Strategic biomodification for raw plant-based pretreatment biorefining toward sustainable chemistry.

Plant-based pretreatment biorefining is the initial triggering process in biomass-conversion to bio-based chemical products. In view of chemical sustainability, the raw plant-based pretreatment biorefining process is more favorable than the fossil-based one. Its direct use contributes to reducing CO2 emissions and the production cost of the target products by eliminating costly steps, such as the separation and purification of intermediates. Three types of feedstock plant resources have been utilized as raw plant feedstock sources, such as: lignocellulosic, starchy, and inulin-rich feedstock plants. These plant sources can be directly used for bio-based chemical products. To enhance the efficiency of their pretreatment biorefining process, well-designed biomodification schemes are discussed in this review to afford important information on useful biomodification approaches. For lignocellulosic feedstock plants, the enzymes and regulatory elements involved in lignin reduction are discussed using: COMT, GAUT4, CSE, PvMYB4 repressor, etc. For inulin-rich feedstock plants, 1-SST, 1-FFT, 1-FEH, and endoinulinase are illustrated in relation with the reduction of chain length of inulin polymer. For starchy feedstock plants, their biomodification is targeted to enhancing the depolymerization efficiency of starch to glucose monomer units. For this biomodification target, six candidates are discussed. These are SBE I, SBE IIa, SBE IIb, GBSS I, PTSTI, GWD 1, and PTSTI. The biomodification strategies discussed here promise to be conducive to enhancing the efficiency of the plant-based pretreatment biorefining process.

Seagrass-based platform strategies for sustainable hydroxymethylfurfural (HMF) production: toward bio-based chemical products.

Today, sustainable chemistry is a key trend in the chemical manufacturing industry due mainly to concerns over the global environment and resource security. In sustainable chemical manufacture, the choice of a bio-based feedstock plays a pivotal pillar. In terms of feedstock utilization for producing HMF, which is a multivalent platform intermediate easily convertible to valuable chemical products; biopolymers, biofuels, and other important chemicals, seagrass biomasses can be more favorable feedstocks compared with land plant resources due primarily to easy availability and no systematic farming. Moreover, seagrass feedstocks could contribute cost-effectively and sustainably producing HMF by exploiting the beach-cast seagrasses on seagrass-prairies with no feedstock cost, indicating that seagrass biomasses could be a most promising biofeedstock source for sustainable HMF production. We afford a platform bioprocessing technology that has not been attempted before for sustainable HMF production using raw seagrass biomass. This bioprocess can be operated by simple reaction conditions using inorganic Brønsted acids (mainly HCl) and ionic liquid solvents at relatively low temperatures (120-130 °C). In addition, some bioengineering strategies for improving the growth of seagrass biomass and the quantity/quality of nonstructural carbohydrates (starch, sucrose) that can be used as the feeding substrates for HMF production are also discussed. The main aim of this review is to provide some important information about breakthrough bio/technologies conducive to cost-effective and sustainable HMF production.

Marine plant-based biorefinery for sustainable 2,5-furandicarboxylic acid production: A review.

Marine plants, including macroalgae and seagrass, show promise as biorenewable feedstocks for sustainable chemical manufacturing. This study explores their potential in producing 2,5-furandicarboxylic acid (FDCA), a versatile platform chemical for commodity polymers. FDCA-based polyethylene 2,5-furandicarboxylate offers a sustainable alternative to petroleum-derived polyethylene terephthalate, commonly used in plastic bottles. Our research pioneers the concept of a marine plant-based FDCA biorefinery, introducing innovative approaches for sustainability and cost-effectiveness. This review outlines the use of ionic liquid-based solvents (ILS) and deep eutectic solvent (DES) systems in FDCA production. Additionally, we propose biomodification strategies involving target enzyme-encoding genes to enhance the depolymerization of non-structural storage glucans in marine plants. Our findings pave the way for eco-friendly biorefineries and biorenewable plastics.

Conversion of inulin-rich raw plant biomass to 2,5-furandicarboxylic acid (FDCA): Progress and challenge towards biorenewable plastics.

