scholarly journals Transient Expression of Chi42 Genes from Trichoderma asperellum in Nicotiana benthamiana by Agroinfiltration

2021 ◽  
Vol 26 (01) ◽  
pp. 177-184
Author(s):  
Nguyen Quang Duc Tien

The present study reports the transient expression of chi42 genes encoding 42 kDa chitinase from T. asperellum SH16 in N. benthamiana via agroinfiltration. The efficacy of agroinfiltration for chi42 genes including a wild-type gene (Chi42) and two synthetic genes (syncodChi42-1 and syncodChi42-2) was determined. Accordingly, coinfiltration of two vectors pMYV719 carrying one of three genes chi42 and pMYV508 carrying gene p19 expedited the higher expression of recombinant enzymes whose genes were optimized for codon usage in plant tissues. The highest chitinolytic activity of about 290 U/mL was found in plants containing the gene syncodChi42-2 after 7 days of injection, 1.7 and 2.6 times higher than that of genes syncodChi42-1 and chi42. Recombinant chitinase has also exhibited activity against the pathogenic fungus Sclerotium rolfsii under in vitro condition. A higher expression level of syncodChi42-2 gene in N. benthamiana and its antifungal activity promise potential applications in the field of transgenic crops against phytopathogenic fungi. © 2021 Friends Science Publishers

2012 ◽  
Vol 7 (12) ◽  
pp. 1934578X1200701 ◽  
Author(s):  
Vivek Ahluwalia ◽  
Nandini Garg ◽  
Birendra Kumar ◽  
Suresh Walia ◽  
Om P. Sati

Vanillin oxime- N-O-alkanoates were synthesized following reaction of vanillin with hydroxylamine hydrochloride, followed by reaction of the resultant oxime with acyl chlorides. The structures of the compounds were confirmed by IR, 1H, 13C NMR and mass spectral data. The test compounds were evaluated for their in vitro antifungal activity against three phytopathogenic fungi Macrophomina phaseolina, Rhizoctonia solani and Sclerotium rolfsii by the poisoned food technique. The moderate antifungal activity of vanillin was slightly increased following its conversion to vanillin oxime, but significantly increased after conversion of the oxime to oxime- N-O-alkanoates. While vanillin oxime- N-O-dodecanoate with an EC50 value 73.1 μg/mL was most active against M. phaseolina, vanillin oxime- N-O-nonanoate with EC50 of value 66.7 μg/mL was most active against R. solani. The activity increased with increases in the acyl chain length and was maximal with an acyl chain length of nine carbons.


Author(s):  
Amanda P. Mattos ◽  
Fabricio P. Povh ◽  
Bruna B. Rissato ◽  
Vítor V. Schwan ◽  
Kátia R. F. Schwan-Estrada

Aims: This study is aimed to evaluate the in vitro antifungal activity effect of the crude aqueous extract (CAE), hydrolate (HY) and essential oil (EO) of Corymbia citriodora, Cymbopogon citratus, Cymbopogon flexuosus and Curcuma longa against the phytopathogenic fungi Alternaria steviae, Botryosphaeria dothidea, Colletotrichum gloeosporioides and Sclerotium rolfsii, and assess, in situ, the effectiveness of CAE of medicinal plants in reducing the severity of the cucumber anthracnose. Methodology: The EOs and HYs were obtained by hydrodistillation. The CAEs were prepared by the turbolysis method. Mycelial growth of the fungi was measured daily, by the diametrically opposite method. In the in vivo test, the CAEs were sprayed on the cotyledon leaves of healthy cucumber plants with three days after were inoculated with C. lagenarium. The severity of assessment of the disease was based on a scale of notes. Results: The medicinal plants studied showed antifungal activity against all or almost all pathogens. In general, treatment with CAE and HY of C. longa revealed the highest inhibition against the fungi tested. With the exception of the EO of C. longa, the other EOs showed total inhibition against all the fungi and in all the concentrations tested. Compared to control, in in vivo assays CAE of C. citratus presents a potential for control of cucumber anthracnose reducing the severity of the disease. Conclusion: The medicinal plants studied produce compounds associated with antimicrobial activity.


