Culture Techniques Research Articles

Impact of Different Plant Densities and Fertilizer Levels on Yield Attributes, Yield and Fibre Quality Characters of Bt Cotton in HDPS

Adopting a high-density planting system in cotton cultivation has resulted in increased yields compared to traditional methods. Despite the clear benefits, farmers face various challenges throughout the cultivation process, from sowing to harvesting. However, selecting appropriate plant spacings, fertilizer levels and implementing tailored cultivation techniques can significantly enhance productivity. A field experiment was conducted at Siddapur research farm, Regional Agricultural Research Station, Warangal, Telangana, India during kharif 2023 to investigate the “Impact of different plant densities and fertilizer levels on yield attributes, yield and fibre quality characters of cotton in high density planting system (HDPS)”. The experiment was laid out in randomized block design (factorial) and replicated thrice, consisting of 12 treatment combinations comprising of three plant spacings S1: 80 x 20 cm (62,500 plants ha-1), S2: 90 x 15 cm (74,074 plants ha-1), S3: 90 x 60 cm in factor I and four fertilizer levels (F1: 100%RDF, F2: 125%RDF, F3: 100%RDF + Microbial consortia, F4: Control) in factor II. Both the factors significantly influenced the number of bolls per plant, number of bolls m-2 and seed cotton yield per hectare. The results revealed that higher no. of bolls plant-1 were recorded in 90 x 60 cm (18,518 plants ha-1) however, higher no. of bolls m-2, seed cotton yield (2233 kgha-1) obtained from higher plant density (74,074 plants ha-1) with the spacing 90 x 15 cm. Boll weight was not significantly influenced by varied plant spacings and fertilizer levels. Among the different levels of fertilizer, 125%RDF recorded highest no. of bolls per plant, no. of bolls m-2 and seed cotton yield (2362 kgha-1). Fibre quality characters viz., Upper half mean length(mm), micronaire(µginch-1), tenacity(gtex-1) and uniformity index (%) were not significantly influenced by both plant spacings and fertilizer levels as they are primarily governed by genetic makeup of cotton genotypes. For optimum seed cotton yield, cotton should be sown at closer spacing 90 x 15 cm and the application of 125%RDF was economically feasible in high density planting system.

Gold-antibody-aptamer complexed electrochemical sensing surface for septic arthritis biomarker determination

Septic arthritis (SA) is more severe in patients with rheumatoid arthritis, joint surgical issues, or a weakened immune system. Timely diagnosis of SA is crucial for effective treatment. Traditional diagnostic methods such as ELISA, white blood cell counting, blood culture, qPCR, and imaging techniques are often less accurate and time-consuming. Researchers are focusing on developing highly sensitive biosensors for SA using blood-based biomarkers. Procalcitonin is a protein and a well-established biomarker for SA. This research focuses on developing a procalcitonin interdigitated electrode (IDE) biosensor using a probe made of an aptamer and antibody-modified gold nanoparticle (AuNP) complex. The probe was attached to the IDE through an amine linker and then interacted with procalcitonin. AuNPs increased the attachment of the aptamer and antibody to the IDE, enabling the detection of procalcitonin at levels as low as 10 ng/mL, with a linear regression curve ranging from 10 to 100 ng/mL [y = 4.0691x - 2.1887; R2 = 0.9937]. Furthermore, procalcitonin-spiked serum elevated the current level with increasing procalcitonin concentrations, while control performances did not enhance the current, indicating the selective and specific detection of procalcitonin. This AuNP-aptamer-antibody complexed biosensor effectively identifies procalcitonin at low levels and aids in the diagnosis of SA.

