Article Archive: Volume 2 Issue 1 2023

Effect of Storage Time and Room Temperature on Physicochemical and Geometric Properties of Banana (Musa Spp.) Fruit

  • Tolcha Techane Alemu

1. Introduction Banana(Musa spp.) fruit is the most popular fresh fruit in all over the world. It is the major fruit crop grown in many developed and developing countries [1]. Bananas is the most commonly consumed fruits and it has great importance to small-scale farmers in the developing countries of the tropics and subtropics [2]. [3] stated that banana is the world’s second most important fruit crop after oil palm. About 87% of the entire bananas grown worldwide are produced by small-scale farmers for consumption or sale to local and regional markets [4]. This makes banana to be the prime leading fruit crop in terms of volume and value in the world market [5]. In most world countries both developed and developing production of banana fruits becoming more due to its benefit. In 2020, bananas production for southwest Asia continent countries like Viet Nam was 2.19 million tonnes and its production of increased from 470,000 tonnes in 1971 to 2.19 million tonnes in 2020 growing at an average annual rate of 3.37% [6]. According to [7], in Ethiopia about 107,890.60 hectares of land is covered by fruit crops. From these bananas contributed about 58.59% of the fruit crop area followed by avocados which contributed 16.53% of the area. Fruits and vegetables are universally considered vital elements in a healthy diet. Like other fruits and vegetables, banana fruits are the most popular fruit and one of the world’s most important staple foods, along with rice, wheat, and maize [8]. Not only they provide crucial vitamins and proteins, but also have health-giving qualities [9]. It is a good source of potassium, magnesium, copper, manganese, vitamin C and B6. Banana is also a good source of energy, low in protein and fat content, and has several medicinal properties [10]. Due to its attractive texture, good glucose sugar and flavor bananas are popular by consumers. Banana (Musa spp.) is a highly perishable fruit. The perishability of the banana fruit is attributed to immense physiological changes after harvest [11]. The port of total postharvest loss on bananas is considered to be 30-40 % [12]. Postharvest pest and diseases affect a wide variety of banana fruit that lack appropriate postharvest storage facilities, particularly in developing countries. Infection by fungi and bacteria may occur during the growing season, at harvest time, during handling, storage, transport, and marketing, or even after purchase by the consumer 26.5% of which 56% of the loss occurred at the retail level due to rotting before reaching to consumers in Ethiopia [13]. Temperature is the most important environmental factor that influences the deterioration of harvested climacteric fruit. It has been shown that temperature has a profound effect on the rates of biological reactions [14]. Physical injuries to the produce must be avoided wherever possible during the handling and distribution of fruits [15]. Postharvest losses are one of the concerns of food security and poverty reduction strategies in many developing countries including Ethiopia and Vietnam [16]. Banana fruit contains low fat, an excellent source of dietary fiber, vitamin C, vitamin B6, and manganese[17]. The presence of potassium and fiber in large amounts in bananas also helps to combat atherosclerosis, which can lead to heart attack and stroke [18]. Since banana fruits are highly perishable in nature they undergo many physiological and biochemical changes that lead to ripening and senescence. According to [19] in developing and developed countries banana fruit is considered to be the best of all fruit crops because of its excellent flavor, attractive fragrance, beautiful peel color, delicious taste, and nutritional value. Despite its nutritional and health importance, there is a huge postharvest loss of this fruit due to its perishable nature. However properties and shelf life of bananas can be extended by controlling storage temperature and food industries can use it during scarcity to consume and produce different products. To reduce postharvest loss and to increase the shelf life, efforts are needed to develop postharvest technologies to manage storage temperatures that are not health hazardous and would suit the climatic and socio-economic conditions of countries [20]. Therefore, appropriate temperature management is the most important tool for extending shelf life and maintaining the quality of banana fruit as well preservephysico-chemical and geometric properties. Therefore this study aims to investigate the effect of storage time and room temperature on physico-chemical and geometric properties of banana fruits. 2. MATERIALS AND METHODS Description study of the area The experiment was conducted in the Viet Nam national University of Agriculture (VNUA) at the laboratory of Food Processing and technology. Sample collection and preparation Unripe and free from any defects banana fruits were purchased from the supermarket around VNUA in University Viet Nam from the producer and it was transported into the food science laboratory at VNUA. After it was transported into the laboratory, it put at room temperature and relative humidity (22°C–28°C and 82%–85% RH) according to (Opara et al,.2012; Ahmad et al.,2006). During the determination of physicochemical and geometric properties, banana fruits were cut at 2 cm and hardness and penetration were determined. Experimental design and treatment application The experiment was laid out in a completely randomized design (CRD) with four replications considering storage at room temperature and storage time by using Minitab software version 16. The factors consisted of (1) storage time and (2) room temperature and eighteen banana fruits were used. Data were collected Physiological Weight loss The percentage of physiological weight loss was determined by using the methods of [21]. Weight loss of fruits was calculated from the initial weight of fruits per treatment and at each storage interval period. It was determined using a sensitive balance (type JD2000-2). Fruit weight daily variation from the initial measurement was expressed as a weight loss %. Weight loss (%) = Initial weight − final weight × 100  Initial Weight Hardness and penetration The hardness and penetration with peel and without peel were measured by using a textural analyzer instrument according to [22]. During the measurement of hardness and penetration of banana fruits with … Read more

