Phytochemicals Screening and Anti-proliferative Activities of Anacardium occidentale Shoot Extract in Breast Cancer Cells

@#Introduction: Anacardium occidentale or cashew are popular traditional food and have raised research interest for complementary cancer treatment. Cancer has become leading cause of death and treatment involved severe side effects. In present study, we aim to study the anti-proliferative effects of cashew shoots in breast cancer (MDAMB-231), colorectal cancer (HT-29) and liver cancer (HepG2) cell lines. Methods: Cell lines were treated with 70% ethanolic cashew extract for cytotoxicity test with MTT assay. AO/PI dual fluorescent assay and RNase/PI staining were used to determine apoptosis induction effects. Phytochemicals screening was carried out by using gas chromatography mass spectrometry (GCMS) and liquid chromatography mass spectrometry (LCMS). Results: The cytotoxicity assay of cashew shoot extract demonstrated IC50 of 81.1 ± 0.11 μg/ml for MDA-MB-231, 307.5 ± 2.31 μg/ml for HT-29 and 272.6 ± 1.91 μg/ml for HepG2 cell lines. The apoptotic bodies include chromatin condensation, cell blebbing and nuclear fragmentation and apoptosis induction were shown by AO/PI staining. There was significant increase of cell count in sub-G0 phase in MDA-MB-231 cell lines treated with cashew shoot extract. It was demonstrated that cashew shoot extract contained 38 compounds from GCMS such as sitosterol, tannin, pyrogallol, phenol and 20 compounds from LCMS such as citric acid, gallic acid, myricetin and hinokiflavone that may give rise to its anti-cancer effect. Conclusion: Cashew shoot extract demonstrated potential anti-cancer properties thus further study is required to investigate its mechanism as anti-cancer agent.

Organogenesis on apical buds in common fig (Ficus carica) var. Black Jack

BACKGROUND Plant tissue culture involves the use of explants obtained from plants to induce organogenesis with the help of plant growth regulators (PGRs). Micropropagation techniques provide a faster and economical solution to the limitations associated with traditional methods of plant cultivation. The present study focuses on the multiple shoot induction and proliferation of Ficus carica var. Black Jack. Factors that influence the growth of in vitro multiple shoots on the apical buds, which include growth media and PGRs, were investigated in this study. Different concentrations of cytokinins like 6-benzylaminopurine (BAP), Thidiazuron (TDZ), and Kinetin (Kin) were used on woody plant medium (WPM) for the optimization of media for multiple shoot induction and proliferation. RESULTS Apical buds of Ficus carica var. Black Jack growing in WPM supplemented with BAP produced the healthiest plantlets, with the highest number of multiple shoots. The most efficient medium composition which produced the highest number of multiple shoots (37.8) per growing explant was WPM supplemented with 20 mM BAP. Proliferated multiple shoots were efficiently rooted using WPM + 20 mM BAP + 8 mM indole-3-acetic acid (IAA). This optimized medium composition significantly enhanced the production of multiple, disease-free plantlets using single apical bud explants of Ficus carica var. Black Jack. CONCLUSIONS In the present study the observations indicate that WPM supplemented with 20 mM BAP is the best-suited medium for organogenesis and multiple shoot culture of Ficus carica var. Black Jack, and this technique can be potentially applied for commercialization of the plant

An efficient protocol for in vitro regeneration of peanut (Arachis hypogaea L ) cultivar L14

The present work aims to establish an efficient protocol for in vitro regeneration of peanut (Arachis hypogaea) cultivar L14. The study showed that de-embryonated cotyledon was a suitable explant for shoot multiplication on MS medium containing 4 mg/L BAP. The highest number of shoots per explant obtained after 4 weeks of culture was up to 6.8 shoots. Shoots in vitro were able to produce a large number of approximately 11 roots on MS medium supplemented with 0.5 mg/L NAA. These results will be very useful in establishing an in vitro regeneration protocol for peanut cultivar L14 during gene transfer in the next studies to improve their disease resistance.

