Biotechnological Tools Research Articles

Linking the role of melatonin in plant stress acclimatization

Melatonin is a derivative of tryptophan and consists of a conserved domain present ubiquitously from bacteria to higher organisms. Melatonin is an emerging pleiotropic plant growth hormone that is pivotal in coping with various biotic and abiotic stress conditions. It regulates downstream signaling transduction that modulates plants' physiological and biochemical pathways. Melatonin can be externally applied and could strengthen the stress tolerance mechanism of plants by enhancing ROS-scavenging enzymatic and non-enzymatic antioxidant defense systems. Which consequently exhibits lower lipid peroxidation, MDA content, greater enzyme activity, metabolic pool, ion homeostasis, and plasma membrane integrity under stress conditions. In addition, melatonin protects photosynthetic machinery by maintaining cell membrane stability and chlorophyll recovery. It also stimulates the electron transport chain during photosynthesis and D1 protein biosynthesis and maximizes the photochemistry efficiency (Fv/Fm) of photosystem II and photochemical quenching. Besides abiotic stress, melatonin plays a significant role in biotic stress tolerance. This review emphasizes Melatonin as a stress protectant under biotic and abiotic stress. Further, we have highlighted integrating advanced biotechnology tools with recent information for the possible application of melatonin in the current scenario of changing climate.

Square wave voltammetry based electrochemical determination of affinity of cholesterol triethylene glycol modified DNA-aptamers for protoporphyrin IX

Recent advancement in molecular medicine has seen applications of advanced biotechnology tools such as aptamer technology in therapeutics and diagnostics. Aptamer technology has witnessed various approaches including “Click-Chemistry” towards modifying aptamer structure to improve its potentials, but limited studies have reported the influence of such alteration on aptamer's specificity and affinity for their targets. Here, we utilized square wave voltammetry (SWV) electrochemical sensing based on heme to show the effects of cholesterol-triethylene-glycol (COL-TEG) modification of protoporphyrin-IX DNA-aptamers (OKA_24 and OKA_26) on their affinity for heme. Binding was evaluated by immobilizing 5 μM of heme onto cysteamine-glutaraldehyde-coated gold-electrode to construct electrochemical biosensor. Sensing of native/modified-aptamer was achieved by incubating their varying concentrations (9.76 nM - 10 μM) with heme-coated gold-electrode in HKSCM buffer pH 5, for 15 min. Chloroquine (2.5 μM) and non-binding HPIX-aptamer (2.5 μM) served as controls. Ferrocene was the redox solution used for SWV analysis. Protoporphyrin-IX DNA-aptamers specificity for heme was not tarnish by lipid conjugation. Selective binding of 2.5 μM of COL-TEG-OKA_24 and COL-TEG-OKA_26 to heme induced peak-current reduction by 30.68% and 24% respectively. Incubation of OKA_24 and OKA_26 aptamers produced resistance to current flow through the heme-coated gold-electrode by 23.21% and 14.4 8% respectively. Affinity SWV reveals that cholesterol conjugation decreases the affinity of COL-TEG-OKA_24 (KD = 4 7.13 ± 3.767 nM) and COL-TEG-OKA_24 (KD = 84.6 ± 8.7 nM) by 3- fold. There is a need to check the impact of such alteration on inhibition of heme to hemozoin polymerization, a process mediated by Plasmodium falciparum.

Bioprinting technology for the management of diabetic foot disease: Emerging applications, challenges, and prospects

Most conventional therapies have limitations in the repair of complex wounds caused by chronic inflammation in patients with diabetic foot ulcers (DFUs). In response to the demand for more biotechnology strategies, bioprinting has been explored in the regeneration field in recent years. However, challenges remain regarding the structure of complex models and the selection of proper biomaterials. The purpose of this review is to introduce the current applications of bioprinting technology in chronic diabetic foot wound healing. First, the most common application of bioprinting in producing skin equivalents to promote wound healing is introduced; second, functional improvements in the treatment of chronic and difficult-to-heal DFU wounds facilitated by bioprinting applications are discussed; and last but not least, bioprinting applications in addressing unique diabetic foot disease characteristics are summarized. Furthermore, the present work summarizes material selection and correlations between three-dimensional (3D) bioprinting and a variety of biomimetic strategies for accelerating wound healing. Novel, biotechnological tools such as organoids for developing new biomaterials for bioprinting in the future are also discussed.  

