{"title":"Assessment of the genetic fidelity of true-to-type regenerants of medicinal plant Rheum emodi using RAPD and ISSR molecular markers","authors":"Sweta Upadhyay, Anjali Uniyal, Vijay Kumar, Sanjay Gupta","doi":"10.25303/1809rjbt1980204","DOIUrl":"https://doi.org/10.25303/1809rjbt1980204","url":null,"abstract":"Rheum emodi commonly known as rhubarb is mainly found in Northern Himalayas. It is a valuable medicinal plant having major pharmacological activities such as antimicrobial, anticancer, antioxidant, anti-inflammatory and is used extensively as purgative, stomachic and astringent tonic and improves gastro related problems. This herb is used by the local communities for medicinal as well as common eating purpose. This leads to its immense decline in its natural habitat and now this herb falls under threatened species and demands conservation. In this prospective, an efficient in vitro propagation method from callus culture has been achieved using leaf explants excised from the juvenile plant of R. emodi. Murashige and Skoog (MS) basal medium was used for regeneration procedure with different concentration of phytohormones. Maximum frequency of callus formation (84.44±0.27%) was observed in MS+36.19μM (2, 4-D) in combination with 11.10μM (BAP). The highest percentage of adventitious shoot regeneration was observed as 75.56±0.27% and the maximum number of shoots per explant that is 3.67±0.27 was achieved on MS basal medium containing BAP (35.5 μM) and Kn (11.61 μM). The maximum frequency of rooting was observed in MS full strength media + IAA (28.55 μM) + BAP (8.88 μM). The highest frequency of roots per shoot was observed as 5.0±0.47 with an average root length of 11±1.25mm. For ascertaining the clonal fidelity, 20 ISSR markers and 15 RAPD markers were assayed and employed to validate the true-to-type regenerants of Rheum emodi. Out of 15 RAPD and 20 ISSR markers, 7 markers and 15 markers produced distinct, clear and scorable bands with an average of 4.5 bands and 4.4 bands per marker respectively among the tissue cultured progenies. For each primer, the banding pattern was uniform and comparable to mother plant and showed about 99% homology. All the markers produce the monomorphic bands and no variation was detected among the micropropagated plants. Thus, the analysis of ISSR and RAPD patterns revealed that the bands were shared by both in vitro raised plants and parent clump confirming the genetic stability. DNA based molecular markers have proved to be versatile tools in diverse fields of biology. These markers proved to be model tools for routine analysis of clonal fidelity of micropropagated plants prior to commercialization.","PeriodicalId":21091,"journal":{"name":"Research Journal of Biotechnology","volume":"40 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-08-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"135162880","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Phung Thi Bich Hoa, Nguyen Hoang Tue, Hoang Lan Phuong, Nguyen Xuan Huy, Nguyen Hoang Loc
{"title":"Investigation on growth and development of 42 kDa chitinase transgenic peanuts (Arachis hypogaea L.) cultivar L14 under in vivo condition","authors":"Phung Thi Bich Hoa, Nguyen Hoang Tue, Hoang Lan Phuong, Nguyen Xuan Huy, Nguyen Hoang Loc","doi":"10.25303/1809rjbt1750181","DOIUrl":"https://doi.org/10.25303/1809rjbt1750181","url":null,"abstract":"The purpose of this study was to assess the growth and development of chitinase transgenic peanuts against phytopathogenic fungi including lines WTA-2 containing the Chi42 gene, S1A-15 containing the syncodChi42-1 gene and S2A-12 containing the syncodChi42-2 gene. The present results show that the peanut lines containing two optimized genes derived from the Chi42 wild-type gene of Trichoderma asperellum SH16, S1A-15 (syncodChi42-1) and S2A-12 (syncodChi42-2) seemed to grow stronger and produce a higher number of mature pods, weight of 100 pods and weight of 100 seeds than line WAT-2 with the Chi42 gene and non-transgenic control. Moreover, lines S1A-15 and S2A-12 also resulted in higher seed quality in terms of lipid and protein content than in comparison to the WTA-2 line and the control. These findings suggest that chitinase transgenic peanuts containing one of two synthetic genes in peanut roots could be a promising candidate for peanut production against phytopathogenic fungi.","PeriodicalId":21091,"journal":{"name":"Research Journal of Biotechnology","volume":"26 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-08-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"135162883","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"7, 12- Dimethylbenz[a]anthracene: A potent and multivariant Chemical carcinogen","authors":"S. Mirunalini, R. Susmitha","doi":"10.25303/1809rjbt2470258","DOIUrl":"https://doi.org/10.25303/1809rjbt2470258","url":null,"abstract":"Cancer is currently the second major cause of mortality globally. It is a collection of disorders categorized as abnormal cell development which can infiltrate and can spread to countless sections of the body. It has been essentially brought about by genetic and environmental sources mainly, countless