Plant growth analysis describing the soybean plants response on dryland field to seed co-inoculation / Análise de crescimento de plantas descrevendo a resposta do cultivo de soja no campo à coinoculação de sementes

ABSTRACT: Plant growth analysis can be used for soybean plants evaluation to identify morphologic changes caused by soil microbes after seed inoculation. The objective was to measure changes of inoculated soybean plants grown under regular field Brazilian production conditions. The experiment was carried out to compare 5 inoculation treatments: T1 (Bradyrhizobium japonicum and B. diazoefficiens), T2 (T1 and Azospirillum brasilense), T3 (T1 and Trichoderma asperellum), T4 (T1 and T. virens, and Bacillus amyloliquefaciens) and T5 (T1 and Penicillum bilaiae). Leaf area, shoot and root dry matter were measured at vegetative and reproductive growth stages. Results of this study validate the contribution of plant growth analysis of soybeans plants for identifying their responses promoted by the combined inoculation of their seeds with selected microbes. Also, the microbial effects on plant growth vary not only between microbes but also between parts of the plants and through the plant development stages. The introduction of different microbes in soybean rhizosphere combined with Bradyrhizobium sp. strains can contribute to increase crop dry matter productivity during its growing cycle.

RESUMO: Análise de crescimento de plantas pode ser utilizada para avaliação de plantas e identificação de mudanças causadas por microrganismos de solo, depois de serem inoculados nas sementes. O objetivo deste estudo foi medir as mudanças no crescimento de plantas de soja inoculadas em condição de cultivos extensivos de campo no Brasil. O experimento foi conduzido para comparar cinco tratamentos inoculados: T1 (Bradyrhizobium japonicum e B. diazoefficiens), T2 (T1 e Azospirillum brasilense), T3 (T1 e Trichoderma asperellum), T4 (T1 and T. virens, e Bacillus amyloliquefaciens) e T5 (T1 e Penicillum bilaiae). A área foliar e a massa seca da parte aérea e de raízes foram medidas nos estádios vegetativos e reprodutivos. Os resultados validam a contribuição da análise de crescimento de plantas de soja para identificar as respostas causadas pela inoculação de sementes com combinação de microrganismos selecionados. Os efeitos dos microrganismos no crescimento das plantas não variam apenas entre os microrganismos, mas também entre as partes das plantas e ao longo do seu desenvolvimento. A introdução de diferentes micróbios na rizosfera de soja, combinados com cepas de Bradyrhizobium sp. pode aumentar a produtividade de massa seca das culturas durante o seu ciclo de crescimento.

Improving micropropagation of some grape cultivars via boron, calcium and phosphate

BACKGROUND: An efficient regeneration protocol is a priority for the successful application of plant biotechnology. Grape nodal explants were used to develop a micropropagation protocol for Thompson Seedless and Taify cvs. Explants were cultured on MS medium supplemented with Kinetin or benzylaminopurine (BA) and indolebutyric acid (IBA). RESULTS: For both cultivars, axillary buds were grown, only, on a medium enriched with kinetin, moreover, shoot tip necrosis and callus formation were observed on Thompson Seedless cv. cultures grown on a medium with BA. Supplementing the growth medium with 100 mM (boron) B and 2.5 mM (calcium) Ca successfully help overcome these phenomena. The highest regenerated shoot numbers (14 and 6.2 explant 1 ) for Taify and Thompson Seedless cvs., respectively, were on media supplemented with 13.2 mM BA + 4.9 mM IBA and BA 13.2 mM + 5.8 mM IBA, respectively. Moreover, these media supported the developing shoots to have the heaviest dry weights (1.46 and 0.72 mg explant 1 ) for Taify and Thompson Seedless cvs., respectively. Thompson Seedless cv. regenerated shoot numbers and their dry weights were significantly increased by increasing the MS medium PO4 concentration. However, these two parameters were significantly decreased for Taify cv. Developing shoots were elongated and rooted on MS medium enriched with 4.9 mM, IBA 100 mM B and 2.5 mM Ca. Plantlets were acclimatized and successfully transferred to the greenhouse conditions. CONCLUSIONS: A novel promising protocol for Thomson Seedless and Taify cvs. micropropagation using single nodes has been developed.

