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Functional analysis of a GWAS pleiotropic hotspot suggests an auxin biosynthesis gene (AhPDS1), regulating pod development in peanut (Arachis hypogaea L.).

Peanut productivity and quality improvement rely on understanding the genetic factors influencing pod and seed size. This study aims to identify genetic factors and regulatory mechanisms influencing pod and seed size in peanuts. Herein, a genome-wide association study (GWAS) was conducted using 390 accessions from 15 peanut growing regions to analyze pod and seed traits across multiple planting seasons. A significant phenotypic variation was observed, with broad-sense heritability ranging from 53.6 to 85.4%. Strong correlations between pod and seed traits further suggest potential for co-selection in breeding efforts. A pleiotropic hotspot on chromosome B06 was strongly associated with six pod and seed traits. A peanut pod size regulator AhPDS1 (PODSIZE-1, Ahy_B06g085516) homolog of Arabidopsis thaliana YUCCA4 (AtYUC4, AT5G11320), involved in auxin biosynthesis, was selected as a candidate regulating pod and seed size. Quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) confirmed higher AhPDS1 expression in large pod as compared with the small pod genotypes. Subcellular localization showed AhPDS1 to be predominantly cytoplasmic, and GUS reporter assays indicated widespread expression in roots, stems, leaves, flowers, and pods, suggesting a broad functional role. Further overexpression of AhPDS1 in Arabidopsis and rice enhanced pod, seed, and grain sizes via the indole-3-pyruvic acid pathway in transgene lines. These findings highlight AhPDS1 as a potential target for peanut molecular breeding, offering opportunities to enhance pod size via auxin biosynthesis and support sustainable crop improvement.

Arachis

Proteomic characterization of acidic aqueous extracts from Vicia faba L. pod valves identifies chitinase as a major co-extracted protein macromolecule.

Naturally acidic aqueous extracts from Vicia faba L. pod valves are being explored as sustainable, L-DOPA-oriented plant preparations. Pod valves represent an underutilized processing by-product reported to contain L-DOPA, a compound widely used in Parkinson's disease therapy, while acidic aqueous media may help preserve its physicochemical stability. However, the protein macromolecules co-extracted from V. faba pod valves under these conditions remain poorly characterized. This information is relevant because persistent plant proteins may influence extract composition, stability, susceptibility to degradation, and downstream processing requirements. Here, we characterized co-extracted V. faba protein macromolecules in aqueous pod-valve extracts prepared in ultrapure water or naturally acidic media, including 2% Phyllanthus emblica, 5% Punica granatum, and 2% Ribes rubrum. Protein profiles were first evaluated by SDS-PAGE and subsequently analyzed by nanoflow liquid chromatography coupled to high-resolution tandem mass spectrometry (nLC-MS/MS). Protein identifications were complemented with Gene Ontology annotation and a descriptive semi-quantitative assessment of relative protein representation across extraction media. Chitinase was the most represented V. faba-assigned protein macromolecule across the extracts, with additional highly represented proteins including glucan endo-1,3-beta-D-glucosidase, pathogenesis-related proteins, and polyphenol oxidase A1. These co-extracted proteins are mainly associated with plant defense, stress responses, cell-wall remodeling, and oxidative processing, suggesting that they may be relevant for extract quality attributes during handling and storage. This study provides a compositional proteomic reference for the co-extracted protein macromolecules present in acidic aqueous extracts from V. faba pod valves, supporting future studies on extract stability, processing optimization, and the development of standardized plant-based preparations.

Vicia faba

Carob pod (Ceratonia siliqua) meal in geese diets.

1. The apparent and true metabolisable energy values of carob pods meal for geese were measured to be 6.1 MJ/kg and 6.6 MJ/kg respectively. 2. Performance from 5 to 12 weeks was examined in geese fed on four diets containing 0, 100, 200 and 300 g/kg of carob pods meal. 3. The inclusion of carob pods meal up to 200 g/kg in geese diets did not affect the performance. 4. At 300 g/kg performance was highly depressed. 5. The digestibility of protein in the diets decreased linearly with an increase in the level of inclusion of carob pods meal. 6. The length of small intestine, large intestine and caeca and the weight of gizzard expressed per kg of body weight increased with an increase in the level of carob pods meal, which is rich in fibre, in the diets.

Animal Feed

Electron microscopic studies in cultivated plants. I. Green pods of Phaseolus vulgaris var. nanus L.