patients have impacted by ecological factors like polycyclic aromatic hydrocarbons (PAHs). However, many PAHs in the ecosystem are generated from natural factors like direct combustion, petroleum and volcanic activity. Among these, 7,12 dimethylbenz[a]anthracene (DMBA) has been widely recognized for its capability to induce carcinogenesis in animals and humans. The carcinogenic activity of DMBA indicates the impact of well-differentiated tumors and it is recognized for generating DNA-reactive species that increase the oxidative damage in cells through their metabolism. Additionally, the configuration of DNA adducts and oxidative intermediates acquired from DMBA metabolism impairs prominent cellular activities by harming lipid and protein barriers. DNA adduct is made up of highly reactive intermediates. DMBA-induced cancer displayed higher expression of aryl hydrocarbon receptor (AhR), c-Myc, cyclinD1, activation of upstream regulatory mechanisms and frequent detection of NF-κB pathway components. In this study, we emphasized the growth of several cancers and the kinetic impact of DMBA to probable chemical carcinogenic activities.","PeriodicalId":21091,"journal":{"name":"Research Journal of Biotechnology","volume":"188 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-08-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"135163057","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"A study to determine effect of metal ions for optimization of L-Asparaginase producers for bioprocessing","authors":"Kakumanu Satishbabu, Prasuna Ravi Gyana","doi":"10.25303/1809rjbt088097","DOIUrl":"https://doi.org/10.25303/1809rjbt088097","url":null,"abstract":"The present study encompasses isolation of marine bacteria from marine soil samples collected from Chirala, Coastal area of Andhra Pradesh. Work includes isolation, screening, morphological and molecular characterization of selected L- Asparaginase producers. Purified enzyme was used for enzyme kinetic studies. Marine soil sample was collected in sterile bottle and brought to the lab bench and a total of 12 morphologically distinct bacterial colonies were isolated from collected marine soil samples and coded as Chirala Marine Bacteria. All 12 Chirala Marine Bacteria isolates were qualitatively screened for L-Asparaginase production. Out of 12 Chirala marine bacteria isolates, 9 isolates show positive response to L-Asparaginase activity. Effect of inducers i.e. carbon, nitrogen, aminoacids, phosphates and metal ions was studied.","PeriodicalId":21091,"journal":{"name":"Research Journal of Biotechnology","volume":"40 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-08-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"135162648","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Sangeeta Rani Tripathy, Shaikh Ameeruddin, Susmita N. Pradhan, Sarita Das
{"title":"In vitro Antioxidant, Antibiotic Complementing or Supplementing Potential and Urobactericidal Activity of Leaf Extract of Marsilea quadrifolia L.: The Water Fern","authors":"Sangeeta Rani Tripathy, Shaikh Ameeruddin, Susmita N. Pradhan, Sarita Das","doi":"10.25303/1809rjbt1200131","DOIUrl":"https://doi.org/10.25303/1809rjbt1200131","url":null,"abstract":"From generations, most of the communities of India are dependent on locally available ethnomedicines for tackling urogenital related issues. The efficacies of these herbal supplements for prophylactic and curative purposes need immediate scientific validation. For this, the accurate antibacterial potential of methanolic extracts of leaves of Marsilea quadrifolia L. (MMq) are investigated via each possible method of estimation including the supplementary and complementary antibacterial effects on pathogenic strains. The phytochemical constituents like carbohydrates, flavonoids, glycosides, phenolics, steroids and terpenoids are tested positive in the MMq. The total phenolic content (TPC) and total flavonoid content (TFC) are calculated to be 16.48 ± 0.4 mg/g of gallic acid equivalent and 96.33±4.4 mg/g of rutin equivalent respectively from the standard calibration curves. The IC50 value of MMq is found to be 258.55μg/ml in 1, 1 diphenyl-2-picrylhydrazyl radical (DPPH) assay by taking ascorbic acid as the standard. The extract gives significant complementary or supplementary inhibitory effects when added to certain antibiotic discs specifically against Enterococcus faecalis and Escherichia coli. The efficacy of MMq is verified and confirmed through different antibacterial assays against all the test uropathogenic bacteria. A reduction of 52%, 20.9%, 39.7% and 86.6% in number of colonies is noted in cfu/ml on treating the bacterial stains of E. coli, Staphylococcus aureus, E. faecalis and Proteus vulgaris respectively with a higher dose of MMq (1200μg) as compared to a lower dose of (200μg) in spread plate method. Present study recorded a minimum inhibitory concentration (MIC) of 2.5mg/ml of MMq each against E. faecalis, E. coli, Pseudomonas aeruginosa and 1.25mg/ml against S. aureus and P. vulgaris respectively. The preliminary reports of this study on MMq affirm an in-depth analysis of the usefulness of this plant as a miracle