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PubMed2005

A protective endophyte of maize: Acremonium zeae antibiotics inhibitory to Aspergillus flavus and Fusarium verticillioides.

The maize endophyte Acremonium zeae is antagonistic to kernel rotting and mycotoxin producing fungi Aspergillus flavus and Fusarium verticillioides in cultural tests for antagonism, and interferes with A. flavus infection and aflatoxin contamination of preharvest maize kernels. Chemical studies of an organic extract from maize kernel fermentations of Acremonium zeae (NRRL 13540), which displayed significant antifungal activity against Aspergillus flavus and F. verticillioides, revealed that the metabolites accounting for this activity were two newly reported antibiotics pyrrocidines A and B. Pyrrocidines were detected in fermentation extracts for 12 NRRL cultures of Acremonium zeae isolated from maize kernels harvested in Illinois (4/4 cultures), North Carolina (5/5), Georgia (1/2) and unrecorded locations within the USA (2/2). Pyrrocidine B was detected by LCMSMS in whole symptomatic maize kernels removed at harvest from ears of a commercial hybrid that were wound-inoculated in the milk stage with A. zeae (NRRL 13540) or (NRRL 13541). The pyrrocidines were first reported from the fermentation broth of an unidentified filamentous fungus LL-Cyan426, isolated from a mixed Douglas Fir hardwood forest on Crane Island Preserve, Washington, in 1993. Pyrrocidine A exhibited potent activity against most Gram-positive bacteria, including drug-resistant strains, and was also active against the yeast Candida albicans. In an evaluation of cultural antagonism between 13 isolates of A. zeae in pairings with A. flavus (NRRL 6541) and F. verticillioides (NRRL 25457), A. zeae (NRRL 6415) and (NRRL 34556) produced the strongest reaction, inhibiting both organisms at a distance while continuing to grow through the resulting clear zone at an unchanged rate. Maximum colony diameters for A. zeae (NRRL 6415) and (NRRL 13540), on potato dextrose agar after 14 d, were attained within the range of 25-30 degrees C, with less growth recorded at 15 degrees and 37.5 degrees and no growth at 5 degrees. Potential interactions between A. zeae and other maize endophytes are considered and the significance of these interactions relative to the aflatoxin and fumonisin contamination of preharvest maize is presented. This is the first report of natural products from Acremonium zeae.

Acremonium↗

PubMed2006

[A strategy based on comparative genomics to align ESTs of maize].

In this study, a new strategy to locate ESTs on maize linkage groups was described. In the strategy, the rice (Oryza sativa L.) genomic sequence database of was employed to locate maize EST on rice linkage groups, and then to locate on maize linkage group by comparative genetics mapping between rice and maize genome. The aligned ESTs information should available for further study on genomics and gene cloning. As an example, 139 ESTs of maize were assayed, and 96 maize ESTs (69%) were homologous with rice genomic sequence, 55% (77/139) ESTs were located on maize linkage groups based on the strategy, indicating that the locating approach of ESTs is feasible and available.

Expressed Sequence Tags↗

PubMed1980

Field outbreaks of hyperoestrogenism (vulvo-vaginitis) in pigs consuming maize infected by Fusarium graminearum and contaminated with zearalenone.

During the spring and summer of 1979 field outbreaks of porcine hyperoestrogenism characterized by swelling and reddening of the vulva and teats in prepubertal gilts and enlargement of the mammae in young males occurred in the mistbelt of the Natal Midlands. On a farm in the Ixopo district, pigs were fed a mixed ratio containing home-grown yellow maize stored on the cob in a crib and all the young pigs with a mass of 25-85 kg (=a total of 350) were affected. Maize ears from the crib had a high percentage (91%) of kernels infected by Fusarium graminearum and contained 10 mg/kg of zearalenone. The mixed ration contained 0,95 mg/kg zearalenone. After dilution of the contaminated maize with good quality white maize, a dramatic decrease in both the incidence and severity of clinical signs occurred within 3-4 days. On a farm in the Winterton district, approximately 80% of the growing pigs with a mass of more than 40 kg fed a mixed ration containing shelled yellow maize from the silo of the local Agricultural Co-operative were affected. The incidence as well as the severity of the swelling of the vulvas and/or treats were lower than on the farm at Ixopo, thus indicating a lower level of oestrogen intake. A sample of maize from the silo delivered to this farm contained only 0,06 mg/kg zearalenone while the mixed ration did not contain chemically detectable levels of zearalenone. This is the first published record of field outbreaks of porcine hyperoestrogenism associated with the ingestion of F. graminearum infected maize contaminated with zearalenone in the Republic of South Africa.

