scholarly journals In-vitro Screening for Disease Resistance in Wheat Genotypes against Bipolaris sorokiniana Using Callus Culture Method

2020 ◽  
pp. 82-87
Author(s):  
. Deepti ◽  
Swati Rani ◽  
Kumari Anjani ◽  
Rajiv Kumar ◽  
Vinay Kumar Sharma
Keyword(s):  
Callus Culture ◽  
Culture Method ◽  
Bipolaris Sorokiniana ◽  
In Vitro Screening ◽  
Susceptible Genotype ◽  
Resistant Genotype ◽  
Maximum Area ◽  
Wheat Genotypes ◽  
Least Area

The present investigation was done to identify the efficacy of callus culture method for in-vitro screening to identify resistant and susceptible wheat genotypes. The study led to establishment of protocol for in-vitro screening of susceptible and resistant wheat genotypes against Bipolaris sorokiniana. The Bipolaris sorokiniana crude toxin was used at various concentrations to supplement the callusing medium and the response of twelve genotypes was studied. The susceptible genotype Agra local showed maximum area prone to death of the callus because of effect of toxin in medium supplemented with MS+2,4-D (4.0 mgl-1) + NAA (2 mgl-1) with B. sorokiniana toxin whereas the resistant genotype Yangmai#6 showed least area affected by the toxin and the cream/ white callus observed in the case of controlled medium supplemented with toxin. The genotypes which were found to be resistant and susceptible in the field condition were clearly identified as the same using this method.

scholarly journals In Vitro Screening of Falcataria moluccana (Miq.) with Gall Rust (Uromycladium tepperianum (Sacc.) Filtrate as Media Selection

2016 ◽  
Vol 19 (2) ◽  
pp. 111 ◽  
Author(s):  
Asri Insiana Putri ◽  
Mohammad Na'iem ◽  
Sapto Indrioko ◽  
Sri Rahayu ◽  
Ari Indrianto
Keyword(s):  
Phenolic Compound ◽  
Total Phenolics ◽  
Culture Method ◽  
Dry Weight ◽  
Total Phenolic ◽  
Media Selection ◽  
In Vitro Screening ◽  
Tissue Culture Method ◽  
The Mean

In vitro screening of Falcataria moluccana (Miq.) was conducted by tissue culture method. Seeds fromtwo different site of community forest, 400 m (S1) and 800 m (S2) above sea level, were used as material.Double concentration of MS (Murashige & Skoog, 1962) with 40 mg/l gall rust (Uromycladium tepperianum(Sacc.) fi ltrate were used for media selection. The results of this research showed that 66 % axenic plantlets invitro from S1 and 27 % from S2 were still survived after 3 months incubation without subculture. The meanof fresh weight (2. 21 ± 0. 26 g) and dry weight (1. 97 ± 0. 12 g) from S1 plantlets lower than the mean of freshweight (2. 87 ± 0,18 g) and dry weight (2. 16 ± 0. 14 g) from S2 plantlets. Qualitative of terpenes, saponins andquantitative of total phenolics were analyzed from those gall rust extract, as source of fi ltrate media, attackedand un-attacked of F. moluccana. They all qualitatively have capability to produce terpenoid and saponin. Itis notice that U. tepperianum, un-cultured pathogen, contain of those compound that may play a role as codeterminantsof pathogenecity. While the highest total phenolic compound were contained in gall rust extract(2. 35 %), followed by attacked F. moluccana branches (1. 18 %) and un-attacked F. moluccana branches (0. 44%). This indicated that phenolic compound in gall rust has higher activity as a response of F. moluccana to U.tepperianum pathogen pressures and result of this study suggest the great value of gall rust fi ltrate for use asmedia selection in vitro.

scholarly journals Whole Genome Sequencing and Expression Analysis of ToxA in Bipolaris Sorokiniana Provides Discernment of Pathogenicity Causing Spot Blotch of Wheat

2021 ◽  
Author(s):  
Rashmi Aggarwal ◽  
Shweta Agarwal ◽  
Sapna Sharma ◽  
Malkhan Singh Gurjar ◽  
Bishnu Maya Bashyal ◽  
...  
Keyword(s):  
Expression Analysis ◽  
Gc Content ◽  
Bipolaris Sorokiniana ◽  
Spot Blotch ◽  
Secretory Proteins ◽  
Whole Genome ◽  
In Planta ◽  
Wheat Genotypes ◽  
Average Gene Density