Campylobacter: understanding its role as the primary bacterial cause of food-borne illnesses – current state of knowledge

Introduction Campylobacteriosis, caused primarily by Campylobacter jejuni and Campylobacter coli, is a significant public health concern worldwide. Since 2007 it has been the most frequently reported zoonotic disease in humans across the European Union. Campylobacteriosis is also the leading cause of bacterial diarrhea. Aim of the study This review aims to provide a comprehensive overview of the epidemiology, clinical manifestations, transmission dynamics, diagnostic approaches, and prevention strategies associated with Campylobacter infections. Brief description of the state of knowledge The transmission of Campylobacter spp. typically occurs via the fecal-oral route, with contaminated food, especially poultry, and water serving as common sources of infection. Notably, Campylobacter infections exhibit distinct seasonal patterns and demographic trends, with children and young adults being particularly susceptible. Clinical manifestations range from mild gastrointestinal symptoms to more severe complications, such as reactive arthritis, and Guillain-Barré syndrome. Diagnosis usually relies on microbiological testing. Some of these methods require specialized cultivation techniques, with challenges posed by the phenotypic diversity of Campylobacter species. Summary Effective surveillance and prevention strategies are essential for mitigating the burden of campylobacteriosis and its associated sequelae on both individual and population health. Current research focuses on improving preventive measures, underscoring the importance of advancing public health strategies, and further studies into the epidemiology, pathogenesis, and treatment options. These efforts are crucial for effectively addressing Campylobacteriosis and reducing its effects on human health and agriculture.

Designer cellular spheroids with DNA origami for drug screening.

Current in vitro models struggle to balance the complexity of human diseases with suitability for large-scale drug tests. While 3D cultures simulate human tissues, they lack cellular intricacy, and integrating these models with high-throughput drug screening remains a challenge. Here, we introduce a method that uses self-assembling nucleic acid nanostructures decorated living cells, termed NACs, to create spheroids with a customizable 3D layout. To demonstrate its uniqueness, our method effectively creates designer 3D spheroids by combining parenchymal cells, stromal cells, and immune cells, leading to heightened physiological relevance and detailed modeling of complex chronic diseases and immune-stromal interactions. Our approach achieves a high level of biological fidelity while being standardized and straightforward to construct with the potential for large-scale drug discovery applications. By merging the precision of DNA nanotechnology with advanced cell culture techniques, we are streamlining human-centric models, striking a balance between complexity and standardization, to boost drug screening efficiency.

Sweet cherry forms obtained with embryo culture techniques and their study in the Crimean steppe

Sweet cherry forms obtained with embryo culture techniques and their study in the Crimean steppe

Candidiasis in Choloepus sp.-A Review of New Advances on the Disease.

Candidiasis is a significant fungal infection caused by various species of the genus Candida, posing health challenges to a wide range of animals, including Choloepus species (two-toed sloths). This review article aims to provide a comprehensive understanding of candidiasis in Choloepus sp., highlighting the etiology, epidemiology, pathogenesis, clinical manifestations, diagnosis, treatment, and prevention strategies. This article begins by examining the causative agents, primarily focusing on Candida albicans, which is the most commonly implicated species in candidiasis. The epidemiological aspects are discussed, emphasizing the prevalence of candidiasis in wild and captive Choloepus populations and identifying predisposing factors, such as immunosuppression, stress, poor nutrition, and environmental conditions. Pathogenesis is explored, detailing the mechanisms through which Candida species invade host tissues and evade immune responses. Clinical manifestations in Choloepus sp. are described, including oral thrush, cutaneous lesions, and gastrointestinal infections, and their impact on the health and behavior of affected individuals. Diagnostic methods, including culture techniques, histopathology, and molecular assays, are reviewed to highlight their roles in accurately identifying Candida infections. This article also covers treatment options, focusing on antifungal therapies and supportive care tailored to the unique physiology of Choloepus sp. Finally, prevention and management strategies are discussed, emphasizing the importance of maintaining optimal husbandry practices, regular health monitoring, and early intervention to reduce the incidence and impact of candidiasis in Choloepus populations. This review underscores the need for further research to enhance our understanding of candidiasis and improve health outcomes for these unique and vulnerable animals.