drug against certain uropathogens and urinary tract infections.","PeriodicalId":21091,"journal":{"name":"Research Journal of Biotechnology","volume":"50 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-08-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"135162650","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Efficient plant regeneration via indirect organogenesis and genetic stability assessment by molecular markers in an endangered medicinal plant Operculina turpethum (L.) Silva Manso","authors":"Bhagyashree Biswal, Biswajit Jena, Alok Kumar Giri, Reena Parida, Laxmikanta Acharya","doi":"10.25303/1809rjbt029036","DOIUrl":"https://doi.org/10.25303/1809rjbt029036","url":null,"abstract":"Operculina turpethum (L.) Silva Manso is reported to be effective against several diseases including tumor, jaundice, gastrointestinal disease etc. The plant is facing extinction due to overexploitation and inefficient conservation strategies. In this work, an effective approach for micropropagation of O. turpethum by indirect organogenesis from stem explant is devised. Induction of callus culture was carried out on Murashige and Skoog medium enriched with benzyl adenine (3mg/L). Subsequently, the callus cultured on MS medium with 1mg/L BA + 1mg/L kinetin generated a maximum of 3.20 ± 0.7 number of shoots/0.5gm callus within 35 days of incubation. On MS medium with Indole-3-acetic acid (IAA)(0.2mg/L), 98.4% of the shoots were successfully rooted. The in vitro regenerated plantlets were acclimatised and shifted to outdoor condition with a success rate of 95%. The genetic stability of micro propagated plantlets was assessed using PCR-based molecular markers, random amplified polymorphic DNA (RAPD) and inter simple sequence repeat (ISSR) which yielded a total of 27 and 31 scorable bands respectively. The monomorphic banding pattern of in vitro propagated plants confirms their genetic homogeneity. This work is report on in vitro plant regeneration through indirect organogenesis and evaluation of genetic fidelity of micropropagated plantlets of O. turpethum.","PeriodicalId":21091,"journal":{"name":"Research Journal of Biotechnology","volume":"29 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-08-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"135162651","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Phylogenomics based Identification of Microbial Biocontrol agent against Alternaria solani","authors":"Karun Wilson, Arunachalam Sathiavelu","doi":"10.25303/1809rjbt044057","DOIUrl":"https://doi.org/10.25303/1809rjbt044057","url":null,"abstract":"Alternaria solani is a fungal pathogen causing early blight infection in several solanaceous crops resulting in an annual yield loss up to 79%. Understanding the ecological niche and microbial interactions can help us contemplate new biocontrol agents against A. solani. This article combines the existing knowledge on ecology and biocontrol of A. solani, with the antagonist Bacillus subtilis ZD01 to identify potential biocontrol properties in its phylogenetically related species. Various physical, chemical and biological factors influencing the growth of A. solani were analyzed along with retrieving the whole genome sequence of B. subtilis ZD01. Using bioinformatic tool for Phylogenomics, genomes that share high similarity with B. subtilis ZD01 were identified. Resulting genomes were compared for the similarity in pathways, biosynthesis of secondary metabolites and biosynthetic gene clusters using Patric 3.6.12 and antiSMASH 6.0 web tools. Based on these, two new strains of bacteria were identified which share a similar ecological niche with A. solani and shares significantly similar properties with B. subtilis ZD01. Both strains were found to produce known clusters of bacillibactin, fengycin, surfactin and terpene which further strengthens its biocontrol potential. This study provides evidence suggesting new biocontrol agents and need for further exploration to identify unexplored microbes for plant-protection.","PeriodicalId":21091,"journal":{"name":"Research Journal of Biotechnology","volume":"15 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-08-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"135162657","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Reshma N. Sirasagikar, Ustad Bushra, Ashok Sudarshan, Agsar Dayanand
{"title":"Detection of actinobacteria from sediment soil: Exploration of metabolites as fungal plant pathogen inhibitors and plant growth promoters","authors":"Reshma N. Sirasagikar, Ustad Bushra, Ashok Sudarshan, Agsar Dayanand","doi":"10.25303/1809rjbt1550169","DOIUrl":"https://doi.org/10.25303/1809rjbt1550169","url":null,"abstract":"The rhizosphere is an area with dense microbial activity. Actinobacteria are one among the rhizomicroflora inhabiting in the plant rhizosphere by protecting plants from pathogens by producing secondary metabolites, as plant growth promoters and by producing hydrolytic enzymes. But in the modern era of agriculture, the outrageous use of chemical fertilizers in the agriculture fields is causing major environmental pollution and the agricultural soil is losing its texture and fertility. So, in the present study, we aimed to isolate and screen indigenous actinobacterial strains which are capable of producing antifungal metabolites, plant growth promoting ability and hydrolytic enzymes. A total of 23 isolates of actinobacteria have been isolated. Among all the other actinobacterial isolates, Streptomyces sp. DRPG-15, which was isolated from the sediment soil of the Tamankal River, revealed a noticeable antagonistic activity against Macrophomina phaseolina for Sorghum and Sclerotium rolfsii for Chickpeas respectively. 