Genes involved in the deformations of the shoot apical meristem in somatic embryos of Capsicum chinense Jacq.

Somatic embryos (SE) of habanero pepper (Capsicum chinense Jacq.) represent persistent deformations in the shoot apical meristem (SAM), which inhibits their capacity to form organs and subsequently plants. In dicotyledonous plants, SAM is formed in the apex, between cotyledons and it plays a central role in postembryonic shoot organ formation. Based on the previous knowledge on the role of some families of gene in the formation, organization and maintenance of the SAM, the expression patterns of WUS, WOX2, NAM, STM, PIN1 and PIN7 genes were analysed, which would allow us to elucidate the possible implication of these genes in SAM deformations in the SE of C. chinense. The results show that the expression patterns of STM and PIN1 in the SE were completely opposite to the respective expression pattern obtained in zygotic embryos (ZE). Moreover, NAM and PIN7 showed an over accumulation of transcripts in SE, compared with ZE. This is the first time in the genus Capsicum that alterations in the expression pattern of key genes of the SE development are reported, as well as its possible implication in the persistent deformations of the SAM.

Elicitation effect on the production of asiaticoside and asiatic acid in shoot, callus, and cell suspension culture of Centella asiatica

Centella asiatica is an important medicinal plant which contains various phytocompounds. Asiatic acid and asiaticosideare two major compounds which are responsible for its various pharmaceutical activities. The present study analyzesthe effect of elicitor, i.e., methyl jasmonate on the synthesis of asiaticoside and asiatic acid (ATA) in shoot, callus, andcell suspension cultures of C. asiatica. A high-performance liquid chromatography analysis showed that the elicitationwith 100 µM concentration of methyl jasmonate enhanced asiaticoside content by 69-fold in callus culture, 39-fold inshoot cultures, and ATA by 1.9-fold in cell suspension culture. Thus, elicitation with methyl jasmonate is an effectivemethod of increasing the rate of biosynthesis of asiaticoside and ATA in plant cell cultures of C. asiatica

Genetic parameters of growth, and resistance to the shoot borer, in young clones of the tree Cedrela odorata (Meliaceae)

Damage by Hypsipyla grandella Zeller restricts the success of plantations of Cedrela odorata. The most critical and vulnerable infection period by H. grandella is during the first three years. The aim was to estimate the expected genetic gain for the selection of clones of fast growth and resistant to the attack by this insect. A trial with 40 clones produced by grafting was planted in Veracruz, Mexico. Heritability, genetic correlations and genetic gain of clones were estimated with data at two years-old. The survival rate of the trial was of 97 % (466 living ramets in total), 2.84 m in total high and 2.32 cm of average normal diameter. Aditionally, 9.9 % of the ramets did not present attacks, indication of evasion, and 0.6 percent without response to attack. With response to attack 89.5 % of the ramets (several degrees of tolerance): 29 % with a single shoot, 14.2 % with several shoots and 46.3 % with a dominant shoot of several shoots formed. The clonal heritability of normal diameter, height, volume, stem taper index, number of attacks and response to the attack was H2c = 0.81, 0.80, 0.81, 0.61, 0.34, and 0.26. The genetic correlations between height, diameter and volume were ≥ 0.95, and the correlation of the volume with the incidence and the response to the attack was rg = -0.31 and rg = 0.62, that is a decrease in number of attacks and moderate increase in tolerance. With a selection intensity of 10 %, selecting the four clones with higher volume will produce a genetic gain of 82 % for volume, a decrease of 10.9 % of number of attacks and 6.3 % of better recovery from damage; this is more tolerance to the insect attack. The genetic gain justifies the use of the best clones in commercial plantations in Veracruz, Mexico.