The current commercial plastic manufactures have been produced using petroleum-based resource. However, due to concerns over the resource depletion and the environmental sustainability, bioresource-based manufacturing processes have been developed to cope against these concerns. Bioresource-derived 2,5-furandicarboxylic acid (FDCA) can be utilized as a building block material for plastic manufactures. To date, numerous technologies have been developed for the production of FDCA using various types of bio-based feedstocks such as hydroxymethylfurfural (HMF), 6-C sugars, and polysaccharides. The commercial companies produce FDCA using HMF-based production processes due to their high production efficiency, but the high price of HMF is a problem bottleneck. Our review affords important information on breakthrough approaches for the cost-efficient and sustainable production of FDCA using raw plant feedstocks rich in inulin. These approaches include bioprocessing technology based on the direct use of raw plant feedstocks and biomodification of the target plant sources. For the former, an ionic liquid-based processing system is proposed for efficient pretreatment of raw plant feedstocks. For the latter, the genes encoding the key enzymes; sucrose:sucrose 1-fructoyltransferase (1-SST), fructan:fructan 1-fryuctosyltransferase (1-FFT), fructan 1-exohydrolase (1-FEH), and microbe-derived endoinulinase, are introduced for biomodification conducive to facilitating bioprocess and improving inulin content. These approaches would contribute to cost-efficiently and sustainably producing bio-based FDCA.

Toward Sustainable Hydroxymethylfurfural Production Using Seaweeds.

Chemical manufacturing involves carbon sources releasing CO2 into the atmosphere. By contrast, seaweeds are carbon sinks that can absorb released CO2 and therefore have great potential for use as feedstocks in sustainable chemical manufacturing. In particular, seaweeds could contribute to mitigating vast amounts of global CO2 emissions. Accordingly, seaweeds could be an excellent candidate biomaterial for sustainable production of hydroxymethylfurfural (HMF), called a 'sleeping giant' platform chemical due to its wide versatility in chemical manufacturing. HMF is produced through sugar dehydration mechanisms, and seaweed storage glucans comprised of glucose can be appropriate feeding substrates for its production. This opinion article introduces a new opportunity for sustainable production of HMF using storage glucan-rich seaweeds.

Raw plant-based biorefinery: A new paradigm shift towards biotechnological approach to sustainable manufacturing of HMF.

Unlike petrorefinery, biorefinery uses carbon-based biomaterials, such as plant feedstocks, as the major feeding input materials in chemical manufacturing. To date, petroleum-based resources have been used for the production of wide spectrums of chemical products. However, petrorefinery is currently associated with a variety of issues, i.e., concerns over adverse impacts on the environment and human society. As an alternative technology, the sustainable biorefinery is a matter of great importance in industrial chemical manufacturing due primarily to its sustainability. As carbon-based resources, plants are paramount biomaterials for biorefinery process required in sustainable chemical manufacturing. In particular, raw plant-based biorefinery is a breakthrough technology for chemical manufacturing due mainly to its sustainable benefits. Nowadays, numerous biorefinery technologies have been developed for the production of industrially valuable chemicals. HMF, a versatile platform chemical, can be produced by dehydrating hexose sugars using raw plant feedstocks such as inulin-rich, starch-rich, and lignocellulosic plants and now, it is generally recognized as a chemical feedstock for future chemical manufacturing and bioenergy production. In this review article, this emerging hybrid technology is discussed in relation to the production of HMF from raw plant feedstocks mentioned above. In addition, the plant candidates useful for biorefinery processing of raw plant feedstocks are introduced and bioengineering strategy for their genetic modification is together described to provide current knowledge on sustainable biorefinery.

Purification and characterization of cellulase produced by Bacillus amyoliquefaciens DL-3 utilizing rice hull.

A microorganism hydrolyzing rice hull was isolated from soil and identified as Bacillus amyloliquefaciens by analysis of 16S rDNA and partial sequences of the gyrA gene, and named as B. amyloliquefaciens DL-3. With the analysis of SDS-PAGE, the molecular weight of the purified cellulase was estimated to be 54kDa. The purified cellulase hydrolyzed avicel, caboxymethylcellulose (CMC), cellobiose, beta-glucan and xylan, but not p-Nitrophenyl-beta-D-glucopyranoside (PNPG). Optimum temperature and pH for the CMCase activity of the purified cellulase were found to be 50 degrees C and pH 7.0, respectively. The CMCase activity was inhibited by some metal ions, N-bromosuccinimide and EDTA in the order of Hg(2+)>EDTA>Mn(2+)>N-bromosuccinimide>Ni(2+)>Pb(2+)>Sr(2+)>Co(2+)>K(+). The open reading frame of the cellulase from B. amyloliquefaciens DL-3 was found to encode a protein of 499 amino acids. The deduced amino acid sequence of the cellulase from B. amyloliquefaciens DL-3 showed high identity to cellulases from other Bacillus species, a modular structure containing a catalytic domain of the glycoside hydrolase family 5 (GH5), and a cellulose-binding module type 3 (CBM3).

