Plant Tissue Culture Techniques Research Articles

Pomegranate plant tissue culture technique - a review

‌Background: The pomegranate, or Punica granatum L., is a member of the "Punicaceae" family, which also includes Punica protopunica and Punica granatum. Originating in Afghanistan and Iran, it has spread over Asia, Africa, and Europe's Mediterranean region. The traditional method of pomegranate propagation is laborious and time-consuming. It does not guarantee healthy, disease-free plants. Its shortcomings include poor success rates, extremely sluggish replication, and a one-year establishing period for new plants. Tissue culture is a technique that can be used to grow plants in all seasons without restrictions. Several pomegranate-growing nations have tried in vitro propagation to generate virus-free, similar-type plants, and it has been widely used for propagating many fruit trees. Research Purpose: The goal of this essay was to examine how to do plant tissue culture method and get familiar with the techniques. with an emphasis on the pomegranate micropropagation. The only aspect of plant tissue culture that may be able to get around these issues is in vitro technology. Therefore, there is plenty of room for employing micropropagation to multiply desired genotypes on a wide scale. Research Method: This review article was conducted based on scientific websites in Persian and English languages, including SID, Magiran, Google Scholar, Web of Science, and Library Studies, Results: Pomegranate plants multiply quickly through micropropagation, producing plants (clones) that are genetically similar. In a short amount of time, a single explant can yield thousands of plants. These plants match exactly. The study provides a standardized protocol for in vitro regeneration of nodal explants, which could be useful in large-scale production of uniform and disease-free plantlets. The process of plant tissue culture reduces the need for importing expensive plant seeds.

Efficacy of Plant Tissue Culture Techniques for Eliminating Black Mulberry Idaeovirus (BMIV) from Infected Black Mulberry (Morus nigra).

Obtaining virus-free plants is a crucial step in disease management that enables reliable and profitable fruit farming. The present study applied various in vitro virus elimination protocols, including apical shoot culture, chemotherapy, thermotherapy, cryotherapy, and their combination, to eliminate black mulberry Idaeovirus (BMIV) from sour black mulberry. First, a shoot tip (0.5-2 mm) culture protocol was optimized, and four ribavirin concentrations (0, 10, 20, and 30 mg/L) were investigated over five weeks as a form of chemotherapy (ch). For the first thermotherapy treatment (Ch + Th1st), chemotherapy treatment was followed by a gradual increase in the temperature (24-33 °C). In another experiment (Th2nd + Ch), in vitro shoots were incubated in the dark for two weeks at two different temperatures (35 ± 1 °C and 37 ± 1 °C, for one week each). Subsequently, the shoot tips were incubated with various ribavirin doses. Finally, cryotherapy (Cr) was used with or without immersing the shoot tips in liquid nitrogen. A two-step RT-PCR was performed to assess the presence of the virus in 7-8-week-old in vitro plants. Th2nd + Ch significantly increased the shoot tip burst and plant survival/morphogenesis compared to the other treatments. Except for the application of cryotherapy, the protocols eliminated BMIV in different proportions, and the highest virus elimination rate (50%) was obtained by applying 30 mg/L ribavirin during the Ch + Th1st treatment. These findings are essential in preventing the dissemination of the virus and enabling the safe movement of germplasm around the world.

Development of In Vitro Regeneration Protocol for Sweet Pepper (Capsicum annuum L.) using Cotyledon as Explant

Plant tissue culture techniques offer a promising avenue for the efficient propagation of sweet pepper (Capsicum annuum L.), a valuable crop with significant agricultural importance. An effective and repeatable in vitro plant regeneration protocol was developed for the CKN-1 (Lemon yellow) and CKN-8 (Red) genotypes of sweet pepper. Utilizing cotyledonary leaf and cotyledonary node explants, various concentrations and combinations of growth regulators were tested for shoot rejuvenation. For both explants, MS medium supplemented with 8 mg/L BAP, 0.02 mg/L NAA, and 0.5 mg/L IAA was found to be most effective for achieving high rates of response and shoot induction. Notably, cotyledonary node explants outperformed cotyledonary leaf explants, displaying a remarkable 80% response within 11-12 days. Additionally, the supplementation of 1 mg/L IBA with MS medium significantly improved root initiation, with cotyledonary node explants exhibiting the highest responsiveness at 83.77%. Throughout all stages, the genotype CKN-8 (Red) consistently outperformed CKN-1 (Lemon yellow), including in the percentage of regenerated plant establishment. J Bangladesh Agril Univ 22(3): 310–316, 2024