The role of biotechnological tools for mitigating abiotic stress in a changing climate – preface

The role of biotechnological tools for mitigating abiotic stress in a changing climate – preface

An improved in-vitro regeneration protocol using scutellum of mature and immature embryos of wheat

Generally, conventional practices take a long time to improve agronomic traits in common wheat (Triticum aestivum L.). However, biotechnological tools provide the best opportunity to enhance crop potential within a limited time. A simple and repeatable protocol for wheat regeneration is the priority for transforming the genetic makeup of elite wheat cultivars. Among 10 types of callus induction media (CIM), CIM4 with supplemented Murashige and Skoog (MS) media+ dicamba (2 mg/l) has 98.22 and 97.33% callus induction efficiency (CIE) in immature and mature scutellum, respectively. Precocious germination, considered a key barrier during callus induction, was fully controlled by excising the embryonic axis. Among 29 different shoot induction media (SIM) assessed in the study, SIM 29 containing zeatin 5 mg/l, 6-benzylaminopurine (BAP) 5 mg/l, 2,4-dichlorophenoxyacetic acid (2,4-D) 0.25 mg/l,1-naphthaleneacetic acid (NAA) 0.25 mg/l, and copper sulfate (CuSO4) 20 mg/l was found most responsive for shoot induction. The combined effect of cytokinin and CuSO4 resulted in an improved wheat regeneration response in two popular Indian wheat varieties, namely HD2967 and HD3171. Rooting was observed at 100% using IAA and IBA @ 0.2 mg/l concentration in root induction media (RIM). The whole in-vitro regeneration protocol from embryo to transfer in the greenhouse took 8 to 9 weeks. The in-vitro regeneration protocol developed in the present study could further be utilized to improve the genetic makeup of elite wheat cultivars utilizing genetic engineering tools.

GOLDEN2‐LIKE transcription factors: A golden ticket to improve crops?

Societal Impact StatementThe human population is expected to reach 9.7 billion in the next 30 years, increasing the strain on our already precarious food system. Climate change is shifting weather patterns, leading to unpredictable and catastrophic events that further threaten the agronomic sector. Plant scientists are implementing biotechnological tools to sustainably increase both the production and nutritional content of our crops. Engineering GOLDEN2‐LIKE (GLK) transcription factors is a promising route to improve photosynthesis, as well as other important agronomical traits, to achieve food security for a growing population under an unpredictable climate.SummaryUsing agricultural biotechnology to increase the photosynthetic efficiency of crops has been a focus of plant science research over the last two decades. Transcription factors coordinate the expression of gene networks that are the basis of plant development and physiological responses and, as such, are good targets to help improve photosynthesis. Among the known plant transcriptional regulators, GOLDEN2‐LIKE transcription factors (GLKs) may be ideal candidates to improve photosynthesis in crops, as they are master regulators of genes associated with photosynthesis and chloroplast biogenesis across a broad diversity of plant lineages. Moreover, recent work has revealed their involvement in environmental response, pathogen defence and development regulation across the plant's whole life cycle. Thus, manipulating GLK expression and activity, alone or likely in combination with other modifications, has clear potential to improve plant development and growth. Here, we review the research into GLK function and discuss the potential of these key transcription factors as biotechnological tools to enhance photosynthetic efficiency and stress tolerance in crops. Additionally, we take advantage of the vast plant genome and transcriptome datasets available to explore the evolutionary history of GLKs across the plant kingdom and discuss the implications for their adoption into crop engineering projects.