Scanning (SEM) and transmission (TEM) electron microscopic studies in the legumen of Phaseolus vulgaris var. nanus L. (kidney beans) were carried out. In this work emphasis was laid on the analysis of the morphological structure of the parenchyma tissue, being the chief component of the pod at edible maturity. The leaf character of the pod becomes especially evident by the occurence of stomata, trichomes, and typical cuticle structures on the outside of the pod (SEM). The cells of the mesophyll-like parenchyma tissue are distinguished by a great range of variation of their plastids (chloroplasts--chloroamyloplasts--amyloplasts) and some special cytological features such as a strongly developed rough ER, big nuclei, and numerous plasmodesmata (TEM).

Cell Nucleus

[Antienzymatic activities of two Leguminosae: bean pods and bean bulls. Antiamylase activity].

Amylase inhibitory activities have been sought in the powders of Phaseolus vulgaris pods and of Vicia faba bean hulls. Unlike to the bean, Phaseolus vulgaris pod has no amylase inhibitory activity and the presence of tannin could not be determined. Contrary, Vicia faba bean hull shows an obvious amylase inhibitory activity. It contains an appreciable content of tannins which may be responsible of this activity. The quantities of amylase whose are inhibited increase in some measure with the quantities of enzyme added. They are not in direct ratio to the quantities of extract, then, inhibition may be of no competitive type.

Amylases

[Enzyme inhibitory activities of two Leguminosae: Phaseolus vulgaris L. pods and Vicia faba L. hulls. Trypsin inhibitory activity].

Trypsin inhibitory activities of Phaseolus vulgaris L. pods and Vicia faba L. bean hulls have been studied according to the extraction conditions of the powder: solvent pattern, temperature and pH. The two vegetal powders have a trypsin inhibitory activity quite similar, weak but appreciable. The trypsin inhibitor of Phaseolus vulgaris L. pods is thermosensitive but stable at low pH. Inversely the Vicia faba L. bean hulls one is thermostable but it feels low pH.

Fabaceae

Fine structural analysis of the morphological changes involved in the blanching, cooking, dehydration and rehydration of green bean pod tissue.

The influence of blanching and cooking on green bean pod tissue (Phaseolus vulgaris var.nanus L.) was studied by transmission electron microscopy and characteristic histological and cytological effects were recorded. The analysis of dehydraded and rehydrated samples (dehydration by hot air, rehydration by boiling) showed that much of the shrinkage after dehydration can be compensated by water absorption during rehydration. The morphological appearance of boiled raw tissue, however, is not reached.

Cooking

Analysis of the nucleotide sequence of bean pod mottle virus middle component RNA.

The complete nucleotide sequence of the middle component RNA (M RNA) of the comovirus bean pod mottle virus (BPMV) has been determined. The sequence consists of 3662 nucleotides and contains a single long open reading frame sufficient to code for a protein of 113,353 Da. The proteolytic processing sites within this protein have been identified by comparison with the known three-dimensional structure of the virion and cleavage at these sites would lead to a range of products consistent with those observed during processing of the M RNA-encoded polyproteins in vitro. We have performed computer-aided searches for reiterated sequences within BPMV M RNA which might explain why ordered RNA is visible in the electron density map of BPMV middle component particles (Chen, Z., Stauffacher, C. V., Li, Y., Schmidt, T., Bomu, W., Kamer, G., Shanks, M., Lomonossoff, G., and Johnson, J. E., 1989, Science 245, 154-159). These searches revealed both the presence of overrepresented pentameric sequences and a consensus sequence which was repeated 15 times within the RNA sequence.

Amino Acid Sequence

Molecular characterization of vitellogenin and its receptor with CRISPR-based sgRNA validation in the legume pod borer, Maruca vitrata (Geyer) (Lepidoptera: Crambidae).