Effects of permanent waving on changes of protein and physicomorphological properties in human head hair.

Two types of permanent waving [digital perm (DP) and croquignole winding perm (CWP)] and two waving lotions [cysteamine-HCl, pH 9.31, liquid type (lotion A) and sodium thioglycolate, pH 9.97, cream type (lotion B)] were used for this study. The protein content was decreased by permanent waving treatments on the whole, and the degree of reduction was dependent on the hair styling and waving lotion used. The greatest decrease (by 58%) was found in hairs processed with the three-treatment performance of DP using lotion B. SDS-PAGE identified the presence of the two most abundant polypeptides, with approximately 48 kDa and 60 kDa, and two large polypeptides, with approximately 200 kDa and 210 kDa, which would belong to the keratin family. Some physical properties of the hairs (tensile strength, diameter, swelling, and elongation) were examined with the permanent waving treatments. In general, the repeated waving treatment and the use of lotion B showed more negative effects on hair care than other treatments. Some morphological changes were observed with a scanning electron microscope (SEM). The most prominent change in the hair surface was observed in the hair specimen with the three-treatment performance of DP using lotion B. More severe signs of damage appeared on the hair with lotion B than with lotion A. As the numbers of permanent waves increased, the degree of damage to the hair surface increased on the whole. However, there was no indication of changes to the hair surface with one permanent waving treatment, as determined by SEM analysis.

Molecular and biochemical characterizations of dehydroascorbate reductase from sesame (Sesamum indicum L.) hairy root cultures.

Dehydroascorbate reductase (DHAR) is a biotechnologically or physiologically important reducing enzyme in the ascorbate-glutathione recycling reaction for most higher plants. A DHAR cDNA was isolated from sesame (Sesamum indicum L.) hairy roots, and its structure and biochemical properties were characterized to provide some information about its expressional and biochemical profiles in the hairy root cultures. The cDNA contained a catalytic motif CXXS, which may be indicative of a thiol-dependent redox function. A fusion DHAR expressed in an Escherichia coli expression system was purified with four purification steps until a homogeneous single band signal was seen in an acrylamide gel, and its antibody was prepared for Western blot analyses. The biochemical results showed that the purified recombinant DHAR had an optimal pH of around 6.0, which was different from those (pH 7.8-8.2) of other plant species. The temperature optimal for the DHAR activity was in a relatively wide range of 30-60 degrees C. It was proved by a real-time RT-PCR technique that the transcription activity of the DHAR was about 2-5-fold higher during the first 3 week cultures than during the latter 3 week ones. The highest activity of the sesame DHAR was detected in the 4 week cultures of the hairy roots, after which its activity was rapidly decreased to approximately 80%, suggesting that the most active DHAR occurred in this culture period. Western blot analyses confirmed that the presence of DHAR enzyme was identified in both cultures of the fused E. coli and the sesame hairy roots.

A simple improved method for protein extraction from human head hairs.

This study was conducted to establish a simple efficient method for extracting the protein from human head hair materials, which can be a useful tool for the protein analysis applicable to various types of human head hairs. The method developed saves extraction time and effort considerably. The method includes four steps: cutting the hair samples into small pieces 1-2 mm in length, washing them with distilled water, incubating the hair samples in a buffer solution at 50 degrees C for 24 hr, and finally filtering the incubated mixtures through three layers of nylon mesh. This method is reproducible and reliable. SDS-PAGE analysis of the hair protein extracted by this method shows a clear protein profile on the gel, which is frequently observed in other hair sources. Two smaller sizes of molecular weights are also detected with the SDS-PAGE analysis. Not commonly found in other hair sources, they seem to be other types of human hair proteins.

Production of high molecular weight pullulan by Aureobasidium pullulans HP-2001 with soybean pomace as a nitrogen source.