2021 ◽  
Vol 21 (1) ◽  
pp. 72
Author(s):  
Muhammad Hasan Basri ◽  
Lalu Zulkifli ◽  
Abdul Syukur

Plant damage by pathogenic fungi is often found in plants, one of which is caused by Sclerotium rolfsii. Biological control strategy offers a promising alternative for managing disease in plants because they are environmental friendly compared to pesticides application. One of the biological control offered is by using endophytic fungi isolated from Vitex trivolia L. The aim of the study was to isolate, to identify macroscopic and microscopic endophytic fungi from Vitex trifolia L and to test their antagonism potency against the pathogenic fungus Sclerotium rolfsii in vitro. The isolation obtained 7 endophytic fungi isolates identified based on their genus characteristics, nsmely Periconia sp, Aspergillussp, Dendrophoma  sp, Geotrichum  sp, Ampulliferina  sp, Chalara  sp, dan Bispora sp and 2 isolates have not been identified. The Antibacterial test of the fungi isolate on the 4 tested bacteria showed that of all the fungi isolate have low activity. The antagonism test using the direct opposition method with the PIRG formula, showed that the 3 isolates had high percentage of growth inhibition, in which ALJ1, BLJ5, and ALJ3 isolate has  85%, 90%, and 100% respectively. This potency could be used as biological agents on the pathogenic fungus Sclerotium rolfsii.


2014 ◽  
Vol 70 (a1) ◽  
pp. C799-C799
Author(s):  
Byron DeLaBarre ◽  
Fang Wang ◽  
Jeremy Travins ◽  
Stefan Gross ◽  
Erin Artin ◽  
...  

A number of human cancers harbor somatic point mutations in the genes encoding isocitrate dehydrogenases- 1 and -2 (IDH1, IDH2)[1]. These mutations alter residues in the enzyme active sites and confer a gain-of-function in cancer cells, resulting in the accumulation and secretion of the oncometabolite R (-)-2-hydroxyglutarate (2HG). 2HG is a potent inhibitor of DNA methylating enzymes such as TET2[2]. This suggests a connection between cancer related IDH mutations and aberrant epigenetics. As such, IDH represents an important new druggable target in the pursuit of novel cancer therapies. We have developed a small molecule, AGI-6780, that potently and selectively inhibits the tumor-associated mutant IDH2/R140Q. A crystal structure of AGI-6780 complexed with IDH2/R140Q revealed that the inhibitor binds in an allosteric manner at the dimer interface[3]. While structures of IDH1 and IDH2 were known, this is the first ever structure of an inhibited IDH protein and shows a novel conformation of IDH2. The results of steady-state enzymology analysis were consistent with allostery and slow-tight binding by AGI-6780. Treatment with AGI-6780 induced differentiation of TF-1 erythroleukemia and primary human acute myelogenous leukemia (AML) cells in vitro. These data provide proof-of- concept that inhibitors targeting mutant IDH2/R140Q could have potential applications as a differentiation therapy for cancer.


2016 ◽  
Vol 5 (07) ◽  
pp. 4714
Author(s):  
Smrity Prabha ◽  
Bapat U. C.* ◽  
Jyoti Kumar

The anti-fungal activity of ethanolic and petroleum ether extracts of eight medicinal plants, viz., Acorus calamus Linn, Justicia adhatoda L. Lawsonia Inermis L., Lantana camara Linn., Pongamia pinnata (L.) Pierre, Solanum nigrum Linn., Vitex negundo L. and Wedelia chinensis (Osbeck) Merr. were tested in vitro against phytopathogenic fungus Sclerotium rolfsii Sacc.was evaluated using poison food method. The results obtained showed that the Petroleum ether extracts of leaves and flowers of L.camara, leaves of Lawsonia inermis and S.nigrum did not inhibit the growth of S. rolfsii. However, it was observed that the ethanolic extracts of the rhizome of A. calamus and the leaves of L. Camara showed 61% and 50% inhibition of the growth of S. Rolfsii respectively. The most promising results were obtained with the petroleum ether extract of A. calamus, which exhibited 100% inhibition of S. Rolfsii with MIC of 10.4 mg/ ml. 


