Agrobacterium rhizogenes mediated genetic transformation and its applications

Agrobacterium rhizogenes mediated transformation is a powerful technique in plant biotechnology that enables the rapid and efficient production of transformed hairy roots. This process is particularly advantageous for various plant species, spanning angiosperms (both dicotyledonous and monocotyledonous plants), gymnosperms, and even moss. The key element in this transformation is the root-inducing (Ri) plasmid carried by A. rhizogenes. The process begins with the introduction of A. rhizogenes to wounded plant tissues. The bacterium, being Gram-negative, harbors the Ri plasmid, which contains the transfer-DNA (T-DNA) fragment. This T-DNA integrates into the host plant genome, triggering the formation of hairy roots. The Ri T-DNA can also transmit through meiosis, ensuring that the genetic modifications are heritable. The A. rhizogenes-mediated transformation system is particularly valuable for species that are challenging to transform using Agrobacterium tumefaciens. It offers several advantages over A. tumefaciens-mediated transformation, including a faster process and a higher transformation frequency

EFFICACY OF WATTAKAKA VOLUBILIS AGAINST ALUMINIUM SULPHATE INDUCED IN LIVER TOXICITY

Objective: The modulation of membrane-bound ATPases, carbohydrate metabolizing enzymes and mitochondrial TCA cycle enzymes in Wattakaka volubilis on aluminium sulphate induced liver toxicity.Methods: Medicinal therapy requires careful assessment of effective treatment offering an acceptable safety over human health. Experimental animals were divided into five groups (Untreated, negative and positive control, hepatic group and the hepatic group fed on Wattakaka volubilis), Metabolising enzymes level, estimation of DNA, RNA and quantification of DNA fragmentation and gene expression were investigated. These altered enzyme levels were ameliorated significantly by administration of Wattakaka volubilis at the concentration of 200 mg/kg in drug-treated animals.Results: Results showed that treatment with methanol extract of Wattakaka volubilis normal level of enzymes which are compared with silymarin. This was evident from the significant increase in p<0.05, p<0.01, p<0.001 enzyme levels. Aluminium sulphate induced rats showed decreased the activities of metabolising enzymes and increased DNA fragmentation in the liver. This clearly explained the reason for the hepatoprotective activity of Wattakaka volubilis leaf extract.Conclusion: The methanolic leaf extract of W. volubilis showed high protective activity against aluminium sulphate-induced hepatotoxicity

Crucial stages and process for successful banana micropropagation through plant tissue cultutre techniques

Banana is a vegetatively propagated plant in which tissue culture is employed vastly. The apical shoot tip culture technique is capable to mass produce disease-free planting material for commercial propagation of banana seedlings. Healthy planting material serves as a major input for successful farming of year round crop. The success of tissue culture depends largely on the selection of mother suckers, in vitro culture and rooting and hardening of plants. The crucial stages for successful micropropagation are discussed in detail

Successful intracytoplasmic sperm injection with testicular spermatozoa using pentoxifylline and HOS from a male with necrozoospermia: a case report

The study was done to compare fertilization, cleavage, and pregnancy outcome of a man affected with necro zoospermia  using pentoxifylline and a hypoosmotic swelling (HOS) test to select viable spermatozoa from testicular biopsy specimens. A patient was diagnosed with necrozoospermia by semen analysis, and testicular spermatozoa were injected intracytoplasmically, using pentoxifylline and HOS (Hypo osmotic swelling test) and the following ICSI results were observed. Significantly higher fertilization rates (pentoxifylline 100% vs HOS 80%) were observed. There was no significant difference observed in cleavage rates among both groups. Finally, a healthy female infant with birth weight 3 kg was delivered at the second frozen embryo transfer cycle.it was found that selecting viable spermatozoa using pentoxifylline was more effective in fertilization and pregnancies than obtaining it through a HOS test. ICSI is effective for necrozoospermic man

Review on Agni w.s.r. to Jatharagni and its role in Amotpatti

Agni have been told as process of Paka (digestion) etc. Hence in Shareera, the metabolic process from rasa to Shukra and the development of the fertilized egg all these refer that the ancient Rushis had a full-fledged knowledge about Agni, and have presented it in a very well form. The knowledge about Agni was well developed during ancient time. Agni has been expressed in mantras clearly. There is an elaborated description regarding qualities and functions of Agni in classics. There are elaborated references regarding Agni in Kriya Shaarira and Vikruti Vijnyana. The treatment principle in Ayurveda depends on Agni; hence acharya Bhoja has opined that Kayachikitsa is nothing but Kayagni Chikitsa. Hence the present review highlights the importance of Agni w.s.r. to Jatharagni and its role in Amotpatti

Over expression of anti-apoptotic gene in banana cv Rasthali enhances resistance against Fusarium oxysporum f. sp. cubense Race 1