The production of pullulan by Aureobasidium pullulans HP-2001 was enhanced by yeast extract as a nitrogen source as well as soybean pomace. The highest production of pullulan by A. pullulans HP-2001 with yeast extract was 5.5 g/l whereas that of pullulan with soybean pomace was 7.5 g/l. The gas chromatogram of pullulan produced by A. pullulans HP-2001 with soybean pomace as a nitrogen source showed that the major and minor components were glucose and mannose. The FTIR spectra of pullulans produced with yeast extract, a mixture of yeast extract and soybean pomace, and soybean pomace alone exhibited similar features. The increase in content of reducing sugars after pullulanase treatment of pullulans produced with different nitrogen sources indicated that all the pullulans had alpha-(1,6) glucosidic linkages of alpha-(1,4) linked maltotriose units. The average molecular weights of pullulans produced with various concentrations of yeast extract and soybean pomace ranged from 0.17 to 1.32x10(6) and from 1.32 to 5.66x10(6), respectively. All pullulans produced by A. pullulans HP-2001 in this study had the same basic structures, but their ratios of monomeric components were a little different, which might result in the production of pullulans with different molecular weights.

Enhanced Production of carboxymethylcellulase by a marine bacterium, Bacillus velezensis A-68, by using rice hulls in pilot-scale bioreactor under optimized conditions for dissolved oxygen.

The optimal conditions for the production of carboxymethylcellulase (CMCase) by Bacillus velezensis A-68 at a flask scale have been previously reported. In this study, the parameters involved in dissolved oxygen in 7 and 100 L bioreactors were optimized for the pilot-scale production of CMCase. The optimal agitation speed and aeration rate for cell growth of B. velezensis A-68 were 323 rpm and 1.46 vvm in a 7 L bioreactor, whereas those for the production of CMCase were 380 rpm and 0.54 vvm, respectively. The analysis of variance (ANOVA) implied that the highly significant factor for cell growth was the aeration rate, whereas that for the production of CMCase was the agitation speed. The optimal inner pressures for cell growth and the production of CMCase by B. velezensis A-68 in a 100 L bioreactor were 0.00 and 0.04 MPa, respectively. The maximal production of CMCase in a 100 L bioreactor under optimized conditions using rice hulls was 108.1 U/ml, which was 1.8 times higher than that at a flask scale under previously optimized conditions.

Enhanced carboxymethylcellulase production by a newly isolated marine bacterium, Cellulophaga lytica LBH-14, using rice bran.

The aim of this work was to establish the optimal conditions for production of carboxymethylcellulase (CMCase) by a newly isolated marine bacterium using response surface methodology (RSM). A microorganism producing CMCase, isolated from seawater, was identified as Cellulophaga lytica based 16S rDNA sequencing and the neighborjoining method. The optimal conditions of rice bran, ammonium chloride, and initial pH of the medium for cell growth were 100.0 g/l, 5.00 g/l, and 7.0, respectively, whereas those for production of CMCase were 79.9 g/l, 8.52 g/l, and 6.1. The optimal concentrations of K2HPO4, NaCl, MgSO4·7H2O, and (NH4)2SO4 for cell growth were 6.25, 0.62, 0.28, and 0.42 g/l, respectively, whereas those for production of CMCase were 3.72, 0.54, 0.70, and 0.34 g/l. The optimal temperature for cell growth and the CMCase production by C. lytica LBH-14 were 35 degrees C and 25 degrees C, respectively. The maximal production of CMCase under optimized condition for 3 days was 110.8 U/ml, which was 5.3 times higher than that before optimization. In this study, rice bran and ammonium chloride were developed as carbon and nitrogen sources for the production of CMCase by C. lytica LBH-14. The time for production of CMCase by a newly isolated marine bacterium with submerged fermentations reduced to 3 days, which resulted in enhanced productivity of CMCase and a decrease in its production cost.

Chromium halides mediated production of hydroxymethylfurfural from starch-rich acorn biomass in an acidic ionic liquid.