Multipurpose Impacts of Silver Nitrate on Direct Organogenesis of Begonia rex cv. DS-EYWA via Transverse Thin Cell Layering (tTCL) Technique

Begonia rex cv. DS-EYWA is an important plant for indoor and outdoor cultivation, and cv. DS-EYWA is a rare unique cultivar with curly, colorful leaves. Due to their importance, applying plant tissue culture techniques for mass and healthy production in a short period of time without seasonal limitation is of immense economic value. Applying several concentrations of silver nitrate (AgNO3) in combination with varied concentrations of cytokinins including 6-benzylaminopurine (BAP), thidiazuron (TDZ) (0, 0.5, 1, 1.5 mgL−1), and 1-naphthaleneacetic acid (NAA) auxin (0, 0.5, 1 mgL−1) via focusing on transverse thin cell layer (tTCL) petiole explants for high-scale production was used to establish an efficient in vitro propagation protocol. Our results showed that even low concentrations (25 mgL−1) can control internal bacterial infection and increase shoot direct regeneration efficiency. A combination of 1.5 mgL−1 BAP, 0.5 mgL−1 NAA, and 25 mgL−1 AgNO3 was the best treatment to increase the number of direct regenerated shoots, and a lower concentration of BAP (0.5 mgL−1) can be suggested for shoot elongation. Elongated shoots were successfully rooted in MS basal medium and acclimatized in a 1:1 peat moss/perlite sterilized pot mixture.

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.

An update on biotechnological intervention mediated by plant tissue culture to boost secondary metabolite production in medicinal and aromatic plants

AbstractSince prehistoric times, medicinal and aromatic plants (MAPs) have been employed for various therapeutic purposes due to their varied array of pharmaceutically relevant bioactive compounds, i.e. secondary metabolites. However, when secondary metabolites are isolated directly from MAPs, there is occasionally very poor yield and limited synthesis of secondary metabolites from particular tissues and certain developmental stages. Moreover, many MAPs species are in danger of extinction, especially those used in pharmaceuticals, as their natural populations are under pressure from overharvesting due to the excess demand for plant‐based herbal remedies. The extensive use of these metabolites in a number of industrial and pharmaceutical industries has prompted a call for more research into increasing the output via optimization of large‐scale production using plant tissue culture techniques. The potential of plant cells as sources of secondary metabolites can be exploited through a combination of product recovery technology research, targeted metabolite production, and in vitro culture establishment. The plant tissue culture approach provides low‐cost, sustainable, continuous, and viable secondary metabolite production that is not affected by geographic or climatic factors. This study covers recent advancements in the induction of medicinally relevant metabolites, as well as the conservation and propagation of plants by advanced tissue culture technologies.

Enhancing salinity tolerance in Salix linearistipularis through advanced plant tissue culture techniques

Salinity is a major environmental challenge that hampers agricultural sustainability by affecting vast expanses of arable land globally, including significant portions of the Songnen Plain, which hosts substantial areas of saline-sodic soil detrimental to most plant life. This study focuses on Salix linearistipularis, a naturally salt-tolerant woody plant endemic to the region, to explore its potential in ecological restoration and soil improvement. Through extensive laboratory experiments, we employed various plant tissue culture methods to ascertain the most effective techniques for regenerating Salix linearistipularis, thereby facilitating its use in combating soil salinization. The research utilized several culture media and hormonal treatments to determine optimal conditions for seed germination, callus induction, and subsequent plant regeneration. Key findings indicate that specific combinations of growth hormones significantly enhance the rate of callus formation and adventitious bud differentiation, essential for the plants survival and growth in saline conditions. Furthermore, the study demonstrates the potential for these methods to produce genetically stable plants capable of thriving in hostile environments. The findings provide a valuable foundation for future applications in ecological restoration projects aimed at mitigating soil salinity issues.