El daño por Hypsipyla grandella limita el éxito de las plantaciones de Cedrela odorata. El periodo más crítico y vulnerable es durante los primeros tres años. El objetivo del estudio fue estimar la ganancia genética esperada en la selección de clones de mayor crecimiento y resistentes al ataque del insecto. Un ensayo de 40 clones producidos por injertos fue plantado en Veracruz, México. Con datos a la edad de dos años se estimaron: la heredabilidad, correlaciones genéticas y ganancia genética de los clones. El ensayo presentó una supervivencia del 97 % (466 rametos vivos en total), con 2.84 m de altura y 2.32 cm de diámetro normal promedio. El 9.9 % de los rametos no presentaron ataques, indicando evasión, y se estimó un 0.6 % sin respuesta al ataque. Con respuesta al ataque 89.5 % de los rametos (diferentes niveles de tolerancia): 29 % con un solo brote, 14.2 % con varios brotes, y 46.3 % con un brote dominante de varios brotes formados. La heredabilidad clonal del diámetro normal, altura, volumen, índice de conicidad, número de ataques y respuesta al ataque fue de H2 c = 0.81, 0.80, 0.81, 0.61, 0.34 y 0.26, respectivamente. Las correlaciones genéticas entre la altura, diámetro y volumen fueron ≥ 0.95, y del volumen con la incidencia y la respuesta al ataque de rg = -0.31 y rg = 0.62, es decir se presentó una disminución en el número de ataques y el aumento moderado en la tolerancia. Con una intensidad de selección del 10 %, seleccionando los cuatro clones de mayor volumen se obtuvo una ganancia genética de 82 % para el volumen, una disminución de 10.9 % de ataques y 6.3 % mejor recuperación del daño, esto es más tolerancia al ataque del insecto. La ganancia genética justifica la utilización de los mejores clones en plantaciones comerciales para Veracruz.

Anti-proliferative and Apoptosis-Inducing Effects of Morinda citrifolia L. Shoot on Breast, Liver, and Colorectal Cancer Cell Lines

@#Introduction: Morinda citrifolia or also known as noni is commonly consumed raw or blanched as side dishes or ‘ulam’. As cancer is one of the most leading causes of death in the world, we aimed to evaluate the anti-proliferative potential of noni shoot against various types of cancer cell lines. Methods: The breast cancer (MDA-MB-231), liver cancer (HepG2), and colorectal cancer (HT-29) cell lines were treated with 70% ethanol extract of noni shoot for cytotoxicity testing using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Apoptosis induction effects were examined using AO/PI dual fluorescent assay and cell cycle analysis using flow cytometry. Gas chromatography-mass spectrometry (GCMS) was also carried out to characterize the active compounds in noni shoot. Results: The cytotoxicity assay demonstrated noni shoot had IC50 of 49.72 µg/mL, 307.5 µg/mL and 65.43 µg/ mL against MDA-MB-231, HepG2, and HT-29 cell lines, respectively. The AO/PI staining showed apoptotic bodies such as cell blebbing, chromatin condensation, and nuclear fragmentation was markedly induced in the selected cancer cell lines-treated with noni shoot extract. Apoptosis induction by noni shoot was showed by a significant increase in sub G0/G1 phase in MDA-MB-231 and HT-29 cell lines of cell cycle analysis. It was found that noni shoot extract contained mostly acetic acid and ethriol that may contribute to its anti-cancer properties. Conclusion: These findings showed the potential anticancer properties of noni shoot extract thereby, further studies are needed to understand the mechanism of noni as anti-cancer agent and possibility to be developed as a nutraceutical or functional food products.