Abstract Background: Spot blotch disease of wheat caused by Bipolaris sorokiniana Boerma (Sacc.) is an emerging problem in South Asian countries. In this study, whole genome of highly virulent isolate of Bipolaris sorokiniana (BS112) was sequenced, pathogenicity related gene(s) were identified and role of ToxA gene in spot blotch disease development was established.Results: Bipolaris sorokiniana isolate BS112 infecting wheat was sequenced using hybrid assembly approach. The assembly size of the genome was 35.64Mb (GenBank accession number RCTM00000000) with GC content of 50.2%, providing coverage of 97.6% on reference ND90Pr genome. Average gene density predicted was 250-300 genes/Mb. A total of 235 scaffolds were obtained using pyScaf assembler with N50 of 16,54,800 bp. In addition, 152 transcription factors involved in various biological processes were identified and a total of 682 secretory proteins were predicted using secretome analysis. ToxA gene (535bp) was analyzed and identified in the genome of B. sorokiniana which revealed 100% homology with ToxA gene of Pyrenophora tritici repentis. Further, ToxA gene was amplified, sequenced and validated in the 39 isolates of B. sorokiniana which confirmed the presence of ToxA gene in all the isolates of B. sorokiniana. All these ToxA sequences were submitted in NCBI database (MN601358-MN601396). As ToxA gene interacts with Tsn1 gene of host, 13 wheat genotypes were evaluated for the Tsn1 gene and five genotypes (38.4%) were found to be Tsn1 positive with more severe necrotic lesions compared to Tsn1 negative wheat genotypes. In vitro expression analysis of ToxA gene in B. sorokiniana isolate (BS112) using qPCR revealed maximum upregulation (14.67 fold) at 1st day after inoculation (DAI). Further, in planta expression analysis of ToxA gene in Tsn1 positive and Tsn1 negative genotypes, Agra local and Chiriya 7 respectively was also conducted. Results revealed maximum expression (7.89 fold) of ToxA gene in Tsn1 positive genotype, Agra local at 5th DAI compared to Tsn1 negative genotype Chiriya 7 which showed minimum expression (0.048 fold) at 5th DAI. Conclusions: Full genome of B. sorokiniana was sequenced; secreted proteins and virulence genes were identified in the genome. ToxA gene was validated in thirty nine isolates of B. sorokiniana. In planta ToxA-Tsn1 interaction studies established that spot blotch disease is more severe in Tsn1 positive genotypes. This genomic resource will provide a new insight into better understanding and management of spot blotch disease and B. sorokiniana of wheat.

In Vitro Screening of Azalea for Resistance to Azalea Lace Bug

HortScience ◽  
1998 ◽  
Vol 33 (3) ◽  
pp. 506b-506
Author(s):  
Carol D. Robacker ◽  
S.K. Braman
Keyword(s):  
Leaf Damage ◽  
In Vitro Screening ◽  
Screening Assay ◽  
Delaware Valley ◽  
In Vitro Techniques ◽  
Culture Tube ◽  
Lace Bug ◽  
Lace Bugs ◽  
Laboratory Bioassays

Azalea lace bug (Stephanitis pyrioides) is the most serious pest on azalea. Results of laboratory bioassays and field evaluations of 17 deciduous azalea taxa have identified three resistant taxa: R. canescens, R. periclymenoides, and R. prunifolium. Highly susceptible taxa are `Buttercup', `My Mary', R. oblongifolium, and the evergreen cultivar `Delaware Valley White'. To determine whether in vitro techniques would have potential value in screening or selecting for resistance, or for the identification of morphological or chemical factors related to resistance, an in-vitro screening assay was developed. In-vitro shoot proliferation was obtained using the medium and procedures of Economou and Read (1984). Shoots used in the bioassays were grown in culture tubes. Two assays were developed: one for nymphs and one for adult lace bugs. To assay for resistance to nymphs, `Delaware Valley White' leaves containing lace bug eggs were disinfested with 70% alcohol and 20% commercial bleach, and incubated in sterile petri plates with moistened filter paper until the nymphs hatched. Five nymphs were placed in each culture tube, and cultures were incubated for about 2 weeks, or until adults were observed. To assay for resistance to adults, five female lace bugs were placed in each culture tube and allowed to feed for 5 days. Data collected on survival and leaf damage was generally supportive of laboratory bioassays and field results. Adult lace bugs had a low rate of survival on resistant taxa. Survival of nymphs was somewhat reduced on resistant taxa.

