Daya Antibakteri Estrak Kulit Dan Biji Buah Pulasan (Nephelium Mutabile) Terhadap Staphylococcus Aureus Dan Escherichia Coli Secara in Vitro

Traditional medicine from drug crop has more than a pharmacological effect so that its use should be acurate and correct. The mistake in traditional medicine USAge and or drug crop can be dangerous for health. Therefore, it is required a complete scientific information to avoid it. It had been done the research of anti-bacteria activity test from crude extract of ethyl acetate and ethanol from skin and seed of Pulasan to bacterium Staphylococcus aureus and Escherichia coli with dilution method to determine Minimum Inhibitory Concentration (MIC) and Minimum Bakterisidal Concentration (MBC). The biggest MIC and MBC was respectively obtained from ethyl acetate extract that was 0,76 mg/ml, and ethanol extract that was 156,13 mg/ml. In general, skin and fruit seed extract of pulasan (Nephelium mutabile) has bigger resistance ones to bacterium Staphylococcus aureus compared to Escherichia coli

Molecular Cloning, Characterization and Expression Analysis of the <i>SAMS</i> Gene during Adventitious Root Development in IBA-Induced Tetraploid Black Locust

<div><p>S-Adenosylmethionine synthetase (SAMS) catalyzes the synthesis of S-adenosylmethionine (SAM), a precursor for ethylene and polyamine biosynthesis. Here, we report the isolation of the 1498 bp full-length cDNA sequence encoding tetraploid black locust (<i>Robinia pseudoacacia</i> L.) SAMS (<i>TrbSAMS</i>), which contains an open reading frame of 1179 bp encoding 392 amino acids. The amino acid sequence of TrbSAMS has more than 94% sequence identity to SAMSs from other plants, with a closer phylogenetic relationship to SAMSs from legumes than to SAMS from other plants. The TrbSAMS monomer consists of N-terminal, central, and C-terminal domains. Subcellular localization analysis revealed that the TrbSAMS protein localizes mainly to in the cell membrane and cytoplasm of onion epidermal cells and <i>Arabidopsis</i> mesophyll cell protoplasts. Indole-3-butyric acid (IBA)-treated cuttings showed higher levels of <i>TrbSAMS</i> transcript than untreated control cuttings during root primordium and adventitious root formation. <i>TrbSAMS</i> and its downstream genes showed differential expression in shoots, leaves, bark, and roots, with the highest expression observed in bark. IBA-treated cuttings also showed higher SAMS activity than control cuttings during root primordium and adventitious root formation. These results indicate that <i>TrbSAMS</i> might play an important role in the regulation of IBA-induced adventitious root development in tetraploid black locust cuttings.</p></div

Amino acid sequence alignment of TrbSAMS with SAMS sequences from other plant species.

<p>GenBank accession numbers for nucleotide sequences: <i>Glycine max</i>, XP_003550837.1; <i>Cajanus cajan</i>, AEY85025.1; <i>Glycine soja</i>, ABY25855.1; <i>Ricinus communis</i>, XP_002512570.1; <i>Medicago truncatula</i>, XP_003609861.1; <i>Vitis vinifera</i>, XP_002266358.1; <i>Populus trichocarpa</i>, XP_002312296.1; <i>Cucumis sativus</i>, XP_004168041.1; <i>Prunus persica</i>, AGF95108.1. The same and similar amino acid residues are highlighted in black and gray respectively; the underlined sequences indicate NADP-binding sites and substrate specificity domains. Three conserved motifs are indicated in red. Two SAM synthetase signature motifs are underlined.</p

Relative expression levels of <i>TrbSAMS</i> (A), <i>TrbSAMDC</i> (B), <i>TrbPAO</i> (C), and <i>TrbACS</i> (D) during different IBA-induced and untreated rooting phases in softwood cuttings of tetraploid black locust.

<p>Initiation phase (I). Callus induction phase (C). Root primordia formation phase (RP). Adventitious root formation and elongation phase (AR). Error bars represent the standard deviation (SD) calculated from three biological replicates with IBA, CK (n = 3). CK, control treatment; IBA, indole-3-butanoic acid.</p

Morphological changes in tetraploid black locust cuttings undergoing adventitious root development in a sand bed.

<p>a. Softwood cuttings before cutting. b. A white callus appeared at 10 days after cutting. c. A yellow callus appeared and tiny adventitious roots emerged (root primordium) at 15 days after cutting. d. Adventitious root formation and elongation at 20 days after cutting.</p

Predicated tertiary structure of TrbSAMS protein established by SWISS-MODE.

<p>The amino and carboxylic termini are labeled N and C, respectively. The entrances of substrate binding sites are marked with white arrows.</p

SAMS activity (A), polymines contents (B), ACS activity (C), and ethylene production (D) during the different IBA-induced and untreated rooting phase in softwood cuttings of tetraploid black locust.

<p>Initiation phase (I), callus induction phase (C), root primordia formation phase (RP), adventitious root formation and elongation phase (AR). Error bars represent the standard deviation (SD) calculated from three biological replicates with IBA, CK (n = 3). CK, control treatment; IBA, indole-3-butanoic acid.</p

Additional file 2: of De novo sequencing and comparative transcriptome analysis of adventitious root development induced by exogenous indole-3-butyric acid in cuttings of tetraploid black locust

Summary of the blastx results for the tetraploid black locust transcriptome against five databases. (DOCX 13 kb

Phylogenetic tree of TrbSAMS and other plant SAMS proteins.

<p>GenBank accession number for nucleotides sequences: <i>Glycine max</i>, XP_003550837.1; <i>Cajanus cajan</i>, AEY85025.1; <i>Glycine soja,</i> ABY25855.1; <i>Ricinus communis</i>, XP_002512570.1; <i>Medicago truncatula</i>, XP_003609861.1; <i>Vitis vinifera</i>, XP_002266358.1; <i>Populus trichocarpa</i>, XP_002312296.1; <i>Cucumis sativus</i>, XP_004168041.1; <i>Theobroma cacao</i>, EOY06891.1; <i>Gossypium hirsutum</i>, ADN96174.1; <i>Prunus persica</i> AGF95108.1; <i>Lycoris radiata</i> AFC88125.1; <i>Arabidopsis thaliana</i>, NP_188365.1; <i>Oryza officinalis</i> CAJ45561.1; <i>Gladiolus grandiflorus</i>, ADM18304.1. Underlining indicates the amino acid sequence of TrbSAMS cloned in this study. The phylogenetic tree was constructed using the program MEGA 5.0. The numbers shown at internal nodes indicate the occurrence of these nodes in 1000 replicates, and the bar represents 20% sequence divergence.</p

Anatomical changes in tetraploid black locust softwood cuttings undergoing IBA-induced adventitious root development.

<p>a. Cross-section of the stem before cutting. b. Parenchyma cells of callus at 10 days after cutting. c. Root primordium from the parenchyma cells at the junction between the pith rays and cortex at 15 days after cutting. d. Adventitious roots appearing at 20 days after cutting. Scale bars, 20 µm.</p