Plant Lines Research Articles

Apple pomace as an alternative substrate for butanol production

Butanol-producing strains Clostridium sp. UCM B-7570 and C. acetobutylicum UCM B-7407 were used for research from “Collection of strains of microorganisms and plant lines for food and agricultural biotechnology” of the Institute of Food Biotechnology and Genomics of the National Academy of Sciences of Ukraine, glycerol (BASF, Germany) and apple pomace (total moisture 4%) after apple juice production. The aim of this work was to study the possibility of using apple pomace by domestic butanol-producing strains of Clostridium sp. UCM B-7570 and C. acetobutylicum UCM B-7407 as a substrate. Producers were cultured on medium with different concentrations of apple pomace, glycerol was used for the inoculation. The presence of ethanol, acetone, and butanol in the culture liquid was determined using a gas chromatograph. It was determined that a significant part of the macrocomponent composition of the extracts can be used in bioconversion by producing strains of the genus Clostridium. It was determined that the highest concentration of butanol (10 g/dm3) was at a concentration of 120 g/dm3 in the extracts. The obtained data showed the possibility of using apple pomace as a substrate in biobutanol technology.

Biberde (Capsicum annuum L.) Haploid Bitki Eldesinde Anter Kültürü Yöntemi

Pepper is from the Capsicum genus of the Solonaceae family and is one of the vegetable species that is loved and grown in the world and in our country. After China and Mexico, Turkey ranks third in pepper production in the world. It is a great advantage for us that the climate structure of our country is suitable for vegetable cultivation. For this reason, it has caused a significant variety of pepper in the country, as it adapts to every region in its pepper. In this way, it has also ensured the enrichment of our gene resources. The richness of gene resources is an important factor in the success of breeding studies. One of the purposes of breeding studies is to develop varieties. It is used in breeding studies with anther culture method in pepper, and new varieties are developed in this way. Because of obtaining homozygous lines in plants, haploid plant production has an important place in plant breeding. In this study, the definition and application of anther culture and the factors affecting success, the researches made with the anther culture method in the world and in our country and the developments in recent years are discussed.

Study of the Effect of Trifloralin Herbicide, Seed Rate and Planting Distances in Controlling Broad-Leaf Weeds Associated with Wheat (Triticum aestivum L.) Under the Conservation Agriculture System

The study was conducted in the fields of one of the farmers in order to control the broad Weeds associated with the wheat crop by chemical and agricultural methods and with a conservative cultivation system for the seasons 2018/2019 and 2019/2020, and the study included three factors: the first factor is the Trifloraline herbicide and at two levels (0, 1248) cm3 / ha, the second factor rates the effectiveness of the herbicide, and the third factor rates the effectiveness of the cultivation system. Important findings are summarized below. The study was conducted using the system of global experiments in the split-split plots, with seeding at three levels (80, 100, and 120) kg / ha, and the third factor, planting distances at two levels (15 and 30) cm between the lines, with design of complete random sectors and with three replications; Herbicide treatment reduced all traits significantly compared to comparison treatment. Seeding rate increased broad-leaf weeds and side branches and decreased bush susceptible leaf dry weight. All traits improved at 30 cm between planting lines. Interaction achieved. In the second season, the herbicide treatment and 120 kg/ha seed rate reduced broadleaf weed dry weight. The herbicide treatment and 15-cm distance reduced all traits and seasons. In the second season, the interaction between the seed rate of 120 kg/ha and the distance of 15 cm between the planting lines reduced the dry weight of the broad-leaf weed, while the herbicide treatment and the seed rate of 80 kg/ha did the same. In the second season, broad-leaved weeds decreased in dry weight by 15 cm.

A Possible Alternative to Mining in Valea Jiului

Abstract Starting from the interdependence of tourism - mountain environment - rural environment - sustainable development, four solutions are proposed and developed in the work as an alternative to mining in the area: tourist village, medicinal plant and forest fruit processing line, milk processing line, cardboard packaging line.