Substrate and fertirigation for acclimatization of Fragaria x ananassa vitroplants

The in vitro plant tissue culture technique is based on the isolation of organs, tissuesor cells, which, by establishing them in aseptic conditions, and culture medium ofdefined composition and incubation, induce physiological responses, cell divisionand morphogenesis. An application of the tissue culture technique ismicropropagation that consists of stages: 0) selection of the stock plant; I) surfacedisinfection of plant material to establish it in culture medium; II) multiplication ofpropagules; III) rooting of shoots, growth of seedlings and preparation fortransplanting to substrate; IV) transfer of micropropagated plants to substrate foracclimatization in greenhouse. The objective was to evaluate the acclimatization ofmicropropagated strawberry Fragaria x ananassa Duch. plants that were establishedin substrates containing perlite and peat alone or combined in different proportions,and that were fertigated with different dilutions of nutrient solution. After 34 days ofacclimatization from a total of 210 plants evaluated in the various substrate andfertigation conditions, all were acclimatized, and the plants that reached the biggestsize were those established in substrates with 50 to 100% peat and were fertigatedwith any dilutions of nutrient solution.

Floriculture and Transgenic

“Genetic engineering has revolutionized floriculture by enabling the modification of plant species and the creation of new flower color varieties (Chandler, Stephen et al, 2010). Despite the global cultivation of transgenic crops, floriculture has been slow to adopt transgenic breeding. "Floriculture, with its cut flower market as a significant component, is on track to achieve a market size of $43.8 billion by 2027, according to Markets and Markets. This industry plays a crucial role in uplifting rural economies through international trade. For instance, floriculture products from Africa find their way to markets in the U.S. and Europe. Similarly, Asian nations such as Japan, China, and India are strengthening their financial positions through the import and export of floriculture commodities (Tanaka, Yoshizu et al., 2009). One of the notable advancements in floriculture has been the creation of new flower color varieties by manipulating pigment metabolic pathways in plants. However, there are challenges. Some flowering plants, despite their high market value, do not realize their full economic potential due to issues like prolonged flowering seasons and diseases." These challenges can be addressed through the production of transgenic plants using genetic engineering and plant tissue culture techniques. The study aims to explore the potential of genetic engineering associated with plant tissue culture techniques in enhancing floriculture, particularly in developing new flower color varieties and improving the production of high-value flowering plants, thereby highlighting the economic impact of floriculture globally.

Unfolding phenolics biosynthetic plasticity of Artemisia alba Turra through plant tissue culture techniques

Unfolding phenolics biosynthetic plasticity of Artemisia alba Turra through plant tissue culture techniques

Influence of Irrigation Water Acidity, Chemical Fertilizer and Propagation Method on the Contents of Nutrient Elements of Strawberry (Fragaria x ananassa Duch.) cv. Festival

The experiment was conducted to study the effect of the reproduction method and acidity of irrigation water and chemical fertilization plus their interactions on the strawberry fruit of the cv. Festival indicators. The percentage of total soluble solids, total acidity, total sugars, ascorbic acid content, and nitrate content were estimated. Results showed the superiority of the method of propagation using the plant tissue culture technique. The ascorbic acid content was 32.73 mg.100 g-1 fresh mass for the method of propagation by runners. Also, the water treatment with pH 5.5 was superior for the total sugars content in fruits 6.72% and the content of ascorbic acid 32.71 to 73 mg.100 g-1 fresh mass compared to the control treatment. As for the triple interaction, the method of propagation with runners and pH water 5.5 and the fertilizer recommendation of macro-elements outperformed in the percentage of T.S.S and carbohydrate content in the strawberry fruits reached 11.70% and 10.41%, respectively, but the total acidity decreased to 2.37 mg.g-1 for the same combination. In contrast, the highest content of ascorbic acid in fruits 44.81 mg.100 g -1 of fresh mass was recorded by using the propagation method of tissue culture technique. Moreover, the lowest nitrate content was 11.19 mg. 100 g -1 dry mass when using the propagation method of runners and pH of irrigation water at 6.5 plus the fertilizer recommendation of macro-elements.