Strategies for enhancing plant disease resistance: A review of molecular plant pathology insights

  • Gangadhara Donggali1
  • Santhoshini Elango2
  • Sarvesh Kumar3
  • Wajid Hasan4
  • Kiran5

Introduction Emerging technologies and approaches have paved the way for ground breaking advancements in plant disease resistance [1]. The review highlights the transformative potential of tools like CRISPR-Cas9, RNA interference, and the power of bioinformatics in deciphering the complexities of plant-pathogen interactions [2-3]. However, amidst these scientific triumphs lie challenges and ethical considerations. The review addresses the hurdles in implementing these strategies, including regulatory and biosafety concerns, and the imperative need for ethical stewardship in genetic modification and biotechnological research. Throughout the review, we draw upon compelling case studies and practical applications that demonstrate the real-world impact of molecular plant pathology in fortifying crops against diseases. From blight-resistant potatoes to virus-resistant papayas, these success stories underscore the potential of science to safeguard our agricultural systems, this review provides a comprehensive overview of the multifaceted landscape of strategies for enhancing plant disease resistance [4]. We explore the recent advancements and future prospects in the field, with an overarching goal of contributing to the sustainability and resilience of global agriculture. Background Plant diseases have long plagued agriculture, posing a continuous threat to the world’s food production and security. These insidious adversaries, often in the form of bacteria, fungi, viruses, and nematodes, have the capacity to devastate entire crop yields, leading to substantial economic losses and food shortages. The ever-evolving nature of plant pathogens demands a proactive and adaptable approach to disease management. In the quest to mitigate the impact of plant diseases, Molecular Plant Pathology has emerged as a critical scientific discipline [5-7]. This interdisciplinary field marries molecular biology, genetics, microbiology, and plant physiology to unravel the intricate mechanisms that govern plant-pathogen interactions. By deciphering the molecular dialogues between plants and their assailants, researchers have uncovered novel strategies to bolster plant immunity and mitigate the damage caused by pathogens. Challenges Plant disease management is fraught with challenges. Traditional methods, such as the application of chemical pesticides, while effective to some extent, raise concerns about environmental sustainability and the development of pesticide-resistant strains of pathogens. Additionally, global climate change and increased global trade facilitate the spread of plant diseases to new regions, intensifying the need for innovative solutions to protect our crops [8]. The path to devising effective strategies for enhancing plant disease resistance is further complicated by the diverse and rapidly evolving nature of pathogens. As plants and pathogens engage in a perpetual arms race, understanding the intricacies of this biological warfare becomes paramount. This review aims to explore the arsenal of strategies at our disposal, with a particular focus on the molecular-level insights derived from Molecular Plant Pathology. Molecular Mechanisms of Plant Pathogen Interaction Gene-for-Gene Hypothesis At the core of plant-pathogen interactions is the gene-for-gene hypothesis, which posits a specific interaction between a resistance gene in the plant and an avirulence gene in the pathogen. This model, first conceptualized by Harold Flor in the 1940s, has been instrumental in understanding the genetic basis of disease resistance. Plants with specific resistance genes can recognize the products of avirulence genes from pathogens, triggering a defense response [9]. Effector-Triggered