Transcriptome architecture of the three main lineages of agrobacteria.

Agrobacteria are a diverse, polyphyletic group of prokaryotes with multipartite genomes capable of transferring DNA into the genomes of host plants, making them an essential tool in plant biotechnology. Despite their utility in plant transformation, genome-wide transcriptional regulation is not well understood across the three main lineages of agrobacteria. Transcription start sites (TSSs) are a necessary component of gene expression and regulation. In this study, we used differential RNA-seq and a TSS identification algorithm optimized on manually annotated TSS, then validated with existing TSS to identify thousands of TSS with nucleotide resolution for representatives of each lineage. We extend upon the 356 TSSs previously reported in Agrobacterium fabrum C58 by identifying 1,916 TSSs. In addition, we completed genomes and phenotyping of Rhizobium rhizogenes C16/80 and Allorhizobium vitis T60/94, identifying 2,650 and 2,432 TSSs, respectively. Parameter optimization was crucial for an accurate, high-resolution view of genome and transcriptional dynamics, highlighting the importance of algorithm optimization in genome-wide TSS identification and genomics at large. The optimized algorithm reduced the number of TSSs identified internal and antisense to the coding sequence on average by 90.5% and 91.9%, respectively. Comparison of TSS conservation between orthologs of the three lineages revealed differences in cell cycle regulation of ctrA as well as divergence of transcriptional regulation of chemotaxis-related genes when grown in conditions that simulate the plant environment. These results provide a framework to elucidate the mechanistic basis and evolution of pathology across the three main lineages of agrobacteria. IMPORTANCE Transcription start sites (TSSs) are fundamental for understanding gene expression and regulation. Agrobacteria, a group of prokaryotes with the ability to transfer DNA into the genomes of host plants, are widely used in plant biotechnology. However, the genome-wide transcriptional regulation of agrobacteria is not well understood, especially in less-studied lineages. Differential RNA-seq and an optimized algorithm enabled identification of thousands of TSSs with nucleotide resolution for representatives of each lineage. The results of this study provide a framework for elucidating the mechanistic basis and evolution of pathology across the three main lineages of agrobacteria. The optimized algorithm also highlights the importance of parameter optimization in genome-wide TSS identification and genomics at large.

The potential of Lecane rotifers in microplastics removal

Dealing with hard-to-degrade plastics pollution of terrestrial and aquatic environments is one of the most urgent problems of the modern world. The smallest fraction (<5 mm) called micro-plastics (MP) has been found everywhere from ice in Greenland, streams, rivers, soil and even in the human placenta. The goal of our research was to assess the ability of rotifers Lecane inermis to remove micro-plastics suspended in the water column. In the experiments we investigated specific interactions between MP, biofilm and rotifers specialized in feeding on biofilm. We hypothesized that MP adhere to the biofilm and after ingestion by rotifers could be extracted from the water in the form of compact conglomerates excreted with fecal pellets. In these experiments, we demonstrated that: (i) the rotifers preferentially ingest microplastics embedded in biofilm, (ii) the presence of microplastics does not affect growth and fecundity of rotifers, and (iii) that MP aggregation is significantly improved by the presence of biofilm, additionally enhanced in the presence of rotifers. Our findings will help to understand the role of micro-grazers, such as L. inermis feeding on biofilm, in the fate of MP in nature. In the longer term, our results could help to develop biotechnological tools for MP removal from the aquatic environment.

Is winter coming? Impact of the changing climate on plant responses to cold temperature.