De novo in vitro shoot morphogenesis from shoot tip-induced callus cultures of Gymnema sylvestre (Retz.) R. Br. ex Sm

BACKGROUND: Gymnema sylvestre is a medicinal woody perennial vine known for its sweetening properties and antidiabetic therapeutic uses in the modern and traditional medicines. Its over-exploitation for the therapeutic uses and to meet the demand of pharmaceutical industry in raw materials supply for the production of anti-diabetic drugs has led to considerable decline in its natural population. RESULTS: An efficient system of shoot bud sprouting from nodal segment explants and indirect plant regeneration from apical meristem-induced callus cultures of G. sylvestre have been developed on Murashige and Skoog (MS) medium amended with concentrations of cytokinins. Of the three growth regulators tested, N6-benzylaminopurine (BAP) was the most efficient and 2.0 mg L-1 gave the best shoot formation efficiency. This was followed by thidiazuron (TDZ) and kinetin (Kin) but, most of the TDZ-induced micro shoots showed stunted growth. Multiple shoot formation was observed on medium amended with BAP or TDZ at higher concentrations. The produced micro shoots were rooted on half strength MS medium amended with auxins and rooted plantlets acclimatized with 87% survival of the regenerates. CONCLUSIONS: The developed regeneration system can be exploited for genetic transformation studies, particularly when aimed at producing its high yielding cell lines for the anti-diabetic phytochemicals. It also offers opportunities for exploring the expression of totipotency in the anti-diabetic perennial vine.

In Vitro Organogenesis from Root Explants of Passiflora miniata Mast, an Amazonian Species with Ornamental Potential

ABSTRACT The present study reports a shoot organogenesis-based system for in vitro regeneration of Passiflora miniata, an Amazonia passion fruit species. Root segments were cultured in Murashige and Skoog (MS) medium supplemented with different concentrations (range 2-9 µM) of 6-benzyladenine (BA); thidiazuron (TDZ) or kinetin (KIN). Plant growth regulators were not added to the control treatment. Root explants have showed a high regenerative potential. After 30 days of in vitro culture, the root explants showed several shoots formed direct and indirectly. TDZ provided the best response in the differentiation adventitious shoots, mainly in the presence of 6.8 µM. The cytokinins BA and KIN responded producing a reduced number of shoots. After 120 days, rooted regenerated plants were transferred to a greenhouse for acclimatization. This regeneration system opens new perspectives for micropropagation and conservation of this wild Amazonic passion fruit species.

Critical vitrification steps towards survival of Garcinia hombroniana (Clusiaceae) shoot tips after cryopreservation / Etapas críticas de vitrificación para la supervivencia de ápices caulinares de Garcinia hombroniana (Clusiaceae) después de crioconservación

Abstract A cryopreservation protocol was developed for in vitro shoot tips of Garcinia hombroniana using the vitrification technique. Four critical steps in the technique were investigated, namely preculture, loading, dehydration with Plant Vitrification Solution 2 (PVS2), and unloading. Shoot tips precultured for 48 h gave significantly higher survival (75 %) compared to 24 h preculture (50 %) after cryopreservation. Treatment with 1 M glycerol plus 0.4 M sucrose as a loading solution gave higher survival (45.83 %) compared to the other treatments (0.4 M sucrose + 2 M glycerol; 0.4 M sucrose). Shoot tips dehydrated with PVS2 for 25 min gave the highest survival after immersion in liquid nitrogen. Stepwise PVS2 treatment for 15 min with 50 % PVS2 followed by 10 min with 100 % PVS2 solution improved survival of the shoot tips after cryopreservation (41.67 %). Murashige and Skoog medium with 0.4 M sucrose gave significantly higher survival (66.67 %) than MS with 1.2 M sucrose (25 %) as an unloading solution. Water content was shown to decrease throughout the whole vitrification steps from 6.83 ± 1.66 g g-1 dw for fresh shoot tips down to 2.93 ± 0.28 g g-1 dw after PVS2 treatment. Further study on each step including recovery medium is required to improve the survival. Nevertheless, the present study showed the potential of using the vitrification technique for cryopreservation of G. hombroniana. Rev. Biol. Trop. 66(3): 1314-1323. Epub 2018 September 01.

