Absence Of BAP Research Articles

The impact of sucrose and 6-benzylaminopurine on shoot propagation and vitrification in Aroniamelanocarpa (black chokeberry)

Vitrification is one of the most significant issues encountered in plant tissue culture applications. It diminishes the quality of in vitro plants, causing their leaves and stems to appear watery and translucent, and it may impede the success of the acclimatization step. In this respect, this study investigates the impact of sucrose and 6-Benzylaminopurine (BAP) concentrations on shoot regeneration and vitrification development in Aronia melanocarpa, known for its high antioxidant content and health benefits. Initially, the presence of BAP, in combination with varying sucrose concentrations, leads to a substantial increase in shoot number, and the largest number (7 shoots per nodal explant) was observed in the Murashige and Skoog (MS) medium containing 3% sucrose and 5.0 mg/L BAP. Furthermore, sucrose concentration plays a crucial role in shoot growth, with higher concentrations promoting more extensive shoot development. However, when 3% sucrose was combined with higher BAP (from 1.0- to 5.0 mg/L), an increased incidence of vitrification was observed over time. Interestingly, lower sucrose concentrations (1% or 2%) combined with 0.5 mg/L or 2.5 mg/L BAP initially delayed vitrification but eventually led to its occurrence. Microscopic analysis of leaf samples with varying levels of vitrification indicates significant differences in the density of stomata, further confirming the detrimental impact of vitrification on cellular structures and physiological processes. The recovery of vitrified plants was evaluated using different growth media combinations. The absence of BAP in the medium led to higher recovery percentages (min 96%) without necrosis, while the addition of 0.5 mg/L BAP promoted shoot growth but potentially inhibited root development. It has been found that media with 1 mg/L BAP and either 10 g/L or 20 g/L sucrose, as well as media with 30 g/L sucrose and 0.5 mg/L BAP, are the most suitable for efficient shoot regeneration with minimal vitrification risk. However, increasing BAP levels for faster shoot regeneration also raises the risk of vitrification. During acclimatization, vitrified plants exhibited stunted shoot growth, shorter and narrower leaves, reduced root numbers and lengths, and decreased survival rates, particularly under lower humidity conditions. The cultivation period required for the recovery of the crop was determined to be 6 weeks under greenhouse conditions for a sustainable plant propagation.

Enhancing resistance and cell survival in Acipenser ruthenus liver, gill, and kidney cells: The potential of heat shock protein inducers against PAH-benzo[a]pyrene stress.

The Caspian Sea has faced many environmental challenges, such as oil pollution. Heat shock proteins (HSPs) play a critical role in stress conditions and physiological changes caused by disease or injury. By evaluating the effects of various HSP inducers (HSPi), including Pro-Tex® (NOP: 800 mM), amygdalin (AMG: 80 mM), and a novel synthetic compound derived from pirano piranazole (SZ: 80 µm) on isolated cells from Sterlet Sturgeon (Acipenser ruthenus) treated with 75% IC50 PAH-benzo[a]pyrene (BaP; B75). This study examines whether there is a correlation between exposure to the BaP pollutant and HSPs in fish. In vitro, after culturing cells from the liver, kidney, and gills, they were treated with HSPi compounds in the presence and absence of BaP. Western blotting was used to assess HSP27, HSP70, and HSP90 expression patterns. A variety of enzyme activities were measured before (without treatment) and after treatment with HSPis and HSPi + B75, including cytochrome P450 (CYP450) activity, specific enzyme activity for acetylcholinesterase (AChE), antioxidant capacity, liver indicator enzymes, cortisol levels, and immunity parameters. When compared to the control group, cells treated with B75 showed the lowest AChE enzyme activity (p < 0.0001). CYP450 activity was highest in group B75, while HSPi caused the opposite effect (p < 0.0001). HSPi + B75 increased HSP levels and antioxidant parameters while decreasing cortisol and liver indicator enzymes (p < 0.0001). HSPi may be a powerful and reliable method for enhancing the resistance of A. ruthenus to BaP stresses before exposure. Treating cells with HSP-inducing compounds, such as NOP, AMG, and SZ, can assist them in managing stress and increase HSP (27, 70, and 90) protein expression. Furthermore, the study findings suggest that HSPis can also mitigate the adverse effects of stress, ultimately increasing cell survival and resistance.