Immunity (ETI) Effector-triggered immunity is a critical concept in plant pathology. Pathogens secrete effector proteins to manipulate plant cellular processes and promote infection. However, plants have evolved resistance proteins that can recognize these effectors and activate robust immune responses, often leading to localized cell death to confine the pathogen. This form of immunity is typically race-specific, providing high levels of resistance against particular pathogen strains [10]. Systemic Acquired Resistance (SAR) Another key aspect of plant immunity is systemic acquired resistance (SAR), a whole-plant resistance response that occurs following an initial infection. SAR involves the production of signaling molecules such as salicylic acid, which activates defense genes throughout the plant, conferring resistance against a broad spectrum of pathogens. This mechanism is critical for developing long-lasting and broad-spectrum disease resistance. Genetic Engineering and Plant Disease Resistance Modifying Plant Genomes Advances in genetic engineering have provided powerful tools for enhancing plant disease resistance. Techniques such as gene editing using CRISPR-Cas9 allow for precise modifications of plant genomes. By either introducing new resistance genes or editing existing ones, scientists can create plant varieties with improved resistance to specific pathogens [11]. In this review, we embark on a journey through the multifaceted world of enhancing plant disease resistance. We will dissect the molecular mechanisms that underpin plant-pathogen interactions, highlighting the breakthroughs in our understanding of how plants detect and respond to invaders. Moreover, we will delve into the realm of genetic engineering, where scientists have harnessed the power of molecular biology to fortify plants against a spectrum of diseases. Our exploration extends to the often-overlooked but immensely promising arena of plant microbiomes, where beneficial microbes play a pivotal role in bolstering plant health and immunity. We will also scrutinize emerging technologies and approaches, showcasing their potential to revolutionize disease resistance strategies [12]. The review will not shy away from the ethical and practical considerations surrounding these strategies, including regulatory frameworks and biosafety precautions. Through case studies and practical applications, we will illustrate how these strategies have been put into practice, emphasizing their real-world impact. Role of Microbiomes in Disease Resistance Symbiotic Relationships Plants are not isolated entities but rather exist in complex ecosystems, interacting with a diverse array of microorganisms [13]. These plant-associated microbiomes, including bacteria, fungi, and other microorganisms, can play a significant role in enhancing disease resistance. Symbiotic relationships, such as those between legumes and nitrogen-fixing bacteria, not only provide nutritional benefits but can also strengthen the plant’s defense mechanisms. Harnessing Microbiomes Recent studies have focused on harnessing the potential of these microbiomes to boost plant immunity. For instance, certain soil bacteria can induce systemic resistance in plants, providing broad-spectrum protection against a range of pathogens [14]. Understanding and manipulating these interactions open up new avenues for sustainable disease management strategies. Challenges and Ethical Considerations Biosafety and Regulatory Issues While the potential of these strategies is immense, they come with their own set of challenges. Biosafety concerns, particularly regarding the release of genetically modified organisms (GMOs) into … Read more

Innovations in Sustainable Agriculture: Integrating Technology and Traditional Practices for Crop Improvement

  • Sulochna1
  • Md Zeeshan2
  • Avdhesh Kumar Patel3
  • Nitin Kumar4
  • Meeniga Venkateswarlu5