Animal Feed↗

PubMed1984

Anaerobic expression of maize glucose phosphate isomerase I.

Anaerobic treatment of maize seedlings results in the selective expression of 10 major and 10 minor polypeptides designated as anaerobic polypeptides (ANPs) (Sachs, M. M., Freeling, M., and Okimoto, R. (1980) Cell 20, 761-767). Enzymatic functions have been ascribed to two of these polypeptides, alcohol dehydrogenases I and II. This report identifies a third anaerobic polypeptide, a maize glucose phosphate isomerase. Glucose phosphate isomerase specific activity increased 60% in 24 h of anaerobiosis and to more than twice its original level in 72 h. Antiserum to spinach cytoplasmic glucose phosphate isomerase 1) was monospecific against maize protein, 2) reduced glucose phosphate isomerase activity in maize extracts by 60%, and 3) recognized a single polypeptide selectively labeled with [35S]methionine during anaerobiosis. This polypeptide co-purified with the major glucose phosphate isomerase of maize. Maize glucose phosphate isomerase co-migrated on two-dimensional polyacrylamide gels with ANP55, the anaerobic polypeptide with a molecular weight of 55,000 on sodium dodecyl sulfate-polyacrylamide gels. Electrophoretically distinguishable alleles of Phi 1 (phosphohexoisomerase-1 gene located at maize chromosome 1L-140) predictably altered the electrophoretic behavior of ANP55, thus proving that Phi 1 encodes ANP55 and that ANP55 is a glucose phosphate isomerase.

Alleles↗

PubMed1994

Expression of branching enzyme II of maize endosperm in Escherichia coli.

A cDNA clone encoding maize branching enzyme II (BEII) has been independently isolated from a maize endosperm cDNA library. The deduced protein sequence of maize BEII was compared with that of BE from diverse sources. The gene encoding mature BEII of maize endosperm has been expressed in E. coli using the T7 promoter. The expressed BEII was purified to near homogeneity so that amylolytic activity and bacterial BE could be completely eliminated from the BE preparation. The expressed enzyme showed very similar properties to those of BEII purified from developing maize endosperm. This result confirmed our earlier report that BEII had a lower rate of branching amylose and the rate of branching amylopectin was twice that of branching amylose. This study also showed a greater advantage of purifying BEII from the bacterial expression system than from developing maize endosperm. Most importantly, this study has established a useful tool to study the structure-function relationships of the maize BE using site-directed mutagenesis.

1,4-alpha-Glucan Branching Enzyme↗

PubMed1996

Inhibition of UDP-glucose: protein transglucosylase by a maize endosperm protein factor.

UDP-glucose: protein transglucosylase (UPTG, EC 2.4.1.112) catalyzes the first step of protein-bound alpha-glucan synthesis in potato tuber and developing maize endosperm. The presence of a non-dialyzable, heat labile protein responsible for low levels of UPTG activity in developing maize endosperm was investigated. UPTG activity in 5-day old maize seedlings and potato tuber solubilized preparations was also reduced by the endosperm preparation. FPLC-Mono Q column chromatography of developing maize endosperm was effective in separating the inhibitor protein (IP) from UPTG. After gel filtration on Superose 12, IP yielded a major polypeptide of about 80 kDa on SDS-PAGE. IP was purified by gel filtration on Superose 12 and preparative SDS-PAGE, and specific antibodies were prepared. Polyclonal antibodies reacted specifically with an 80 kDa polypeptide of developing maize endosperm on Western blot. They also recognized a similar band in 5-day old maize seedlings, but not in potato tubers. The identification of a factor that regulates the level of UPTG activity in developing maize endosperm may help to elucidate the functional role of the enzyme in the initiation of starch synthesis during seed development.

Enzyme Inhibitors↗

PubMed2026

Evolution of maize recombination landscape during domestication.

