakteri indigenous  di Kabupaten Pasaman Barat, (2) Pengujian isolat rizobakteri indigenous  (RBI) sebagai plant growth promoting rihzobacteria (PGPR), dan untuk pengendalian G.boninense di pre-nursery kelapa sawit terdiri dari 29 perlakuan (27 isolat RBI, tanpa inokulasi G. boninense sebagai kontrol positif, dan inokulasi G. boninense sebagai kontrol negatif) dengan masing-masing 5 ulangan, serta (3) Pengujian aktivitas antagonisme isolat RBI terhadap G. boninense. Data dianalisis dengan sidik ragam, apabila berbeda nyata dilanjutkan dengan uji Least Significance Different (LSD) pada taraf 5%. Hasil penelitian menunjukkan diperoleh tiga isolat terbaik (R10 2.2, R9 2.1, dan R10 2.3) yang mampu meningkatkan pertumbuhan kelapa sawit dan menekan perkembangan penyakit busuk pangkal batang G.boninense secara in planta dan in vitro.ABSTRACTRhizobacteria is a group of bacteria that actively colonize plant roots, increase growth and control plant pathogen. The objective of the research was to obtain indigenous rhizobacteria isolate (RBI) to increase growth and control basal stem rot on oil palm seedlings in in planta and characterize of antagonistic ability in in vitro. Experimental research consisted of 3 stages by using Completely Randomized Design (CRD): (1) Isolation of indigenous rhizobacteria in West Pasaman region, (2) Indigenous rhizobacteria isolate testing as a plant growth promoting rhizobacteria (PGPR) and to control of G. boninense on pre nursery of oil palm consisted of 29 treatments (27 RBI isolates, without G. boninense inoculation as positive control, and G. boninense inoculation as negative control) with 5 replications each. (3) Testing of RBI isolate antagonism activity towards G. boninense. Data were analyzed by variance, if the result significantly different, it was continued by using Least Significance Different (LSD) at 5% level. The results showed that best three isolates (R10 2.2, R9 2.1 and R10 2.3) were able to increase growth of palm oil and to suppress the development of G.boninense basal stem rot in in planta and in in vitro.

scholarly journals In vitro evaluation of grapevine endophytes, epiphytes and sap micro-organisms for potential use to control grapevine trunk disease pathogens

2021 ◽  
Author(s):  
Robert Blundell ◽  
Molly Arreguin ◽  
Akif Eskalen
Keyword(s):  
Biological Control ◽  
Mycelial Growth ◽  
Fungal Pathogens ◽  
Dual Culture ◽  
Biological Control Agents ◽  
In Planta ◽  
Eutypa Dieback ◽  
Fungal Isolates ◽  
Wound Protection

SummaryGrapevine trunk diseases (GTDs) threaten the economic sustainability of viticulture worldwide causing a significant reduction of both yields and quality of grapes. Biological control presents a promising sustainable alternative to cultural and chemical methods to mitigate the effects of pathogens causing GTDs, including Botryosphaeria dieback, Eutypa dieback and Esca. This study aimed to identify naturally occurring potential biological control agents from a variety of grapevine tissues, including sap, cane and pith and evaluate their antagonistic activity against selected fungal pathogens responsible for GTDs in vitro. Bacterial and fungal isolates were preliminary screened in vitro to determine their antifungal activity via a dual culture assay against Neofusicoccum parvum and Eutypa lata. Among the fungal isolates, Trichoderma spp. inhibited E. lata mycelial growth up to 64% and N. parvum mycelial growth up to 73% with overgrowth and stopped growth being the likely antagonistic mechanisms. Among the bacterial isolates, Bacillus spp. inhibited E. lata mycelial growth up to 20% and N. parvum mycelial growth up to 40%. Select antagonistic isolates of Trichoderma, Bacillus and Aureobasidium spp. were subject to further dual culture antifungal analysis against Diplodia seriata and Diaporthe ampelina, with Trichoderma isolates consistently causing the greatest inhibition. Volatile organic compound antifungal analysis revealed that these Trichoderma isolates resulted significantly inhibited mycelial growth of N. parvum, E. lata and D. ampelina causing up to 20.11%, 60.55% and 70.9% inhibition respectively (P≤0.05). Multilocus sequence analysis revealed that the Trichoderma isolates are most closely related to Trichoderma asperellum and Trichoderma hamatum. This study identifies grapevine sap as a novel source of potential biological control agents for control of GTDs to support existing efforts to control GTDs. Further testing will be necessary to fully characterize these microbes mode of antagonism and assess their efficacy for pruning wound protection in planta.

scholarly journals Trichoderma dan Gliocladium untuk Mengendalikan Penyakit Busuk Akar Fusarium pada Bibit Kelapa Sawit