Pengaruh Penambahan Kombinasi Benzyl Amino Purine dan Pepton Terhadap Pertumbuhan Planlet Anggrek Cattleya sp.

Orchidaceae is one type of ornamental plant which is a leading commodity. Cattleya sp. are a potential type of orchid in terms of productivity and high economic value. Conventional Cattleya sp. cultivation is considered less effective because the number of tillers produced is very limited. Therefore an alternative Cattleya sp. cultivation is needed using plant tissue culture techniques or in vitro. The research objective is to determine the effect of the addition of the combination of BAP and peptone on the growth of orchid plantlets Cattleya sp. This experiment was conducted in the Plant Tissue Culture Laboratory of the Faculty of Mathematics and Natural Science Universitas Negeri Malang from August to November 2016. This study used a completely randomized design. The treatment was in the form of ½ MS medium added with a combination of Benzyl Amino Purine (BAP) and Peptone. The explant used was the protocorm of Cattleya sp. 1 month old with a size of ± 0.5 cm. Protocorm was inoculated as much as 10 protocorm on media treatment. The results showed that Peptone can affected the growth of Cattleya sp. plantlets. The ½ MS treatment with the addition of peptone produced the highest number of leaves and the highest plantlet height, while the ½ MS treatment without the addition of BAP and peptone produced the highest number of roots.

Application of plant tissue culture techniques for genetic improvement of Dendrocalamus strictus Nees

Application of plant tissue culture techniques for genetic improvement of Dendrocalamus strictus Nees

Application of plant tissue culture techniques for genetic improvement of Dendrocalamus strictus Nees

Application of plant tissue culture techniques for genetic improvement of Dendrocalamus strictus Nees

Pembekalan Teknik Propagasi Kultur Jaringan Tumbuhan bagi Guru SMA/Sederajat di Samarinda untuk Mendukung Kurikulum Merdeka

Background: The transition of the 2013 curriculum to the Merdeka curriculum requires teachers to be more creative, applicable, and able to stimulate students' critical powers in understanding selected subjects. However, students' elective subjects in Modern Biotechnology, such as Plant Tissue Culture, are poorly understood in practice. This activity aims to improve practical skills in plant propagation techniques by providing plant tissue culture techniques for high school/equivalent teachers in the field of Biology in Samarinda and surrounding areas. Method: This service was carried out by Biology lecturers at FMIPA Mulawarman University and was attended by 22 Biology and agriculture teachers in Samarinda and surrounding areas. Service activities consist of 4 stages, namely the initial stage, theoretical provision stage, practicum stage, and evaluation stage. Results: There was an increase in participants' understanding of plant tissue culture from 'poor' to 'good' level. Evaluation of material presentation and suitability of theory and practice reached the very good category with percentages of 80% and 92%, respectively. Reflections on debriefing showed that 80% of participants thought this activity added insight and was fun. Conclusion: Activities were carried out according to the target and can contribute to the student learning process at school, especially in plant tissue culture.

Optimization of Different Auxin and Cytokinin Combination in Nutrient Medium for Establishment of Optimal in vitro Multiple Plantlet in Ficus carica L. cv Siyah Orak

