Expression analysis of biosynthetic pathway genes vis-à-vis podophyllotoxin content in Podophyllum hexandrum Royle

Kumar, Pawan; Pal, Tarun; Sharma, Neha; Kumar, Varun; Sood, Hemant; Chauhan, Rajinder Singh

doi:10.1007/s00709-015-0757-x

Cited by 44 publications

(12 citation statements)

References 39 publications

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“…The lack of Amarogentin in undifferentiated callus cultures and less amount of Mangiferin in callus culture of S. chirayita, might be due to absence of cell machinery programming along with the absence of balanced cell organization to occur in tissue culture condition for metabolite synthesis. As already reported by Kumar et al 2015 wherein callus cultures of S. chirayita accumulates less amount of Amarogentin i.e. 0.05 and 0.09% of Mangiferin and further Kumar et al 2014 reported negligible amount of Mangiferin and very less amount of Amarogentin, when they have already added Adenine sulphate for enhanced production of secondary metabolites, such findings were in relevance to our study, where we have reported 8.57 µg/mg of Mangiferin and negligible amount of Amarogentin at 15 °C in callus initiation stage.…”

Section: Discussionsupporting

confidence: 76%

Optimizing nutrient media conditions for continuous production of shoot biomass enriched in major medicinal constituents, amarogentin and mangiferin of endangered medicinal herb, Swertia chirayita

Gupta

Sood

2022

Vegetos

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Biosynthesis and accumulation of Amarogentin and Mangiferin from shoot culture of endangered herb Swertia chirayita helped in rescuing its natural population along with continuous production of quality rich herbal material. Although, presence of Amarogentin and Mangiferin had already been reported, but such studies did not elaborate the significant developmental stages at two varying temperature (15 ± 1 °C and 25 ± 1 °C) in shoot cultures of S. chirayita . Different developmental stages involved throughout from callus induction to complete regeneration of plant by using shoot cultures of S. chirayita, reveal different amounts of significant medicinal compounds having high pharmacological importance like bearing anti-diabetic and anti-cancerous properties. So in the present study, different developmental stages i.e. plant segment as leaf disc explants, initiation of callus formation, callus mass development, shoots primordial, manifold shoot formation and shoot elongation with complete growth were explored for accumulation of Amarogentin and Mangiferin. The Amarogentin content was 4.72 µg/mg at 15 ± 1 °C and 4.41 µg/mg at 25 ± 1 °C whereas Mangiferin content was 15.54 µg/mg at 15 ± 1 °C and 9.70 µg/mg at 25 ± 1 °C in leaf discs provided with the medium MS + 2,4D = 1 mg/L, 6BAP = 0.5 mg/L, TDZ = 0.5 mg/L, respectively. The accumulation of Amarogentin and Mangiferin started from callus cultures differentiating into shoots and reached to the detectable amount equivalent to actual leaf explants in fully grown shoots with content of 5.79 µg/mg at 15 ± 1 °C and 5.35 µg/mg at 25 ± 1 °C whereas 15.56 µg/mg at 15 ± 1 °C and 13.15 µg/mg at 25 ± 1 °C provided with the medium MS + IBA = 3 mg/L, KN = 1 mg/L, respectively. Maximum accumulation of bioactive compounds was observed in ≈3 months old in-vitro grown shoots at 15 ± 1˚ C wherein, the content of Amarogentin was ≈8.51 folds higher and Mangiferin was ≈4.09 folds higher than the ≈3 months old green house grown shoots. So, the in-vitro raised shoots of S. chirayita enriched with marker medicinal compounds would be utilized as ready to use raw material for pharmaceutical industries for herbal drug formulations and can be utilized to transfer under natural habitats for conserving its diminishing population.