Low levels of flavonoids in Arabidopsis thaliana during phenanthrene exposure suggest new roles for a flavanone 3-hydroxylase

Polycyclic aromatic hydrocarbons (PAHs) contamination impacts the well-being of fauna, flora, and the environment. PAHs are a group of compounds consisting of two or more benzene rings. Due to their structure, PAHs are very stable, generally non-reactive and resistant to biodegradation; yet these compounds are cytotoxic in biological systems. In plants, PAHs induce oxidative stress and cell death, and inhibit growth. Previous PAH exposure studies from our laboratory showed an up-regulation of the Arabidopsis thaliana gene At3g51240. At3g51240 encodes a flavanone 3-hydroxylase (F3H), an enzyme catalyzing the hydroxylation of flavanones, key intermediates in the flavonoid biosynthesis pathway. Considering that flavonoids may act as antioxidants and protect cells against oxidative stress, we hypothesized that F3H was important in alleviating PAH-induced stress symptoms. Analysis of flavonoid content by high performance liquid chromatography showed low levels of flavonoids in phenanthrene-treated plants. Analysis of At3g51240 loss-of-function mutant plant lines growing on phenanthrene-containing growth media showed reduced phenotypic stress responses. Interestingly, low levels of H2O2 were observed in the mutant plants, suggesting that this putative flavanone 3-hydroxylase may be involved in PAH oxidation, with the consequent H2O2 generation. Furthermore, in vitro assays with purified recombinant 6xHis tagged AT3G51240 showed enzyme activity on the PAH when phenanthrene was used as a substrate. Together, these data suggest that F3H is involved in PAH detoxification in planta with the subsequent accumulation of toxic metabolites. This research also indicates the potentiality of using specialized metabolism enzymes such as oxidoreductase in the genetic engineering of plants to remediate PAH-contaminated soils.

KERAGAAN GENOTIPE MUTAN BAAS SELEM GENERASI KEDUA (M2) HASIL IRADIASI SINAR GAMMA

Research aims to determine changes in genotype characters that appear in the second generation Baas Selem (Oryza sativa L.) mutant (M2) resulting from gamma irradiation. This research was conducted from May to September 2021 in Nyur Lembang Village, Narmada District, West Lombok Regency, West Nusa Tenggara using an Expansion Randomized Block Design (RAK). The treatments used were 26 treatments consisting of 24 mutant plant genotypes and 2 control plants were tested using analysis of variance at the 5% level, then the significant results were tested further using the partition design RAK model. If there is a significant difference between the treatments, a follow-up test is carried out using the BNT test at the 5% level. Based on the results of a one-way analysis of variance for the characters tested from the 26 treatments (24 mutant plant lines namely G23, G87, G46, G65, G15, G34, G39, G5, G61, G54, G22, G74, G41, G72, G85 , G82, G39, G7, G141, G75, G62, G42, G81, and G21, 1 Baas Selem parent and 1 comparison variety, Inpago Unram, showed differences in genotypic characters on parameters of panicle length and number of empty grain per panicles.
 Keywords: Genotype Performance; Black Rice (Baas Selem), mutation induction

Antioxidants of Non-Enzymatic Nature: Their Function in Higher Plant Cells and the Ways of Boosting Their Biosynthesis.

Plants are exposed to a variety of abiotic and biotic stresses leading to increased formation of reactive oxygen species (ROS) in plant cells. ROS are capable of oxidizing proteins, pigments, lipids, nucleic acids, and other cell molecules, disrupting their functional activity. During the process of evolution, numerous antioxidant systems were formed in plants, including antioxidant enzymes and low molecular weight non-enzymatic antioxidants. Antioxidant systems perform neutralization of ROS and therefore prevent oxidative damage of cell components. In the present review, we focus on the biosynthesis of non-enzymatic antioxidants in higher plants cells such as ascorbic acid (vitamin C), glutathione, flavonoids, isoprenoids, carotenoids, tocopherol (vitamin E), ubiquinone, and plastoquinone. Their functioning and their reactivity with respect to individual ROS will be described. This review is also devoted to the modern genetic engineering methods, which are widely used to change the quantitative and qualitative content of the non-enzymatic antioxidants in cultivated plants. These methods allow various plant lines with given properties to be obtained in a rather short time. The most successful approaches for plant transgenesis and plant genome editing for the enhancement of biosynthesis and the content of these antioxidants are discussed.

Stochastic organelle genome segregation through Arabidopsis development and reproduction.