PRODUCTION RESPONSE OF CUCUMBER (CUCUMIS SATIVUS L.) PLANTS DUE TO APPLICATION OF Sp-36 AND MANURE

The cucumber plant (Cucumis sativus L.) is a vegetable plant that can be consumed in fresh or processed form. Cucumber production data from 2019-2022 continues to decline. One of the factors causing the decline in cucumber production is cultivation techniques that have not been carried out properly. To increase cucumber production, it is necessary to pay attention to the availability of nutrients. The fertilizer used is SP 36 fertilizer and organic fertilizer. The aim of the research is: To determine the interaction of SP36 fertilizer and manure on the growth and production of the best cucumber plants. The research was carried out in July - August 2023. The design used was a factorial Randomized Block Design (RAK). First factor: SP-36 fertilizer dose consisting of; P1: Control, P2: 150 kg ha-1 equivalent to 60 gpetak-1, P3: 250 kg ha-1 equivalent to 100 gpetak-1, Factor II: manure consisting of: N1: Control, N2: Cow manure 10 tonha-1 is equivalent to 4 kgpetak-1, N3: 10 tonha-1 chicken manure is equivalent to 4 kgpetak-1, Based on the results obtained: The interaction of SP-36 fertilizer and manure has a very real influence on the number of leaves, fruit length, plant height, fruit diameter and fruit weight Keywords: Production, Cucumis sativus L, SP-36, Cow manure

Agrobacterium transformation of Rhodiola sp.: current status and limitations

The study of secondary metabolites has led to the discovery of new drugs for treating human diseases. However, consistent plant supply can be challenging, leading to the use of plant tissue culture techniques such as hairy root culture. Hairy roots have stable genetics, lateral branching, and can produce secondary metabolites, including alkaloids, flavonoids, and terpenoids. Research on hairy roots as a subject began in the late 19th century, and for the last four decades, hairy roots have been utilized for producing secondary metabolites and recombinant proteins. This article focuses on Rhodiola species - genus of perennial plants that belongs to the family Crassulaceae - and its potential as a source of secondary metabolites using hairy root culture techniques. Rhodiola sp. is widely distributed throughout the Arctic regions of the Northern Hemisphere, with several species having significant medicinal properties. The article discusses the possible use of hairy root cultures for the production of Rhodiola secondary metabolites, including salidroside and rosavins, which have demonstrated significant pharmacological activity in various studies. The use of elicitation and genetic engineering techniques to boost secondary metabolite production in Rhodiola hairy roots is also explored. Overall, the article highlights the potential of Rhodiola hairy root cultures as a valuable source of secondary metabolites with medicinal properties. However, despite some studies Rhodiola hairy root induction and culturing still remains highly unexplored.

Spore Production and Inoculum Formulation of Claroideoglomus Etunicatum and its Application in Maize (Zea Mays)

Recently, the demand for AMF (Arbuscular Mycorrhiza Fungi) fertilizer for agriculture, plantation, and forestry in Indonesia has been very high. Meanwhile, the unavailability of an applicable AMF inoculum that's easy to use and inexpensive is one of the constraints on AMF application as a fertilizer in the field. It indicates that the best formulated inoculum must be found. The study aimed to produce AMF spores using conventional and hydroponic system, formulate the best AMF inoculum, and measure the growth response in maize (Zea mays) after applying the formulated inoculum. In the formulation stage, the study was designed using a Completely Randomized Design (CRD) with 1 factor. The factor was the type of formulation. Data were analyzed using SAS software version 9 and further tested using Duncan's Multiple Range Test (DMRT) at 5% alpha (α) level. The results reported that the conventional pot culture technique (CVNE2) treatment that had been inoculated by Claroideoglomus etunicatum produced the highest spore density amount of 350 ± 6.97 per 10 g of zeolites (P<0.05). On the other hand, the hydroponic NFT (NFTE2) treatment that had been inoculated by C. etunicatum obtained the highest AMF percent colonization amount of 98% ± 2.66% (P<0.05). Analysis of variance in the formulation stage showed TZ60F1 treatment had a significant effect (P<0.05) in increasing plant height, the number of leaves, and dry weight of Zea mays, respectively 120 cm ± 1.7 cm; 15 ± 0.84; 26.3 g ± 2.46 g in 3 months after application.