Despite the plethora of knowledge about the benefits of meiotic recombination and numerous theoretical studies examining how recombination rates evolve, there is a general lack of empirical support and consensus across species. To fill this knowledge gap, we characterized the evolution of recombination landscape in maize during its domestication from teosinte and related the observed changes to established theoretical frameworks. Through examining recombination in experimental populations of maize and teosinte and the population genomics approach of identifying historical recombination events using ancestral recombination graph inference to generate saturated maize and teosinte recombination maps, we found that during domestication, maize experienced a 12% increase in its genome-wide recombination rate. Furthermore, maize evolved higher recombination rates on the long arms of chromosomes in regions closer to centromeres, where recombination is generally very low. The repatterning of crossover events came from changes in global crossover positioning rather than alterations in cis-acting chromatin factors. Consequently, we found evidence of selection acting on trans-acting recombination modifiers affecting crossover interference and controlling the interference-dependent class I crossover pathway. We show that CO repatterning was likely beneficial for maize fitness, as significant recombination rate increases were predominantly in gene-rich regions, which harbor domestication-related variation. This work suggests genomic and mechanistic processes leading to the evolution of meiotic recombination landscape in response to directional selection pressure and provides evidence for the evolutionary advantage of recombination.

Zea mays↗

PubMed2007

Inhibition of maize streak virus (MSV) replication by transient and transgenic expression of MSV replication-associated protein mutants.

Maize streak disease is a severe agricultural problem in Africa and the development of maize genotypes resistant to the causal agent, Maize streak virus (MSV), is a priority. A transgenic approach to engineering MSV-resistant maize was developed and tested in this study. A pathogen-derived resistance strategy was adopted by using targeted deletions and nucleotide-substitution mutants of the multifunctional MSV replication-associated protein gene (rep). Various rep gene constructs were tested for their efficacy in limiting replication of wild-type MSV by co-bombardment of maize suspension cells together with an infectious genomic clone of MSV and assaying replicative forms of DNA by quantitative PCR. Digitaria sanguinalis, an MSV-sensitive grass species used as a model monocot, was then transformed with constructs that had inhibited virus replication in the transient-expression system. Challenge experiments using leafhopper-transmitted MSV indicated significant MSV resistance--from highly resistant to immune--in regenerated transgenic D. sanguinalis lines. Whereas regenerated lines containing a mutated full-length rep gene displayed developmental and growth defects, those containing a truncated rep gene both were fertile and displayed no growth defects, making the truncated gene a suitable candidate for the development of transgenic MSV-resistant maize.

Gene Expression↗

PubMed1986

The structure and expression of maize genes encoding the major heat shock protein, hsp70.

We have isolated and sequenced two maize genomic clones that are homologous to the Drosophila hsp70 gene. One of the maize hsp70 clones contains the entire hsp70 coding region and 81 nucleotides of the 5' nontranslated sequence. The predicted amino acid sequence for this maize protein is 68% homologous to the hsp70 of Drosophila. The second maize hsp70 clone contains only part of the coding sequence and 1.1 kb of the 5' flanking sequence. This 5' flanking sequence contains two sequences homologous to the consensus heat-shock-element sequence. Both maize genes are thermally inducible and each contains an intron in the same position as that of the heat-shock-cognate gene, hsc1, of Drosophila. The presence of an intron in the maize genes is a distinguishing feature in that no other thermally inducible hsp70 genes described to date contain an intron. We have constructed a hybrid hsp70 gene containing the entire hsp70 coding sequence with an intron, and 1.1 kb of the 5' flanking sequence. We demonstrate that this hybrid gene is thermally inducible in a transgenic petunia plant and that the gene is expressed from its own promoter.

Journal Article↗

PubMed1992

Zmhox1a, the product of a novel maize homeobox gene, interacts with the Shrunken 26 bp feedback control element.

A new maize homeobox gene was isolated by screening a lambda gt11 expression library with the 26 bp Shrunken feedback control element. Zmhox1a (Zea mays homeobox) is an unidentified maize gene mapping to the long arm of chromosome 8. It is a member of a new class of maize homeobox genes only distantly related to the Knotted class. The 3.1 kb Zmhox1a transcript can be detected in different maize tissues and encodes a polypeptide of 719 amino acids. Western blotting experiments detect the native 112 or 115 kDa protein in nuclear protein extracts, the nuclear localization being compatible with a function in transcriptional control. No Zmhox1a protein is detected in maize roots despite the presence of the Zmhox1a transcript; this may indicate a post-transcriptional control mechanism. A highly acidic central region of the Zmhox1a polypeptide implies a transcriptional activator function. The carboxy-terminal part of the maize homeodomain protein is related to the human Oct2 transcription factor, but homology to the POU specific domain is restricted to the POU-B subdomain. It was confirmed by DNase I footprinting experiments that DNA binding of the Zmhox1a homeodomain was at three sites flanking the TATA-box of the Shrunken promoter.