2012 ◽  
Vol 52 (4) ◽  
pp. 458-462 ◽  
Author(s):  
Abdul Aziz A. Al-Askar

Abstract The antifungal activities of ethanolic extracts of three Saudi plants; camel thorn (Alhagi maurorum Medic.), caper (Capparisspinosa L.), and pomegranate (Punica granatum L.) were investigated in vitro against Alternaria alternata, Fusarium oxysporum, Phomadestructiva, Rhizoctonia solani, and Sclerotium rolfsii at concentrations of 0, 3, 6, and 9% (v/v). All tested plant extracts; seeds, roots, and rinds had different degrees of antifungal activity against the tested fungi. When compared with the control, the highest antifungal activity was recorded for camel thorn seeds extract at a concentration of 9%, while, pomegranate rinds extract at 9% came in second. Camel thorn rinds extract came in last even when used at a high concentration. The ethanolic extract of camel thorn seeds may be recommended as a potent bio-fungicide. Extensive studies should be undertaken for the ethanolic extract of camel thorn seeds as a strong antifungal agent against fungal plant diseases.


2015 ◽  
Vol 3 (1) ◽  
pp. 16-19 ◽  
Author(s):  
H. J. Parmar ◽  
Mohamed M. Hassan ◽  
N. P. Bodar ◽  
V. V. Umrania ◽  
S.V. Patel ◽  
...  

Six Trichoderma strains (collected from IARI, New Delhi and MTCC, Chandigarh) were tested for their ability to inhibit soil born pathogenof groundnut mainly Sclerotium rolfsii (causing stem rot on groundnut). Morphological observations of Trichoderma strains as well asphytopathogenic fungi S. rolfsii were made from culture grown at 28 0C for about one week on PDA media. The radial growth, fastest growthand coiling of test fungus of various Trichoderma strains on plant pathogenic fungi S. rolfsii were examined under LEICA phase contrastbinocular light microscope. In vitro percent growth inhibition of S.rolfsii by various Trichoderma strain was recorded at 5 days after inoculationat 28 oC in the 90 cm petriplates. Results obtained from the antagonism study indicated that Trichoderma viride (NBAII Tv 23) inhibited 61%growth of phytopathogenic fungi S. rolfsii followed by T. harzianum (NBAII Th1) (55% growth inhibition of pathogen). This suggested thatamong different Trichoderma strains, T. viride was the best bio-control agent to inhibit in vitro growth of phytopathogen S.rolfsii whichotherwise cause stem rot disease in groundnut.DOI: http://dx.doi.org/10.3126/ijasbt.v3i1.11845  Int J Appl Sci Biotechnol, Vol. 3(1): 16-19      


2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Martha María Arevalos-Sánchez ◽  
Adrián Omar Maynez-Perez ◽  
Felipe A. Rodríguez-Almeida ◽  
José Alfredo Martínez-Quintana ◽  
Fidel Alejandro Sanchez-Flores ◽  
...  

