Giridhar Parvatam

AgNO3: a potential regulator of ethylene activity and plant growth modulator

The aim of this review is to critically analyze the role of silver nitrate (AgNO 3 ) in modulating plant growth and development. In recent years, basic studies on ethylene regulation opened new vistas for applied research in the area of micro-propagation, somatic embryogenesis, in vitro flowering, growth promotion, fruit ripening, and sex expression. Silver nitrate has proved to be a very potent inhibitor of ethylene action and is widely used in plant tissue culture. Few properties of silver nitrate such as easy availability, solubility in water, specificity and stability make it very useful for various applications in exploiting plant growth regulation and morphogenesis in vivo and in vitro . Silver ion mediated responses seem to be involved in polyamines, ethylene- and calcium- mediated pathways, and play a crucial role in regulating physiological process including morphogenesis. The molecular basis for regulation of morphogenesis under the influence of silver nitrate is completely lacking. This review compiles published reports of silver nitrate-mediated in vitro and in vivo studies and focuses on fundamental and applied aspects of plant growth modulation under the influence of silver nitrate.

Biochemical characterization of a key step involved in 2H4MB production in Decalepis hamiltonii

Decalepis hamiltonii is widely known for its flavour molecule 2-Hydroxy-4-Methoxy Benzaldehyde (2H4MB), a structural isomer of vanillin. As the biosynthetic pathway of 2H4MB is not known, we hypothesised 2H4MB origins could be from phenylpropanoid pathway (PPP). Accordingly, a study was conducted using PPP inhibitors (viz. piperonylic acid, MDCA and propanil) against in vitro root cultures of D. hamiltonii to find the branch of PPP which catalyses the 2H4MB formation. HPLC analysis was carried out to quantify 2H4MB levels in control and respective inhibitor treated root cultures in vitro. The results obtained revealed that piperonylic acid did not inhibit 2H4MB biosynthesis in the given period, whereas MDCA and propanil had the marked inhibitory effect. The inhibitory effect was evident with 13.2, 33.6 and 37.9% decrease in 2H4MB levels at 50, 100 and 150mM concentration of MDCA respectively in comparison with control roots. Similarly, the inhibitory effect of propanil on 2H4MB biosynthesis was obvious with 23.7, 49.5 and 57.9% decrease in 2H4MB levels at 50, 100 and 150μM concentration of inhibitor respectively when compared with control roots. Propanil showed a greater slow down effect on 2H4MB biosynthesis compared to MDCA. Incorporation of 0.1, 0.5 and 1.0mM ferulic acid as a precursor to in vitro root cultures of D. hamiltonii showed an increase in 2H4MB levels at the rate of 3.1, 107 and 94.1% respectively as quantified by HPLC analysis. However, ferulic acid in conjunction with propanil did not show any increase in 2H4MB levels. This clearly explains that ferulic acid is channelled through the 4-CL (4-coumarate CoA ligase) enzyme, where it would be converted to feruloyl-CoA and could be further converted to 2H4MB in D. hamiltonii.

PGPR’s mix treatment to Moringa improved plant growth and iron content in foliage as substantiated by biochemical and molecular methods

ABSTRACT In this study, Bacillus pumilus SE34 and B. pumilus T4 were combined with Bacillus subtilis GBO3, B. pumilus INR7 and Pseudomonas fluorescens UOM14 to form COM1, COM2 and COM3, respectively. All combinations were used to find their synergistic effect on Moringa oleifera for growth promotion and Fe accumulation efficiency in foliage. The results indicate a significant increase in Fe content in foliage using COM3 (405.70%) followed by COM2 (105.83%) in comparison to the control with a simultaneous decrease in the soil Fe content. Increased expression of iron-transport-related genes like iron-phytosiderophore oligopeptide transporter and natural-resistance-associated macrophage protein, in foliage of Moringa using real-time PCR correlates with the enriched iron content in foliage of treated plants. Increase in citric acid was in direct relation to the Fe accumulation in foliage. Growth promotion and Fe enhancement using plant-growth-promoting rhizobacteria’s combination was significantly higher, proving its synergistic effect as a great source for sustainable development in agriculture and nutrition.

Recombinant pharmaceutical protein production in plants: unraveling the therapeutic potential of molecular pharming

There is an increasing demand for the generation of recombinant pharmaceutical proteins for a wide array of therapeutic applications. In comparison to bacterial, yeast and animal cells, the production of recombinant proteins in plants with economic and therapeutic importance has only started recently. The most important prerequisite of any expression systems is that it should be simple and inexpensive. In this regard, plant-based expression has emerged an as accepted alternative to conventional expression platforms due to economic feasibility, rapid scalability, higher stability of recombinant proteins, safety due to lack of harmful substances (human, animal pathogens and pyrogens) and capability of producing proteins with desired secondary modifications. Heterologous expression using plants has played a pivotal role in the development of a myriad of recombinant proteins, including neutraceuticals and monoclonal antibodies being utilized in various therapeutic approaches. This paper presents an overview about the current status, various strategies and advantages of pharmaceutical protein production in plant expression systems. We also present a summary of expression of therapeutic monoclonal antibodies, vaccines, clinical trials and the regulatory aspects of plant-based expression. Furthermore, the challenges encountered in plant expression such as costs associated with existing purification strategies are discussed.

Culture Conditions for Production of Biomass, Adenosine, and Cordycepin from Cordyceps sinensis CS1197: Optimization by Desirability Function Method

Background: Cordyceps sinensis (CS) is a traditional Chinese medicine contains potent active metabolites such as nucleosides and polysaccharides. The submerged cultivation technique is studied for the large scale production of CS for biomass and metabolites production. Objective: To optimize culture conditions for large-scale production of CS1197 biomass and metabolites production. Materials and Methods: The CS1197 strain of CS was isolated from dead larvae of natural CS and the authenticity was assured by the presence of two major markers adenosine and cordycepin by high performance liquid chromatography and mass spectrometry. A three-level Box-Behnken design was employed to optimize process parameters culturing temperature, pH, and inoculum volume for the biomass yield, adenosine and cordycepin. The experimental results were regressed to a second-order polynomial equation by a multiple regression analysis for the prediction of biomass yield, adenosine and cordycepin production. Multiple responses were optimized based on desirability function method. Results: The desirability function suggested the process conditions temperature 28°C, pH 7 and inoculum volume 10% for optimal production of nutraceuticals in the biomass. The water extracts from dried CS1197 mycelia showed good inhibition for 2 diphenyl-1-picrylhydrazyl and 2,2-azinobis-(3-ethyl-benzo-thiazoline-6-sulfonic acid-free radicals. Conclusion: The result suggests that response surface methodology-desirability function coupled approach can successfully optimize the culture conditions for CS1197. SUMMARY Authentication of CS1197 strain by the presence of adenosine and cordycepin and culturing period was determined to be for 14 days Content of nucleosides in natural CS was found higher than in cultured CS1197 mycelium Box-Behnken design to optimize critical cultural conditions: temperature, pH and inoculum volume Water extract showed better antioxidant activity proving credible source of natural antioxidants.