Organelle DNA (oDNA) in mitochondria and plastids is vital for plant (and eukaryotic) life. Selection against damaged oDNA is mediated in part by segregation - sorting different oDNA types into different cells in the germline. Plants segregate oDNA very rapidly, with oDNA recombination protein MSH1 a key driver of this segregation, but we have limited knowledge of the dynamics of this segregation within plants and between generations. Here, we reveal how oDNA evolves through Arabidopsis thaliana development and reproduction. We combine stochastic modelling, Bayesian inference, and model selection with new and existing tissue-specific oDNA measurements from heteroplasmic Arabidopsis plant lines through development and between generations. Segregation proceeds gradually but continually during plant development, with a more rapid increase between inflorescence formation and the next generation. When MSH1 is compromised, the majority of observed segregation can be achieved through partitioning at cell divisions. When MSH1 is functional, mtDNA segregation is far more rapid; we show that increased oDNA gene conversion is a plausible mechanism quantitatively explaining this acceleration. These findings reveal the quantitative, time-dependent details of oDNA segregation in Arabidopsis. We also discuss the support for different models of the plant germline provided by these observations.

Mutation of MYB36 affects isoflavonoid metabolism, growth, and stress responses in Lotus japonicus

AbstractIsoflavonoids are mostly produced by legumes although little is known about why and how legumes are able to regulate the biosynthesis of these particular compounds. Understanding the role of potential regulatory genes of the isoflavonoid biosynthetic pathway constitutes an important topic of research. The LORE1 mutation of the gene encoding the transcription factor MYB36 allowed the identification of this gene as a regulator of isoflavonoid biosynthesis in Lotus japonicus plants. The levels of several isoflavonoid compounds were considerably lower in two lines of Ljmyb36 mutant plants compared to the WT. In addition, we found that Ljmyb36 mutant plants were significantly smaller and showed a substantial decrease in the chlorophyll levels under normal growth conditions. The analysis of plants subjected to different types of abiotic stress conditions further revealed that mutant plants presented a higher sensitivity than WT plants, indicating that the MYB36 transcription factor is also involved in the stress response in L. japonicus plants.

Breeding of high seed oil oleate levels into Upland cotton from wild Gossypium barbadense L. germplasm

AbstractA high seed oil oleate trait identified in wild Gossypium barbadense L. cotton accession GB713 (PI 608139) was bred into Upland cotton (Gossypium hirsutum L.) cultivar Sure‐Grow 747 (SG747, PVP 9800118). Starting from the F2 generation of an initial cross between GB713 and SG747, plants were selected by measuring the seed oil fatty acid distributions by gas chromatography. Selected F3 plants were crossed to M Rk‐Rn 6, a line previously created from GB713 and SG747 for improved nematode resistance. High‐oleate plants were selected in the F2 population. Selected plants were then crossed to HOa1 (a ∼35% oleate line derived from M Rk‐Rn 1, a sibling line to M Rk‐Rn 6), and high‐oleate plants were again selected in the F2 population. F3 plants were re‐selected for the high‐oleate trait, and four selected F4 lines were evaluated in 2022 field plots. The four lines had oleate levels between 52% and 57%, approximately triple that found in normal cottonseed oil (15%–19%) and higher than in either the high‐oleate wild GB713 parent (∼40%) or in the previously released HOa cotton plant lines (33%–35%). The increased oleate levels occurred with an approximately equal reduction in linoleic acid. Other seed and fiber properties were largely like those of SG747, although some residual GB713 character existed within the lines. The resulting oil composition should be ideal for use in deep fat frying, with the high level of oleic acid improving the oil's oxidative stability but with sufficient linoleic acid to maintain frying flavor.

Application rate and hydraulic tips used in remotely piloted aircraft affect the phytosanitary products in coffee plant canopies

Most coffee (Coffea arabica) phytosanitary management techniques are performed using ground-based equipment, and remotely piloted aircraft are a recent alternative. Therefore, this study evaluates the effect of different application rates and hydraulic tips used for spreading phytosanitary products on coffee crops using a remotely piloted aircraft, assisted by artificial targets and dye tracing. The experiment was a 4 × 3 factorial randomized block design with four tips (XR 110-01, TT 110-01, AIXR 110-015, and TTJ60 110-02) and three application rates (8, 12, and 16 L ha-1). Hydrosensitive paper was used to analyze the droplet spectrum, and the Brilliant Blue tracer was used to detect spray deposition. The DJI Agras T20 remotely piloted aircraft was used to apply the phytosanitary product. Speed, flight height, and application range were maintained at 5.56 m s-1, 2 m, and 5 m, respectively. The flight direction was perpendicular to the crop planting lines. The application rate and hydraulic tip jointly controlled the accumulation of droplets on the target according to its position in the plant canopy. Therefore, remotely piloted aircraft can be used in coffee phytosanitary management, particularly to control targets that predominately occur in the upper third of the plant canopy.