Climate change is causing alterations in annual temperature regimes worldwide. Important aspects of this include the reduction of winter chilling temperatures as well as the occurrence of unpredicted frosts, both significantly affecting plant growth and yields. Recent studies advanced the knowledge of the mechanisms underlying cold responses and tolerance in the model plant Arabidopsis thaliana. However, how these cold-responsive pathways will readjust to ongoing seasonal temperature variation caused by global warming remains an open question. In this review, we highlight the plant developmental programmes that depend on cold temperature. We focus on the molecular mechanisms that plants have evolved to adjust their development and stress responses upon exposure to cold. Covering both genetic and epigenetic aspects, we present the latest insights into how alternative splicing, noncoding RNAs and the formation of biomolecular condensates play key roles in the regulation of cold responses. We conclude by commenting on attractive targets to accelerate the breeding of increased cold tolerance, bringing up biotechnological tools that might assist in overcoming current limitations. Our aim is to guide the reflection on the current agricultural challenges imposed by a changing climate and to provide useful information for improving plant resilience to unpredictable cold regimes.

A Guide to Cannabis Virology: From the Virome Investigation to the Development of Viral Biotechnological Tools

Cannabis sativa cultivation is experiencing a period of renewed interest due to the new opportunities for its use in different sectors including food, techno-industrial, construction, pharmaceutical and medical, cosmetics, and textiles. Moreover, its properties as a carbon sequestrator and soil improver make it suitable for sustainable agriculture and climate change mitigation strategies. The increase in cannabis cultivation is generating conditions for the spread of new pathogens. While cannabis fungal and bacterial diseases are better known and characterized, viral infections have historically been less investigated. Many viral infection reports on cannabis have recently been released, highlighting the increasing threat and spread of known and unknown viruses. However, the available information on these pathogens is still incomplete and fragmentary, and it is therefore useful to organize it into a single structured document to provide guidance to growers, breeders, and academic researchers. This review aims to present the historical excursus of cannabis virology, from the pioneering descriptions of virus-like symptoms in the 1940s/50s to the most recent high-throughput sequencing reports. Each of these viruses detected in cannabis will be categorized with an increasing degree of threat according to its potential risk to the crop. Lastly, the development of viral vectors for functional genetics studies will be described, revealing how cannabis virology is evolving not only for the characterization of its virome but also for the development of biotechnological tools for the genetic improvement of this crop.

Genome-wide identification and expression profiling of APX gene family under multifactorial stress combinations and melatonin-mediated tolerance in pitaya

Pitaya (Selenicereus undatus L.), an important tropical plant belonging to the Cactaceae family, is rich in antioxidants, dietary fibers and low in calories. Pitaya shows higher tolerance against abiotic stresses; however, various environmental factors lead to excessive production of reactive oxygen species (ROS) persistently that cause injury to the cells. Under vanadium, drought, and heat conditions, plant cells regulate the expression of ascorbate peroxidase (APX, a vital antioxidant enzyme) genes, which play a significant role in detoxifying ROS by catalyzing hydrogen peroxide. In this study, we have identified 65 APX genes (HuAPX) in the pitaya genome and clustered them into 13 subfamilies. Gene structure and motif analysis revealed that HuAPX genes contained well-preserved intron-exon structure and their motif domains also shared the same order within one cluster. Phytohormones-related cis-acting elements including abscisic acid, gibberellin, jasmonic acid, salicylic acid, and auxin were found in abundance in the promoter regions of the HuAPX genes. RNA-seq-based expression analysis of the HuAPX genes unveiled that few genes showed significantly higher expression under single and multifactorial stress combinations of drought, heat, and vanadium. Exogenous applications of melatonin attenuate the vanadium, heat, and drought effects, thus enhancing the growth and development of the pitaya plant, by inducing the HuAPX gene expression efficiently. Moreover, the expression pattern of HuAPX genes were validated through qRT-PCR analysis. Our findings suggest that three candidate genes (HuAPX1/3/6) may play a key role against abiotic stresses by encoding key enzymes related to glutathione metabolism and phenylpropanoid biosynthesis. These results provide valuable information for organizing a new strategy to design the climate-smart pitaya genotypes through conventional and modern biotechnological tools.