Resumen Se desarrolló un protocolo de crioconservación in vitro para ápices caulinares de Garcinia hombroniana mediante la técnica de vitrificación, con cuatro etapas críticas: precultivo, carga de crioprotector, deshidratación con solución de vitrificación vegetal 2 (PVS2), y descarga. Ápices precultivados por 48 h sobrevivieron más (75 %) que los de 24 h (50 %) después de la crioconservación. El tratamiento con glicerol 1 M más sacarosa 0.4 M como solución de carga permitió mayor sobrevivencia (45.83 %) que los otros tratamientos (sacarosa 0.4 M + glicerol 2 M; sacarosa 0.4 M). Los ápices deshidratados con PVS2 por 25 min registraron la mayor sobrevivencia tras inmersión en nitrógeno líquido. El tratamiento gradual por 15 min con solución de PVS2 al 50 %, seguido por 10 min al 100 %, mejoró la sobrevivencia de ápices tras la crioconservación (41.67 %). El medio Murashige-Skoog (MS) con sacarosa 0.4 M produjo una sobrevivencia significativamente mayor (66.67 %) que MS con sacarosa 1.2 M (25 %) como solución de descarga. El contenido de agua disminuyó a lo largo del proceso de vitrificación desde 6.83 ± 1.66 g g-1 peso seco, en ápices frescos, hasta 2.93 ± 0.28 g g-1 peso seco después del tratamiento con PVS2. Se requiere de más investigación sobre cada etapa, incluyendo el medio de recuperación, para mejorar la tasa de sobrevivencia. Sin embargo, este estudio muestra el potencial de la vitrificación para la crioconservación de G. hombroniana.

Organogenesis of Abelmoschus moschatus Medik. Using Aseptic Seedling Explants

Aim: Abelmoschus moschatus have been extensively used in traditional medicine as well as in perfume industries. The primary goal of the present research was to develop an efficient plant regeneration protocol of Abelmoschus moschatus from aseptic seedling explants such as cotyledon, internode, leaf and root. Methodology: The seeds of Abelmoschus moschatus were surface sterilized with 0.1% mercuric chloride and 70% ethanol were cultured on ½ MS basal media for developing aseptic seedlings Aseptic seedling explants were cultured on different concentrations of auxins for callus induction. Later callus was transferred on to different concentrations of cytokinins for shoot regeneration and for in vitro, rooting different concentrations of auxins were used. Finally, such in vitro developed plantlets were acclimatized. Results: Half strength MS medium with 1% sucrose was used for raising aseptic seedlings. Maximum of 92% response of callus induction was obtained from leaf explants on MS medium + 2 mg/L 2, 4-dichlorophenoxyacetic acid. An average of 2.4 shoots per callus were observed on MS + 2 mg/L benzyl-6-aminopurine from leaf explant. The regenerated shoots were best rooted on 1/2 MS + 0.5 mg/L indole-3-butyric acid. The regenerated plantlets were established with 70% survival. Conclusion: An efficient plant regeneration protocol of Abelmoschus moschatus was developed.

