Growing Uses of 2A in Plant Biotechnology

Modern biotechnology enables an organism to produce a totally new product which the organism does not or cannot produce normally through the incorporation of the technology of ‘Genetic engineering’. Biotechnology shows its technical merits and new development prospects in breeding of new plants varieties with high and stable yield, good quality, as well as stress tolerance and resistance. Some of the most prevailing problems faced in agricultural ecosystems could be solved with the introduction of transgenic crops incorporated with traits for insect pest resistance, herbicide tolerance and resistance to viral diseases. Plant biotechnology has gained importance in the recent past for increasing the quality and quantity of agricultural, horticultural, ornamental plants, and in manipulating the plants for improved agronomic performance. Recent developments in the genome sequencing will have far reaching implications for future agriculture. From this study, we can know that the developing world adopts these fast-changing technologies soon and harness their unprecedented potential for the future benefit of human being.

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Plant Biotechnology Workshop for High School Students

HortScience ◽

10.21273/hortsci.33.3.504e ◽

1998 ◽

Vol 33 (3) ◽

pp. 504e-504

Author(s):

Erika Szendrak ◽

Paul E. Read ◽

Jon S. Miller

Keyword(s):

High School ◽

High School Students ◽

Medical Science ◽

Plant Biotechnology ◽

Plant Science ◽

High School Biology ◽

School Students ◽

Subsequent Transformation ◽

Set Up

Modern aspects of many subjects (e.g., computer science and some aspects of medical science) are now taught in many high schools, but the plant sciences are often given short shrift. A collaboration was therefore established with a high school biology program in which pilot workshops could be developed to enable advanced students to gain insights into modern plant science techniques. A successful example is the workshop on plant biotechnology presented in this report. This workshop is simple and flexible, taking into account that most high school biology laboratories and classrooms are not set up for sophisticated plant science/biotechnology projects. It is suitable for from 10 to 30 students, depending upon space and facilities available. Students work in pairs or trios, and learn simple disinfestation and transfer techniques for micropropagation and potential subsequent transformation treatments. Students gain insights into: sterile technique and hygiene; plant hormones and their physiological effects; plant cell, tissue and organ culture; the influence of environmental factors on response of cells and tissues cultured in vitro; and an understanding of the phenomenon of organogenesis and resulting plant growth and development. This workshop has been tested on several classes of students and following analysis, several refinements were included in subsequent iterations. Results of the students' experiments have been positive and instructive, with student learning outcomes above expectations. Further details of the workshop techniques and approach will be presented.

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On the Absence of a Plant Biotechnology Cluster (Dutch)

SSRN Electronic Journal ◽

10.2139/ssrn.1795068 ◽

2011 ◽

Author(s):

Geert Vissers ◽

Ben Dankbaar

Keyword(s):

Plant Biotechnology

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Plant-Derived Nano and Microvesicles for Human Health and Therapeutic Potential in Nanomedicine

Pharmaceutics ◽

10.3390/pharmaceutics13040498 ◽

2021 ◽

Vol 13 (4) ◽

pp. 498

Author(s):

Mariaevelina Alfieri ◽

Antonietta Leone ◽

Alfredo Ambrosone

Keyword(s):

Therapeutic Potential ◽

Biological Activities ◽

Plant Biotechnology ◽

Bioactive Metabolites ◽

Anticancer Activities ◽

Long Distance ◽

In Vivo Models ◽

Therapeutic Tools

Plants produce different types of nano and micro-sized vesicles. Observed for the first time in the 60s, plant nano and microvesicles (PDVs) and their biological role have been inexplicably under investigated for a long time. Proteomic and metabolomic approaches revealed that PDVs carry numerous proteins with antifungal and antimicrobial activity, as well as bioactive metabolites with high pharmaceutical interest. PDVs have also been shown to be also involved in the intercellular transfer of small non-coding RNAs such as microRNAs, suggesting fascinating mechanisms of long-distance gene regulation and horizontal transfer of regulatory RNAs and inter-kingdom communications. High loading capacity, intrinsic biological activities, biocompatibility, and easy permeabilization in cell compartments make plant-derived vesicles excellent natural or bioengineered nanotools for biomedical applications. Growing evidence indicates that PDVs may exert anti-inflammatory, anti-oxidant, and anticancer activities in different in vitro and in vivo models. In addition, clinical trials are currently in progress to test the effectiveness of plant EVs in reducing insulin resistance and in preventing side effects of chemotherapy treatments. In this review, we concisely introduce PDVs, discuss shortly their most important biological and physiological roles in plants and provide clues on the use and the bioengineering of plant nano and microvesicles to develop innovative therapeutic tools in nanomedicine, able to encompass the current drawbacks in the delivery systems in nutraceutical and pharmaceutical technology. Finally, we predict that the advent of intense research efforts on PDVs may disclose new frontiers in plant biotechnology applied to nanomedicine.