16S rRNA sequence analysis showed that the Streptomyces sp. DRPG-15 exhibited 100% homology with Streptomyces enissocaesilis DRPG-15 OP985046. All 23 isolates were screened for the production of indole acetic acid, hydrogen cyanide, ammonia production and nitrate reduction and were also screened for various hydrolytic enzymes like Caseinase, Protease, Cellulase, Amylase, Chitinase, Pectinase, Gelatinase, L-asparaginase and Streptomyces sp. DRPG-15 was the only isolate that showed positive results for antagonistic activity, plant growth-promoting ability and enzymatic activity. The entire study indicates that the selected strain of Streptomyces sp. DRPG-15 is implying its possible utilization as a natural bioinoculant for sustainable agriculture.","PeriodicalId":21091,"journal":{"name":"Research Journal of Biotechnology","volume":"206 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-08-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"135162658","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
K.V.N. Rajeshwari, B. Srilatha, Reddy V. Srilekha, Prasad G. Shyam
{"title":"Non-Steroidal Anti-Inflammatory Drugs (NSAIDS) as potential inhibitors of tankyrase-2 for colon cancer prevention: an in silico study","authors":"K.V.N. Rajeshwari, B. Srilatha, Reddy V. Srilekha, Prasad G. Shyam","doi":"10.25303/1809rjbt2160221","DOIUrl":"https://doi.org/10.25303/1809rjbt2160221","url":null,"abstract":"Colon cancer is one of the most lethal and common cancers worldwide. The potential risk factor recognized for tumor development was chronic inflammation. Therefore, targeting inflammatory pathways and tankyrases which control Wnt pathway involved in cancer pathogenesis has proved effective in preventing the formation of colon cancer and its progression. Hence, in the present investigation, to find the potential anti-inflammatory drugs (NSAIDs) capable of inhibiting tankyrases, 12 NSAIDs were selected as ligands targeting the proteins tankyrases which play a major role in cancer progression. An advanced docking software Auto dock was selected for the study. Among the 12 anti-inflammatory drugs selected, mefanemic acid was found to be potential inhibitor of tankyrases followed by Indomethacin, Piroxicam, Ketorolac and Etoricoxib. Rest of the drugs under study showed moderate degree of inhibition. Least inhibition potential was recorded with diclofenac. Hence, from the above results, it is clear that Mefanemic acid, Indomethacin, Piroxicam, Ketorolac have the potential of preventing colon cancer by inhibiting tankyrases-2. However, further in vitro and in vivo investigations are required for confirmation.","PeriodicalId":21091,"journal":{"name":"Research Journal of Biotechnology","volume":"17 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-08-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"135162881","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Implementation of Design of Experiment (DoE) for process development and optimization of biosimilars","authors":"Vinay Rewaria, Mahendran Botlagunta, Pardhasaradhi Mathi","doi":"10.25303/1809rjbt1370146","DOIUrl":"https://doi.org/10.25303/1809rjbt1370146","url":null,"abstract":"Selection of a suitable basal media, feed, feeding strategy and glycan structures has a pivotal role during the process development and optimization of monoclonal antibodies. Instead of using one-factor-at-time approach or classical approach for media and feed selection, a DoE approach was implemented. A Simplex centroid mixture design was applied to identify the suitable media/media combinations by evaluating cell growth and productivity followed by a full factorial design for screening of the best suitable feed that enhanced the productivity. Optimization of the process was performed using Response surface methodology with central composite design to determine the optimum interaction between the selected media and feeds. Statistically designed experiments were performed to determine the factors influencing glycan. Uridine, MnCl2 and galactose were added as per the factorial design for evaluating their effect on modulating glycosylation. Prediction of the best combination was made with the maximum response titer by Design-Expert software 12.0. A threefold increase in titer (1909 mg/L) and significant improvement in glycosylation were obtained by statistical approach.","PeriodicalId":21091,"journal":{"name":"Research Journal of Biotechnology","volume":"42 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-08-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"135162887","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}