The most popular banana cv Rasthali was transformed with anti-apoptotic gene, AtBAG4 regulated with two different promoters viz., ZmBgl and ubiquitin to enhance the tolerance levels to Fusarium oxysporum f. sp. cubense Race 1 (FOC1). The differences in gene expression driven by two promoters revealed that stronger expression of AtBAG4 gene under the ubiquitin promoter suppressed the infection and spreading processes of FOC1 in transgenic banana under standard bioassay systems. Analysis using the real time PCR showed the varying levels of AtBAG4 gene expression under two promoters. It was evident that ZmBgl driven AtBAG4 lead to lower gene expression in leaves which correlated with lesser levels of resistance to FOC1. Constitutive expression of AtBAG4 under the control of ubiquitin promoter showed increased transgene transcripts which directly correlated with the enhanced tolerance against FOC1 from seedlings stage to active vegetative phases. This study reveals the importance of constitutive expression of anti-apoptotic gene showing enhanced tolerance against the most dreaded FOC1 in highly susceptible variety Rasthali

Standardization of sterilization protocol for explants and its suitability for direct organogenesis in tuberose cv. Arka Vaibhav

A study was carried out to standardize the sterilization protocol for different explants (terminal stem scale,immature flower bud and tepal segment) and to select the suitable explant for the direct organogenesis of tuberose cv. Arka Vaibhav. The highest survival per cent (100) and uncontaminated cultures (0.00) of terminal stem scale explant was observed in pre-treatment with overnight soaking of terminal stem scale in the solution comprising carbendazim (0.1%), chlorothalonil (0.05%) and myristyl trimethyl ammonium bromide (cetrimide) (0.05%) and subsequently surface sterilization with 70% ethanol (1 min), 4% sodium hypochlorite (10 min) followed by 0.1% HgCl2 (15 min). The explant immature flower bud recorded the highest survival per cent (100) and maximum aseptic cultures in the treatment T1 comprised of 1.0 drop Tween-20 + 70% ethanol (30 sec) and 1% sodium hypochlorite (3 min). Pre-treatment of tepal segment explant in 0.1% carbendazim (30 min) solution followed by surface sterilization with combination of 1.0 drop Tween-20 + 70% ethanol (30 sec) followed by 1% sodium hypochlorite (3 min) registered 91.66% of survival with the minimum contamination (10%) in the treatment. Among the three explants used, the terminal stem scale was found suitable for direct organogenesis with early greenness (5.72 days) and highly responsive to shoot induction (100%) in MS medium supplemented with 4 mg/L BAP + 0.1mg/L IAA. Other two explants viz., immature flower bud and tepal segment failed to respond for direct organogenesis by shoot induction instead produced profuse callus

COMPARATIVE STUDY OF Î’-CYCLODEXTRIN PRODUCTION FROM THE CGTASE PRODUCING NOVEL STRAINS- ITS CHARACTERIZATION AND HOST-GUEST COMPLEX FORMATION

The enzymatic synthesis of β-CD by CGTase produced from different strains of alkaliphilic bacterial culture which was isolated from the cultivated sugarcane fields and standard MTCC cultures using starch as substrate. An alkalophilic bacteria were grown for six days at static conditions at pH 10.5. The time course of CGTase activity was studied with the maximum activity observed on 6th day. The activity check and its confirmation were done by Dextrinising and Phenolphthalein assay. The soluble starch was best substrate to produce the cyclodextrin. Extraction of β-CD was done using complexing agents, these agents binds only to β-CD and forms complex, the complex thus formed was recovered. The obtained β-CD was made inclusion complex with guest molecule and was further characterized using UV absorption spectrophotometer, FT-IR and melting point.   Keywords: Cyclodextrin glycosyltransferases, Cyclodextrins, Bacillus licheniformis

Oxygen Activation and Radical Transformations in Heme Proteins and Metalloporphyrins

As a result of the adaptation of life to an aerobic environment, nature has evolved a panoply of metalloproteins for oxidative metabolism and protection against reactive oxygen species. Despite the diverse structures and functions of these proteins, they share common mechanistic grounds. An open-shell transition metal like iron or copper is employed to interact with O_2 and its derived intermediates such as hydrogen peroxide to afford a variety of metal–oxygen intermediates. These reactive intermediates, including metal-superoxo, -(hydro)peroxo, and high-valent metal–oxo species, are the basis for the various biological functions of O_2-utilizing metalloproteins. Collectively, these processes are called oxygen activation. Much of our understanding of the reactivity of these reactive intermediates has come from the study of heme-containing proteins and related metalloporphyrin compounds. These studies not only have deepened our understanding of various functions of heme proteins, such as O2 storage and transport, degradation of reactive oxygen species, redox signaling, and biological oxygenation, etc., but also have driven the development of bioinorganic chemistry and biomimetic catalysis. In this review, we survey the range of O_2 activation processes mediated by heme proteins and model compounds with a focus on recent progress in the characterization and reactivity of important iron–oxygen intermediates. Representative reactions initiated by these reactive intermediates as well as some context from prior decades will also be presented. We will discuss the fundamental mechanistic features of these transformations and delineate the underlying structural and electronic factors that contribute to the spectrum of reactivities that has been observed in nature as well as those that have been invented using these paradigms. Given the recent developments in biocatalysis for non-natural chemistries and the renaissance of radical chemistry in organic synthesis, we envision that new enzymatic and synthetic transformations will emerge based on the radical processes mediated by metalloproteins and their synthetic analogs