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The politics of genetic technoscience for conservation: The case of blight-resistant American chestnut

Environment and Planning E Nature and Space ◽

10.1177/25148486211024910 ◽

2021 ◽

pp. 251484862110249

Author(s):

Jessica C Barnes ◽

Jason A Delborne

Keyword(s):

Participant Observation ◽

Historical Analysis ◽

Plant Biotechnology ◽

American Chestnut ◽

Non Profit ◽

Socially Constructed ◽

Neoliberal Capitalism ◽

Political Outcomes ◽

Genetic Technologies ◽

Genetics And Genomics

Innovations in genetics and genomics have been heavily critiqued as technologies that have widely supported the privatization and commodification of natural resources. However, emerging applications of these tools to ecological restoration challenge narratives that cast genetic technoscience as inevitably enrolled in the enactment and extension of neoliberal capitalism. In this paper, we draw on Langdon Winner’s theory of technological politics to suggest that the context in which genetic technologies are developed and deployed matters for their political outcomes. We describe how genetic approaches to the restoration of functionally extinct American chestnut trees—by non-profit organizations, for the restoration of a wild, heritage forest species, and with unconventional intellectual property protections—are challenging precedents in the political economy of plant biotechnology. Through participant observation, interviews with scientists, and historical analysis, we employ the theoretical lens provided by Karl Polanyi’s double movement to describe how the anticipations and agency of the developers of blight-resistant American chestnut trees, combined with chestnut biology and the context of restoration, have thus far resisted key forms of the genetic privatization and commodification of chestnut germplasm. Still, the politics of blight-resistant American chestnut remain incomplete and undetermined; we thus call upon scholars to use the uneven and socially constructed character of both technologies and neoliberalism to help shape this and other applications of genetic technoscience for conservation.

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Whole-genome sequencing reveals rare off-target mutations in CRISPR/Cas9-edited grapevine

Horticulture Research ◽

10.1038/s41438-021-00549-4 ◽

2021 ◽

Vol 8 (1) ◽

Author(s):

Xianhang Wang ◽

Mingxing Tu ◽

Ya Wang ◽

Wuchen Yin ◽

Yu Zhang ◽

...

Keyword(s):

Whole Genome Sequencing ◽

Genome Editing ◽

Genome Sequencing ◽

Plant Biotechnology ◽

High Specificity ◽

Fruit Trees ◽

Whole Genome ◽

Nucleotide Polymorphisms ◽

Indel Mutation ◽

Target Sites

AbstractThe CRISPR (clustered regularly interspaced short palindromic repeats)-associated protein 9 (Cas9) system is a powerful tool for targeted genome editing, with applications that include plant biotechnology and functional genomics research. However, the specificity of Cas9 targeting is poorly investigated in many plant species, including fruit trees. To assess the off-target mutation rate in grapevine (Vitis vinifera), we performed whole-genome sequencing (WGS) of seven Cas9-edited grapevine plants in which one of two genes was targeted by CRISPR/Cas9 and three wild-type (WT) plants. In total, we identified between 202,008 and 272,397 single nucleotide polymorphisms (SNPs) and between 26,391 and 55,414 insertions/deletions (indels) in the seven Cas9-edited grapevine plants compared with the three WT plants. Subsequently, 3272 potential off-target sites were selected for further analysis. Only one off-target indel mutation was identified from the WGS data and validated by Sanger sequencing. In addition, we found 243 newly generated off-target sites caused by genetic variants between the Thompson Seedless cultivar and the grape reference genome (PN40024) but no true off-target mutations. In conclusion, we observed high specificity of CRISPR/Cas9 for genome editing of grapevine.

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Shoot Regeneration Is Not a Single Cell Event

Plants ◽

10.3390/plants10010058 ◽

2020 ◽

Vol 10 (1) ◽

pp. 58

Author(s):

Patharajan Subban ◽

Yaarit Kutsher ◽

Dalia Evenor ◽

Eduard Belausov ◽

Hanita Zemach ◽

...