Autocrine TGF-alpha is associated with Benzo(a)pyrene-induced mucus production and MUC5AC expression during allergic asthma

ObjectsBenzo(a)pyrene (BaP), an environmental pollutant, is present in high concentrations in urban smog and cigarette smoke and has been reported to promote high mucin 5AC (MUC5AC) expression. Epithelium-derived inflammatory cytokines are considered an important modulator of mucus oversecretion and MUC5AC overexpression. Here, we investigated whether the effect of BaP on MUC5AC overexpression was associated with cytokine autocrine activity in vivo and in vitro. MethodsIn vivo, BALB/c mice were treated with ovalbumin (OVA) in the presence or absence of BaP. Allergy-induced mucus production was assessed by Alcian Blue Periodic acid Schiff (AB-PAS) staining. The human airway epithelial cell line NCI-H292 was used in vitro. MUC5AC and transforming growth factor (TGF)-α mRNA levels were assessed with real-time quantitative PCR. The concentration of cytokines was measured by ELISA. The MUC5AC, p-ERK, ERK, p-EGFR and EGFR proteins were detected by Western blotting in cells or by immunohistochemistry in mouse lungs. Small-interfering RNAs were used for gene silencing. ResultsTGF-α was overproduced in the supernatant of NCI-H292 cells treated with BaP. Knockdown of TGF-α expression inhibited the BaP-induced increase in MUC5AC expression and subsequent activation of the EGFR-ERK signalling pathway. Knocking down aryl hydrocarbon receptor (AhR) expression or treatment with an ROS inhibitor (N-acetyl-L-cysteine) could relieve the TGF-α secretion induced by BaP in epithelial cells. In an animal study, coexposure to BaP with OVA increased mucus production, MUC5AC expression and ROS-EGFR-ERK activation in the lung as well as TGF-α levels in bronchoalveolar lavage fluid (BALF). Furthermore, the concentration of TGF-α in BALF was correlated with MUC5AC mRNA levels. Additionally, TGF-α expression was found to be positively correlated with MUC5AC expression in the airway epithelial cells of smokers. Compared with non-smoker asthma patients, TGF-α serum levels were also elevated in smoker asthma patients. ConclusionAutocrine TGF-α was associated with BaP-induced MUC5AC expression in vitro and in vivo. BaP induced TGF-α secretion by activating AhR and producing ROS, which led to activation of the EGFR-ERK pathway

In vitro culture of Mentha viridis: quality and intensity of light on growth and production of volatiles

Mentha viridis (Lamiaceae) has a great potential for economic exploration due to its essential oil production, which is widely used in chemical and pharmaceutical industries. The aim of this work was to study the effect of light spectra, in the presence and absence of growth regulators (BAP), and light intensity on M. viridis growth and production of volatile compounds under in vitro conditions. Nodal segments from prior established in vitro cultured plants were placed on solidified MS medium in the presence and absence of BAP under different light-emitting diodes (LED): blue; red; blue:red at proportions of (2.5:1) and (1:2.5); and fluorescent light. The explants were also exposed to a photosynthetic photon flux density (PFD) of 28, 51, 64, 76 and 113 μmol m(-2) s(-1) using a cool fluorescent light. After 30/40 days of culture, growth and production of volatiles of the plants was verified by Head-Space - GC/MS. Blue and red LED combinations promoted the growth of plantlets significantly. The presence of BAP negatively affected plantlet growth. PFD affected plantlet growth and morphological characteristics of M. viridis. The rise in light intensity promoted a higher accumulation of dry matter of leaves and total plant. The production of volatile compounds was affected both in quality and quantity by light conditions applied in this experiment. Therefore, the manipulation of spectrum and light intensity in in vitro culture environment, might be a strategy for the production of compounds of interest to industries.

Probing the action of a novel anti-leukaemic drug therapy at the single cell level using modern vibrational spectroscopy techniques

Acute myeloid leukaemia (AML) is a life threatening cancer for which there is an urgent clinical need for novel therapeutic approaches. A redeployed drug combination of bezafibrate and medroxyprogesterone acetate (BaP) has shown anti-leukaemic activity in vitro and in vivo. Elucidation of the BaP mechanism of action is required in order to understand how to maximise the clinical benefit. Attenuated total reflectance Fourier transform infrared (ATR-FTIR) spectroscopy, Synchrotron radiation FTIR (S-FTIR) and Raman microspectroscopy are powerful complementary techniques which were employed to probe the biochemical composition of two AML cell lines in the presence and absence of BaP. Analysis was performed on single living cells along with dehydrated and fixed cells to provide a large and detailed data set. A consideration of the main spectral differences in conjunction with multivariate statistical analysis reveals a significant change to the cellular lipid composition with drug treatment; furthermore, this response is not caused by cell apoptosis. No change to the DNA of either cell line was observed suggesting this combination therapy primarily targets lipid biosynthesis or effects bioactive lipids that activate specific signalling pathways.

