Plant-based Production Research Articles

Plant Flavonoid Production in Bacteria and Yeasts

Flavonoids, a major group of secondary metabolites in plants, are promising for use as pharmaceuticals and food supplements due to their health-promoting biological activities. Industrial flavonoid production primarily depends on isolation from plants or organic synthesis, but neither is a cost-effective or sustainable process. In contrast, recombinant microorganisms have significant potential for the cost-effective, sustainable, environmentally friendly, and selective industrial production of flavonoids, making this an attractive alternative to plant-based production or chemical synthesis. Structurally and functionally diverse flavonoids are derived from flavanones such as naringenin, pinocembrin and eriodictyol, the major basic skeletons for flavonoids, by various modifications. The establishment of flavanone-producing microorganisms can therefore be used as a platform for producing various flavonoids. This review summarizes metabolic engineering and synthetic biology strategies for the microbial production of flavanones. In addition, we describe directed evolution strategies based on recently-developed high-throughput screening technologies for the further improvement of flavanone production. We also describe recent progress in the microbial production of structurally and functionally complicated flavonoids via the flavanone modifications. Strategies based on synthetic biology will aid more sophisticated and controlled microbial production of various flavonoids.

Opportunities and Challenges for Market Oriented Lupin (Lupinus spp.) Production in Ethiopia

Over recent years, the demand for livestock products in Ethiopia is increasing; most of this demand is coming from urban centers, especially from Addis Ababa, as the buyers are demanding better quality and more hygienic produce. This has fueled interest in better quality animal feed, mainly from larger-scale commercial livestock producers. With this background, there has been interest in stimulating plant-based protein production that can be incorporated into livestock feed. Small quantities of lupins are grown (less than 20,000 ton per year) in the North Western part of Ethiopia for human consumption. These lupins are bitter due to their alkaloid concentration and are unsuitable for livestock feed. If non-bitter (sweet) varieties are grown, then they could be a source of protein for animals as is the case in some countries, such as Australia. The Ethiopian market for livestock feed was surveyed and segmented to evaluate the opportunities for lupins. The survey revealed that the livestock sector in Ethiopia is suffering from feed shortage both in terms of quality and quantity. Lupin, as a potential protein source feed has an opportunity to be incorporated into the livestock feed industry. Currently it is estimated that if sweet lupins were available, the short-term demand from commercial livestock producers and feed processors market segment is 20,000 t/year. In the medium-term, it is expected that as the national poultry production expands, there will be an increased demand from the large feed mill factories to around 35,000 to 40,000 t/year grain for high quality protein such as lupin. It is recommended that the already started sweet lupin promotion in the country has to be done at scale. In addition, production of sweet lupin in Ethiopia should be focused on the commercial farming sector and the marketing should be through the large-scale processors or direct to large-scale livestock producers.

Food Processing Technologies Aspects on Plant-Based Milk Manufacturing: Review

The emergence of immune and non-immune related diseases such as lactose intolerance and milk allergy has contributed to the milk consumption pattern changes over the last decades. Moreover, the problem with calorie concern, a saturated fatty acid in dairy milk, and the emergence of vegan diets have further shifted consumer choices to dairy milk alternatives. Plant-based milk is the liquid part extracted from plants such as legumes, nuts, cereals, seeds, and pseudo cereals. It is regarded as a replacement for dairy milk and perceived as a healthier product due to the presence of bioactive compounds. However, the nutritional content, sensory properties, and stability of the products are the main drawbacks of plant-based milk, as these properties are very different compared to dairy milk. Application of food processing technologies in plant-based milk production have been applied to ensure the quality of the final product. New technologies such as non-thermal processing have been applied to address issues related to the quality of plant-based milk. This article reviewed the technological aspects of plant-based milk production, including advanced processing technologies.

Protein Extraction from Spirulina Platensis with The Cellulase Enzyme Assistance

Microalgae is a relatively new, sustainable source of protein supplements. Among various microalgae strains, Spirulina platensis has shown an effective protein potential source compared with others. A high-performance protein recovery technology is required for this prospect. When the cellulase enzyme supported the recovery process, the condition to get the highest protein recovery efficiency (40.13±2.87%) were 50 UI/g dry algae for enzyme activity, 1:20 for the ratio of dry algae: solvent, 7.0 for pH value with a process temperature of 50oC for 90 minutes. This study succeeded in using cellulase enzymes to support protein extraction with high recovery efficiency from Spirulina platensis grown in Vietnam, opening up an additional source of protein in the trend of plant-based meat production.