Effects of Leucaena leucocephala (lamk. ) shoot tips plus young leaf extract containing mimosine on reproductive system of male rats / Efectos de los brotes de Leucaena leucocephala (lamk. ) y del extracto de hojas jóvenes que contiene mimosina en el sistema reproductivo de ratas macho

This study aimed to determine the mimosine level and examine the male reproductive toxicity effects of Leucaena leucocephala (LL) shoot tips plus young leaf extract. Mimosine level in LL extract was determined by thin layer chromatography before administration in animals. Male rats were divided into control and LL (1,500 mg/KgBW) groups (n = 6). After 60 days of experiment, serum sex hormones, sperm quality, and testicular histopathology were assayed and observed. Malondialdehyde (MDA) level and expressions of steroidogenic acute regulatory (StAR) and phosphorylated proteins in testicular lysate were examined by western blotting. Results showed that mimosine levels in LL extract was 17.35 ± 1.12 % of dry weight. LL significantly decreased FSH & LH levels, sperm qualities, and seminiferous tubule diameter compared to the control (p<0.05). Seminiferous tubular atrophies, germ cell sloughing, and degenerations were observed in LL group. In addition, testicular MDA level and StAR protein expression were significantly decreased in LL group. LL extract could increase the expression of a 50 kDa phohorylated protein in testicular lysate. In conclusion, LL extract has mimosine and reproductive toxicity effects on males.

Este trabajo tuvo como objetivo determinar el nivel de mimosina y examinar los efectos de la toxicidad reproductiva de los brotes de Leucaena leucocephala (LL), más el extracto de hojas jóvenes, en ratas macho. El nivel de mimosina en el extracto de LL se determinó mediante cromatografía en capa fina antes de la administración en animales. Las ratas se dividieron en grupos de control y LL (1,500 mg / kgBW) (n = 6). Después de 60 días, se analizaron y observaron las hormonas sexuales séricas, la calidad de los espermatozoides y la histopatología testicular. A través de Western Blot se examinaron el nivel de malondialdehído (MDA), las expresiones de reguladores agudos esteroidogénicos (StAR) y las proteínas fosforiladas en el lisado testicular. Los resultados mostraron que los niveles de mimosina en el extracto de LL fueron 17.35 ± 1.12 % del peso seco. LL disminuyó significativamente los niveles de FSH y LH, la calidad de los espermatozoides y el diámetro de los túbulos seminíferos en comparación con el control (p <0,05). Se observaron atrofias en los túbulos seminíferos, desprendimiento de células germinales y degeneraciones en el grupo LL. Además, el nivel de MDA testicular y la expresión de la proteína StAR se redujeron significativamente en el grupo LL. El extracto de LL podría aumentar la expresión de la proteína fosforilada de 50 kDa en el lisado testicular. En conclusión, el extracto de LL tiene mimosina y efectos de toxicidad reproductiva en los hombres.

A Review on In vitro Regeneration of Ginger: Tips and Highlights

Ginger (Zingiber officinale) is one of the oriental spices, widely used worldwide for multiple purposes. It is applied as an important ingredient in Ayurvedic preparations from time immemorial. Tissue culture is a practice that is utilized to propagate plants from cells or tissue under sterile conditions. This study is directed to create a review of the successful and reproducible convention for in vitro recovery of ginger with emphasis on effective initial culture establishment. Furthermore, it has dealt with the appropriate explant size and effectiveness of media quality on micropropagation of ginger. The current study recommends that the medium containing Benzyl Amino Purine (BAP) can be used for inducing shoot development of ginger. Among the diverse explants, shoot tips give the quickest response for starting development and the highest number of multiple shoots are produced. As well, it is demonstrated that a survival rate and proper shoot/root expansion can be obtained through the tissue culture methods. Emphasizing the tips and recommendations, this study would be a route-map towards time and cost saving for producing a better quality of ginger.

Fagodisuasión de extractos deBrugmansia candida(Solanaceae) en larvas deHypsipyla grandella(Lepidoptera: Pyralidae) / Phagodeterrence of extracts ofBrugmansia candida(Solanaceae) onHypsipyla grandella(Lepidoptera: Pyralidae) larvae