English

This study determined the therapeutic effect of ethanol leaf extract of Combretum zenkeri on kidney function of rats exposed to benzo[a]pyrene. Male Wistar albino rats were&nbsp; treated with ethanol leaf extract of C. zenkeri (400 mg/kg) in the presence or absence of BaP (200 mg/kg) for four weeks. The blood obtained at the end of the treatment was processed into serum and used for biochemical analyses and kidney tissues for histology. The serum creatinine, urea, sodium ion and potassium ion concentrations of the group exposed to benzo[a]pyrene without treatment with C. zenkeri extracts were significantly higher (p&lt;0.05) as compared to the normal control group. The kidney histology indicated severely damaged glomeruli. The groups treated with the leaf extract pre- or post- benzo[a]pyrene exposure showed significant reduction (p&lt;0.05) in serum concentration of urea, creatinine and sodium ion, while the pre-treated group showed no significant reduction (p&lt;0.05) in serum potassium ion concentration. The biochemical and histological studies indicated that, C. zenkeri can ameliorate benzo[a]pyrene induced oxidative kidney damage. Key words: Combretum zenkeri, benzo[a]pyrene, PAHs, oxidative damage, kidney functions.

Propagação in vitro de copo-de-leite: sulfato de adenina e 6-benzilaminopurina

&lt;p&gt; O copo-de-leite [&lt;em&gt;Zantedeschia aethiopica &lt;/em&gt;(L.) Spreng.], pertencente à família Araceae, é bastante apreciado tanto como flor de corte quanto na composição de jardins. Entretanto, a propagação convencional dessa espécie apresenta baixo rendimen­to. A cultura de tecidos, além de permitir rápida propagação clonal, também proporciona a obtenção de plantas uniformes e sadias. Sendo assim, o objetivo do trabalho foi estudar a influência de diferentes concentrações de 6-benzilaminopurina (BAP) e sulfato de adenina (SA) sobre o desenvolvimento &lt;em&gt;in vitro &lt;/em&gt;de explantes de copo-de-leite. As culturas foram manti­das em meio MS acrescido de BAP (0,0; 8,9; 17,8 e 26,7 μM) e SA (0, 54, 108 e 162 μM) em arranjo fatorial (4x4). O ma­terial vegetal foi cultivado em sala de crescimento com temperatura de 27 ± 1ºC, fotoperíodo de 16 horas, sob intensidade luminosa de 35 μmol m-2 s-1, por período de 60 dias. O delineamento experimental utilizado foi inteiramente casualizado, com quatro repetições, e a unidade experimental composta de três tubos de ensaio, sendo cada tubo constituído por um explante. A interação significativa entre os fatores BAP e SA ocorreu para número de folhas por explante (4,8) e massa fresca da parte aérea (0,73 g) com 26,7 μM de BAP e 108 μM de SA. Para número de brotos obtidos por explante (2,6) apenas o BAP apresentou significância na concentração de 22,19 μM. Os brotos apresentaram maior comprimento (5,0 cm) na ausência de BAP. A adição de BAP incrementou o número de brotos por explante. O emprego de sulfato de adenina em combinação com BAP teve efeito positivo para o acumulo de massa fresca e numero de folhas no cultivo &lt;em&gt;in vitro&lt;/em&gt;.&lt;/p&gt;

Multiplicação in vitro de segmentos apicais caulinares de segurelha (Satureja hortensis L.)

O presente estudo teve por objetivo avaliar o efeito de BAP, na presença e ausência de ANA, sobre a multiplicação in vitro de segmentos apicais caulinares de Satureja hortensis. Os explantes foram isolados de plântulas germinadas in vitro e cultivados em meio nutritivo MS. O delineamento experimental utilizado foi o inteiramente casualizado, em esquema fatorial 2x5, correspondendo às concentrações de ANA (0 e 1 µM) e de BAP (0; 5; 10; 15 e 20 µM), com seis repetições, cada uma composta por três explantes. Para a porcentagem de explantes com brotações houve efeito significativo para o fator BAP, aumentando à medida que cresce a concentração da citocinina. A variável "número de brotos por explante" apresentou interação entre os fatores, havendo a maior formação de brotos na presença de ANA na faixa de 10 - 15 µM de BAP. Na presença da auxina, o maior valor ocorreu com 15 µM de BAP tendendo a diminuir independente da presença de ANA. O enraizamento dos segmentos apicais foi maior na presença de ANA e ausência de BAP, diminuindo com o aumento da citocinina. O número de folhas foi influenciado pela maior concentração de BAP sendo que a 15 µM foi observado o maior número, tendendo à queda com aumento da concentração. A concentração de 15 µM de BAP, independente de ANA, proporcionou os melhores resultados na multiplicação da espécie produzindo aumento de brotações e número de folhas, à exceção do enraizamento que foi influenciado pela auxina.