Chromium halides were introduced for the sustainable production of hydroxymethylfurfural (HMF) from raw acorn biomass using an acidic ionic liquid. The free sugars (glucose and maltose) released by the acidic hydrolysis of the biomass were confirmed by the FT-IR absorption bands around 995-1014cm(-1) and HPLC. FESEM analysis showed that the acorn biomass contains various sizes of starch granules and their structures were severely changed by the acidic hydrolysis. An optimal concentration of HCl for the HMF yields was 0.3M. The highest HMF yield (58.7+1.3dwt%) was achieved in the reaction mixture of 40% [OMIM]Cl+10% ethyl acetate+50% 0.3M HCl extract containing a mix of CrBr(3)/CrF(3). The combined addition of two halide catalysts was more effective in the synthesis of HMF (1.2-fold higher on average) than their single addition. The best productivity of HMF was found at 15% concentration of the biomass and at 50%, its relative productivity declined down to ca. 0.4-fold.

Kinetic study on the pretreatment and enzymatic saccharification of rice hull for the production of fermentable sugars.

The production of fermentable sugars from rice hull was studied by dilute acid pretreatment and enzymatic saccharification. Rice hull (15%, w/v) was pretreated by 1% (v/v) sulfuric acid at high temperature (120 approximately 160 degrees C) for 15, 30, 45, and 60 min, respectively. The maximum sugar concentration from rice hull in the prehydrolysate was obtained at 140 degrees C for 30 min, but the enzymatic saccharification yield from the corresponding pretreated rice hull is not high. To another aspect, the maximum enzymatic saccharification yield was achieved at 160 degrees C for 60 min, while the recovery of fermentable sugars was the poorest. To take account of fermentable sugars from pretreatment and enzymatic saccharification, the maximum yield of sugars was obtained only when rice hull was treated at 140 degrees C for 30 min. Under this condition, 72.5% (w/w) of all sugars generated from the raw material can be recovered. The kinetic study on the enzymatic saccharification of dilute acid pretreated rice hull was also performed in this work by a modified Michaelis-Menten model and a diffusion-limited model. After calculation by a linear and a non-linear regression analysis, both models showed good relation with the experimental results.

Seed-specific expression of sesame microsomal oleic acid desaturase is controlled by combinatorial properties between negative cis-regulatory elements in the SeFAD2 promoter and enhancers in the 5'-UTR intron.

The regulation of genes involved in primary lipid metabolism in plants is much less well understood than that in many other pathways in plant biology. In the investigation reported here, we have characterized transcriptional regulatory mechanisms controlling seed-specific FAD2 expression in sesame (Sesamum indicum). FAD2 codes for extra-plastidial FAD2 desaturase, which catalyzes the conversion of oleic acid to linoleic acid. Promoter analysis of the sesame FAD2 gene (SeFAD2) using the beta-glucuronidase (GUS) reporter system demonstrated that the - 660 to - 180 promoter region functions as a negative cis-element in the seed-specific expression of the SeFAD2 gene. Sesame and Arabidopsis FAD2 genes harbor one large intron within their 5'-untranslated region. These introns conferred up to 100-fold enhancement of GUS expression in transgenic Arabidopsis tissues as compared with intron-less controls. Prerequisite cis-elements for the SeFAD2 intron-mediated enhancement of gene expression and the promoter-like activity of SeFAD2 intron were identified. SeFAD2 transcripts were induced by abscisic acid (ABA) in developing sesame seeds, and the - 660 to - 548 and - 179 to - 53 regions in the SeFAD2 promoter were implicated in ABA-responsive signaling. Theses observations indicate that an intron-mediated regulatory mechanism is involved in controlling not only the seed-specific expression of the SeFAD2 gene but also the expression of plant FAD2 genes, which are essential for the synthesis of polyunsaturated fatty acids.

Expression and characterization of extracellular fungal phytase in transformed sesame hairy root cultures.

A recombinant fungal phytase was produced by cultures of sesame hairy roots transformed with Agrobacterium rhizogenes, purified and its molecular properties were characterized. Its transcription level and the phytase production were rapidly increased after 4 weeks of the cultures, suggesting that its transcription and protein synthesis might concur. Western blot analysis provided evidence that the recombinant fungal phytase was secreted into the liquid culture medium of the hairy roots. The phytase enzyme secreted was purified by three steps of ultrafiltration, DEAE-Sepharose ion exchange chromatography, and Sephadex G-100 size-exclusion chromatography. As a result, one single band signal was observed with SDS-PAGE, indicating that the purification step was reasonable. The positive signs of both the zymogram and the PAS staining on SDS-PAGE suggested that the activity of the final product phytase was active and glycosylated. The optimal reaction temperature of the phytase was between 50 and 60 degrees C and at over 60 degrees C its activity was reduced by 30-90%, depending on the temperatures applied. Pre-incubation at temperatures of 20-50 degrees C showed stable catalytic activity, while at over 50 degrees C the phytase activity was gradually decreased by 90%. The optimal pH was between 4 and 5 pH values for the recombinant fungal phytase, while for native phytase it was at pH 5.0. Addition of iron ion inhibited the phytase activity but treatments of some cations, EDTA, and PMSF showed no effect on the activity or slightly stimulated it positively.