High Burden of Carbapenem-Resistant Organisms Screened by Xpert Carba-R Assay Versus Culture from Intensive Care Patients in Indonesia

The World Health Organization (WHO) considers carbapenem-resistant organisms (CROs) to be critical-level pathogens. Regular screening for high-risk CRO colonization is essential, especially in the ICU. Direct detection of carbapenem-resistant genes is possible using the FDA-approved Xpert Carba-R assay. This study evaluated its reliability compared with the culture technique at a tertiary hospital in Indonesia. A high number of CRO colonization was found using the culture technique and the Xpert Carba-R assay with about 31 and 26 positive results out of 100 total samples, respectively. Both methods detected blaNDM in 11 samples, and the Xpert Carba-R assay detected one sample co-presenting with blaVIM that was not detected by PCR. The Xpert Carba-R assay did not detect the gene in 73 samples following negative results with the culture technique. Fifteen samples were detected gene by the Xpert Carba-R assay though there was no gene by the culture method, showing that the Xpert Carba-R assay demonstrated a high degree of sensitivity in identifying carbapenem-resistance genes. Carbapenem-resistance genes common in Indonesia other than those examined by Xpert Carba-R assay in this study (i.e., blaOXA-23 and blaOXA-24) or non-enzymatic mechanisms may also produce resistance in many colonies without the examined genes. Finally, the Xpert Carba-R assay produced faster findings than the culture technique.

Modification in Automated Aeroponic Indoor Air Purifier (Pavana) and its Feasibility Analysis

Indoor air pollution is becoming an increasingly genuine issue with the progression of chemical-based building materials as they tend to exert pollutants like benzene, formaldehyde, CO, CO2 etc. Lot of researchers have produced various ideas of air purifiers concentrated on removing only one or in some cases more than one type of pollutants. But the issue with them is cost, aesthetics, and efficiency in removing multiple pollutants simultaneously. The easiest solution for this can be found in nature. There are several species of plants capable of removing different air pollutants efficiently. Here in this project, we are trying to design an eco-friendly indoor air purifier using bamboo.in this purifier we will be using an aeroponic system for providing water and nutrients to all the plants, which will be removing the pollutants from the air. Aeroponic is a plants cultivation technique in which the roots hang suspended in the air while nutrient solution is delivered to them in the form of fine mist. Depending upon the observed result

Spheroids and organoids: Their implications for oral and craniofacial tissue/organ regeneration

Spheroids are spherical aggregates of cells. Normally, most of adherent cells cannot survive in suspension; however, if they adhere to each other and grow to a certain size, they can survive without attaching to the dish surface. Studies have shown that spheroid formation induces dedifferentiation and improves plasticity, proliferative capability, and differentiation capability. In particular, spontaneous spheroids represent a selective and efficient cultivation technique for somatic stem cells. Organoids are considered mini-organs composed of multiple types of cells with extracellular matrices that are maintained in three-dimensional culture. Although their culture environment is similar to that of spheroids, organoids consist of differentiated cells with fundamental tissue/organ structures similar to those of native organs. Organoids have been used for drug development, disease models, and basic biological studies. Spheroid culture has been reported for various cell types in the oral and craniofacial regions, including salivary gland epithelial cells, periodontal ligament cells, dental pulp stem cells, and oral mucosa-derived cells. For broader clinical application, it is crucial to identify treatment targets that can leverage the superior stemness of spheroids. Organoids have been developed from various organs, including taste buds, oral mucosa, teeth, and salivary glands, for basic biological studies and also with the goal to replace damaged or defective organs. The development of novel immune-tolerant cell sources is the key to the widespread clinical application of organoids in regenerative medicine. Further efforts to understand the underlying basic mechanisms of spheroids and organoids will lead to the development of safe and efficient next-generation regenerative therapies.

Low-Potassium Fruits and Vegetables: Research Progress and Prospects.