Technological profiles, upgrading and the dynamics of growth: Country-level patterns and trajectories across distinct stages of development

We investigate the dynamic and qualitative nature of technological change in 96 countries between 1980 and 2021 from a structuralist technology upgrading perspective. First, drawing from patent data, we map the dynamics of technological knowledge by exploring the growth rates and significance of technology clusters. Second, we explore whether there is a relationship between specific technology clusters and economic growth. Third, we examine whether countries at different levels of development share similar or heterogeneous technology upgrading profiles and how patterns of technology upgrading have changed over time. We use a long-term, technology-level, cross-country patent dataset, and to address the issues identified we apply two complementary analytical methods: Generalized Method of Moments (GMM) panel data analysis and Fuzzy Set Qualitative Comparative Analysis (fs/QCA). We find a significant association between growth dynamics and country-level specific technology clusters that is driven by the ongoing ICT-based technological revolution and enabling nanotechnology, biotechnology and automation tools. Heterogenous trajectories in technological profiles allowed us to distinguish between more productive and less productive technology upgrading profiles at different income levels. Our results suggest that innovation policy should go beyond mission oriented policies focused solely on newly emerging technologies. Instead, it should develop policy mixes conceived as portfolios of missions focused on technology clusters with disparate objectives, requirements and institutional setups.

An inducible CRISPR-Kill system for temporally controlled cell type-specific cell ablation in Arabidopsis thaliana.

The application of the CRISPR/Cas system as a biotechnological tool for genome editing has revolutionized plant biology. Recently, the repertoire was expanded by CRISPR-Kill, enabling CRISPR/Cas-mediated tissue engineering through genome elimination by tissue-specific expression. Using the Cas9 nuclease from Staphylococcus aureus (SaCas9), CRISPR-Kill relies on the induction of multiple double-strand breaks (DSBs) in conserved repetitive genome regions, such as the rDNA, causing cell death of the targeted cells. Here, we show that in addition to spatial control by tissue-specific expression, temporal control of CRISPR-mediated cell death is feasible in Arabidopsis thaliana. We established a chemically inducible tissue-specific CRISPR-Kill system that allows the simultaneous detection of targeted cells by fluorescence markers. As proof of concept, we were able to eliminate lateral roots and ablate root stem cells. Moreover, using a multi-tissue promoter, we induced targeted cell death at defined time points in different organs at select developmental stages. Thus, using this system makes it possible to gain new insights into the developmental plasticity of certain cell types. In addition to enabling tissue engineering in plants, our system provides an invaluable tool to study the response of developing plant tissue to cell elimination through positional signaling and cell-to-cell communication.

The potentialities of omics resources for millet improvement.

Millets are nutrient-rich (nutri-rich) cereals with climate resilience attributes. However, its full productive potential is not realized due to the lack of a focused yield improvement approach, as evidenced by the available literature. Also, the lack of well-characterized genomic resources significantly limits millet improvement. But the recent availability of genomic data and advancement in omics tools has shown its enormous potential to enhance the efficiency and precision faced by conventional breeding in millet improvement. The development of high throughput genotyping platforms based on next-generation sequencing (NGS) has provided a low-cost method for genomic information, specifically for neglected nutri-rich cereals with the availability of a limited number of reference genome sequences. NGS has created new avenues for millet biotechnological interventions such as mutation-based study, GWAS, GS, and other omics technologies. The simultaneous discovery of high-throughput markers and multiplexed genotyping platform has aggressively aided marker-assisted breeding for millet improvement. Therefore, omics technology offers excellent opportunities to explore and combine useful variations for targeted traits that could impart high nutritional value to high-yielding cultivars under changing climatic conditions. In millet improvement, an in-depth account of NGS, integrating genomics data with different biotechnology tools, is reviewed in this context.