Amino Acid Sequence↗

PubMed2006

Proteome analysis of maize pollen for allergy-relevant components.

Over the last few decades, the cultivation of maize (Zea mays) has strongly increased in Central Europe. We therefore decided to study the allergen composition and the allergenic potency of its pollen in comparison with pollen from timothy grass (Phleum pratense), a typical representative of the native grasses. We found that 65% of the sera reactive to timothy pollen also bound to maize pollen proteins. By using 2-DE immunoblotting, followed by incubation with mAbs directed against known allergens or protein sequencing, those IgE-reactive components were further classified. Although novel, maize-specific pollen allergens could not be found, the presence of crossreacting allergens belonging to groups 1 and 13 (Zea m 1 and 13), both having high IgE prevalence, as well as the presence of the less important group 3 and 12 allergens was found. The structural variability of Zea m 1 and Zea m 13 was determined by sequencing clones isolated from a maize pollen cDNA library. This revealed sequence identities of 72 and 70%, respectively, to the corresponding Phl p 1 and Phl p 13 allergens of timothy grass pollen. IgE-crossreactivity was further studied using immunoblot inhibition tests. Here, timothy pollen extract completely blocked IgE binding to maize, whereas maize pollen extract blocked IgE reactivity to only some timothy pollen allergens.

Allergens↗

PubMed2006

Persistence and efficacy of two diatomaceous earth formulations and a mixture of diatomaceous earth with natural pyrethrum against Tribolium confusum Jacquelin du Val (Coleoptera: Tenebrionidae) on wheat and maize.

Laboratory tests were conducted to assess the insecticidal and residual effects of three diatomaceous earth (DE) formulations, Insecto, PyriSec and SilicoSec, against Tribolium confusum Jacquelin du Val on wheat and maize. Quantities of wheat and maize were treated with the above formulations at 500, 1000 and 1500 mg kg(-1) and stored at 25 degrees C and 55% relative humidity (RH). Samples were taken on the day of storage and every 30 days until completion of a 360 day period of storage. Adults of T. confusum were exposed to these samples at 25 degrees C and 55% RH and the mortality was measured after 24 and 48 h and 7 and 14 days of exposure. Adult mortality was higher on wheat than on maize. At the beginning of the storage period, mortalities after 14 day exposure on maize treated with the highest rate were 60, 63 and 81% for Insecto, PyriSec and SilicoSec respectively, while on wheat the mortality was 100% for all DEs. On the same commodities 360 days after treatment the respective mortality figures for wheat were 99, 98 and 100%, while in the case of maize they did not exceed 7%. Generally, mortality at exposures < or =48 h decreased with increasing storage time. Furthermore, although mortality on wheat increased with dose, the increase in DE dose from 1000 to 1500 mg kg(-1) resulted in only a small increase in mortality. Thus a DE treatment of 1000 mg kg(-1) was shown to provide long-term protection of wheat against T. confusum, although higher DE application rates and exposure intervals are needed for a satisfactory level of protection of maize against this pest.

Animals↗

PubMed1994

Identification of a maize root transcript expressed in the primary response to nitrate: characterization of a cDNA with homology to ferredoxin-NADP+ oxidoreductase.

