Plant Breeding Techniques Research Articles

Role of Biotechnology in Improving Crop Yield and Enhancing Food Security

Biotechnology has had a huge impact on agriculture and ultimately enhanced our quality of life by studying, altering, and cloning various crops. Hybrid seeds, synthetic seeds, photosynthesis boosters, stress-resistant plants and crops, bio-fertilizers, and bio-pesticides are a few examples of potential applications. Biotechnology may be useful for managing livestock, preserving agricultural goods, and sustaining present crop yields while using fewer pesticides, herbicides, and fertilisers. In addition to improving traditional plant breeding techniques, biotechnology also provides a better replacement for food that is produced. When used in conjunction with other cutting-edge agricultural technologies, it offers a creative and environmentally friendly solution to meet consumer demand for sustainable agriculture. Biotechnology significantly increases agricultural output by enhancing disease prevention and drought and flood resistance. This not only satisfies the rising need for food but also aids in the financial success of farmers. Techniques based on biotechnology provide effective, cheap solutions to pest control problems. Producers have created a protein that effectively eliminates pest issues with cotton, maize, and potato crops.

Long-distance transport of the pear HMGR1 mRNA via the phloem is associated with enhanced salt tolerance

Grafting is the main asexual propagation method for horticultural crops and can enhance their resistance to biotic or abiotic stress. Many mRNAs can be transported over long distances through the graft union, however, the function of mobile mRNAs remains poorly understood. Here, we exploited lists of candidate mobile mRNAs harboring potential 5-methylcytosine (m5C) modification in pear (Pyrus betulaefolia). dCAPS RT-PCR and RT-PCR were employed to demonstrate the mobility of the 3-hydroxy-3-methylglutaryl-coenzyme A reductase1 (PbHMGR1) mRNA in grafted plants of both pear and tobacco (Nicotiana tabacum). Overexpressing PbHMGR1 in tobacco plants enhanced salt tolerance during seed germination. In addition, both histochemical staining and GUS expression analysis showed that PbHMGR1 could directly respond to salt stress. Furthermore, it was found that the relative abundance of PbHMGR1 increased in heterografted scion, which avoided serious damage under salt stress. Collectively, these findings established that PbHMGR1 mRNA could act as a salt-responsive signal and move through the graft union to enhance salt tolerance of scion, which might be used as a new plant breeding technique to improve resistance of scion through a stress-tolerant rootstock.

Increasing genomic prediction accuracy for unphenotyped full-sib families by modeling additive and dominance effects with large datasets in white spruce.

Genomic selection is becoming a standard technique in plant breeding and is now being introduced into forest tree breeding. Despite promising results to predict the genetic merit of superior material based on their additive breeding values, many studies and operational programs still neglect non-additive effects and their potential for enhancing genetic gains. Using two large comprehensive datasets totaling 4,066 trees from 146 full-sib families of white spruce (Picea glauca (Moench) Voss), we evaluated the effect of the inclusion of dominance on the precision of genetic parameter estimates and on the accuracy of conventional pedigree-based (ABLUP-AD) and genomic-based (GBLUP-AD) models. While wood quality traits were mostly additively inherited, considerable non-additive effects and lower heritabilities were detected for growth traits. For growth, GBLUP-AD better partitioned the additive and dominance effects into roughly equal variances, while ABLUP-AD strongly overestimated dominance. The predictive abilities of breeding and total genetic value estimates were similar between ABLUP-AD and GBLUP-AD when predicting individuals from the same families as those included in the training dataset. However, GBLUP-AD outperformed ABLUP-AD when predicting for new unphenotyped families that were not represented in the training dataset, with, on average, 22% and 53% higher predictive ability of breeding and genetic values, respectively. Resampling simulations showed that GBLUP-AD required smaller sample sizes than ABLUP-AD to produce precise estimates of genetic variances and accurate predictions of genetic values. Still, regardless of the method used, large training datasets were needed to estimate additive and non-additive genetic variances precisely. This study highlights the different quantitative genetic architectures between growth and wood traits. Furthermore, the usefulness of genomic additive-dominance models for predicting new families should allow practicing mating allocation to maximize the total genetic values for the propagation of elite material.