Reguladores de crescimento na propagação in vitro de abacaxizeiro ornamental

The ornamental pineapple plant Ananas comosus (L.) Merr. var. ananassoides (Baker) Coppens &amp; F. Leal., belongs to the Bromeliaceae family, standing out for its ornamental potential, which is little explored economically. In vitro propagation allows a high production of pathogens-free plants. The objective of this work was to evaluate the different concentrations of 6-benzilaminopurine (BAP) and naphthalene acetic acid (ANA) in the process of in vitro propagation of Ananas comosus var. ananassoides. A completely randomized experimental design was used, with a 4 x 4 factorial scheme, consisting of four BAP concentrations (0; 0.5; 1.0 and 2.0 mg L-1) and four ANA concentrations (0; 0.25; 0.5 and 1.0 mg L-1). According to the results, the addition of 1.0 mg L-1 of BAP promoted the highest number of sprouts. The highest average sprout length was obtained in the absence of BAP and the highest root number was obtained with the addition of 0.25 mg L-1 of ANA, in the absence of BAP.

Direct OrgaNogenesis from Cotyledons in Cultivars of Citrus clementina Hort. Ex Tan

An efficient protocol to induce shoot buds regeneration in Citrus clementina cultivars (‘Monreal’, ‘SRA 63’ and ‘SRA 64’) by direct orgaNogenesis has been developed using cotyledons as explants. Cotyledons transversely cut in three segments and entire ones were cultured on Murashige and Skoog (1962) solidified medium containing vitamins, 500 mg l-1 malt extract, 50 g l-1 sucrose and supplemented with three different concentrations of BAP (8.8, 13.2 and 17.6 μM). In all three cultivars the entire cotyledons showed more shoot morphogenic potential than transversely cut ones and after 60 incubation days the optimum BAP concentration was 17.6 μM in ‘Monreal’ (50% ± 2.89% of frequency regeneration) and 13.2 μM in ‘SRA 63’ (33.33% ± 3.33%) and ‘SRA 64’ (25.93% ± 1.85%). In absence of BAP No mor-phogenesis occurred, demonstrating the absolute requirement of this hormone for shoots induction. The young shoots showed a regular growth in the culture tubes containing the basal medium without hormones, and the rooted plantlets survived after acclimatization. This protocol may find application in Citrus genetic improvement programs.

Regeneration of Blackgram (Vigna mungo L.) on Changes of Hormonal Condition

The study comprised of experiments for shoot regeneration and plantlet formation from cotyledonary node of Vigna mungo by culturing them on low concentration of BAP followed by transfer to hormone free MS medium. Cotyledonary node explants were cultured on different concentrations of BAP (0, 1, 2.5 and 50 mg l-1). Shoot regeneration occurred from cotyledonary nodes irrespective of the presence or absence of BAP in the medium. However, culture of cotyledonary node explants for 10 days on medium containing 1.0 mg l-1 BAP followed by transfer to hormone free medium gave higher number of shoots (9.33/explant) compared to culture of the explants on hormone free medium for 15 days followed by transfer to medium containing 1.0 mg l-1 BAP (8.33/explants). The regenerated shoots were transferred to rooting medium supplemented with different concentrations of IBA and NAA. The high frequency (100.0%) of rooting was observed with MS medium supplemented with 0.5 mg l-1 IBA. The rooted plants were transferred to pots for hardening.

Ácido giberélico (ga3) e 6-benzilaminopurina (bap) no crescimento in vitro de macela [Egletes viscosa (L.) Less.

A micropropagação é uma ferramenta promissora que pode ser utilizada para propagação e preservação de muitas espécies de plantas medicinais que vêm sendo exploradas de forma indiscriminada e que estão em vias de extinção. Conduziu-se este trabalho com o objetivo de induzir o crescimento in vitro de brotações de macela, em meio de cultura MS com diferentes concentrações de GA3 (0; 0,1 e 0,5 mgL-1) e BAP (0; 1 e 2 mgL-1). Foram utilizadas brotações de macela com diâmetro médio de 1,5 cm, retiradas de plantas preestabelecidas in vitro, inoculadas em frascos contendo 40 ml do meio de cultura e mantidas em sala de crescimento com temperatura média de 26 ± 1oC, fotoperíodo de 16 h e intensidade luminosa em torno de 2000 lux fornecida por lâmpada tipo fluorescente branca fria. Utilizou-se como delineamento experimental um fatorial 3 x 3, num total de nove tratamentos com 15 explantes por tratamento. Aos 30 dias, verificou-se o maior crescimento em altura das plantas, no tratamento com 0,5 mgL-1 de GA3 na ausência de BAP. O maior número de brotações ou crescimento em diâmetro foi observado com maiores concentrações dos reguladores. Porém, na presença dos reguladores ocorreu um maior número de plantas com folhas hiperhídricas e malformadas.