To more fully understand the biochemical and molecular events which occur in plants exposed to nitrate, cDNAs whose accumulation was enhanced in nitrate- and cycloheximide-treated maize (Zea mays L. W64A x W182E) roots were isolated. The 340 bp Zmrprn1 (for Zea mays root primary response to nitrate) cDNA also hybridized with a probe enriched for nitrate-induced sequences, and was characterized further. Sequence analysis of a near full-length cDNA (Zmrprn1A) showed strong homology (> 90% amino acid identity) with a root ferredoxin-NADP+ oxidoreductase (FNR) of rice, and 45-50% amino acid identify with leaf FNR genes. When expressed in Escherichia coli, the Zmrprn1A cDNA produced a protein with NADPH: ferricyanide reductase activity, consistent with the enzymatic properties of an FNR. The Zmrprn1 cDNA hybridized with a 1.4 kb transcript which was expressed in the maize root primary response to nitrate. That is, mRNA levels in roots increased rapidly and transiently in response to external nitrate, and low levels of nitrate (10 microM) induced transcript accumulation. The accumulation of the Zmrprn1 transcript was not prevented by cycloheximide, indicating that the cellular factor(s) required for expression were constitutively present in maize roots. The Zmrprn1 mRNA accumulated specifically in response to nitrate, since neither K+ nor NH4+ treatment of roots caused transcript accumulation. Maize leaves had about 5% of the transcript level found in roots, indicating a strong preference for expression of Zmrprn1 in roots. Analysis of maize genomic DNA indicated the presence of only a single gene or very small gene family for the Zmrprn1. Together, the data indicate that Zmrprn1A encodes a nitrate regulated maize root FNR.

Amino Acid Sequence↗

PubMed1996

Regulation of cytosolic pyruvate, orthophosphate dikinase expression in developing maize endosperm.

Pyruvate orthophosphate dikinase (PPDK, E.C. 2.7.9.1) is an abundant enzyme in the leaves of C4 plants associated with the dicarboxylic acid pathways of CO2 fixation in the dark. PPDK activity has also been detected in the seeds of maize and other, non-C4 cereals, where its role has yet to be established. Using an anti-PPDK serum, two cross-reacting species of M(r) close to 90 000 were detected in developing maize endosperm of wild-type plants. In two independent opaque-2 mutant lines, one of the polypeptides was absent and the other was reduced in level. Similarly, endosperm PPDK mRNA levels were greatly reduced in the opaque-2 maize lines compared to wild type, suggesting that endosperm PPDK gene expression is under Opaque-2 control. However, a low level of PPDK mRNA could still be detected in these mutants, indicating that PPDK gene expression is not absolutely dependent on Opaque-2 but rather can be modulated by it. This interpretation was reinforced by the demonstration that the distribution of PPDK transcripts is not affected in o2 mutants, although the level is reduced, and that PPDK mRNA is detectable prior to 02 mRNA during the maturation of wild-type maize endosperm. Using oligonucleotides specific for the different maize PPDK genes, the o2 mutations were shown to affect only cyPPDKZml gene expression in maize line A69Y.

Base Sequence↗

PubMed1994

Differential transcription of plastome-encoded genes in the mesophyll and bundle-sheath chloroplasts of the monocotyledonous NADP-malic enzyme-type C4 plants maize and Sorghum.

The transcription of plastome-encoded genes in mesophyll and bundle-sheath chloroplasts of the monocotyledonous NADP-malic enzyme-type C4 species Zea mays L. (maize) and Sorghum bicolor (L.) Moench. was investigated. RNA accumulation and transcription were assayed starting from isolated mesophyll and bundle-sheath chloroplasts and using quantitative northern and run-on transcription analysis. Determination of the mesophyll to bundle-sheath ratios of transcript abundance in maize and Sorghum chloroplasts showed that the mRNAs of the plastome-encoded photosystem II genes analysed (psbA, psbB, psbD, psbH and psbE/F) varied from 2.5- to 4.0-fold (maize) and 3.1- to 5.2-fold (Sorghum), respectively. The rbcL transcript, in contrast, was more abundant in bundle-sheath chloroplasts of both species, about 3-fold in maize and more than 10-fold in Sorghum. On the other hand, transcripts of genes encoding the 16S ribosomal RNA (r16) and subunits of photosystem I (psaA) and the cytochrome b/f complex (petB, petA) accumulated to similar levels in both types of chloroplasts. Determination of absolute transcript levels for rbcL and psbA in chloroplasts from maize and Sorghum demonstrated that for both genes, the mesophyll to bundle-sheath differences in transcript abundance were more pronounced in Sorghum. Measurements of the transcriptional activities of rbcL and psbA showed that the transcription rate of rbcL is higher in bundle-sheath chloroplasts while psbA is more actively transcribed in mesophyll chloroplasts. The differences in the transcription rates between the two chloroplast types were again more pronounced in Sorghum, thus reflecting the differences between maize and Sorghum in the relative levels of the rbcL and psbA transcripts. However, although transcription rate and mRNA abundance are correlated, they did not exactly match one another. This indicates additional regulation of transcript abundance at the level of RNA stability.