Quality Enhancement in Forage Crops of Cereals and Legumes

This article focus on quality improvement in forage crops, specifically grasses, cereals, and legumes. As milk and many other by-products obtained from dairy farms, poultry, etc are getting into demand, high-quality forage for livestock production is well recognized, and efforts to improve forage quality can have significant economic and environmental benefits. And an overview of current strategies for quality improvement, including breeding and genetic selection, management practices, and how these practices affect the quality of forage crops such as silica, lignin, and other phenolic components. Use of different breeding methods more over like synthetic cultivars, recurrent selection, etc. the use of biotechnology tools such as RNAi interference, tissue culture, and marker-assisted selection in the particular crop that has been developed for increasing the nutritional value, proteins that present in forage crops, and to decrease the harmful chemicals. Additionally, the paper discusses the challenges associated with quality improvement in forage crops and potential solutions. And about different types of grasses used for different types of cattle as supplements. Limitations that cause obstruction to improving the quality of forage crops. digestibility and how to increase the quality and protein content in milk and cattle. And nutrition that needs to be present in prescribed quantities in forage crops. Overall, this paper highlights the need for continued research and innovation in the field of forage crop quality improvement to support sustainable agriculture and meet the increasing demand for high-quality livestock feed.

Inoculation with extreme endophytes improves performance and nutritional quality in crop species grown under exoplanetary conditions.

Technological advances have made possible long space travels and even exoplanetary colonies in the future. Nevertheless, the success of these activities depends on our ability to produce edible plants in stressful conditions such as high radiation, extreme temperatures and low oxygen levels. Since beneficial microorganisms, such as fungal endophytes from extreme environments, have helped agriculture cope with those difficulties, endophytic fungi may be a putative tool to ensure plant growth under exoplanetary conditions. Additionally, growing crops in polyculture has been shown to increase productivity and spatial efficiency, which is essential given the likely space restrictions in such conditions. We evaluated the effect of the inoculation with a mix of two fungal endophytes from the Atacama Desert on performance (survival and biomass) and nutritional quality of three crop species (lettuce, chard and spinach) grown under exoplanetary conditions. In addition, we measured the amount of antioxidants (flavonoids and phenolics) as possible mechanisms to cope with such abiotic conditions. The exoplanetary conditions were; high UV radiation, low temperature, low water availability, and low oxygen levels. These crops were put in growing chambers in monoculture, dual culture and polyculture (the three species in the same pot) for 30 days. Our results show that inoculation with extreme endophytes improved survival by ca. 15 - 35% and biomass by ca. 30 - 35% in all crop species. The most evident increase was when grown in polyculture, except for survival in spinach, where inoculated plants had higher survival only in dual culture. Nutritional quality and the amount of the antioxidant compounds antioxidants increased in all crop species when inoculated with the endophytes. Overall, fungal endophytes isolated from extreme environments such as the Atacama Desert, the driest desert in the world, could be a key biotechnological tool for future space agriculture, helping plants cope with environmental stress. Additionally, inoculated plants should be grown in polyculture to increase crop turnover and space-use efficiency. Lastly, these results provide useful insights to face the future challenges of space-farming.

Challenges in Establishing Sustainable Innovation in the Biotechnology Sector: A Case of DNA Labs, Nigeria

Entrepreneurship progresses through challenges, and the trend is even more striking in upcoming industries like medical biotechnology in Nigeria. The modern-day shift from conventional therapeutic approaches to advanced biomedical practices through medical biotechnology is becoming increasingly rapid. While showing outstanding promises and sustainable development, the cost that comes along with the employment, operation, expertise, and maintenance of medical biotechnological tools presents a difficult hurdle that needs curtailment in developing countries. This research used ‘DNA Labs, Nigeria’ as a case study to discover associated barriers affecting sustainable entrepreneurship and innovation in Nigeria's medical biotechnology sector. Finance was identified as the major constraint due to the expensive nature of tools and expertise required to run the services; regulatory policies and arbitrary public perception of biotechnology-based diagnoses were identified, amongst others, as the variable hindrances. DNA Labs, Nigeria picked out alternative revenue-generating schemes and strategized partnerships as possible measures of overcoming the industry's challenges.