Introduction In an era marked by the pressing challenges of climate change, population growth, and resource depletion, the imperative for sustainable agriculture has taken center stage on the global agenda. The traditional methods that have sustained agricultural practices for centuries are now being reimagined and complemented by cutting-edge technologies [1-2 ]. The fusion of age-old wisdom with modern innovation has become a beacon of hope, offering a promising solution to the need for resilient and environmentally conscious farming practices. This article delves into the dynamic landscape of sustainable agriculture, where the integration of technology and traditional farming methods intertwines to bring about transformative changes [3]. As the demand for food production rises to meet the needs of a growing population, it is crucial to explore avenues that not only enhance crop yields but also mitigate the environmental impact of agricultural practices. The synergy between technology and tradition holds the key to striking this delicate balance, ensuring that agriculture becomes a sustainable and adaptive force in the face of unprecedented challenges. Throughout the following sections, we will unravel the various innovations that characterize this harmonious integration [4]. From precision farming and sensor technologies that offer real-time insights into crop health to organic farming practices that prioritize ecological balance, the marriage of tradition and technology is reshaping the agricultural landscape. These innovations go beyond mere modernization; they represent a holistic approach to crop improvement, drawing on the strengths of both the past and the present. As we navigate the intricate web of advancements in sustainable agriculture, the goal is clear: to forge a path that not only addresses the immediate challenges faced by farmers but also establishes a foundation for a resilient and sustainable agricultural future. The following sections will illuminate how these integrated approaches contribute to crop improvement, emphasizing the potential for a more sustainable and environmentally conscious global agriculture. Precision farming stands as a beacon of innovation in modern agriculture, leveraging cutting-edge technologies to revolutionize traditional farming practices [5]. At its core, precision farming integrates global positioning systems (GPS), advanced sensors, and sophisticated data analytics to optimize the efficiency of various agricultural processes. This section explores how these technologies, working in tandem, enable farmers to make informed decisions, enhance crop yields, and reduce the environmental footprint of farming operations. a. Real-time Monitoring: Precision farming relies on real-time monitoring to gather crucial data on soil conditions, crop health, and weather patterns. GPS technology provides accurate geospatial information, allowing farmers to precisely locate and understand the nuances of their fields. This information is vital for making data-driven decisions that optimize resource utilization. b. Data-Driven Decision Making: The integration of sensors in precision farming facilitates data collection on a granular level. Soil sensors measure moisture levels and nutrient content, while crop health sensors monitor factors like chlorophyll levels and plant stress. This wealth of data empowers farmers to make informed decisions regarding irrigation, fertilization, and pest control. By tailoring these interventions to the specific needs of each area within a field, farmers can maximize efficiency and minimize resource wastage. c. Optimizing Resource Use: One of the key benefits of precision farming is its ability to optimize the use of resources such as water, fertilizers, and pesticides. By understanding the variability in soil conditions across a field, farmers can implement variable rate applications, delivering resources precisely where and when they are needed. This not only improves crop yields but also reduces the overall environmental impact by minimizing excess use of inputs. d. Environmental Sustainability: The environmental impact of traditional farming practices, such as excessive use of water and chemicals, has raised concerns about long-term sustainability. Precision farming addresses these concerns by promoting a more targeted and efficient use of resources. By minimizing wastage and runoff, precision farming contributes to the conservation of water and the preservation of soil health, aligning agricultural practices with broader sustainability goals. In conclusion, precision farming and sensor technologies represent a paradigm shift in agriculture, enabling farmers to navigate the complexities of modern farming with unprecedented precision [6-9]. The seamless integration of GPS, sensors, and data analytics not only improves crop yields but also positions agriculture as a more environmentally sustainable endeavor, paving the way for a future where technology and tradition work hand in hand to meet the challenges of a changing world. In the pursuit of sustainable agriculture, a harmonious blend of traditional wisdom and modern techniques is exemplified by organic farming and agroecology. Rooted in the principles of ecological balance and environmental stewardship, these practices offer a holistic approach to crop cultivation. This section explores the tenets of organic farming and agroecology, highlighting their emphasis on natural alternatives, crop rotation, and the integration of traditional practices with contemporary innovations. a. Ecological Balance and Biodiversity: Central to organic farming and agroecology is the emphasis on maintaining the ecological balance within agricultural systems. These practices prioritize biodiversity by fostering a symbiotic relationship between crops and the surrounding environment. By eschewing synthetic pesticides and fertilizers, organic farming promotes the health of ecosystems, supporting natural pollinators, beneficial insects, and soil microorganisms. b. Avoidance of Synthetic Inputs: Organic farming rejects the use of synthetic pesticides, herbicides, and fertilizers, relying instead on natural alternatives. This commitment to avoiding harmful chemicals not only safeguards the health of consumers but also prevents the contamination of soil and water. Organic farmers often employ methods such as companion planting and the release of natural predators to manage pests, showcasing the potential for nature-based solutions. c. Crop Rotation and Soil Health: Crop rotation, a practice embedded in traditional farming wisdom, is a cornerstone of organic agriculture. By rotating crops seasonally, the depletion of specific nutrients in the soil is mitigated, and the risk of pest and disease outbreaks is minimized. This cyclical approach to planting contributes to the long-term health and fertility of the soil, promoting sustainable agricultural practices. d. Integration with Modern Techniques: A key strength of organic farming and agroecology lies in their adaptability to modern techniques. Farmers practicing organic methods can integrate technology for … Read more