With the increasing number of patients with chronic kidney disease (CKD) and the improved recognition of nutritional therapy, research on low-potassium (LK) fruits and vegetables for CKD patients has gained global attention. Despite its already commercial availability primarily in Japan, public awareness remains limited, and cultivation methods lack a comprehensive strategy. This review offers an extensive examination of the developmental significance, current cultivation techniques, and existing limitations of functional LK fruits and vegetables with the objective of providing guidance and inspiration for their exploitation. Additionally, this review investigates various factors influencing K content, including varieties, temperature, light, exogenous substances, harvest time, and harvest parts, with a focus on optimizing production methods to enhance potassium utilization efficiency (KUE) and decrease the K content in plants. Finally, the review outlines the shortcomings and prospects of research on LK fruits and vegetables, emphasizing the importance of interdisciplinary research (in agriculture technology, medicine, and business) for patients with CKD and the future development of this field.

Short-term cultured tumor fragments to study immunotherapy combinations based on CD137 (4-1BB) agonism

ABSTRACT Biomarkers for cancer immunotherapy are an unmet medical need. The group of Daniela Thommen at the NKI recently reported on novel methodologies based on short-term cultures of patient-derived tumor fragments whose cytokine concentrations in the supernatants and activation markers on infiltrating T cells were associated with clinical response to PD-1 blockade. We set up a similar culture technology with tumor-derived fragments using mouse tumors transplanted into syngeneic immunocompetent mice to test an agonist anti-CD137 mAb and its combinations with anti-PD-1 and/or anti-TGF-β. Increases in IFNγ concentrations in the tissue culture supernatants were detected upon in-culture activation with the anti-CD137 and anti-PD-1 mAb combinations or concanavalin A as a positive control. No other cytokine from a wide array was informative of stimulation with these mAbs. Interestingly, increases in Ki67 and other activation markers were substantiated in lymphocytes from cell suspensions gathered at the end of 72 h cultures. In mice bearing bilateral tumors in which one was excised prior to in vivo anti-CD137 + anti-PD-1 treatment to perform the fragment culture evaluation, no association was found between IFNγ production from the fragments and the in vivo therapeutic outcome in the non-resected contralateral tumors. The experimental system permitted freezing and thawing of the fragments with similar functional outcomes. Using a series of patient-derived tumor fragments from excised solid malignancies, we showed IFNγ production in a fraction of the studied cases, that was conserved in frozen/thawed fragments. The small tumor fragment culture technique seems suitable to preclinically explore immunotherapy combinations.

High-Throughput Mining of Novel Compounds from Known Microbes: A Boost to Natural Product Screening.

Advanced techniques can accelerate the pace of natural product discovery from microbes, which has been lagging behind the drug discovery era. Therefore, the present review article discusses the various interdisciplinary and cutting-edge techniques to present a concrete strategy that enables the high-throughput screening of novel natural compounds (NCs) from known microbes. Recent bioinformatics methods revealed that the microbial genome contains a huge untapped reservoir of silent biosynthetic gene clusters (BGC). This article describes several methods to identify the microbial strains with hidden mines of silent BGCs. Moreover, antiSMASH 5.0 is a free, accurate, and highly reliable bioinformatics tool discussed in detail to identify silent BGCs in the microbial genome. Further, the latest microbial culture technique, HiTES (high-throughput elicitor screening), has been detailed for the expression of silent BGCs using 500-1000 different growth conditions at a time. Following the expression of silent BGCs, the latest mass spectrometry methods are highlighted to identify the NCs. The recently emerged LAESI-IMS (laser ablation electrospray ionization-imaging mass spectrometry) technique, which enables the rapid identification of novel NCs directly from microtiter plates, is presented in detail. Finally, various trending 'dereplication' strategies are emphasized to increase the effectiveness of NC screening.