Novel insights regarding the safety and efficacy of pyrethroid-coated nanoparticles against Hyalomma ticks

Nanoparticles have been shown to inhibit major life cycle stages of ticks, indicative of the promising application of nanomaterials against hard ticks. The study thus probed into one of the alternative options to curtail Hyalomma by employing nanocomposites consisting of pyrethroids (cypermethrin and deltamethrin) coated nanoparticles of iron oxides and iron sulfides keeping alongside the evaluation of their toxicity through plant and mammalian cell lines. The nanoparticles used in this study were roughly spherical in morphology and exhibited various size dimensions upon characterization using SEM, EDX, and FTIR. The application of nanomaterials on female ovipositioning tick showed a decline up to 15% (females ovipositioned) in deltamethrin-coated FeO NPs, whereas this decline was up to 18% in Cyp-FeS NPs and up to 5% in Cyp-FeO NPs. Similarly, the larval hatching was also impacted, leading to a hatching percentage of 5% and only 1% by application of Cyp-FeS NPs and Cyp-FeO NPs, respectively. Similarly, the larval groups had LC90 of 4.1 and 4.73 mg/L for the Cyp-FeO NPs and Cyp-FeS NPs groups. The delta-FeO NPs and delta-FeS NPs demonstrated a promising effect against adult ticks, showing LC50= 3.5 mg/L, LC90= 6.7 mg/L and LC50= 3.8 mg/L, LC90= 7.9 mg/L, respectively. MTT assay revealed that the pyrethroids coupled with iron oxide nanoparticles showed the least cytotoxicity even at the highest concentration (10–1 µL) among other nanomaterials. The study thus concluded a safer spectrum of non-target effects of pyrethroids-coated nanomaterials in addition to their significant anti-tick activity.

Linear mixed models with two longitudinal factors in the study of sugarcane dry root mass

There are agronomic experiments where measurements of a response variable are carried out over more than one longitudinal factor, for example, at different depths over time. These observations, made systematically in each experimental unit, can be correlated and might have heterogeneous variances at the different levels of the longitudinal factor. It was possible to model this correlation between repeated measures and the heterogeneity of variances by using mixed models. Thus, it was necessary to adapt some covariance structures that are common in experiments with only one longitudinal factor. The objective of this study was to use the class of linear mixed models to study sugarcane root dry mass. The experiment was the randomized complete blocks design and the parcels received four nitrogen doses. Repeated measurements were made over two longitudinal factors, one being qualitative ordinal (depths) and one being quantitative (distances from the planting line). It was possible to select a parsimonious covariancestructure and another one to explain the average behavior of the responses through likelihood ratio tests, Wald tests, and using the AIC and BIC information criteria. The adjustment of the selected model was verified by using residual diagnostics graphs.

Sources of Resistance to Powdery Mildew in Wild Barley (Hordeum vulgare subsp. spontaneum) Collected in Jordan, Lebanon, and Libya

Barley powdery mildew (BPM) is caused by the pathogen Blumeria hordei (Bh) and can lead to severe yield loss. Plant pathologists are looking for new sources of resistance to BPM. Barley accessions, including the wild subspecies Hordeum vulgare subsp. spontaneum (Hvs), are stored in many gene banks and are often a valuable source of economically important characteristics. The wild barley Hvs could be a valuable resistance source for BPM. The aim of the presented investigation was to detect new sources of BPM resistance in 81 accessions of Hvs collected in Jordan (46), Lebanon (24), and Libya (11). European differential isolates of BPM were used, and resistant single plant lines were selected for use from fifteen accessions from Jordan and Libya. These resistant single plant lines were tested for the presence of specific resistance genes using a differential set of Bh isolates. Hypotheses about the presence of specific resistance genes were made by comparing the reaction spectra of the tested lines with those of differential lines. After an analysis of the obtained results, it was concluded that all 31 tested single plant lines of Hvs had genes for resistance that are not represented in the barley differential set for resistance genes to Bh. Twenty-six lines of Hvs selected from accessions originated in Jordan and Libya showed resistance reactions to all isolates used. These lines will be further tested as new sources of effective resistance and used in barley prebreeding programs.