Review on the Strategies for Enhancing Plant Disease Resistance: Insights from Molecular Plant Pathology

  • Gangadhara Doggalli1
  • Yumkhaibam Sonia Shahni2
  • M. Sekhar3
  • S. Jaya Prabhavathi4
  • G. Malathi5

Introduction Agriculture, the backbone of global food production, faces relentless challenges from various plant pathogens that threaten crop health and productivity. In the quest for sustainable and resilient agriculture, understanding and enhancing plant disease resistance have become imperatives. The discipline of molecular plant pathology has emerged as a powerful tool, providing unprecedented insights into the intricate mechanisms governing plant-pathogen interactions. This article explores strategies for augmenting plant disease resistance by drawing upon the wealth of knowledge derived from molecular plant pathology. The vulnerability of crops to diseases can have profound consequences on food security and economic stability. Plant diseases, caused by bacteria, fungi, viruses, and other pathogens, lead to yield losses, reduced crop quality, and increased reliance on chemical interventions. The escalating global demand for food necessitates innovative approaches to protect crops from the ever-evolving threat of diseases [1-3]. Molecular plant pathology unravels the molecular intricacies of plant-pathogen interactions, providing a foundation for developing targeted strategies for disease management. At the forefront of plant defense is the intricate immune system, which operates at the molecular level. Understanding how plants recognize and respond to pathogens is essential for designing effective strategies for disease resistance. To devise effective strategies for disease resistance, a comprehensive understanding of how pathogens breach plant defenses is crucial [4]. As we embark on this exploration of strategies for enhancing plant disease resistance, it is evident that molecular plant pathology holds the key to unlocking the secrets of plant immunity. It delve into genetic strategies, molecular breeding approaches, omics technologies, and biotechnological interventions that leverage this knowledge to bolster plant defenses and pave the way for a more resilient and sustainable agricultural future [5].  Understanding Molecular Plant Pathology In the intricate dance between plants and pathogens, the field of molecular plant pathology serves as a guiding light, unraveling the molecular intricacies that govern the dynamics of plant diseases. Molecular plant pathology is a multidisciplinary science that employs advanced molecular and genetic techniques to dissect the mechanisms underlying plant-pathogen interactions [6]. The arsenal of tools and techniques in molecular plant pathology empowers researchers to probe the molecular landscape of plant-pathogen interactions with unprecedented precision. Polymerase chain reaction (PCR), next-generation sequencing, and functional genomics are among the key methodologies that have revolutionized our ability to decipher the genetic makeup of plants and pathogens. The advent of genomics has catalyzed a paradigm shift in our understanding of plant diseases. Whole-genome sequencing of plants and pathogens has unearthed crucial insights into the genetic basis of resistance and susceptibility [7]. Understanding the language of genes and proteins is essential for deciphering the molecular dialogue between plants and pathogens. Transcriptomics and proteomics enable the comprehensive analysis of gene expression patterns and protein profiles during infection [8]. Functional genomics provides the means to dissect the role of specific genes in plant defense and pathogen virulence. Through techniques