Modification of Pea Starch Digestibility through the Complexation with Gallic Acid via High-Pressure Homogenization.

Pea starch and some legume starches are the side streams of plant-based protein production. Structural modification toward moderate digestibility and desirable functionality is a way to increase the economic values of these side-stream starches. We applied an innovative and sustainable technique, high-pressure homogenization, to alter pea starch structure, which resulted in a high level of complexation with the small phenolic acid molecule, gallic acid, to alter starch digestibility. This study showed a great level of disruption of the compact starch structure represented by the decrease in gelatinization temperature, enthalpy change, and relative crystallinity. The addition of a high concentration (10%) of gallic acid contributed to a typical V-type X-ray diffractometry pattern. Data demonstrated a significant decrease (~23%) in the susceptibility to α-amylase and an increase in resistant starch (~13%). In addition, starch functionality was improved with a reduced retrogradation rate. Pea starch responded to the high-pressure homogenization process well. Compared with the rice and maize starch reported in the literature, pea starch required a reduced amount of gallic acid to form a high level of complexation with a significant delay in starch digestion.

Rational domestication of a plant-based recombinant expression system expands its biosynthetic range.

Plant molecular farming aims to provide a green, flexible, and rapid alternative to conventional recombinant expression systems, capable of producing complex biologics such as enzymes, vaccines, and antibodies. Historically, the recombinant expression of therapeutic peptides in plants has proven difficult, largely due to their small size and instability. However, some plant species harbour the capacity for peptide backbone cyclization, a feature inherent in stable therapeutic peptides. One obstacle to realizing the potential of plant-based therapeutic peptide production is the proteolysis of the precursor before it is matured into its final stabilized form. Here we demonstrate the rational domestication of Nicotiana benthamiana within two generations to endow this plant molecular farming host with an expanded repertoire of peptide sequence space. The in planta production of molecules including an insecticidal peptide, a prostate cancer therapeutic lead, and an orally active analgesic is demonstrated.

A systematic review of natural toxins occurrence in plant commodities used for plant-based meat alternatives production

The ongoing shift from traditional diets to plant-based meat alternatives is governed by the friendly-character related to consumers’ health and environment. However, the beneficial aspects of meat alternatives overshadow the possible adverse effects that accompany them. The present systematic review shows that the contamination of the most common plant-based meat alternatives, soybean, chickpea, pea, and seitan with mycotoxins is understudied or not studied at all. Even though they are toxic and were found in soy-based food, tropane and β-carboline alkaloids contamination data in plant-based meat alternatives is also lacking. Mycotoxin mixtures that can have additive or synergistic toxic effects have been found in multiple soy-based food, revealing the high risk that consumers expose themselves to. To better understand the risks that come along with the shift to plant-based meat diets, future research is needed regarding contamination data of plant-based meat alternatives with natural toxins. Maximum limits for contaminants found in plant-based meat alternatives need to be established by the European Commission in order to ensure consumers’ food safety.

Environmental Impact of Animal-Based Food Production and the Feasibility of a Shift Toward Sustainable Plant-Based Diets in the United States

Evidence consistently suggests that plant-based diets promote human and planetary health. Reducing large-scale animal-based food production generates environmental benefits, as the entire livestock agriculture chain plays an outsized role in greenhouse gas emissions, land change and degradation, and scarcity-weighted water use. However, substituting animal products with their plant-based counterparts must come with consideration of the nutritional quality and resource usage of plant-based food production and processing operations. Several policy reforms have been implemented at the national, state, and municipal levels in the United States to support a transition toward more plant-based diets. Federal programs such as the Supplemental Nutrition Assistance Program and the Dietary Guidelines for Americans generally promote the consumption of unprocessed plant-based foods but include little to no information on sustainability and the harmful environmental impact of animal-based foods. National policies are complemented by state efforts aimed at incentivizing produce purchased from local suppliers and encouraging resource-conserving agriculture. At the local level, public schools are implementing programs to promote plant-based protein on their menus, and urban gardens are sprouting across the country to increase access to organic farming. This mini-review examines these policy reforms and behavioral intervention strategies, based on the social-ecological model, and discuss their capacity and limitations to promote a shift toward sustainably produced plant-based diets in the United States. We conclude that transforming the food systems toward plant-based diets in the animal-centered United States requires multi-sector collaboration and context-specific policy solutions to address diet-related climate concerns without neglecting health, social, and financial constraints.