2019 ◽  
Vol 14 (6) ◽  
pp. 196
Author(s):  
Siti Juariyah ◽  
Efi Toding Tondok ◽  
Meity Suradji Sinaga
Keyword(s):  
Oil Palm ◽  
Biocontrol Agents ◽  
Crown Rot ◽  
Dual Culture ◽  
In Vitro Test ◽  
In Planta ◽  
Fusarium Spp ◽  
Vitro Test ◽  
Rot Disease

Trichoderma and Gliocladium for Controling Fusarium Root Rot Disease of Oil Palm SeedlingsFusarium spp. have been reported as the causal agent of common spear rot and crown rot diseases on oil palm.  An effective strategy to control these diseases is not available yet. This research was aimed to find biocontrol agents for effective control of crown rot disease on oil palm seedlings caused by Fusarium spp. The experiment consisted of 3 parts i.e. pathogenicity test of 3 isolates of Fusarium, identification and in vitro test of biocontrol agents, and in planta test of biocontrol agents against Fusarium spp. In vitro test was done through dual culture test and test for volatile compound produced by the biocontrol agents. In planta test was conducted through inoculation of Fusarium spp. into oil palm seedlings growing on medium containing  selected biocontrol agents i.e. Trichoderma harzianum, T. virens, T. inhamatum, and Gliocladium fimbriatum. In vitro test showed that Gliocladium fimbriatum 1 and 2 were inhibited effectively the growth of Fusarium spp. on the dual culture test, whereas T. harzianum Gadingrejo 2 was inhibited effectively the growth of Fusarium spp. on volatile compound test. The application of biocontrol agents was effective to protect oil palm seedlings from Fusarium spp. infection.

scholarly journals Characterization of In Planta—Induced Rust Genes Isolated from a Haustorium-Specific cDNA Library

1997 ◽  
Vol 10 (4) ◽  
pp. 427-437 ◽  
Author(s):  
Matthias Hahn ◽  
Kurt Mendgen
Keyword(s):  
Cdna Library ◽  
Expression Patterns ◽  
Host Cells ◽  
In Planta ◽  
Developmentally Regulated ◽  
Uromyces Fabae ◽  
Infection Structures ◽  
Genes Encoding ◽  
Peptidyl Prolyl Isomerases

Rust fungi are plant parasites that depend on living host tissue for growth. For invasion of leaves, dikaryotic urediospores differentiate germ tubes and infection structures that penetrate through stomata. Biotrophic growth occurs by intercellular mycelia that form haustoria within host cells. A cDNA library was constructed from haustoria isolated from broad bean leaves infected by Uromyces fabae. Differential screening revealed that a high proportion (19%) of the haustorial cDNAs are specifically expressed in planta but are not expressed, or are much weaker, in germlings or infection structures produced in vitro. A total of 31 different in planta-induced genes (PIGs) were identified. Some of the PIGs are highly expressed in haustoria. The PIGs are single or low copy number genes in the rust genome. A variety of developmentally regulated expression patterns of PIG mRNAs were observed. Sequence analysis of PIG cDNAs revealed similarities to genes encoding proteins involved in amino acid transport, thiamine biosynthesis, short-chain dehy-drogenases, metallothioneins, cytochrome P-450 monooxy-genases, and peptidyl-prolyl isomerases.

scholarly journals Endophytic bacteria isolated from Leptospermum scoparium produce compounds that inhibit Ilyonectria and Neofusicoccum species in vitro

2015 ◽  
Vol 68 ◽  
pp. 445-445
Author(s):  
W.A. Wicaksono ◽  
E.E. Jones ◽  
J. Monk ◽  
H.J. Ridgway
Keyword(s):  
New Zealand ◽  
Essential Oil ◽  
International Research ◽  
Endophytic Bacteria ◽  
Antimicrobial Properties ◽  
Dual Culture ◽  
Antimicrobial Compounds ◽  
In Planta ◽  
Leptospermum Scoparium

Leptospermum scoparium JRForst et GForst var scoparium or m257;nuka is a New Zealand indigenous shrub This shrub is valued for the essential oil and medicinal honey it produces that have antimicrobial properties International research has demonstrated that endophytic bacteria can either directly produce or modify metabolites in planta Therefore bacteria within the m257;nuka endomicrobiome may also produce antimicrobial compounds A total of 192 endophytic bacteria were recovered from surface sterilised leaf stem and root tissue from three different sites These bacteria were assessed in dual culture assays against the grapevine pathogens Ilyonectria spp and Neofusicoccum spp Eleven and three endophytic bacteria showed ability to inhibit I liriodendri and N luteum respectively These endophytic bacteria produced both diffusible and volatile compounds that inhibited the pathogens Isolate W4R11 inhibited the growth of Ilyonectria spp by 4164 after 7 days incubation Isolate W1R33 could inhibit the faster growing Neofusicoccum spp by 2053 after 3 days incubation Microscopy showed that diffusible compound produced by isolate I1R21 caused hyphae and spore abnormality of Ilyonectria spp This study indicated that m257;nuka can potentially provide a new source of microorganisms for use in sustainable agriculture

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