Insecticidal Active Rotenoids from Plant Parts and Callus Culture of Medicago sativa L. from a Semiarid Region of India (Rajasthan)

2020 ◽  
Vol 16 (6) ◽  
pp. 937-941
Author(s):  
Sharad Vats ◽  
Preeti Mehra
Keyword(s):  
Medicago Sativa ◽  
Callus Culture ◽  
Point Of View ◽  
Semiarid Region ◽  
Disease Vectors ◽  
Vector Borne Diseases ◽  
Plant Parts ◽  
Resistant Varieties ◽  
Medicago Sativa L

Background: Vector-borne diseases are quite prevalent globally and are one of the major causes of deaths due to infectious diseases. There is an availability of synthetic insecticides, however, their excessive and indiscriminate use have resulted in the emergence of resistant varieties of insects. Thus, a search for novel biopesticide has become inevitable. Methods: Rotenoids were isolated and identified from different parts of Medicago sativa L. This group of metabolites was also identified in the callus culture, and the rotenoid content was monitored during subculturing for a period of 10 months. Enhancement of the rotenoid content was evaluated by feeding precursors in a tissue culture medium. Results: Four rotenoids (elliptone, deguelin, rotenone and Dehydrorotenone) were identified, which were confirmed using spectral and chromatographic techniques. The maximum rotenoid content was found in the seeds (0.33±0.01%), followed by roots (0.31±0.01%) and minimum in the aerial parts (0.20±0.05%). A gradual decrease in the rotenoid content was observed with the ageing of subcultured tissue maintained for 10 months. The production of rotenoids was enhanced up to 2 folds in the callus culture using amino acids, Phenylalanine and Methionine as precursors as compared to the control. The LC50 value of the rotenoids was found to be 91 ppm and 162 ppm against disease vectors of malaria and Dracunculiasis, respectively. Conclusion: The study projects M. sativa as a novel source of biopesticide against the disease vectors of malaria and Dracunculiasis. The use of precursors to enhance the rotenoid content in vitro can be an effective venture from a commercial point of view.

Comparative Investigation on Formation and Accumulation of Rare Phenylpropanoids in Plants and in vitro Cultures of Pimpinella major

1990 ◽  
Vol 45 (6) ◽  
pp. 602-606 ◽  
Author(s):  
B. Merkel ◽  
J. Reichling
Keyword(s):  
Callus Culture ◽  
Nutrient Medium ◽  
Callus Cultures ◽  
Comparative Investigation ◽  
In Vitro Cultures ◽  
Liquid Nutrient Medium ◽  
Whole Plants ◽  
Plant Seedlings

Abstract Unorganized callus and leaf/root-differentiating callus cultures of Pimpinella major have been established in liquid nutrient medium. Their capacity to accumulate rare phenylpropanoids such as epoxy-pseudoisoeugenol tiglate, epoxy-anol tiglate and anol tiglate was compared with that of seedlings and whole plants. The unorganized callus cultures were not able to accumulate any phenylpropanoids. In comparison, the leaf/root-differentiating callus culture promoted the accumulation of epoxy-pseudoisoeugenol tiglate (up to 90 mg/100 g fr.wt.) but not that of anol-derivatives. The accumulated amount of EPT in PMD-SH was comparable with that in plant seedlings.

scholarly journals The Diversity of Root-Associated Endophytic Fungi from Four Epiphytic Orchids in China

Diversity ◽  
2021 ◽  
Vol 13 (5) ◽  
pp. 197
Author(s):  
Tao Wang ◽  
Miao Chi ◽  
Ling Guo ◽  
Donghuan Liu ◽  
Yu Yang ◽  
...  
Keyword(s):  
Endophytic Fungi ◽  
Southern China ◽  
Culture Method ◽  
Amplicon Sequencing ◽  
Community Diversity ◽  
Mycorrhizal Symbiosis ◽  
Epiphytic Orchids ◽  
Culture Independent ◽  
Almost All

Root-associated endophytic fungi (RAF) are found asymptomatically in almost all plant groups. However, little is known about the compositions and potential functions of RAF communities associated with most Orchidaceae species. In this study, the diversity of RAF was examined in four wild epiphytic orchids, Acampe rigida, Doritis pulcherrima, Renanthera coccinea, and Robiquetia succisa, that occur in southern China. A culture-independent method involving Illumina amplicon sequencing, and an in vitro culture method, were used to identify culturable fungi. The RAF community diversity differed among the orchid roots, and some fungal taxa were clearly concentrated in a certain orchid species, with more OTUs being detected. By investigating mycorrhizal associations, the results showed that 28 (about 0.8%) of the 3527 operational taxonomic units (OTUs) could be assigned as OMF, while the OTUs of non-mycorrhizal fungal were about 99.2%. Among the OMFs, Ceratobasidiaceae OTUs were the most abundant with different richness, followed by Thelephoraceae. In addition, five Ceratobasidium sp. strains were isolated from D. pulcherrima, R. succisa, and R. coccinea roots with high separation rates. These culturable Ceratobasidium strains will provide materials for host orchid conservation and for studying the mechanisms underlying mycorrhizal symbiosis.

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