Abstract Background The production of agricultural wastes still growing as a consequence of the population growing. However, the majority of these residues are under-utilized due their chemical composition, which is mainly composed by cellulose. Actually, the search of cellulases with high efficiency to degrade this carbohydrate remains as the challenge. In the present experiment, two genes encoding an endoglucanase (EC 3.2.1.4) and β-glucosidase (EC 3.2.1.21) were overexpressed in Escherichia coli and their recombinant enzymes (egl-FZYE and cel-FZYE, respectively) characterized. Those genes were found in Trabulsiella odontermitis which was isolated from the gut of termite Heterotermes sp. Additionally, the capability to release sugars from agricultural wastes was evaluated in both enzymes, alone and in combination. Results The results have shown that optimal pH was 6.0 and 6.5, reaching an activity of 1051.65 ± 47.78 and 607.80 ± 10.19 U/mg at 39 °C, for egl-FZYE and cel-FZYE, respectively. The Km and Vmax for egl-FZYE using CMC as substrate were 11.25 mg/mL and 3921.57 U/mg, respectively, whereas using Avicel were 15.39 mg/mL and 2314.81 U/mg, respectively. The Km and Vmax for cel-FZYE using Avicel as substrate were 11.49 mg/mL and 2105.26 U/mg, respectively, whereas using CMC the enzyme did not had activity. Both enzymes had effect on agricultural wastes, and their effect was improved when they were combined reaching an activity of 955.1 ± 116.1, 4016.8 ± 332 and 1124.2 ± 241 U/mg on corn stover, sorghum stover and pine sawdust, respectively. Conclusions Both enzymes were capable of degrading agricultural wastes, and their effectiveness was improved up to 60% of glucose released when combined. In summary, the results of the study demonstrate that the recombinant enzymes exhibit characteristics that indicate their value as potential feed additives and that the enzymes could be used to enhance the degradation of cellulose in the poor-quality forage generally used in ruminant feedstuffs.


2020 ◽  
Vol 7 (03) ◽  
Author(s):  
PREM PANDEY ◽  
G. C. SAGAR ◽  
SUNDARMAN SHRESTHA2 ◽  
HIRAKAJI MANANDHAR ◽  
RITESH K. YADAV ◽  
...  

Nine isolates of Trichoderma spp. were isolated from different agro- ecological regions of Nepal viz; Jumla, Palpa, Chitwan, Tarahara, Banke, Illam and Salyan and screened against Sclerotium rolfsii Sacc. Adreded soil borne phytopathogen causing collar rot of chickpea in chickpea; In-vitro efficacy of nine fungal antagonist (Trichoderma spp.) against Sclerotium rolfsii were screened. Pot experiment was done to find out the effective management of S. rolfsi through Tricoderma using different methods i.e. Seed treatment, soil drenching and soil application. All the tested isolates of Trichoderma spp. were found effective on mycelial growth inhibition and sclerotial parasitization of S. rolfsii. Trichoderma isolated from Palpa district showed maximum growth inhibition (%) of pathogen periodically after 48(93.78%), 72(96.00%), 96(97.96%) and 120(100.00%) hours of inoculation. Parasitized sclerotium showed minimum sclerotial germination on agar plates. Moreover, Trichoderma species isolated from Palpa districts showed second best percent mycelial growth inhibition periodically at 72(25.00%), 120(29.16%), 168(29.16%) and 216(29.16%).In pot experiment at 40 days after sowing, Seedling height was maximum in soil drenching with 30g per 100ml of water (22.27cm) and Mortality percentage of seedlings was least or highest disease control was observed in seed treated with 109cfu/ml (0.000%).


2009 ◽  
Vol 29 (01) ◽  
pp. 17-20 ◽  
Author(s):  
I. Marx ◽  
I. Badirou ◽  
R. Pendu ◽  
O. Christophe ◽  
C. V. Denis

SummaryVon Willebrand factor (VWF) structure-function relationship has been studied only through in vitro approaches. The VWF-deficient mouse model has been extremely useful to examine the in vivo function of VWF but does not allow a more subtle analysis of the relative importance of its different domains. However, considering the large size of VWF and its capacity to interact with various ligands in order to support platelet adhesion and aggregation, the necessity to evaluate independently these interactions appeared increasingly crucial. A recently developed technique, known as hydrodynamic injection, which allows transient expression of a transgene by mouse hepatocytes, proved very useful in this regard. Indeed, transient expression of various VWF mutants in VWF-deficient mice contributed to improve our knowledge about the role of VWF interaction with subendothelial collagens and with platelets receptors in VWF roles in haemostasis and thrombosis. These findings can provide new leads in the development of anti-thrombotic therapies.


Sign in / Sign up

Export Citation Format

Share Document