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Dharmendra Kumar

Publications and source records attributed to Dharmendra Kumar.

5 recordsLinked to original sources

Developmental block in ruminant embryos: Mechanisms, molecular insights and potential interventions.

Developmental block remains one of the major hurdles that makes it hard to develop embryos in vitro more efficiently. In ruminants, it is predominantly observed during the 8-16 cell stage, coinciding with the maternal-to-zygotic transition (MZT) and embryonic genome activation (EGA). In addition, reducing maternal transcripts and initiating embryonic transcription correctly is a major reason for developmental arrest. A broad array of molecular mechanisms has been implicated, encompassing incomplete epigenetic regulation, mitochondrial dysfunction, oxidative stress, improper cell cycle progression, and dysregulated apoptosis. During this process, several key genes, including ZAR1, NPM2, DPPA3, DNMTs, Cyclin B1, BCL2, and antioxidant enzymes (SOD1, GPX1, and CAT) have been recognized as essential regulators of the block. External factors, especially poor in vitro culture conditions, high oxygen levels, and the secretion of harmful metabolites, make developmental failure even worse. Recent research has underscored the significance of antioxidant supplementation, epigenetic modulators, and enhanced culture systems in mitigating developmental barriers. Therefore, the current review summarises the contemporary insights into the factors and molecular mechanisms responsible for ruminant embryonic developmental block, focusing on MZT, oxidative stress, and epigenetic regulation. It also addresses potential strategies to enhance the developmental competence of ruminant embryos in vitro.

Embryo↗

An effective method for isolation and regeneration of Solanum tuberosum mesophyll protoplasts for transgene-free genome editing.

An effective system for isolating and regenerating protoplasts is crucial for research in genome engineering. This study focused on refining a protocol for the isolation and regeneration of mesophyll protoplasts from the leaves of Solanum tuberosum cv. Kufri Jyoti. Key factors influencing protoplast yield and viability, such as dark pretreatment, pre-plasmolysis, enzyme concentrations, and osmoticum levels, were thoroughly assessed and optimized. The highest protoplast yield and viability were achieved with an enzyme mixture of 1.0% cellulase R-10 and 0.5% macerozyme R-10 after 16 h of incubation. Furthermore, culturing on a Murashige and Skoog-based medium (MSPI) without ammonium nitrate, enriched with an osmoticum concentration of 0.4 M and a carefully adjusted auxin-to-cytokinin ratio, successfully facilitated protoplast division, microcalli proliferation, and minicalli formation. Callus proliferation and shoot induction were accomplished on MS13K medium supplemented with naphthaleneacetic acid (NAA) and zeatin riboside. Root initiation and elongation were promoted on MS basal medium supplemented with indole-3-butyric acid (IBA) at 1 mg/L. The regenerated plantlets were subsequently acclimatized and hardened under controlled greenhouse conditions. This robust protoplast-to-plant protocol serves as a crucial resource for the introduction of ribonucleoprotein complexes into plant cells, facilitating accurate, transgene-free genome editing.

Callus induction↗

Variability in Indian isolates of Arthrobotrys dactyloides Drechsler: a nematode-trapping fungus.

Five isolates of Arthrobotrys dactyloides (A, B, C, D, and E) were isolated from different locations of India. Their variability in relation to morphology, radial growth, and nematode capturing or trap-forming ability was observed. All of the five isolates produced two-celled slender conidia, whereas wider two- and three-celled conidia were produced by isolates A, C, D, and E only. The wider two- and three-celled conidia were not observed in cultures of isolate B. The isolate B produced macroconidia as well as microconidia. The microconidia were produced on separate conidiophores of smaller size. Macroconidia and microconidia were never produced on the same conidiophore, but the two types of conidiophores were produced on same or different hyphae. Similar to macroconidia, the microconidia also produced constricting rings of smaller size in presence of Meloidogyne graminicola. The constricting rings formed on microconidia did not capture second-stage juveniles of M. graminicola because of their smaller size. Among all the isolates, isolate B showed slow growth and higher nematode-capturing ability or trap-forming ability.

Animals↗

Effect of cysteine on methionine production by a regulatory mutant of Corynebacterium lilium.

The production of methionine by submerged fermentation using a mutant strain of Corynebacterium lilium was studied to determine suitable conditions for obtaining high productivity. The mutant strain resistant to the methionine analogues ethionine, norleucine, methionine sulfoxide and methionine methylsulfonium chloride produced 2.34 g l(-1) of methionine in minimal medium containing glucose as carbon source. The effect of cysteine on methionine production in a 15 l bioreactor was studied by supplementing cysteine intermittently during the course of fermentation. The addition of cysteine (0.75 g l(-1)h(-1)) every 2 h to the production medium increased the production of methionine to 3.39 g l(-1). A metabolic flux analysis showed that during cysteine supplementation the ATP consumption reduced by 20%. It also showed that the increase in flux from phosphoenol pyruvate to oxaloacetate leads to higher methionine production. Results indicate that controlling the respiratory quotient close to 0.75 will produce the highest amount of methionine and that regulatory mutants also resistant to analogues of cysteine would be better methionine over producers.

Adenosine Triphosphate↗

Methionine production by fermentation.

Fermentation processes have been developed for producing most of the essential amino acids. Methionine is one exception. Although microbial production of methionine has been attempted, no commercial bioproduction exists. Here, we discuss the prospects of producing methionine by fermentation. A detailed account is given of methionine biosynthesis and its regulation in some potential producer microorganisms. Problems associated with isolation of methionine overproducing strains are discussed. Approaches to selecting microorganism having relaxed and complex regulatory control mechanisms for methionine biosynthesis are examined. The importance of fermentation media composition and culture conditions for methionine production is assessed and methods for recovering methionine from fermentation broth are considered.

Corynebacterium↗