Early events of multiple bud formation and shoot development in soybean embryonic axes treated with the cytokinin, 6‐benzylaminopurine

Early events of multiple bud formation and shoot development in germinating soybean embryonic axes treated for 24 hr with the cytokinin, 6‐benzylaminopurine (BAP), were compared to the development of untreated control axes using four different techniques: photomicrography, scanning electron microscopy, histology, and autoradiography. Shoot apex development in BAP‐treated embryonic axes was delayed by about 9 to 15 hr. A transient inhibition of DNA synthesis in the primary apical meristem and axillary buds was observed with subsequent changes in the timing of cell division patterns in these regions. Meristematic regions (supernumerary vegetative buds) were observed in BAP‐treated axes around the perimeter of the apical dome at and above the level of the axillary buds. Cells elongated from some of the BAP‐induced meristematic regions to form four to six shoots. In the absence of BAP, excision of the primary apical meristem and/or axillary buds did not result in multiple bud formation. These results suggest that transient exposure to BAP interrupted chromosomal DNA replication and reprogrammed the developmental fate of a large number of cells in the shoot apex. We postulate that interruption of DNA synthesis, either directly, by interfering with DNA replication, or indirectly, by preventing entry into S‐phase, effected redetermination of the shoot apex cells.

Organic nitrogen stimulates caulogenesis from hypocotyl callus of Phyllanthus fraternus

Factors involved in promoting caulogenesis from hypocotyl explants of Phyllanthus fraternus were studied. Hypocotyl explants were cultured on B5 medium supplemented with 2,4-D or NAA in the presence and absence of BAP (at concentrations 0, 10(-7), 10(-6) and 10(-5)M). Adventitious shoots differentiated from callus developed from the cut ends of 12.5% of the hypocotyl segments cultured on medium supplemented with 10(-6)M BAP in combination with 10(-6)M 2,4-D or 10(-6)M NAA. Profuse rooting occurred from the hypocotyl explants on medium supplemented with 10(-6)M BAP + 10(-6)M NAA. Incorporation of casein hydrolysate in B5 medium along with 10(-6)M BAP + 10(-7)M 2,4-D enhanced the frequency of cultures with adventitious shoots upto 68.0%. Glutamine, glutamic acid or proline could partially substitute for the effect of casein hydrolysate. Amongst the hypocotyls from 3-14 d old seedlings, the best caulogenesis was obtained with hypocotyls from 7 d old seedlings both in presence or absence of casein hydrolysate. Best rooting of shoots was achieved on half-strength B5 medium supplemented with 10(-6)M IBA. After hardening, plantlets were successfully transferred to the soil.

Effect of 6-Benzyl Aminopurine on the Stabilization of Absorption Spectrum and Hill Activity of Isolated Chloroplasts

Summary Beet spinach chloroplasts isolated in the presence or absence of BAP (6-benzyl aminopurine) were suspended in sugar or salt medium and were allowed to age in light (1,000 JM -2 s -1 ). Loss in the Hill activity (H 2 0 to ferricyanide (Fe (CN 6 ) 3- ) due to light-induced ageing could be partly protected by the inclusion of the hormone (BAP). The extent and pattern of protection of chloroplast function by the hormone is similar for both salt and sugar media. During light-induced ageing, chloroplasts also lose the capacity to use the artificial electron donor DPC (diphenyl carbazide) for photoreduction of DCPIP. This loss is fast er for chloroplasts suspended in salt medium than for chloroplasts incubated in sugar medium. BAP could not provide any protection for this loss in DPC supported DCPIP photoreduction ability of chloroplasts. Loss in chlorophyll contents during light induced ageing was found to be negligible. However, absorption spectrum for chloroplasts showed changes in t he blue to red peak ratio upon ageing in light. This was found to be more pronounced in chloroplasts incubated in salt medium wih or without BAP treatment than in chloroplasts incubated in sugar medium. Accompanied this change in peak ratio, there is a small blue shift (1-2 nm) in the position of red absorption peak. BAP seems to arrest this shift.