Ficus carica Linnaeus is a flowering plant under the Moraceae family, usually propagated conventionally from cuttings due to the seeds being non-viable. However, this method is prone to diseases, and pests, time-consuming and space-intensive. Therefore, other methods are needed to overcome these issues. This study was conducted to induce callus and multiple shoots via plant tissue culture techniques enabling mass production of fig plants. Initially, leaf segments of Ficus carica L. cv Siyah Orak were cultured on different MS media strengths (¼, ½, ¾,1 MS) to induce callus. The highest callus means weight was observed on explant cultured in ¾ MS media (875±0.036). Callus was proliferated by subculturing explant into ¾ MS media supplemented with different concentrations of TDZ (0, 0.5, 1.0, 1.5, 2.0, 2.5, 3.0 mg/L). MS media (3/4) supplemented with 2.0 mg/L TDZ (920±0.03) shows the best result for callus proliferation. Callus induction using transverse and longitudinal thin cell layers from nodal segments cultured on different MS media strengths (¼, ½, ¾,1 MS) shows ¼ MS as the optimum media for both tTCL (100±0) and lTCL (96.7±0.15). Friable callus (%) was observed the highest on ½ MS (63.33±0.55) and ¼ MS (76.67±0.50) media for both tTCL and lTCL, respectively. As for the number of leaves produced, both tTCL (0.83±0.0.28) and lTCL (1.00±0.33) explant showed the best results in ¼ MS media. Apical buds produced the highest mean for both the number of leaves and length of the shoot on 1MS media supplemented with 2.0 mg/L BAP (3.5±0.20, 13.73±0.66), respectively. For root formation (%) and number of roots, both show the best results in media supplemented with 2.5 mg/L IAA (10±0.31, 0.83±0.50). It can be concluded that the best shoot growth performance was observed from apical bud cultured on 1MS media supplemented with 2.0 mg/L BAP+ 2.5 mg/L IAA.

Advancement in Tissue Culture Techniques for Fruit Crops

Tissue culture is a highly promising approach that enables the efficient propagation of many plants from tiny fragments of the parent plant within a relatively brief timeframe and confined area. Tissue culture, a contemporary approach, is primarily employed for the efficient and extensive replication of many commercially significant plant species, such as the date palm. Utilizing the tissue culture technique presents a potential approach for generating a substantial quantity of genetically homogeneous palm plants that resemble other plants and yield typical fruit within four years from initial planting. Furthermore, this technique allows to produce date palm plants devoid of diseases, exhibiting an exceptionally high survival rate of nearly 100% when compared to the traditional vegetative propagation of shoots, owing to the robustness of their root system. The process of surface sterilization holds significant importance in the production of explants for in vitro studies, as it effectively addresses the issue of bacterial and fungal contamination originating from field sources, which might vary considerably across different fruit plant species. The efficacy of tissue culture techniques for date palm acclimatization in vitro is contingent upon the observation of leaf count prior to transplantation in the greenhouse. Hence, the primary objective of this study was to investigate the determinants that govern the tissue culture of fruit trees. India is known for being the native land of various fruit crops that are both significant and minor in terms of their importance. These crops include Indian gooseberry (Emblica officinalis Gaertn.), Karonda (Carissa carandas L.), Bael (Aegle marmelos Corr.), Jamun (Syzygium cuminii L.), and jackfruit (Artocarpus heterophyllus L.), etc. These fruits possess considerable nutritional, medicinal, and therapeutic value, making them highly valuable in commercial sectors such as medicine, food, and cosmetics. The limited availability of suitable planting materials imposes constraints on the commercial production process for these crops. Using plant tissue culture techniques holds promise in substantially augmenting the number of novel cultivars or genotypes inside fruit crops. The primary aim of this review study is to consolidate and synthesize the extant body of knowledge about the tissue culture techniques employed in cultivating various fruit crops.

Telosma mosaic virus: An emerging plant RNA virus causing production loss in passion fruit across Asia

AbstractTelosma mosaic virus (TeMV) is a highly infectious and rapidly spreading virus that infects a variety of natural hosts, with passion fruit (Passiflora edulis) being its primary host across parts of Asia. Symptoms of TeMV, which seriously threatens passion fruit production worldwide, include mosaic, deformities and lignification of fruit. We present here a review of the current state of knowledge of TeMV, including its structural characteristics, natural and experimental host range, symptoms and detection, and provide insights into control strategies. The mode of transmission of TeMV remains unclear, but it can be mechanically transmitted through grafting, cuttings and other artificial asexual propagation techniques, which unfortunately constitute the primary means of propagation in the passion fruit industry. Moreover, increasing detection rates of TeMV indicate that this virus probably causes more serious losses to the passion fruit industry than previously reported. No systematic control strategy for TeMV infection currently exists, and no TeMV‐resistant passion fruit varieties are currently available. As a speedy, short‐term solution for dealing with TeMV infection in the absence of a long‐term solution, we propose rapid plant propagation through virus‐free plant tissue culture techniques that can generate a substantial number of virus‐free passion fruit seedlings.