Show me the benefits! Determinants of behavioral intentions towards CRISPR in the United States

Novel plant-breeding techniques such as CRISPR are potentially key tools in the transition to a more sustainable food and agricultural system. For such potential to be realized, greater attention is required to outstanding questions related to public and societal acceptance. The current research thus employed a structural model to elucidate the interplay of factors that influence behavioral intentions towards CRISPR, for a sample of 158 individuals in the United States. By means of partial least-squares structural equation modeling (PLS-SEM), we established the predominance of perceived benefits vis-à-vis perceived risks for explaining behavioral intentions, along with the indirect effects of food technology neophobia, social trust, and environmental worldviews, all transmitted through perceived benefits – and less so, perceived risks. Overall, this offers key insights regarding behavioral intentions towards CRISPR food, highlighting how consumers may be more interested in understanding potential benefits than being dissuaded about any possible risks and signaling ways in which perceptions of benefits might be influenced. Indeed, the strength of the relationship between perceived benefits and behavioral intentions offers a counterpoint to arguments that benefits are not relevant for acceptance as well as marks a potential point of distinction between GM and GE food.

EU regulation of gene-edited plants-A reform proposal.

This article presents a proposal on how the European Union's regulatory framework on genetically modified (GM) plants should be reformed in light of recent developments in genomic plant breeding techniques. The reform involves a three-tier system reflecting the genetic changes and resulting traits of GM plants. The article is intended to contribute to the ongoing debate over how best to regulate plant gene editing techniques in the EU.

Breeding techniques to dispense higher genetic gains.

Plant breeding techniques encompass all the processes aimed at improving the genetic characteristics of a crop. It helps in achieving desirable characteristics like resistance to diseases and pests, tolerance to environmental stresses, higher yield and improved quality of the crop. This review article aims to describe and evaluate the current plant breeding techniques and novel methods. This qualitative review employs a comparative approach in exploring the different plant breeding techniques. Conventional plant breeding techniques were compared with modern ones to understand the advancements in plant biotechnology. Backcross breeding, mass selection, and pure-line selection were all discussed in conventional plant breeding for self-pollination and recurrent selection and hybridisation were employed for cross-pollinated crops. Modern techniques comprise of CRISPR Cas-9, high-throughput phenotyping, marker-assisted selection and genomic selection. Further, novel techniques were reviewed to gain more insight. An in-depth analysis of conventional and modern plant breeding has helped gain insight on the advantages and disadvantages of the two. Modern breeding techniques have an upper hand as they are more reliable and less time consuming. It is also more accurate as it is a genotype-based method. However, conventional breeding techniques are cost effective and require less expertise. Modern plant breeding has an upper hand as it uses genomics techniques. Techniques like QTL mapping, marker assisted breeding aid in selection of superior plants right at the seedling stage, which is impossible with conventional breeding. Unlike the conventional method, modern methods are capable of selecting recessive alleles by using different markers. Modern plant breeding is a science and therefore more reliable and accurate.

Combating hidden hunger caused by wheat and soil-drived zinc deficiency

Essential vitamins, proteins, and microelements are provided by nutrition, but inadequate and nutrient-poor diets can lead to hidden hunger. Zinc deficiency is a significant hidden resource that affects multiple bodily functions, including immune system function, growth, and development. The primary reasons for the prevalence of zinc deficiency in humans are grain-based products with low concentrations and low zinc solubility in the soil. Intensifying plant production and the inability to replace nutrients absorbed in excess from the soil leads to zinc deficiency in the soil. Consequently, substantial reductions in crop yields are observed, along with decreased zinc concentrations in harvested grains. A number of unsustainable strategies, including expensive medical supplements and zinc-enriched flour-based products to address zinc deficiency, are temporary solutions. Additionally, one such strategy is agronomic biofortification, which recommends utilizing water-soluble zinc fertilizers to increase the concentration of zinc in the plant and soil. A more sustainable and cost-effective approach involves employing traditional plant breeding and molecular techniques to develop new zinc-biofortified cultivars. By enriching wheat with zinc, it absorbs 20-40% more zinc from the soil. Here, this paper will discuss the role of zinc deficiency in wheat and soil and its impact on both crop yield and human nutrition, with a particular emphasis on biofortified wheat.