Plants as Sources of Natural and Recombinant Antimalaria Agents.

Malaria is one of the severe infectious diseases that has victimized about half a civilization billion people each year worldwide. The application of long-lasting insecticides is the main strategy to control malaria; however, a surge in antimalarial drug development is also taking a leading role to break off the infections. Although, recurring drug resistance can compromise the efficiency of both conventional and novel antimalarial medicines. The eradication of malaria is significantly contingent on discovering novel potent agents that are low cost and easy to administer. In this context, plant metabolites inhibit malaria infection progression and might potentially be utilized as an alternative treatment for malaria, such as artemisinin. Advances in genetic engineering technology, especially the advent of molecular farming, have made plants more versatile in producing protein drugs (PDs) to treat infectious diseases, including malaria. These recent developments in genetic modifications have enabled the production of native pharmaceutically active compounds and the accumulation of diverse heterologous proteins such as human antibodies, booster vaccines, and many PDs to treat infectious diseases and genetic disorders. This review will discuss the pivotal role of a plant-based production system that expresses natural antimalarial agents or host protein drugs to cure malaria infections. The potential of these natural and induced compounds will support modern healthcare systems in treating malaria infections, especially in developing countries to mitigate human fatalities.

Continuous Piecewise Linear Approximation of Plant-Based Hydro Production Function for Generation Scheduling Problems

An essential challenge in generation scheduling (GS) problems of hydrothermal power systems is the inclusion of adequate modeling of the hydroelectric production function (HPF). The HPF is a nonlinear and nonconvex function that depends on the head and turbined outflow. Although the hydropower plants have multiple generating units (GUs), due to a series of complexities, the most attractive modeling practice is to represent one HPF per plant, i.e., a single function is built for representing the plant generation instead of the generation of each GU. Furthermore, due to the computation time constraints and representation of nonlinearities, the HPF must be given by a piecewise linear (PWL) model. This paper presented some continuous PWL models to include the HPF per plant in GS problems of hydrothermal systems. Depending on the type of application, the framework allows a choice between the concave PWL for HPF modeled with one or two variables and the nonconvex (more accurate) PWL for HPF dependent only on the turbined outflow. Basically, in both PWL models, offline, mixed-integer linear (or quadratic) programming techniques are used with an optimized pre-selection of the original HPF dataset obtained through the Ramer-Douglas-Peucker algorithm. As a highlight, the framework allows the control of the number of hyperplanes and, consequently, the number of variables and constraints of the PWL model. To this end, we offer two possibilities: (i) minimizing the error for a fixed number of hyperplanes, or (ii) minimizing the number of hyperplanes for a given error. We assessed the performance of the proposed framework using data from two large hydropower plants of the Brazilian system. The first has 3568 MW distributed in 50 Bulb-type GUs and operates as a run-of-river hydro plant. In turn, the second, which can vary the reservoir volume by up to 1000 hm3, possesses 1140 MW distributed in three Francis-type units. The results showed a variation from 0.040% to 1.583% in terms of mean absolute error and 0.306% to 6.356% regarding the maximum absolute error even with few approximations.

Unexpected Arabinosylation after Humanization of Plant Protein N-Glycosylation.