Keyword(s):

Gene Family ◽

Single Cell ◽

Shoot Regeneration ◽

Major Gene ◽

Molecular Genetic ◽

Plant Biotechnology ◽

Gene Families ◽

Regeneration Medium ◽

Developmental Processes ◽

Leaf Segments

Shoot regeneration is a key tool of modern plant biotechnology. While many researchers use this process empirically, very little is known about the early molecular genetic factors and signaling events that lead to shoot regeneration. Using tobacco as a model system, we found that the inductive events required for shoot regeneration occur in the first 4–5 days following incubation on regeneration medium. Leaf segments placed on regeneration medium did not produce shoots if removed from the medium before four days indicating this time frame is crucial for the induction of shoot regeneration. Leaf segments placed on regeneration medium for longer than five days maintain the capacity to produce shoots when removed from the regeneration medium. Analysis of gene expression during the early days of incubation on regeneration medium revealed many changes occurring with no single expression pattern evident among major gene families previously implicated in developmental processes. For example, expression of Knotted gene family members increased during the induction period, whereas transcription factors from the Wuschel gene family were unaltered during shoot induction. Expression levels of genes involved in cell cycle regulation increased steadily on regeneration medium while expression of NAC genes varied. No obvious possible candidate genes or developmental processes could be identified as a target for the early events (first few days) in the induction of shoot regeneration. On the other hand, observations during the early stages of regeneration pointed out that regeneration does not occur from a single cell but a group of cells. We observed that while cell division starts just as leaf segments are placed on regeneration medium, only a group of cells could become shoot primordia. Still, these primordia are not identifiable during the first days.

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Plant regeneration and transformation of Trachyspermum ammi using Agrobacterium tumefaciens and zygotic embryos

Journal of Genetic Engineering and Biotechnology ◽

10.1186/s43141-021-00173-8 ◽

2021 ◽

Vol 19 (1) ◽

Author(s):

Masoumeh Nomani ◽

Masoud Tohidfar

Keyword(s):

Agrobacterium Tumefaciens ◽

Plant Regeneration ◽

Genetic Transformation ◽

Zygotic Embryo ◽

Plant Biotechnology ◽

Hplc Method ◽

Zygotic Embryos ◽

Naphthaleneacetic Acid ◽

Transformed Plants ◽

Trachyspermum Ammi

Abstract Background Trachyspermum ammi is one of the key medicinal plant species with many beneficial properties. Thymol is the most important substance in the essential oil of this plant. Thymol is a natural monoterpene phenol with high anti-microbial, anti-bacterial, and anti-oxidant properties. Thymol in the latest research has a significant impact on slowing the progression of cancer cells in human. In this research, embryos were employed as convenient explants for the fast and effectual regeneration and transformation of T. ammi. To regenerate this plant, Murashige and Skoog (MS) and Gamborg's B5 (B5) media were supplemented with diverse concentrations of plant growth regulators, such as 6-benzyladenine (BA), 1-naphthaleneacetic acid (NAA), 2,4-dichlorophenoxyacetic acid (2,4-D), and kinetin (kin). Transgenic Trachyspermum ammi plants were also obtained using Agrobacterium-mediated transformation and zygotic embryos explants. Moreover, two Agrobacterium tumefaciens strains (EHA101 and LBA4404) harboring pBI121-TPS2 were utilized for genetic transformation to Trachyspermum ammi. Results According to the obtained results, the highest plant-regeneration frequency was obtained with B5 medium supplemented with 0.5 mg/l BA and 1 mg/l NAA. The integrated gene was also approved using the PCR reaction and the Southern blot method. Results also showed that the EHA101 strain outperformed another strain in inoculation time (30 s) and co-cultivation period (1 day) (transformation efficiency 19.29%). Furthermore, HPLC method demonstrated that the transformed plants contained a higher thymol level than non-transformed plants. Conclusions In this research, a fast protocol was introduced for the regeneration and transformation of Trachyspermum ammi, using zygotic embryo explants in 25–35 days. Our findings confirmed the increase in the thymol in the aerial part of Trachyspermum ammi. We further presented an efficacious technique for enhancing thymol content in Trachyspermum ammi using Agrobacterium-mediated plant transformation system that can be beneficial in genetic transformation and other plant biotechnology techniques.

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