Consumer and regulatory response to climate change-new plant breeding techniques

Climate change is having a significant impact on the global grape and wine sector. We are seeing earlier and more compressed vintages, more extreme weather events, and a warming of temperatures. These are all leading to management changes in both the vineyard and the winery. Overlaying these physical changes are the mega-consumer trends that are demanding more sustainable production patterns. These trends are changing consumer attitudes to many previously held beliefs. Solutions with a trend toward a sustainable and agrochemical-free agriculture and production chain are needed. Technological advances in plant genetic engineering, coupled with the sequencing of the grapevine genome, has enabled new techniques that can rapidly be used to enhance positive characters in grape vines and wine. Changing consumer attitudes have led to a number of regulators reviewing their existing food regulations for genetically modified (GM) food. The debate around the definitions for GM food and whether these are fit for purpose since the emergence of a range of new techniques for genetic modification has the potential to dramatically change the landscape for grape and wine production internationally. In this paper we explore the current regulatory developments, consumer trends and attitudes and the implications for the grape and wine sector as we seek to cope with the demands of climate change and provide a sustainable future for the planet.

Somatic Embryogenesis, Genetic Modification, and GMOs at the Department of Plant Biotechnology and Cytogenetics at the IHAR

AbstractAt the Plant Breeding and Acclimatization Institute, National Research Institute (IHAR), research on plant tissue cultures began in 1970s. Some in vitro methods, such as anther cultures or isolated microspore cultures, have been directly used to produce homozygous lines in plant breeding. These methods are also a prerequisite for genetic manipulation for both scientific and practical purposes. Here, we describe the achievements of the Department of Biotechnology and Cytogenetics over the last few decades in the area of somatic embryogenesis of cereals, genetic modifications, GMO detection and identification, and GMO legislation. This long-term research on plant regeneration systems has led to the development of the first transgenic triticale plants in the world and the first transgenic plants in Poland, which was followed by a multi-generation study of triticale transgene stability. The gene flow of transgenic triticale and transgenic maize investigated in field studies under Polish environmental conditions, as well as the evaluation of triticale pollen flow, provided scientific data for the development of coexistence measures and GMO risk assessment in case of plant authorization for cultivation. Based on the experience gained over the years, a GMO Controlling Laboratory was established to support the official control of GMOs in Poland and the EU. It was later nominated as one of the National Reference Laboratories collaborating with the Joint Research Center (JRC) of the European Commission, providing scientific and technical support in areas related to health and consumer protection. The GMO Controlling Laboratory is accredited by the Polish Center for Accreditation according to the ISO 17025 standard, ensuring high-quality analyses and flexible accreditation scope. It participates in the ongoing debate in Poland and the EU on the legal aspects of new genomic techniques in plant breeding through publications, lectures at scientific conferences, and by providing expertise to relevant bodies.

Knockout of phytoene desaturase gene using CRISPR/Cas9 in highbush blueberry.

Among the New Plant Breeding Techniques (NPBTs), the CRISPR/Cas9 system represents a useful tool for target gene editing, improving the traits of the plants rapidly. This technology allows targeting one or more sequences simultaneously, as well as introducing new genetic variations by homology-directed recombination. However, the technology of CRISPR/Cas9 remains a challenge for some polyploid woody species, since all the different alleles for which the mutation is required must be simultaneously targeted. In this work we describe improved protocols adapting the CRISPR/Cas9 system to highbush blueberry (Vaccinium corymbosum L.), using Agrobacterium-mediated transformation. As a proof of concept, we targeted the gene encoding for phytoene desaturase, whose mutation disrupts chlorophyll biosynthesis allowing for the visual assessment of knockout efficiency. Leaf explants of in vitro-cultured blueberry cv. Berkeley has been transformed with a CRISPR/Cas9 construct containing two guide RNAs (gRNA1 and gRNA2) targeting two conserved gene regions of pds and subsequently maintained on a selection medium enriched with kanamycin. After 4 weeks in culture on the selection medium, the kanamycin-resistant lines were isolated, and the genotyping of these lines through Sanger sequencing revealed successful gene editing. Some of mutant shoot lines included albino phenotypes, even if the editing efficiencies were quite low for both gRNAs, ranging between 2.1 and 9.6% for gRNA1 and 3.0 and 23.8 for gRNA2. Here we showed a very effective adventitious shoot regeneration protocol for the commercial cultivar of highbush blueberry "Berkeley", and a further improvement in the use of CRISPR/Cas9 system in Vaccinium corymbosum L., opening the way to the breeding mediated by biotechnological approaches.