OsWRKY97, an Abiotic Stress-Induced Gene of Rice, Plays a Key Role in Drought Tolerance.

Drought stress is one of the major causes of crop losses. The WRKY families play important roles in the regulation of many plant processes, including drought stress response. However, the function of individual WRKY genes in plants is still under investigation. Here, we identified a new member of the WRKY families, OsWRKY97, and analyzed its role in stress resistance by using a series of transgenic plant lines. OsWRKY97 positively regulates drought tolerance in rice. OsWRKY97 was expressed in all examined tissues and could be induced by various abiotic stresses and abscisic acid (ABA). OsWRKY97-GFP was localized to the nucleus. Various abiotic stress-related cis-acting elements were observed in the promoters of OsWRKY97. The results of OsWRKY97-overexpressing plant analyses revealed that OsWRKY97 plays a positive role in drought stress tolerance. In addition, physiological analyses revealed that OsWRKY97 improves drought stress tolerance by improving the osmotic adjustment ability, oxidative stress tolerance, and water retention capacity of the plant. Furthermore, OsWRKY97-overexpressing plants also showed higher sensitivity to exogenous ABA compared with that of wild-type rice (WT). Overexpression of OsWRKY97 also affected the transcript levels of ABA-responsive genes and the accumulation of ABA. These results indicate that OsWRKY97 plays a crucial role in the response to drought stress and may possess high potential value in improving drought tolerance in rice.

아르누보(Art Nouveau) 양식을 응용한 아트 업스타일 디자인 연구

This study attempted to use the symbolic and decorative properties of Art Nouveau in hairstyling art through the expression of updo styles, using the organic lines of plant among various styles of Art Nouveau which had a significant influence on many countries early 20th century. First, to examine the formative characteristics of Art Nouveau, this study examined related data and previous studies on diverse artworks. Second, the concept, design factors and shapes of updos were analyzed. Third, 3 different updo styles which emphasize the smoothness of curves were created by adding modern colors to mannequin based on three decorations created with the organic lines of plant in Art Nouveau style as motives. The results found that organic lines in Art Nouveau can be expressed in a diverse fashion in modern design. Updo hairstyles have unlimited potential as art from the perspective of creative art. It is anticipated that studies on the updo hairstyles created with the organic lines of plant would be used in a more diverse and unique way.

Bio-Stimulant for Improving Simmondsia chinensis Secondary Metabolite Production, as Well as Antimicrobial Activity and Wound Healing Abilities.

Simmondsia chinensis is a dioecious, long-lived perennial shrub. Its leaves contain several antioxidant flavonoids that have numerous pharmacological effects. Various strategies have been explored to propagate jojoba with enhanced pharmacological values. This research evaluates the bio-stimulatory impacts of He-Ne laser seed irradiation on seed germination, plantlet growth, and alteration of the composition and bioactivities of phytochemicals in jojoba plants. Jojoba seeds were irradiated for 5, 10, and 15 min before in vitro germination. Germination, growth, and multiplication parameters were recorded during germination, multiple-shoot induction, and rooting stages. The wound healing and antimicrobial activities of methanolic extracts from plant lines obtained from the non-irradiated (control) and 10 min irradiated seeds were compared by excision wound model in Wistar male rats and zone of inhibition assay. Our study revealed that laser irradiation increased seed germination, with the highest percentage observed in seeds irradiated for 10 min. Plant lines from the 10 min irradiated seeds produced more explants with higher explant heights and numbers of leaves, more roots, and higher photosynthetic pigment contents than those of control and other laser testings. By comparing plant extracts from the control and 10 min treatments, we observed that extracts from the 10 min treatment exhibited higher percentages of wound contraction and shorter epithelialization periods. In addition, these extracts also resulted in higher levels of angiogenesis elements (VEGF, TGF-β1, and HIF-1α) and reduced the inflammation regulators (IL-1β, IL-6, TNF-α, and NFκB) in the experimental rats. In concordance, extracts from the 10 min treatment also explained raised antibacterial activities towards Staphylococcus aureus and Escherichia coli. Our findings show that pre-sowing seed treatment with a He-Ne laser (632.8 nm) could be a good technique for stimulating S. chinensis plant growth and increasing the impact compound levels and biological activities.