The role of deadwood substrates in promoting moss growth: Decay class and particle size effects

AbstractPlagiomnium acutum has a high value of landscape application and medicinal value, but there is a lack of related research on propagation and cultivation techniques. The deadwood substrate has rich nutrients and superior water retention properties, which will be conducive to promoting the growth of moss. Nevertheless, the underlying mechanisms by which deadwood influences moss growth are not yet fully unclear. In this study, we pulverized deadwood from five decay classes of Pinus massoniana into three distinct particle sizes. Through a pot experiment, we investigated the effects of decay class and physicochemical properties on the growth and physiology of Plagiomnium acutum, aiming to identify the most suitable growth substrate. The results indicated that both the decay class and particle diameter of deadwood significantly affect the substrate's physicochemical characteristics and the growth indexes of P. acutum, with the decay class exerting a more pronounced effect. The water‐holding porosity, water‐holding capacity, total nitrogen, total phosphorus, total potassium and lignin content of the substrate positively affected the growth of P. acutum, while the bulk density, void ratio, total carbon, carbon‐to‐nitrogen ratio, condensed tannin content and cellulose content had negative impacts. A comprehensive evaluation using a fuzzy membership function indicated that deadwood with higher decay classes (IV and V) was more conducive to the growth of P. acutum. Specifically, substrates from decay class IV with particle sizes of 10–20 mm provided the most favorable conditions for P. acutum and were recommended as the optimal cultivation substrate. The results of this study provide theoretical basis and technical support for the propagation and cultivation of P. acutum, and provide a foundation for further development of the industrial, pharmaceutical and environmental biotechnology potential of P. acutum.

The Effect of Socio-Economic Factors and Crop Cultivation Techniques on the Increase in Shallot Plant Productivity Using SEM Method

The success of farmers in obtaining productive yields of shallot plants from stable and adaptive superior varieties plays a crucial role in increasing plant productivity. Technological innovations are conducted by examining the Socio-Economic Factors and Technical Factors of Shallot Plant Cultivation. However, merely considering the Socio-Economic Aspects and Technical Cultivation Factors (TCF) of shallot plants is insufficient in selecting stable and adaptive superior varieties. Hence, this research combined the Analysis of Moment Structure (AMOS) method and SEM (Structural Equation Modeling). Through SEM-AMOS, the modeling of socioeconomic factors and Crop Cultivation Techniques was conducted while considering the physiological processes of growth and development of genotypes, which explained the interrelation of Socio-Economic Factors and Crop Cultivation Techniques (CCT) with yield components and their effect on the Increase in Shallot Plant Productivity. The results of the AMOS Modeling. If the Probability value (P) > 0.05, then H0 is accepted, and H1 is rejected; if the Probability value (P) < 0.05, then H0 is rejected, and H1 is accepted. Cultivation of Shallot Plants - Socio-Economic Aspect: It was observed that the P-value was 0.094, which is < 0.05. This value is significantly lower than 0.05; therefore, H1 is accepted. Hence, it can be said that socio-economic factors have a positive and significant effect on the cultivation of shallot plants. Cultivation of Shallot Plants - Crop Cultivation Techniques: It was observed that the P-value was 0.019, which is < 0.05. This value is significantly lower than 0.05; therefore, H1 is accepted. Hence, it can be said that there is a positive and significant effect of Crop Cultivation Techniques on Shallot Plants.

Revitalizing gut barrier integrity: role of miR-192-5p in enhancing autophagy via Rictor in enteritis.