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Section: Discussionsupporting

confidence: 76%

Optimizing nutrient media conditions for continuous production of shoot biomass enriched in major medicinal constituents, amarogentin and mangiferin of endangered medicinal herb, Swertia chirayita

Gupta

Sood

2022

Vegetos

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“…Until recently, the knowledge about its biosynthetic pathway was fragmented and key enzymes and corresponding genes were not known (Kumar et al, 2015). A thorough knowledge of complete biosynthetic pathway would ease the access to podophyllotoxin and its natural derivatives which are difficult to produce synthetically (Kamal et al, 2015).…”

Section: Screening For Bioactive Compoundsmentioning

confidence: 99%

Molecular Approaches to Screen Bioactive Compounds from Endophytic Fungi

2016

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Endophytic fungi are capable of producing plant associated metabolites and their analogs with therapeutic value. In order to identify the potential endophytic isolates producing bioactive compounds, one need to screen all isolated endophytes, which may run into hundreds. Isolation of endophytic fungi is relatively a simple process; but screening of the isolated fungi for required metabolite production is a cumbersome process. Endophytic fungi producing plant associated metabolites may contain genes involved in the entire biosynthetic pathway(s). Therefore, ascertaining the presence of key enzymes of a particular biosynthetic pathway could serve as a molecular marker for screening of these endophytes to produce that metabolite. In absence of entire biosynthetic pathways in endophytic fungi, plant genes associated with that metabolic pathway could serve as markers. This review focuses on the impact of molecular approaches to screen the endophytic fungi for the production of bioactive compounds. An attempt has been made on screening of anticancer compounds like taxol (paclitaxel), podophyllotoxin, and camptothecin using molecular markers. The advantages of molecular approaches over conventional methods to screen endophytic fungi and also identification of endophytic fungi are discussed.

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“…Currently, a majority of medicinal plants enhance the bioactivity of their medicinal components by inducing and regulating functional genes present in their biosynthetic pathway [25]. Several reports have shown that in plants, there is a significant correlation between the synthesis and accumulation of secondary metabolites and the expression levels found in the functional pathways of their biosynthetic pathways [26,27].…”

Section: Discussionmentioning

confidence: 99%

Evaluation of Reference Genes for qRT-PCR Normalization in Angelica decursiva under Various Experimental Conditions

He¹,

Bao²,

Wang³

et al. 2020

Preprint

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Angelica decursiva is one of the lending traditional Chinese medicinal plants producing coumarins. Notably, several studies have focused on the biosynthesis and not the qRT-PCR (quantitative real-time reverse transcription polymerase chain reaction) study of coumarins. This qRT-PCR technique has been extensively used to investigate gene expression levels in plants and the selection of reference genes which plays a crucial role in standardizing the data form the qRT-PCR analysis. In our study, 11 candidate reference genes were selected from the existing transcriptome data of Angelica decursiva. Here, four different types of statistical algorithms (geNorm, NormFinder, BestKeeper, and Delta Ct) were used to calculate and evaluate the stability of gene expression under different external treatments. Subsequently, RefFinder analysis was used to determine the geometric average of each candidate gene ranking, and to perform comprehensive index ranking. The obtained results showed that among all the 11 candidate reference genes, SAND family protein (SAND), protein phosphatase 2A gene (PP2A), and polypyrimidine tract-binding protein (PTBP) were the most stable reference genes, where Nuclear cap binding protein 2 (NCBP2), TIP41-like protein (TIP41), and Beta-6-tubulin (TUBA) were the least stable genes. To the best of our knowledge, this work is the first to evaluate the stability of reference genes in the Angelica decursiva which has provided an important foundation on the use of qRT-PCR for an accurate and far-reaching gene expression analysis in this medicinal plant.

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Expression analysis of biosynthetic pathway genes vis-à-vis podophyllotoxin content in Podophyllum hexandrum Royle

Cited by 44 publications

References 39 publications

Optimizing nutrient media conditions for continuous production of shoot biomass enriched in major medicinal constituents, amarogentin and mangiferin of endangered medicinal herb, Swertia chirayita

Optimizing nutrient media conditions for continuous production of shoot biomass enriched in major medicinal constituents, amarogentin and mangiferin of endangered medicinal herb, Swertia chirayita

Molecular Approaches to Screen Bioactive Compounds from Endophytic Fungi

Evaluation of Reference Genes for qRT-PCR Normalization in Angelica decursiva under Various Experimental Conditions

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