Understanding tobamovirus-plant interactions: implications for breeding resistance to tomato brown rugose fruit virus

The genus Tobamovirus comprises a group of single-stranded RNA viruses that affect a wide variety of vegetables of economic importance. Tobamoviruses express a series of proteins that interact with the plant’s cellular machinery, allowing viral infection; during incompatible interactions, active defense is mediated by host proteins encoded by resistance genes. The genes conferring viral resistance and tolerance in non-susceptible hosts have been studied for their ability to transfer desired resistance traits to different crops. The N gene from Nicotiana spp., the repertoire of Tm genes in Solanum spp., the L locus from Capsicum spp., and TOM genes are the most studied genetic sequences for understanding resistance to tobamoviruses. Through classical plant breeding and genetic engineering techniques, it has been possible to introgress these resistance genes (R) into new species. However, new reports highlight the ability of tobamoviruses to overcome R-mediated defense. One of the most notorious recent cases is the tomato brown rugose fruit virus (ToBRFV). The main characteristic of ToBRFV is its capacity to overcome the resistance mediated by the Tm-22 gene, resulting in a limited repertoire of options to combat the virus. To defeat emerging viruses, it is necessary to apply the knowledge from other tobamoviruses-host relationships and use new technologies such as genome-wide association studies (GWAS) to understand and associate the architecture of resistance genes present in the Solanaceae family for the benefit of plant breeding. Although new genomic tools such as CRISPR systems open the possibility of coping with viral diseases, there are no commercial ToBRFV-resistant tomato varieties. Hence, the world’s leading seed suppliers compete to develop and bring these varieties to market.

Ethylene: Management and breeding for postharvest quality in vegetable crops. A review.

Ethylene is a two-carbon gaseous plant growth regulator that involved in several important physiological events, including growth, development, ripening and senescence of fruits, vegetables, and ornamental crops. The hormone accelerates ripening of ethylene sensitive fruits, leafy greens and vegetables at micromolar concentrations, and its accumulation can led to fruit decay and waste during the postharvest stage. Several strategies of crops management and techniques of plant breeding have been attempted in the last decades to understand ethylene regulation pathways and ethylene-dependent biochemical and physiological processes, with the final aim to extend the produce shelf-life and improve the postharvest quality of fruits and vegetables. These investigation approaches involve the use of conventional and new breeding techniques, including precise genome-editing. This review paper aims to provide a relevant overview on the state of the art related to the use of modern breeding techniques focused on ethylene and ethylene-related metabolism, as well as on the possible postharvest technological applications for the postharvest management of ethylene-sensitive crops. An updated view and perspective on the implications of new breeding and management strategies to maintain the quality and the marketability of different crops during postharvest are given, with particular focus on: postharvest physiology (ethylene dependent) for mature and immature fruits and vegetables; postharvest quality management of vegetables: fresh and fresh cut products, focusing on the most important ethylene-dependent biochemical pathways; evolution of breeding technologies for facing old and new challenges in postharvest quality of vegetable crops: from conventional breeding and marker assisted selection to new breeding technologies focusing on transgenesis and gene editing. Examples of applied breeding techniques for model plants (tomato, zucchini and brocccoli) are given to elucidate ethylene metabolism, as well as beneficial and detrimental ethylene effects.

Catabolic reprogramming of Brassica rapa leaf mesophyll protoplasts during the isolation procedure