As biopharmaceuticals, recombinant proteins have become indispensable tools in medicine. An increasing demand, not only in quantity but also in diversity, drives the constant development and improvement of production platforms. The N-glycosylation pattern on biopharmaceuticals plays an important role in activity, serum half-life and immunogenicity. Therefore, production platforms with tailored protein N-glycosylation are of great interest. Plant-based systems have already demonstrated their potential to produce pharmaceutically relevant recombinant proteins, although their N-glycan patterns differ from those in humans. Plants have shown great plasticity towards the manipulation of their glycosylation machinery, and some have already been glyco-engineered in order to avoid the attachment of plant-typical, putatively immunogenic sugar residues. This resulted in complex-type N-glycans with a core structure identical to the human one. Compared to humans, plants lack the ability to elongate these N-glycans with β1,4-linked galactoses and terminal sialic acids. However, these modifications, which require the activity of several mammalian enzymes, have already been achieved for Nicotiana benthamiana and the moss Physcomitrella. Here, we present the first step towards sialylation of recombinant glycoproteins in Physcomitrella, human β1,4-linked terminal N-glycan galactosylation, which was achieved by the introduction of a chimeric β1,4-galactosyltransferase (FTGT). This chimeric enzyme consists of the moss α1,4-fucosyltransferase transmembrane domain, fused to the catalytic domain of the human β1,4-galactosyltransferase. Stable FTGT expression led to the desired β1,4-galactosylation. However, additional pentoses of unknown identity were also observed. The nature of these pentoses was subsequently determined by Western blot and enzymatic digestion followed by mass spectrometric analysis and resulted in their identification as α-linked arabinoses. Since a pentosylation of β1,4-galactosylated N-glycans was reported earlier, e.g., on recombinant human erythropoietin produced in glyco-engineered Nicotiana tabacum, this phenomenon is of a more general importance for plant-based production platforms. Arabinoses, which are absent in humans, may prevent the full humanization of plant-derived products. Therefore, the identification of these pentoses as arabinoses is important as it creates the basis for their abolishment to ensure the production of safe biopharmaceuticals in plant-based systems.

Nutritional Contents and Processing of Plant-Based Milk: A Review

For a long time, animal milk such as cow milk has been regarded as a complete food due to its nutritional content and beneficial health effect. However, the emergence of lactose intolerance and allergy to cow milk, new lifestyles such as vegetarianism and veganism, and sustainability challenges such as water pollution and greenhouse gases emission, have pushed researchers to innovate an alternative product to replace animal milk. Plant-based milk, the aqueous liquid obtained from different plant materials such as cereals, legumes, nuts, seeds, and pseudocereals, has been developed to replace animal milk. Therefore, the knowledge of the nutritional profile and processing steps of plant-based milk production becomes essential. This review describes the information regarding the materials used, the nutritional profiles, and the processing steps of plant-based milk. There is a challenge in developing plant-based milk that has a similar nutrition content to cow milk. Processing technique in terms of formulation such as fortification is needed to produce plant-based milk with acceptable nutritional content and high consumer acceptance.

In Vitro and Clinical Evaluation of Cannabigerol (CBG) Produced via Yeast Biosynthesis: A Cannabinoid with a Broad Range of Anti-Inflammatory and Skin Health-Boosting Properties.

Cannabigerol (CBG) is a minor non-psychoactive cannabinoid present in Cannabis sativa L. (C. sativa) at low levels (<1% per dry weight) that serves as the direct precursor to both cannabidiol (CBD) and tetrahydrocannabinol (THC). Consequently, efforts to extract and purify CBG from C. sativa is both challenging and expensive. However, utilizing a novel yeast fermentation technology platform, minor cannabinoids such as CBG can be produced in a more sustainable, cost-effective, and timely process as compared to plant-based production. While CBD has been studied extensively, demonstrating several beneficial skin properties, there are a paucity of studies characterizing the activity of CBG in human skin. Therefore, our aim was to characterize and compare the in vitro activity profile of non-psychoactive CBG and CBD in skin and be the first group to test CBG clinically on human skin. Gene microarray analysis conducted using 3D human skin equivalents demonstrates that CBG regulates more genes than CBD, including several key skin targets. Human dermal fibroblasts (HDFs) and normal human epidermal keratinocytes (NHEKs) were exposed in culture to pro-inflammatory inducers to trigger cytokine production and oxidative stress. Results demonstrate that CBG and CBD reduce reactive oxygen species levels in HDFs better than vitamin C. Moreover, CBG inhibits pro-inflammatory cytokine (Interleukin-1β, -6, -8, tumor necrosis factor α) release from several inflammatory inducers, such as ultraviolet A (UVA), ultraviolet B (UVB), chemical, C. acnes, and in several instances does so more potently than CBD. A 20-subject vehicle-controlled clinical study was performed with 0.1% CBG serum and placebo applied topically for 2 weeks after sodium lauryl sulfate (SLS)-induced irritation. CBG serum showed statistically significant improvement above placebo for transepidermal water loss (TEWL) and reduction in the appearance of redness. Altogether, CBG’s broad range of in vitro and clinical skin health-promoting activities demonstrates its strong potential as a safe, effective ingredient for topical use and suggests there are areas where it may be more effective than CBD.

