Effect of Exogenously Applied 24-Epibrassinolide and Brassinazole on Xylogenesis and Microdistribution of Cell Wall Polymers in Leucaena leucocephala (Lam) De Wit

Plant growth regulators play a key role in cell wall structure and chemistry of woody plants. Understanding of these regulatory signals is important in advanced research on wood quality improvement in trees. The present study is aimed to investigate the influence of exogenous application of 24-epibrassinolide (EBR) and brassinosteroid inhibitor, brassinazole (BRZ) on wood formation and spatial distribution of cell wall polymers in the xylem tissue of Leucaena leucocephala using light and immuno electron microscopy methods. Brassinazole caused a decrease in cambial activity, xylem differentiation, length and width of fibres, vessel element width and radial extent of xylem suggesting brassinosteroid inhibition has a concomitant impact on cell elongation, expansion and secondary wall deposition. Histochemical studies of 24-epibrassinolide treated plants showed an increase in syringyl lignin content in the xylem cell walls. Fluorescence microscopy and transmission electron microscopy studies revealed the inhomogenous pattern of lignin distribution in the cell corners and middle lamellae region of BRZ treated plants. Immunolocalization studies using LM10 and LM 11 antibodies have shown a drastic change in the micro-distribution pattern of less substituted and highly substituted xylans in the xylem fibres of plants treated with EBR and BRZ. In conclusion, present study demonstrates an important role of brassinosteroid in plant development through regulating xylogenesis and cell wall chemistry in higher plants

Cytotoxic Oleanane-Type Saponins from Albizia inundata

Bioassay-guided fractionation of a CH2Cl2−MeOH extract of the aerial parts of Albizia inundata resulted in the isolation of two new natural oleanane-type triterpene saponins {3-O-[α-l-arabinopyranosyl(1→6)]-2-acetamido-2-deoxy-β-d-glucopyranosyl oleanolic acid (1) and 3-O-[α-l-arabinopyranosyl(1→2)-α-l-arabinopyranosyl(1→6)]-2-acetamido-2-deoxy-β-d-glucopyranosyl acacic acid lactone (2)} along with seven known saponins {3-O-[α-l-arabinopyranosyl(1→6)]-2-acetamido-2-deoxy-β-d-glucopyranosyl echinocystic acid (3), 3-O-[β-d-xylopyranosyl (l→2)-α-l-arabinopyranosyl(l→6)]-2-acetamido-2-deoxy-β-d-glucopyranosyl acacic acid lactone (concinnoside D) (4), 3-O-[β-d-glucopyranosyl(l→2)]-β-d-glucopyranosyl oleanolic acid (5), 3-O-[α-l-arabinopyranosyl(1→2)-α-l-arabinopyranosyl(l→6)]-β-d-glucopyranosyl oleanolic acid (6), 3-O-[β-d-xylopyranosyl(1→2)-α-l-arabinopyranosyl(l→6)]-β-d-glucopyranosyl oleanolic acid (7), 3-O-[α-l-arabinopyranosyl(l→2)-α-l-arabinopyranosyl(1→6)-[β-d-glucopyranosyl(l→2)]-β-d-glucopyranoside echinocystic acid (8), and 3-O-[β-d-xylopyranosyl(l→2)-α-l-arabinopyranosyl(1→6)-[β-d-glucopyranosyl(l→2)]-β-d-glucopyranoside echinocystic acid (9)}. The structures of 1 and 2 were established on the basis of extensive 2D NMR (1H−1H COSY or DQF-COSY, HSQC, HMBC, TOCSY, and HSQC-TOCSY) spectroscopic, ESIMS, and chemical methods. Saponins 1, 3, 6, and 7 showed cytotoxicity against human head and neck squamous cells (JMAR, MDA1986) and melanoma cells (B16F10, SKMEL28) with IC50 values in the range 1.8−12.4 μM, using the MTS assay

Cytotoxic Oleanane-type Triterpenen Saponins with N-acetyl Sugar from Albizia inundata [abstract]

Comparative Medicine - OneHealth and Comparative Medicine Poster SessionBioassay-guided fractionation of a CH2Cl2-MeOH extract of the aerial part of Albizia inundata , collected in Argentina, resulted in the isolation of two new oleanane-type triterpene saponins with N-acetyl sugar (1, 2) and seven known saponins (3-9). Their structures were established on the basis of extensive 2D NMR (HSQC, COSY or DQF-COSY, TOCSY, HMBC), ESIMS and chemical methods. Saponins 1-4 showed cytotoxicity against the in JMAR, MDA 1986, UM-SCC-2 and JHU011 human head and neck squamous cells using the MTS assay