Intestinal inflammation and compromised barrier function are critical factors in the pathogenesis of gastrointestinal disorders. This study aimed to investigate the role of miR-192-5p in modulating intestinal epithelial barrier (IEB) integrity and its association with autophagy. A DSS-induced colitis model was used to assess the effects of miR-192-5p on intestinal inflammation. In vitro experiments involved cell culture and transient transfection techniques. Various assays, including dual-luciferase reporter gene assays, quantitative real-time PCR, Western blotting, and measurements of transepithelial electrical resistance, were performed to evaluate changes in miR-192-5p expression, Rictor levels, and autophagy flux. Immunofluorescence staining, H&E staining, TEER measurements, and FITC-dextran analysis were also used. Our findings revealed a reduced expression of miR-192-5p in inflamed intestinal tissues, correlating with impaired IEB function. Overexpression of miR-192-5p alleviated TNF-induced IEB dysfunction by targeting Rictor, resulting in enhanced autophagy flux in enterocytes (ECs). Moreover, the therapeutic potential of miR-192-5p was substantiated in colitis mice, wherein increased miR-192-5p expression ameliorated intestinal inflammatory injury by enhancing autophagy flux in ECs through the modulation of Rictor. Our study highlights the therapeutic potential of miR-192-5p in enteritis by demonstrating its role in regulating autophagy and preserving IEB function. Targeting the miR-192-5p/Rictor axis is a promising approach for mitigating gut inflammatory injury and improving barrier integrity in patients with enteritis.NEW & NOTEWORTHY We uncover the pivotal role of miR-192-5p in fortifying intestinal barriers amidst inflammation. Reduced miR-192-5p levels correlated with compromised gut integrity during inflammation. Notably, boosting miR-192-5p reversed gut damage by enhancing autophagy via suppressing Rictor, offering a potential therapeutic strategy for fortifying the intestinal barrier and alleviating inflammation in patients with enteritis.

Minor Millets: Miracle Grain of South India

Minor millets are important traditional food crops for dry land farmers in India. India has the third largest area under minor millets cultivation in the world. Minor millets such as finger millet, foxtail millet, proso millet, kodo millet, little millet and barnyard millet are considered as a climate-smart crop. Since these crops are widely divergent for thermo and photoperiods and hence these are known for their climatic resilience and are relatively less prone to biotic stress factors. Added these crops are rich in protein, fat, crude fiber, iron and other minerals and vitamins with low glycemic index in comparison to cereals like wheat and rice. Hence they are labelled as nutri- cereals and act as the source of food, feed and fodder. Minor millets have curative and nutrient - rich properties in the form of antioxidants which prevent weakening of human health such as lowering blood pressure, risk of heart disease, obesity, prevention of cancer, cardiovascular diseases and diabetes etc. Owing to its health benefits, the minor millets output has increased recently due to the availability of high-yielding varieties, pest and disease - resistant varieties, and improved cultivation techniques, rising by 1.05 percent annually. This review aims to document the production status, demand, varietal diversity, current consumption pattern, processing, nutrient composition and therapeutic potential of minor millets for further research on various aspects.

In vitro Propagation of <i>Citrus aurantifolia</i> and its Conservation in Pakistan

Citrus varieties are propagated by both asexual and sexual methods. Generally, rootstocks are propagated sexually through seeds, while most of the commercial varieties are propagated by various asexual methods such as grafting and budding. The technology called plant tissue culture is used for commercial production of plants that are microbe and virus free and to save germplasm of infrequent or threatened species of plants. Fast reproduction techniques have the potential to produce plants free of viruses. Likewise, traditional propagation methods are old, time consuming and are difficult. Tissue culture techniques are assumed for reliable commercial production of economically important plants. In present study shoot tips of Citrus aurantifolia were collected from the field of BCI, NARC. Different disinfectant (Ethanol and NaOCl) with the concentration ranging from 10 to 20 percent were used for culture establishment. Maximum survival percentage was observed in 10% NaOCl. After 5 weeks, established cultures were shifted to hormonal growth media (BAP, GA3 and NAA) at the concentration ranging from 0.1 to 0.8 mg/L. Our finding results that maximum phenotypic characters (plant height, number of leaves and number of shoots were observed on BAP at the concentration of 0.1 to 0.3 mg/L, while maximum number of roots were observed on NAA at the concentration of 0.5mg/L. In present study, for in vitro conservation different conservants like sorbitol and mannitol (10, 20 and 30 g/L) were used. Incubation of cultures was maintained at 21°C under the light intensity of 1000 lux. Old cultures of Citrus aurantifolia were shifted in conservant media. Overall, remarkable difference was observed. In present study mannitol at the concentration of (10, 20 and 30 g/L) showed slow growth after 35 days. In future the plants in conservation media will be used for further propagation and for growing them in fields.