Plant-based engineering for production of high-valued natural products.

Covering: up to March 2022Plants are a unique source of complex specialized metabolites, many of which play significant roles in human society. In many cases, however, the availability of these metabolites from naturally occurring sources fails to meet current demands. Thus, there is much interest in expanding the production capacity of target plant molecules. Traditionally, plant breeding, chemical synthesis, and microbial fermentation are considered the primary routes towards large scale production of natural products. Here, we explore the advances, challenges, and future of plant engineering as a complementary path. Although plants are an integral part of our food and agricultural systems and sustain an extensive array of chemical constituents, their complex genetics and physiology have prevented the optimal exploitation of plants as a production chassis. We highlight emerging engineering tools and scientific advances developed in recent years that have improved the prospects of using plants as a sustainable and scalable production platform. We also discuss technological limitations and overall economic outlook of plant-based production of natural products.

Proteomic Profiling of Interplay Between Agrobacterium tumefaciens and Nicotiana benthamiana for Improved Molecular Pharming Outcomes.

Transient expression of recombinant proteins in plants is being used as a platform for production of therapeutic proteins. Benefits of this system include a reduced cost of drug development, rapid delivery of new products to the market, and an ability to provide safe and efficacious medicines for diseases. Although plant-based production systems offer excellent potential for therapeutic protein production, barriers, such as plant host defense response, exist which negatively impact the yield of product. Here we provide a protocol using tandem mass tags and mass spectrometry-based proteomics to quickly and robustly quantify the change in abundance of host defense proteins produced during the production process. These proteins can then become candidates for genetic manipulation to create host plants with reduced plant defenses capable of producing higher therapeutic protein yields.

Recombinant Protein Production in Plants: A Brief Overview of Strengths and Challenges.

The first recombinant proteins were produced in microbes and animal cells cultivated in bioreactors. These systems have become the standard for industrial-scale recombinant protein manufacturing. Later, the production of recombinant proteins was demonstrated in whole plants, which differ morphologically from cell-based systems and require completely different cultivation conditions. Over time, additional plant-based production platforms were established, including hairy roots and cell suspension cultures, which are more similar to conventional cell-based systems in terms of morphology, procedures, and equipment requirements. In this brief overview of the field, we explain why plant-based systems are becoming increasingly attractive for the production of valuable proteins with scientific and commercial applications, but also highlight the challenges that these systems must overcome to achieve more widespread industrial utilization. We discuss various laboratory protocols and approaches for the production of recombinant proteins in plants, as well as strategies to optimize yields, and the regulatory and legal framework.

Transient recombinant expression of highly immunogenic CagA, VacA and NapA in Nicotiana benthamiana

Interest in the plant-based transient production of recombinant immunogenic antigens has tremendously progressed because plants are cost-effective, easily selectable, free of mammalian contamination, and support complex post-translational modifications. Nicotiana benthamiana is a convenient system for transient expression of recombinant antigens. The present study documented a platform for rapid production of Helicobacter pylori CagA, VacA and NapA antigens three days (first harvest, FH) and six days (second harvest, SH) after agro-infiltration using a syringe. In this study, CagA, VacA and NapA antigen genes from Helicobacter pylori were cloned into the binary vector pBI121 and transformed into Nicotiana benthamiana by the Agrobacterium-mediated process. Leaves of four to five weeks old Nicotiana benthamiana plants were agroinfiltrated with EHA105 subtype of Agrobacterium tumefaciens strain containing cloned CagA (pBI121-CagA), VacA (pBI121-VacA) and NapA (pBI121-NapA) constructs. The transient expression and accumulation of the recombinant genes containing CagA, VacA and NapA expression cassettes were confirmed using qRT-PCR by comparing the relative expression at FH and SH post-infiltration with the non-infiltrated (control) samples and using ELISA at 1/5 and 1/10 dilution ratios. The qRT-PCR findings showed that Agrobacterium-mediated syringe infiltration of leaves of four to five weeks old Nicotiana benthamiana plants produced significantly higher transcript levels of CagA (about 8-fold and 7-fold), VacA (38-fold and 24-fold) and NapA (7-fold and 5-fold) genes at FH and SH compared to the control sample. Besides, the maximum amount of CagA, VacA and NapA antigens were detected at the FH stage compared to the SH stage, when the antibody concentrations of the agro-infiltrated leaf extracts containing these recombinant antigens were diluted in a 1/5 ratio. This study has developed evidence to support that recombinant CagA, VacA and NapA can be transiently produced in Nicotiana benthamiana plants.