Robust estimation of bacterial cell count from optical density

Optical density (OD) is widely used to estimate the density of cells in liquid culture, but cannot be compared between instruments without a standardized calibration protocol and is challenging to relate to actual cell count. We address this with an interlaboratory study comparing three simple, low-cost, and highly accessible OD calibration protocols across 244 laboratories, applied to eight strains of constitutive GFP-expressing E. coli. Based on our results, we recommend calibrating OD to estimated cell count using serial dilution of silica microspheres, which produces highly precise calibration (95.5% of residuals <1.2-fold), is easily assessed for quality control, also assesses instrument effective linear range, and can be combined with fluorescence calibration to obtain units of Molecules of Equivalent Fluorescein (MEFL) per cell, allowing direct comparison and data fusion with flow cytometry measurements: in our study, fluorescence per cell measurements showed only a 1.07-fold mean difference between plate reader and flow cytometry data

Relationship between seasonal cambial activity, development of xylem and phenology in Azadirachta indica growing in different forests of Gujarat State

Seasonal cycle of cambial activity was compared among the trees of Azadirachta indica growing in Moist Deciduous (MDF), Dry Deciduous (DDF) and Scrub land Forest (SF) of Gujarat State. Radial growth occurred in two growth flushes in MDF and DDF. Cambial cell divisions in MDF started in February and June resulting maximal radial growth in August-September when the rains were heavy and ceased in January and May during the drier part of the year. In DDF the first flush of growth commenced in January with maximal xylem development in April and ceased in May. The second flush of cambial activity began in June with the arrival of rains, reached peak in October and ceased in December. Cambium was active throughout the year in SF and attained its peak activity thrice i.e. in February, July and October. With complete maturation of leaves in November, the cell divisions were rather slow in MDF and SF whereas no divisions were encountered in DDF. Cambial rays exhibited large intercellular spaces during drier months in all the three forests. Seasonal behavior of vascular cambium was discussed in relation to phenology and local climatic conditions.Relation entre l'activité saisonnière du cambium, la croissance radiale du xylème et la phénologie chez Azadirachta indica dans différents types d'écosystèmes forestiers de l'état du Gujarat. Les cycles saisonniers de l'activité cambiale chez Azadirachta indica dans différents types d'écosystèmes forestiers de l'état de Gujarat ont été comparés entre des arbres provenant de forêts humide décidue (MDF), sèche décidue (DDF) et de savane arborée (SF). La division des cellules cambiales commence en février et juin pour les MDF, ce qui se traduit par une croissance radiale maximale en août-septembre, au moment des fortes pluies, et un arrêt en janvier et mai pendant les périodes les plus sèches de l'année. Dans les DDF, le premier démarrage de croissance radiale commence en janvier, avec un maximum en avril et un arrêt en mai. Une seconde période d'activité commence en juin avec l'arrivée des pluies, atteint son maximum en octobre et s'arrête en décembre. Dans les SF le cambium reste actif toute l'année avec trois pics d'activité en février, juillet et octobre. Lorsque les feuilles arrivent à maturité complète en novembre, la vitesse de division cellulaire du cambium est relativement faible dans les MDF et le SF et nulle dans les DDF. Dans le cambium, les rayons présentent des espaces intercellulaires importants pendant les mois secs pour les trois types de forêts. Le fonctionnement saisonnier du cambium est ensuite discuté en relation avec la phénologie et les conditions climatiques locales

Chemical Composition in Juvenile and Mature Wood of Branch and Main Trunk of <i>Leucaena leucocephala</i> (Lam.) de Wit

Secondary growth is the most dynamic developmental aspect during the terrestrialization of plants. The development of secondary xylem tissue composed of thick-walled cells with characteristic changes in its structure and chemistry facilitates the growth and development of woody plants. In the present study, the chemical composition of the secondary xylem of juvenile and mature wood from the branch and main trunk of Leucaena leucocephala, has been investigated and the differences established. The biochemical analysis of different cell wall components in the mature wood of the main trunk revealed high holocellulose and α-cellulose and less lignin content in the juvenile wood while its syringyl/guaiacyl (S/G) ratio was less than for the mature wood. As compared to the branch xylem, concentration of cell wall polysaccharides and lignin content was higher in both juvenile and mature wood collected from the main trunk. Thioacidolysis and GC-MS analysis of wood lignin from juvenile and mature wood showed that an increased concentration in lignin content in mature wood is associated with a corresponding increase in S/G ratio. The structural information of the acetylated lignin was investigated by 1H NMR spectroscopy. Our results indicate that the mature wood from the main trunk is superior in pulp yielding and lignin degradability as compared to the juvenile wood of the branch and trunk