Mapping and Functional Analysis of LE Gene in a Lethal Etiolated Rice Mutant at Seedling Stage

An EMS-induced mutant le (lethal etiolated) obtained from a rice variety Zhongjian 100, was characterized by lethal etiolated phenotypes with significantly lower contents of chlorophyll a, chlorophyll b, total chlorophyll and carotenoids, as well as significantly decreased the number of chloroplast grana and irregular and less-stacked grana lamella. The root length, root surface area and root volume of le plants were markedly reduced relative to the wild type. The mutant le showed significantly lower catalase activity and total protein content, significantly higher peroxidase activity,. Using the map-based cloning method, LE gene was mapped to an interval of 48 kb between markers RM16107 and RM16110 on rice chromosome 3. A mutation (from T to C) at the nucleotide position 692 bp of LOC_Os03g59640 (ChlD) occurred, resulting in a change from leucine to proline. By crossing HM133 (a pale green mutant with a single-base substitution of A for G on the exon 10 of ChlD subunit) with a heterozygous line of le (LEle), two plant lines heterozygous at the LE locus and HM133 locus were obtained. Among 15 transgenic plants, 3 complementation lines displayed normal leaf color with significant higher total chlorophyll and chlorophyll a and b contents. The ChlD subunit of the mutant was unable to interact with the ChlI subunit, as demonstrated by a yeast two-hybrid assay. The mutation in le led to lethal etiolated phenotype, but no such phenomenon was observed in the mutants. The interaction between the ChlD and ChlI subunits, chlorophyll synthesis, and chloroplast development may all be impacted by the mutation, which could ultimately result in plant death. The mutation in the AAA+ domain of ChlD blocked the interaction of ChlDle with ChlI, leading to the loss of function of ChlD and hindering chlorophyll synthesis and chloroplast development, and ultimately causing lethal etiolated phenotype of the mutant.

The sonication-assisted whisker method enables CRISPR-Cas9 ribonucleoprotein delivery to induce genome editing in rice

CRISPR/Cas9-based genome editing represents an unprecedented potential for plant breeding. Unlike animal cells, plant cells contain a rigid cell wall, genome editing tool delivery into plant cells is thus challenging. In particular, the delivery of the Cas9-gRNA ribonucleoprotein (RNP) into plant cells is desired since the transgene insertion into the genome should be avoided for industrial applications in plants. In this study, we present a novel RNP delivery approach in rice. We applied the sonication-assisted whisker method, conventionally developed for DNA delivery in plants, for RNP delivery in rice. Combined with marker gene delivery, we successfully isolated OsLCYβ genome-edited lines generated by RNPs. The calli and regenerated shoot of the OsLCYβ mutant showed abnormal carotenoid accumulation. In addition, we also detected, although at a low frequency, genome editing events in rice calli cells by RNP delivery using the sonication-assisted whisker method without any additional. Therefore, the sonication-assisted whisker method could be an attractive way to create RNP-based genome-edited lines in plants.

Microbial analysis of the production line for roasted seaweed snacks

Roasted seaweed snacks are getting popular worldwide. To reveal the main origins of microorganisms present in the roasted seaweed snacks, samples of raw materials, finished products, and environment (both food-contact and nonfood-contact surfaces) were collected from four plants. The total aerobic plate (APC), coliform, and mold counts of these samples were analyzed and the selected samples were also used for 16S rRNA sequencing. Results showed that dried laver contained the highest levels of APC among all raw ingredients, ranging from 2.72 to 6.95 Log CFU/g. Crushed pumpkin seeds and almond flakes had the highest coliform (up to 5.57 Log CFU/g) and mold counts (up to 2.16 Log CFU/g). The addition of seasoning to seaweed sandwich snacks increased the total microbial loads. The roasting process decreased the APC, coliforms, and mold counts of seaweed snacks by 0.22–1.73 Log CFU/g, >0.18- >2.46 Log CFU/g, and >0.18- >0.88 Log CFU/g respectively. The 16S rRNA sequencing analysis showed that the microbiota of roasted seaweed snacks was closely correlated with that of dried laver and the microbial load of roasted sandwich seaweed was primarily from the dried laver while less from microorganisms present in the processing plant environment and line workers.