Maruca vitrata, the legume pod borer, causes yield losses of up to 80% in grain legumes. Increasing insecticide resistance and environmental concerns necessitate sustainable pest management alternatives. In the present study, the complete vitellogenin (Vg) coding sequence (CDS), a key reproductive gene involved in oogenesis and embryonic development, was cloned and molecularly characterised from M. vitrata. The assembled Vg CDS (∼5.3 kb) shared 99.04% sequence identity with the reported M. vitrata Vg sequence (MG799570.1). Phylogenetic analysis demonstrated close evolutionary association with related Lepidopteran species, while protein domain analysis identified three conserved domains, namely LPD_N, DUF1943, and VWD. Among these, the single exon-encoded LPD_N domain was selected as the target region for CRISPR/Cas9-mediated editing. Homology models of Vg and vitellogenin receptor (VgR) (Global Model Quality Estimation (GMQE): 0.58 and 0.51) showed a favourable interaction by protein-protein docking (score: -295.66). Three single-guide RNAs (sgRNAs) were designed, synthesised through in-vitro transcription, and evaluated using in vitro cleavage assays. sgRNA1 targeting the LPD_N domain and sgRNA2 targeting the signal peptide region exhibited efficient site-specific cleavage activity, whereas sgRNA3 failed to induce cleavage because of an unfavourable secondary structure that likely impaired Cas9-sgRNA complex formation. Overall, this study provides the first CRISPR-oriented functional characterisation and sgRNA validation of the M. vitrata Vg gene, together with structural characterisation of VgR and Vg-VgR interaction analysis, providing preliminary molecular resources for future CRISPR/Cas9 studies and supporting future embryo microinjection and heritable genome editing for sustainable management of M. vitrata.

CRISPR/Cas9

Conformations, interactions, and thermostabilities of RNA and proteins in bean pod mottle virus: investigation of solution and crystal structures by laser Raman spectroscopy.

We report and interpret laser Raman spectra of the three virion components of bean pod mottle virus (BPMV). The top component of BPMV is an empty capsid; middle and bottom components package the RNA2 and RNA1 genome segments, respectively. All components were investigated as both single crystals and aqueous solutions, the latter over wide ranges of temperature and ionic strength. The isolated RNA2 molecule of BPMV middle component was also investigated in both H2O and D2O solutions. The results permit assessment of RNA and protein structures, their mutual interactions in the virions, and their conformational thermostabilities and comparison of these structural characteristics for solution and crystal states of the particles. The principal findings of this study are (i) The extent of ordered A-form backbone (74%) and of base pairing (38% AU + 22% GC) in unpackaged (aqueous) RNA2 are significantly altered by packaging. The A-form secondary structure of RNA2 is increased by 12 +/- 4%, and guanine base interactions are also substantially increased with packaging. (ii) The thermostability of Raman-monitored secondary structure of unpackaged RNA2 (Tm approximately 43 degrees C) is greatly increased in the packaged state (Tm approximately 53 degrees C). This increase corresponds to a stabilization of the A-form backbone geometry in 15 +/- 5% of genome nucleotides. (iii) Packaging of RNA2 in the middle component stabilizes subunit-subunit interactions of the capsid, as evidenced by a thermal denaturation temperature Td approximately 65 degrees C for the virion, compared with Td approximately 55 degrees C for the empty capsid. (iv) Raman marker-band shifts implicate the purine 7N sites of RNA2 and aromatic side chains of subunits as the principal targets for RNA-subunit interaction. (v) At the conditions of the present experiments (8 degrees C, pH approximately 7, moderate ionic strength), the subunit secondary structures observed for solutions of the top, middle, and bottom components are indistinguishable by Raman spectroscopy from secondary structures observed for corresponding crystalline samples. (vi) On the other hand, side chains of subunits in the top component (empty capsid) yield significantly different Raman intensities in crystalline and solution states. These differences are interpreted as the result of changes in a small number of side-chain environments between crystal and solution. (vii) Similarly, small differences exist between RNA Raman markers of crystalline and aqueous virions, which are attributed to altered environments of nucleotide residues and to a small increase in the amount of A-form backbone geometry upon going from the crystal to the solution.(ABSTRACT TRUNCATED AT 400 WORDS)

Base Composition

Structural studies of bean pod mottle virus, capsid, and RNA in crystal and solution states by laser Raman spectroscopy.