Biotechnological Tools to Elucidate the Mechanism of Plant and Nematode Interactions.

Plant-parasitic nematodes (PPNs) pose a threat to global food security in both the developed and developing worlds. PPNs cause crop losses worth a total of more than USD 150 billion worldwide. The sedentary root-knot nematodes (RKNs) also cause severe damage to various agricultural crops and establish compatible relationships with a broad range of host plants. This review aims to provide a broad overview of the strategies used to identify the morpho-physiological and molecular events that occur during RKN parasitism. It describes the most current developments in the transcriptomic, proteomic, and metabolomic strategies of nematodes, which are important for understanding compatible interactions of plants and nematodes, and several strategies for enhancing plant resistance against RKNs. We will highlight recent rapid advances in molecular strategies, such as gene-silencing technologies, RNA interference (RNAi), and small interfering RNA (siRNA) effector proteins, that are leading to considerable progress in understanding the mechanism of plant-nematode interactions. We also take into account genetic engineering strategies, such as targeted genome editing techniques, the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR associated protein 9 (Cas9) (CRISPR/Cas-9) system, and quantitative trait loci (QTL), to enhance the resistance of plants against nematodes.

Thin Cell Layer Tissue Culture Technology with Emphasis on Tree Species

An increased dependency on plant-based resources for food, shelter, and medicinal usage has increased their sustainable and unsustainable exploitation. To use this resource sustainably, plant tissue culture (PTC) is one important technology. Among different PTC techniques, thin cell layer (TCL) technology is a relatively simple and easily adaptable technique for in vitro cultures of plants. This technique uses small explants about 0.5–2 mm in thickness excised from different plant organs. It has been successfully used in the large-scale propagation of vegetables, legumes, and plants with medicinal benefits. TCL technology has proven to be effective in stimulating various organogenic responses when combined with various new methods such as nanotechnology or microtome-based explantation, especially in tree species. It is considered an important tool in plant biotechnology. Although the morphogenetic response per explant is usually higher in conventional explants, the appropriate use of plant growth regulators and geometric factors in TCL has the potential to make it more efficient and beneficial. This article provides an overview of the concept of TCL as applied to different plant species, particularly trees, since there are few, if any, summaries of TCL technology, especially in trees. This review will certainly revitalize this important technology so that it can be used effectively for successful mass propagation in the field of plant tissue culture.

Characterization of a novel endo-1,3-fucanase from marine bacterium Wenyingzhuangia fucanilytica reveals the presence of diversity within glycoside hydrolase family 168

Sulfated fucans attract increasing research interests in recent decades for their various physiological activities. Fucanases are indispensable tools for the investigation of sulfated fucans. Herein, a novel GH168 family endo-1,3-fucanase was cloned from the genome of marine bacterium Wenyingzhuangia fucanilytica. The expressed protein Fun168D was a processive endo-acting enzyme. Ultra performance liquid chromatography-high resolution mass spectrum and NMR analyses revealed that the enzyme cleaved the α-1 → 3 bonds between α-l-Fucp(2OSO3−) and α-l-Fucp(2OSO3−) in sulfated fucan from Isostichopus badionotus, and α-1 → 3 bonds between α-l-Fucp(2OSO3−) and α-l-Fucp(2,4OSO3−) in sulfated fucan from Holothuria tubulosa. Fun168D would prefer to accept α-l-Fucp(2,4OSO3−) than α-l-Fucp(2OSO3−) at subsite +1, and could tolerate the absence of fucose residue at subsite +2. The novel cleavage specificity and hydrolysis pattern revealed the presence of diversity within the GH168 family, which would facilitate the development of diverse biotechnological tools for the molecule tailoring of sulfated fucan.