Polyurethane Foam Rafts Supported In Vitro Cultures of Rindera graeca Roots for Enhanced Production of Rinderol, Potent Proapoptotic Naphthoquinone Compound.

Unique phytochemical profile of plants belonging to Boraginaceae family provides a prolific resource of lipophilic pigments from the group of naphthoquinone derivatives. To overcome low compound content, the major obstacle of plant-based production, immobilization of Rindera graeca roots in in vitro cultures was implemented for efficient production of rinderol, novel furanonaphthoquinone derivative with anticancer properties. Chromatographic procedures revealed rinderol presence in extracts of all investigated root lines, derived both from root biomass and post-culture medium. Unexpectedly, in the second stage of the experiment, rinderol production was ceased in control, unmodified culture systems. On the contrary, roots immobilized on PUF rafts uniformly and stably produced rinderol, and its highest amount was noted for transformed root lines after 42 days of cultivation (222.98 ± 10.47 µg/flask). PUF occurred to be the main place of compound accumulation. Moreover, investigation of rinderol biological activity revealed its fast-acting cell death induction in HeLa cervical cancer cells at relatively low concentrations. Presented results revealed successful application of R. graeca roots immobilization on PUF rafts for production and in situ product removal of rinderol, novel lipophilic furanonaphthoquinone with suggested proapoptotic activity.

A plant-biotechnology approach for producing highly potent anti-HIV antibodies for antiretroviral therapy consideration

Despite a reduction in global HIV prevalence the development of a pipeline of new therapeutics or pre-exposure prophylaxis to control the HIV/AIDS epidemic are of high priority. Antibody-based therapies offer several advantages and have been shown to prevent HIV-infection. Plant-based production is efficient for several biologics, including antibodies. We provide a short review on the work by Singh et al., 2020 who demonstrated the transient production of potent CAP256-VRC26 broadly neutralizing antibodies. These antibodies have engineered posttranslational modifications, namely N-glycosylation in the fragment crystallizable region and O-sulfation of tyrosine residues in the complementary-determining region H3 loop. The glycoengineered Nicotiana benthamiana mutant (ΔXTFT) was used, with glycosylating structures lacking β1,2-xylose and/or α1,3-fucose residues, which is critical for enhanced effector activity. The CAP256-VRC26 antibody lineage targets the first and second variable region of the HIV-1 gp120 envelope glycoprotein. The high potency of this lineage is mediated by a protruding O-sulfated tyrosine in the CDR H3 loop. Nicotiana benthamiana lacks human tyrosyl protein sulfotransferase 1, the enzyme responsible for tyrosine O-sulfation. The transient coexpression of the CAP256-VRC26 antibodies with tyrosyl protein sulfotransferase 1 in planta had restored the efficacy of these antibodies through the incorporation of the O-sulfation modification. This approach demonstrates the strategic incorporation of posttranslational modifications in production systems, which may have not been previously considered. These plant-produced CAP256-VRC26 antibodies have therapeutic as well as topical and systemic pre-exposure prophylaxis potential in enabling the empowerment of young girls and women given that gender inequalities remain a major driver of the epidemic.

The Role of Women in Plant-based Textile Production during the Viking Diaspora in Scotland and the Irish Sea Region

Evidence for textiles in viking-age Scotland and the adjacent Irish Sea region derives from small fragments usually surviving as mineralised products associated with metal dress fittings and grave goods such as shield bosses and weaving battens, excavated from the furnished graves of both women and men. Since Scottish viking-age textiles were last reviewed over twenty years ago, this paper collates information from antiquarian finds and more recent excavations which employed considerably enhanced techniques for retrieving fragile archaeological textiles. Evidence is presented for the occurrence and role of plant-based textiles derived from flax and hemp including linen in funerary processes as burial garments, shrouds and wrapping other grave goods, such as weapons and tools. Many richly appointed women's graves in viking-age Scotland were accompanied by tool assemblages used in the manufacture and maintenance of textiles. The presence of these tools raises questions about the status of textile production and the roles women played in it.