The use of leaf mesophyll protoplasts for transformation and genome editing in plants is expected due to the distinctive features of protoplasts, such as cell-wall-free single cells. However, the application of leaf mesophyll protoplasts for molecular breeding is limited because of the recalcitrance of protoplasts to regenerate. We speculated that the primary reason for this recalcitrance is senescence during protoplast isolation before initialization. In this study, we performed profiling, clustering, co-expression analysis, and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis of primary metabolites and transcriptomes of Brassica rapa leaves and leaf mesophyll protoplasts to reveal the reason for senescence. Primary metabolite profiling indicated that the metabolism of B. rapa protoplasts was converted to catabolism. The number of downregulated differentially expressed genes (DEGs) increased during protoplast isolation. An analysis of GO overexpression revealed the activation of genes involved in catabolic and immune system processes and the inactivation of chloroplasts and photosynthetic genes. KEGG pathway analysis showed that the activation of autophagy and proteasomes accounted for senescence-associated proteolysis during protoplast isolation. It also revealed activation of the genes involved in endoplasmic reticulum stress responses and disease resistance responses during the first 3 h of isolation. These results suggest that elicitor receptor-mediated signal transduction stimulates the pathogen-associated molecular patterns (PAMPs), which triggers immunity. There were more downregulated than upregulated transcription factors during protoplast isolation. AT5G39610 (ANAC092, ATNAC2) gene expression was significantly activated throughout the entire period of protoplast isolation. ANAC092 was co-expressed with upregulated AT5G26340 (STP13), which encodes a protein with high affinity, hexose-specific/H + symporter. AT1G50030 encodes target of rapamycin (TOR) proteins, and the expression of TOR decreased during protoplast isolation. The presence of ethylene and the inhibition of photosynthesis-related genes by glucose and sucrose promote senescence-associated gene expression of protoplasts. Since a decrease in glucose downregulates glucose TOR signaling, the inactivated TOR signaling will promote catabolic reprogramming, senescence, and autophagy in protoplasts during protoplast isolation. We concluded that the initialization of protoplasts requires the blocking of these complicated crosstalk signaling pathways to prevent the catabolic reprogramming of protoplasts, which will lead the way to new plant breeding techniques.

Molecular Markers in Breeding of Crops: Recent Progress and Advancements

A convectional plant breeder faces the challenge of how to more effectively and efficiently perform selection and accelerate breeding progress to satisfy the requirements of changing demands for crop cultivars. However, with the development and advancement of molecular marker technology, the fate of plant breeding has shifted from year to year. Recently, different types of molecular markers have been developed, and advancements in sequencing technologies have greatly increased plant improvement. To further our understanding of molecular markers, several reviews have been published in recent decades. However, with the advancement of newly emerging technologies and techniques, the reviewers did not discuss several recently emerged technologies and techniques in plant breeding. Therefore, this article is intended to be reviewed as an overview of recent breakthroughs in DNA markers and their applications in breeding of crops for early and senior researchers with little or no experience with molecular markers. The progress made in molecular plant breeding, genetics, genomic selection, gene pyramiding, MAS, and gene mapping has contributed to a deeper understanding of molecular markers, provided deeper insights into the variability available for crops, and considerably supplemented current breeding techniques. Next-generation sequencing technologies assist in the identification of novel molecular markers for complex and unstructured populations through genotyping-by-sequencing, gene mapping, QTL mapping, and association mapping. Altogether, the classification of molecular markers and their potential application in plants are discussed.

Somatic Embryogenesis of Soybean Glycine Max (L.) Merr.

One of the efforts to increase domestic production is improving superior soybean through plant breeding techniques. Improving superior soybean cannot be separated from the regeneration process. Regeneration via somatic embryo induction has the advantage of being bipolar. Types of explants, genotypes, media composition, and growth regulators play an important role in plant in vitro regeneration. This study aims to obtain a regeneration protocol for two varieties of soybeans, namely, Derap I and Devon I, through induction of somatic embryos. This research was conducted at Tissue Culture Laboratory of the Faculty of Agriculture, Andalas University, Padang, from July to November 2020. This study used a completely randomized design (CRD) that was carried out separately in one experiment with 4 levels of 2,4-D, namely, 10 ppm, 20 ppm, 30 ppm, and 40 ppm, which were repeated 5 times. The results showed that the concentration of 2,4-D 20 ppm was the best concentration for percentage of explants that produced somatic embryos in Derap I (35%), and Devon I (95%).

European consumer and societal stakeholders' response to crop improvements and new plant breeding techniques

AbstractThe global demand for providing nutritious, sustainable, and safe diets for a 10 billion population by 2050 while preserving affordability, reducing environmental impacts, and adapting to climate change will require accelerating the transition to sustainable agri‐food systems. A plausible way to help tackle these challenges is by developing new plant varieties that have improved crop yield, plant nutritional quality, and sustainability (or resilience) traits. However, stakeholders, consumers, and citizens' concerns and appreciation of future‐proofing crops and the acceptability of new plant breeding strategies are not well‐established. These groups are actors in the agri‐food systems, and their views, values, needs, and expectations are crucial in helping to co‐design fair, ethical, acceptable, sustainable, and socially desirable policies on new plant breeding techniques (NPBTs) and the transition to sustainable agri‐food systems. In this study, we engaged with consumer experts and societal stakeholders to consider their perceptions, expectations, and acceptability of improving crops and NPBTs for future‐proofing the agri‐food systems. Our analysis points to a need for governments to take a proactive role in regulating NPBTs, ensure openness and transparency in breeding new crop varieties, and inform consumers about the effects of these breeding programmes and the risks and benefits of the new crop varieties developed. Consumer experts and societal stakeholders considered these strategies necessary to instil confidence in society about NPBTs and accelerate the transition to sustainable agri‐food systems.