like gene silencing and CRISPR-Cas9 genome editing, researchers can manipulate gene expression to elucidate their functions. Bioinformatics in Molecular Plant Pathology The vast amounts of data generated by genomic and omics technologies necessitate sophisticated bioinformatics tools for analysis. Understanding the computational aspects of molecular plant pathology is integral to harnessing the full potential of genomic information [9-10]. In unraveling the molecular intricacies of plant diseases, molecular plant pathology not only deepens our comprehension of fundamental biological processes but also holds the key to developing targeted strategies for enhancing plant disease resistance. As we delve into the next sections, the focus will shift to specific insights gained through molecular plant pathology, paving the way for innovative strategies to bolster plant defenses against pathogens. Plant Immune System In the intricate tapestry of plant-pathogen interactions, the plant immune system stands as the first line of defense, a complex and finely tuned network of molecular responses designed to recognize, resist, and neutralize invading pathogens. This components and mechanisms that constitute the plant immune system, shedding light on the remarkable ways in which plants have evolved to safeguard themselves against an ever-evolving array of microbial threats. At the heart of the plant immune system lies the ability to recognize potential threats. Plants have evolved an elaborate system for detecting pathogen-associated molecular patterns (PAMPs) and responding swiftly to the presence of invaders. Central to the plant immune response are Pattern Recognition Receptors (PRRs), specialized proteins that act as molecular sentinels on the cell surface. These receptors play a pivotal role in recognizing conserved molecular patterns displayed by pathogens. A detailed exploration of PRRs and their signaling cascades provides insights into the initial stages of the plant immune response. As pathogens adapt, plants respond with a more specific and targeted defense mechanism known as Effector-Triggered Immunity (ETI). The co-evolutionary arms race between plants and pathogens unfolds through the dynamics of ETI. Key players in the plant immune system are Resistance (R) genes, genetic elements that confer resistance to specific pathogen and to explores the diversity of R genes and their role in recognizing pathogen effectors, triggering a robust immune response. The intricate genetics of R genes contribute to the variation in plant resistance across different species. Beyond the immediate site of infection, plants have the ability to mount a systemic response throughout their tissues, known as Systemic Acquired Resistance (SAR). [11-12]. The orchestration of the plant immune system involves complex hormonal signaling networks. Salicylic acid (SA), jasmonic acid (JA), and ethylene (ET) are key players in regulating defense responses and understanding the intricacies of the plant immune system is foundational for developing strategies to enhance plant disease resistance. As we delve into subsequent sections, the focus will shift to the mechanisms employed by pathogens to breach plant defenses and the strategies employed by researchers to fortify the molecular guardians of the plant immune system. Mechanisms of Pathogen Attack As plants fortify themselves with intricate defense mechanisms, pathogens, in a relentless evolutionary arms race, deploy an arsenal of strategies to breach these molecular defenses. Pathogens have evolved diverse mechanisms to breach the physical barriers of plants and the strategies employed by pathogens to penetrate plant … Read more