Molecular cloning of low-temperature-inducible ribosomal proteins from soybean.

Three ribosomal protein genes induced by low-temperature treatment were isolated from soybean. GmRPS13 (742 bp) encodes a 17.1 kDa protein which has 95% identity with the 40S ribosomal protein S13 of Panax ginseng (AB043974). GmRPS6 (925 bp) encodes a 28.1 kDa protein which has 94% identity with the 40S ribosomal protein S6 of Asparagus officinalis (AJ277533). GmRPL37 (494 bp) encodes a 10.7 kDa protein which has 85% identity with the 60S ribosomal protein L37 of Arabidopsis thaliana (AF370216). The expression of these ribosomal protein genes started to increase 3 d after low-temperature treatment, whereas the cold-stress protein src1 was highly induced from the first day. Such late response of these ribosomal protein genes may be due to secondary signals during cold adaptation. The induction of ribosomal protein genes might enhance the translation process or help proper ribosome functioning under low-temperature conditions.

Comparative analysis of expressed sequence tags from Sesamum indicum and Arabidopsis thaliana developing seeds.

Sesame (Sesamum indicum) is an important oilseed crop which produces seeds with 50% oil that have a distinct flavor and contains antioxidant lignans. Because sesame lignans are known to have antioxidant and health-protecting properties, metabolic pathways for lignans have been of interest in developing sesame seeds. As an initial approach to identify genes involved in accumulation of storage products and in the biosynthesis of antioxidant lignans, 3328 expressed sequence tags (ESTs) were obtained from a cDNA library of immature seeds 5-25 days old. ESTs were clustered and analyzed by the BLASTX or FASTAX program against the GenBank NR and Arabidopsis proteome databases. To compare gene expression profiles during development of green and non-green seeds, a comparative analysis was carried out between developing sesame and Arabidopsis seed ESTs. Analyses of these two seed EST sets have helped to identify similar and different gene expression profiles during seed development, and to identify a large number of sesame seed-specific genes. In particular, we have identified EST candidates for genes possibly involved in biosynthesis of sesame lignans, sesamin and sesamolin, and also suggest a possible metabolic pathway for the generation of cofactors required for synthesis of storage lipid in non-green oilseeds. Seed-specific expression of several candidate genes has been confirmed by northern blot analysis.

Isolation and characterization of a myo-inositol 1-phosphate synthase cDNA from developing sesame (Sesamum indicum L.) seeds: functional and differential expression, and salt-induced transcription during germination.

A cDNA (SeMIPS1) encoding myo-inositol 1-phosphate synthase (EC 5.5.1.4) (MIPS) has been characterized from sesame (Sesamum indicum L. cv. Dan-Baek) seeds and its functional expression analyzed. The SeMIPS1 protein was highly homologous with those from other plant species (88-94%), while a much lower degree of sequence homology (53-62%) was found with other organisms such as humans, mouse, algae, yeast, Drosophila, bacteria and other prokaryotes. A yeast-based complementation assay in yeast mutants containing a disrupted INO1gene for yeast MIPS confirmed that the SeMIPS1 gene encodes a functional MIPS. Phylogenetic analysis suggested that the SeMIPS1 gene diverged as a different subfamily or family member. Southern hybridization revealed several copies of the SeMIPS1 gene present in the sesame genome and northern blotting indicated that expression of the SeMIPS1gene may be organ specific. Salt stress during sesame seed germination had an adverse influence on transcription of SeMIPS1and greatly reduced transcript levels as the duration of exposure to a saline environment increased and NaCl concentration increased. Germination initiation of sesame seeds was severely delayed as NaCl level increased. These results suggest that expression of SeMIPS1 is down-regulated by salt stress during sesame seed germination.