Structures of protein and RNA components of bean pod mottle virus (BPMV) have been investigated by use of laser Raman spectroscopy. Raman spectra were collected from both aqueous solutions and single crystals of BPMV capsids (top component) and virions (middle and bottom components, which package, respectively, small and large RNA molecules). Analysis of the data permits the assignment of conformation-sensitive Raman bands to viral protein and RNA constituents and observation of structural similarities and differences between solution and crystalline states of BPMV components. The Raman results show that the protein subunits of the empty capsid contain between 45% and 55% beta-strand and beta-turn secondary structure, in agreement with the recently determined X-ray crystal structure, and that this total beta-strand content undergoes a small increase (approximately 5%) with packaging of RNA. The subunits are relatively deficient in alpha-helix secondary structure, estimated at less than 25%, and therefore must contain extensive amounts (greater than 20%) of loops and irregular chain conformations. The Raman spectra also show the following: (1) The molecular environments of as many as four tryptophan residues per subunit are altered upon packaging RNA, resulting in stronger 1N-H hydrogen bonding for two Trp residues and more hydrophobic environments for two indole rings. (2) Hydrogen-bonding states of the seven Tyr residues per subunit do not change detectably when RNA is packaged. At least five tyrosine OH groups are involved exclusively as strong hydrogen bond donors to protein acceptor groups, which suggests restricted access of solvent H2O molecules to these parahydroxyls.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acids

Tannin-rich carob pod for the treatment of acute-onset diarrhea.

Infants aged 3-21 months with acute diarrhea of bacterial and viral origin were treated as inpatients with oral rehydration fluid and randomly received for up to 6 days either a tannin-rich carob pod powder (40% tannins or 21.2% polyphenols and 26.4% dietary fiber), 1.5 g/kg/day (n = 21) to a maximum of 15 g, or an equivalent placebo (n = 20). The duration of the diarrhea from admission was 2.0 +/- 0.27 days in the test group and 3.75 +/- 0.30 days in the placebo group (p less than 0.001). Normalized defecation, body temperature, and weight and cessation of vomiting were reached more quickly by the patients who received the test substance. The test substance was well accepted and tolerated.

Acute Disease

Influence of the C terminus of the small protein subunit of bean pod mottle virus on the antigenicity of the virus determined using monoclonal antibodies and anti-peptide antiserum.

Middle component particles of bean pod mottle virus (BPMV) containing small protein subunits with a cleaved C terminus were used to produce monoclonal antibodies (MAbs). All MAbs were specific for cryptotopes, i.e. epitopes present only on dissociated BPMV protein. The MAbs reacted more strongly with virus protein preparations containing the cleaved form of the small subunit than with preparations containing only the uncleaved form. It seems that the presence of additional residues at the C terminus of the intact small subunit interferes with antibody binding. Antibodies raised against synthetic peptides corresponding to the C terminus of the uncleaved small subunit reacted with both intact virions and dissociated subunits. This C-terminal region seems to play a dominant role in the antigenicity of the virus.

Antibodies, Monoclonal

The molluscicidal activity of seed pods of Swartzia madagascariensis on Marisa cornuarietis.

The molluscicidal potency of ground seed pods of Swartzia madagascariensis was tested in the laboratory against Marisa cornuarietis, a well known competitor and/or predator of pulmonate intermediate hosts of schistosomiasis. Results showed that a minimum concentration of 1.6g ground pods/1 was required to cause 100% mortality in M. cornuarietis. This concentration was by far over and above what is required (ie, 100 mg ground pods/1) to kill 10% of the pulmonate hosts of schistosomiasis. It seems that the application of S. madagascariensis at a concentration inimical to pulmonates (ie, 100 mg ground ponds/1 or slightly higher) in habitats where M. cornuarietis is introduced may not adversely affected to competitor snail population. Hence, the prospect of using S. madagascariensis in conjunction with M. cornuarietis against pulmonate snail hosts is conceived and tentatively recommended.

Animals

[Clinical study of a new preparation from plantago seeds and senna pods].

The lack of fiber in the western diet may contribute to the development of several diseases including gastrointestinal disorders; the clinical effects of a new substance (AGIOLAX) made from plantago seeds and senna pods were studied. 100 patients of both sexes, aged from 40 to 60 years (30 with diabetes mellitus, 40 with obesity and 30 with hyperlipidemia) were treated; everyone complained a slowness, of different degree, of normal intestinal transit time or chronic constipation. The experiment was carried out without the use of a control group. Aim of the present study was to investigate the efficacy and tolerability of the product. In addition to the clinical evaluation of the symptoms, laboratory tests were performed. The patients were treated for 3 months with a daily dose of 2 teaspoons every evening. In the majority of the subjects a good clinical response was obtained; 88% of the patients presented a normalization of the gastrointestinal transit time; only 12% of them did not respond satisfactorily to the substance. Further the drug was well tolerated by 86% of the patients. In conclusion the authors report a good efficacy and tolerability of the product; thus they recommend its use in those disorders characterized by slow intestinal transit time and/or constipation.

Adult