Advancing Quality and Productivity in Floricultural Crops through CRISPR/Cas9Technology

  • Shreedhar Beese
  • Babita Kanwar
  • S R Dhiman
  • Divesh Thakur

INTRODUCTION The scientific community is quite excited about the new DNA editing technology CRISPR-Cas9It is cheaper, faster, better than earlier DNA modification strategies and offers a multitude of potential uses. Researchers investigating genetics and medicine may add, remove or alter certain DNA sequences, altering portions of the genome, thanks to a breakthrough method called CRISPR-Cas9. It is now the simplest, straightforward, adaptable and exact genome editing method available, It’s generating implications in the field of science. The academic community is quite excited about the CRISPR-Cas9 method because it provides a number of benefits over current genome editing methods, including speed, affordability, precision and efficiency. It is estimated that in the next few decades, there will be 9.8 billion people on the planet, and that the need for food would increase by almost 110% from 2005 levels. The most important and challenging issue at hand is how to mitigate the consequences of global warming while simultaneously ensuring safety of food for the world’s expanding population. Global food production needs to be increased immediately, but agriculture is confronted with a number of challenges, including possible environmental risks, the rise of diseases and pests, global variations in temperature and fast population expansion [1]. Remarkable progress in plant genome editing techniques coupled with the availability of genome sequences for numerous crops has enabled the breeding of any desired characteristic. Recent advances in DNA editing using specific-site nuclease have enabled precise, efficient investigations involving selective transgenic placement, genetic manipulation and reverse genetics. Targeted Double Strand Breaks (DSB) DNA, which promotes repairing genetic mechanisms and are introduced using designed nuclease. Depending on the repair procedure and the accessibility of repair templates, various genetic modifications may be accomplished. Homologous Recombination (HR) and Non-Homologous End Joining (NHEJ) are two different types of repair DSB processes that have been discovered. Additionally, a double-stranded DNA template with precise target implementation is mediated by non-homologous end-joining. The disciplines of biotechnology, basic science, and medicine stand to gain much from the use of biological processes and species engineering. Zinc-Finger nucleases (ZNFs), transcription activator-like effector nucleases (TALENs), and Clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9 technology are genome editing techniques which surfaced in recent times [2]. The first two techniques connect DNA-binding proteins with endonuclease catalytic subunits, therefore introducing targeted double-strand DNA breaks (DSBs) at certain sites. Conversely, Cas9 is a nuclease that targets DNA by following short RNAs that have contrasting bases to Watson’s. Compared to ZNFs and TALENs, CRISPR/Cas9 has shown to be a far more precise and effective genome editing technique in biological research. A successful utilization of the CRISPR/Cas9 framework has been demonstrated in the generation of novel varieties with improved features by targeting several genes in different crops. The model from which CRISPR-Cas9 was derived is occurring naturally DNA editing process which bacteria used as immune response. Bacteria infected with virus grab hold of little bits of the virus DNA and rearrange it inside their DNA to create, what are known as CRISPR arrays. Bacteria can “remember” the viruses (or viruses which are closely related to them). In order to create RNA segments that can attach to particular DNA sequences on the viruses in the event that they launch another attack, the bacteria use CRISPR arrays. After then, the bacteria use an enzyme similar to Cas9 to split DNA, rendering the virus dysfunctional. Working Technique?  CRISPR-Cas9 framework is composed of two fundamental parts which alter DNA. Out of these is an enzyme called Cas9. To enable the insertion or removal of DNA segments, which act as set of “molecular scissors” that can split the strands of DNA at a specific site in the genome. RNA segment also known as guide RNA or gRNA, it is composed of 20-base RNA sequence fragment that has been predesigned. It is situated within a bigger RNA structure. While the structural element binds to DNA, the pre-designed nucleotide “guides” Cas9 to the correct location within the genome. This guarantees that the Cas9 enzyme will cut the DNA at the precise spot. The guide RNA’s job is to recognize and bind to a particular DNA sequence. Complementary RNA bases are present in both target DNA sequence of the genome and the guide RNA. It follows from this that, in theory, the guide RNA should only bind to the targeted region and not to any other regions of the genome [3]. Through the use of the DNA repair system, several genes throughout the genome of a target cell. Cas9 cleaves DNA strands at the exact location in the DNA sequence that the guide RNA indicates. At this juncture, cell attempts to repair the damaged DNA after realizing the extent to which damage has been done. Researchers are able to modify many genes within a target cell’s entire genome by utilizing the DNA repair system. The guide RNA typically has a specific 20-base arrangement. These complement the target sequence of the gene that needs to be changed. But for the gRNA to bind, not all 20 bases have to match. This causes an issue since a sequence that includes, let’s say, 19 of 20 complementary nucleotides show up in another part of the genome altogether. This suggests that the target sequence might not be bound there, or the guide RNA may bind instead. The enzyme Cas9 will then cut at the incorrect spot, allowing a mutation to be inserted there. This mutation might not have any effect on the person at all, but it might affect an important gene or another significant component of the chromosome. Researchers are eager to discover a means to guarantee precise binding and cutting of CRISPR-Cas. This can be accomplished in two different manners: first, by using the knowledge of the genome’s DNA sequence to create more precise guide RNAs and second, by taking into account the “off-target” behavior of various Cas9-gRNA complex types employing a Cas9 enzyme, which can only cut the target DNA’s single strand as opposed to its double strand. This implies that for the cut to happen, two Cas9 enzymes and two guide RNAs must be … Read more