Chloroplasts↗

PubMed1994

Molecular cloning of two different cDNAs for maize acetyl CoA carboxylase.

Acetyl CoA carboxylase (EC 6.4.1.2) in plants is a chloroplast-localized, biotin-containing enzyme that catalyses the carboxylation of acetyl CoA to malonyl CoA, the first committed step of the fatty acid biosynthesis pathway. Acetyl CoA carboxylase is the target site for the monocotyledon-specific aryloxyphenoxypropionate and cyclohexanedione groups of herbicides. We have purified a herbicide-sensitive acetyl CoA carboxylase from maize leaves to homogeneity (specific activity 7 mumol min-1 mg-1), separating it during the purification from a minor herbicide-resistant acetyl CoA carboxylase. The purified enzyme is a dimer of 230 kDa subunits. Antibodies raised to the purified acetyl CoA carboxylase detected three cross-reacting clones in a maize leaf cDNA expression library, each having an insert of 4-4.5 kb. Restriction analysis and sequencing showed that the cDNAs were derived from two different transcripts. Comparison of the deduced amino acid sequences with those of chicken and yeast acetyl CoA carboxylases confirmed that both types encoded acetyl CoA carboxylase, corresponding to the C-terminal half of the enzyme. The overall identity of the maize and chicken sequences was 37% (58% similarity) but for some shorter regions was much higher. Analysis of six other acetyl CoA carboxylase clones recovered from the maize cDNA library showed four belonged to one type and two to the other. The nucleotide sequence similarity between the two types of cDNA was approximately 95% in the coding region but considerably less in the 3'-untranslated region. Northern blot analysis of maize RNA showed a single band of 8.2-8.5 kb for acetyl CoA carboxylase mRNA. Southern blot hybridisations indicated that there are probably no more than two genes in maize for acetyl CoA carboxylase. The possible significance of two different cDNAs for acetyl CoA carboxylase is discussed.

Acetyl-CoA Carboxylase↗

PubMed1986

Cloning and characterization of a linear 2.3 kb mitochondrial plasmid of maize.

A linear 2.3 kb DNA molecule found in maize mitochondria was cloned into pUC8. A natural deletion of this plasmid, found in cmsT and some N (fertile) types of maize plants, was mapped to one end of the plasmid. A minor sequence homology to S-2, another linear mitochondrial plasmid, was detected, as well as more significant sequence homology with chloroplast and maize nuclear DNA. Hybridization to teosinte mitochondrial DNA (mtDNA) revealed the presence of part of the maize plasmid in the high molecular weight mtDNA of the maize relatives. RNA dot hybridization indicates that the plasmid is transcribed in mitochondria. The termini of the 2.3 kb linear plasmid contain inverted repeated sequences; of the first 17 nucleotides of the termini, 16 are identical to the terminal inverted repeats of the linear S plasmids found in the mitochondria of cmsS maize plants.

Chromosome Deletion↗

PubMed1994

Consumer acceptability of stiff porridge based on various composite flour proportions of sorghum, maize and cassava.

A study was conducted to determine consumer acceptable proportions of flours in sorghum (var. serena) and maize or sorghum, maize and cassava composite flours. Breeder's serena flour extracted at 75 percent was mixed at various proportions with maize or with maize and cassava to constitute composite flours. The various composite flours were made into stiff porridge and presented to a group of panellists for sensory evaluation. The evaluation was conducted by scoring on a hedonic scale of 0-5 for poor to excellent, respectively, for the parameters colour, texture, flavour, taste and overall acceptability. The responses were analysed statistically. Alternatively the panellists were allowed to eat ad-lib any of the presented samples which appealed to them. The results demonstrated that slight incorporation (< 10%) of sorghum (var. serena) to maize flour leads to reduction in consumer acceptability. However, increased incorporation of sorghum in excess of 10 percent leads to no significant decrease in consumer acceptability until a level of 30 percent incorporation is exceeded. In sorghum, maize, cassava composite flours the formulation of 30:40:30, respectively, was found to be most acceptable. It is suggested that at this combination of flours desirable textural characteristics of the porridge overrides other factors in contributing to its acceptability.

Consumer Behavior↗