Improving Steviol Glycosides Production in Stevia Rebaudian Bertoni by Plant Tissue Culture Technology

Plant tissue culture techniques have made tremendous progress in the production of various active compounds of interest, including Steviol Glycosides in Stevia. Stevia rebaudiana is a perennial herbaceous plant known for its sweet-tasting leaves that contain active substances used as natural low-calorie sweeteners called Steviol glycosides. Because of its high sweetness and low-calorie nature, it is an alternative to sugar. However, conventional cultivation of Stevia plants for the production of Steviol Glycosides faces challenges such as long growth cycles, changes in chemical composition, and environmental determinants. Therefore, plant tissue culture is an alternative for the production of large quantities of Steviol Glycosides by altering various factors such as nutrient media, growth regulators, and response. Photovoltaic and tissue culture methods have been successfully used for plant propagation as well as the production of high-quality glycosides. Through techniques such as callus culture, organogenesis, and somatic embryo culture, large quantities of cotyledons can be rapidly established from small plant parts in a relatively short time. With further advances in tissue culture techniques and an understanding of the physiology and genetics of stevia, the potential for large-scale production of glycosides using Tissue culture methods is a promising technique for the commercial use of this natural sweetener.

Multifaceted Role of Nanomaterials in Modulating In Vitro Seed Germination, Plant Morphogenesis, Metabolism and Genetic Engineering.

The agricultural practices of breeding, farm management and cultivation have improved production, to a great extent, in order to meet the food demands of a growing population. However, the newer challenges of climate change, global warming, and nutritional quality improvement will have to be addressed under a new scenario. Plant biotechnology has emerged as a reliable tool for enhancing crop yields by protecting plants against insect pests and metabolic engineering through the addition of new genes and, to some extent, nutritional quality improvement. Plant tissue culture techniques have provided ways for the accelerated clonal multiplication of selected varieties with the enhanced production of value-added plant products to increase modern agriculture. The in vitro propagation method has appeared as a pre-eminent approach for the escalated production of healthy plants in relatively shorter durations, also circumventing seasonal effects. However, there are various kinds of factors that directly or indirectly affect the efficiency of in vitro regeneration like the concentration and combination of growth regulators, variety/genotype of the mother plant, explant type, age of seedlings and other nutritional factors, and elicitors. Nanotechnology as one of the latest and most advanced approaches in the material sciences, and can be considered to be very promising for the improvement of crop production. Nanomaterials have various kinds of properties because of their small size, such as an enhanced contact surface area, increased reactivity, stability, chemical composition, etc., which can be employed in plant sciences to alter the potential and performance of plants to improve tissue culture practices. Implementing nanomaterials with in vitro production procedures has been demonstrated to increase the shoot multiplication potential, stress adaptation and yield of plant-based products. However, nanotoxicity and biosafety issues are limitations, but there is evidence that implies the promotion and further exploration of nanoparticles in agriculture production. The incorporation of properly designed nanoparticles with tissue culture programs in a controlled manner can be assumed as a new pathway for sustainable agriculture development. The present review enlists different studies in which treatment with various nanoparticles influenced the growth and biochemical responses of seed germination, as well as the in vitro morphogenesis of many crop species. In addition, many studies suggest that nanoparticles can be useful as elicitors for elevating levels of important secondary metabolites in in vitro cultures. Recent advancements in this field also depict the suitability of nanoparticles as a promising carrier for gene transfer, which show better efficiency than traditional Agrobacterium-mediated delivery. This review comprehensively highlights different in vitro studies that will aid in identifying research gaps and provide future directions for unexplored areas of research in important crop species.