The long road to a sustainable banana trade

Societal Impact StatementBananas are the world's most popular dessert fruit and a staple starch crop for millions in low‐ and middle‐income countries. The banana export trade that supplies North America, Europe, and other wealthy nations has a history fraught with exploitation and conflict. The price of cheap bananas has been environmental degradation, violence, and poverty. Only recently have efforts to address the power imbalances in this trade been made. Voluntary certification schemes aim to address multiple sustainability issues, while research into biological control, accelerated plant breeding, and efficient irrigation will help prepare the industry for emerging threats from pests, diseases, and climate change.SummaryBananas are the world's favorite dessert fruit, a staple starch crop for millions, and an important source of income for producers across the tropics and subtropics. Bananas evolved and diversified as giant perennial herbs of open habitats within the humid forests of Southeast Asia and West Oceania and were domesticated around 7000 years BP through a series of hybridization events. This review considers the journey from rainforest riversides to intensively managed monoculture plantations, focussing on the Cavendish banana that comprises nearly the entire global export trade. Climate change increasingly threatens economic sustainability in several major producer regions, requiring responses such as efficient irrigation systems to maintain productivity and water security. Pests and diseases are spreading globally and have severe direct impacts on production as well as indirect impacts via harm to ecological and human health caused by pesticides. New pest and disease management methods employing biological controls and enhancing soil health and new plant breeding techniques must be developed and implemented. The banana production and trade system has been characterized by power imbalances between international firms that own plantations and supply the market and the local agricultural workers who cultivate and harvest the fruit. Voluntary certification schemes have been developed to address the numerous environmental, social, and economic sustainability issues faced by the industry. There are indications, from research on biological disease control to new deals on wages and benefits for banana workers, that change is slowly coming to the global banana trade.

Ethical aspects of GMO regulation in the EU: Regulating new plant breeding techniques as GM has negative effects on sustainability, diversity and inclusion: Regulating new plant breeding techniques as GM has negative effects on sustainability, diversity and inclusion.

Plant breeding plays a crucial role for the EU to live up to its values and promises of sustainability, innovation and diversity and inclusion. The current regulations, however, make it de facto impossible to use new breeding technologies.

Ethical aspects of GMO regulation in the EU: Regulating new plant breeding techniques as GM has negative effects on sustainability, diversity and inclusion

Ethical aspects of <scp>GMO</scp> regulation in the <scp>EU</scp>: Regulating new plant breeding techniques as GM has negative effects on sustainability, diversity and inclusion

Freedom of Choice—Organic Consumers’ Discourses on New Plant Breeding Techniques

In recent years, there have been significant developments in biotechnology, specifically regarding New Plant Breeding Techniques (NPBTs). Such advancements have been driven by the need to develop improved and more sustainable crops while reducing pesticides and fertilisers. NPBTs include a heterogeneous group of methods that allow performing plant mutations more precisely than in genetically modified (GM) technologies, saving time and effort. Although some experts consider NPBTs an opportunity for organic farming expansion, the European Court of Justice in 2018 pronounced against their use in organic farming since all plants obtained by NPBTs should follow the same regulations as Genetically Modified Organisms (GMOs). This study aims to understand consumers’ attitudes and viewpoints towards new breeding techniques. Focus groups and Q methodological approach were used to uncover consensus and divergence among organic consumers in seven selected European countries (Germany, Italy, Latvia, the Netherlands, Spain, Switzerland, United Kingdom). Results of qualitative studies suggest that organic consumers are generally hostile towards NPBTs in organic farming. Using Q methodology, three distinct factors were identified: the “Risk Averse”, the “Technological Optimists”, and the “Socially Concerned”. The results highlight that consumers’ subjective knowledge and understanding of NPBTs diverge from the discourse of NPBTs lobbyists and proponents.