Resumen Hypsipyla grandellaes una plaga forestal clave en América Latina y el Caribe, donde ha impedido todo intento de establecer plantaciones comerciales de caobas (Swieteniaspp.) y cedros (Cedrelaspp.). En la búsqueda de métodos de manejo de sus larvas, que sean preventivos y rentables, así como basados en recursos de la flora tropical, fue evaluada la actividad fagodisuasiva del extracto crudo y de cuatro particiones fitoquímicas (agua, hexano, diclorometano y acetato de etilo) de la flor de reina de la noche,Brugmansia candida(Solanaceae). Se efectuaron bioensayos de laboratorio con concentraciones crecientes del extracto crudo (0.1; 0.3; 1.0; 3.2 y 10.0 % m/v) y con cada una de las particiones (según el rendimiento del proceso de particionamiento). Se utilizó un diseño de bloques completos al azar, con cuatro repeticiones, y se expusieron larvas de instar III de H. grandellaa discos de cedro amargo (Cedrela odorata) impregnados con el respectivo tratamiento, por 24 h. Se midió el porcentaje de consumo de cada disco. El extracto crudo y la partición de hexano, mostraron un evidente efecto fagodisuasivo a partir de concentraciones de 1.0 % y 0.122 % m/v, respectivamente. También se realizó un bioensayo para comparar una sola concentración (0.38 % m/v) del alcaloide escopolamina, el cual redujo en 90% el consumo con respecto a los testigos. Asimismo, se evaluó la mortalidad y los efectos subletales en larvas expuestas al extracto crudo, la partición de hexano y la escopolamina, pero no hubo evidencias de ellos, lo que confirmó su acción fagodisuasiva. Además, se realizó un tamizaje fitoquímico del extracto crudo, y mediante pruebas cualitativas se determinó que los alcaloides, taninos, triterpenos y cumarinas fueron los metabolitos secundarios más importantes. Finalmente, se confirmó la presencia de la escopolamina tanto en el extracto crudo como en la partición de hexano, mediante el método de cromatografía de gases acoplado a un espectrómetro de masas (GC-MS). Se recomienda preparar una formulación de escopolamina más algunos coadyuvantes, y evaluar su eficacia en condiciones de invernadero, como un primer paso para avanzar en el desarrollo de un fagodisuasivo comercial.

Abstract Hypsipyla grandella is a key forest pest in Latin America and the Caribbean, where it has precluded attempts to establish commercial plantations of mahoganies (Swieteniaspp.) and cedars (Cedrelaspp.). In the search for methods to manage its larvae, being both preventative and cost-effective, as well as based upon resources from tropical flora, the crude flower extract of angel's trumpet, Brugmansia candida, as well as four partitions thereof (water, hexane, dichlorometane, and ethyl acetate), were tested for phagodeterrence. Laboratory bioassays involved increasing concentrations of the crude extract (0.1, 0.3, 1.0, 3.2 and 10.0 % w/v) as well as each one of the partitions (in accordance to the yield obtained from the partitioning process). A randomized complete block design, with four replicates, was used. H. grandellainstar III larvae were exposed for 24 h to Spanish cedar (Cedrela odorata) leaf discs dipped into the respective treatment, after which disc consumption was measured. Strong phagodeterrence was detected at concentrations as low as 1.0 % y 0.122 % w/v for the crude extract and the hexane partitions, respectively. In addition, a single bioassay was performed to compare a single concentration (0.38 % w/v) of the alkaloid scopolamine, which reduced consumption in 90 % with respect to the controls. Also, mortality and sublethal effects were assessed in larvae exposed to the crude extract, the hexane partition and scopolamine, with no evidence for them, which confirmed their phagodeterrence. Moreover, the crude extract was submitted to a phytochemical screening by means of a number of qualitative tests, which showed that alkaloids, tannins, triterpenes and cumarins were the most important secondary metabolites. Finally, the presence of scopolamine was confirmed in both the crude extract and the hexane partition, by means of the gas chromatography-spectrometry (GC-MS) analytical method. It is recommended to prepare a formulation of scopolamine plus some adjuvants, in order to test its effectiveness under greenhouse conditions, as a first step to advance in the development of a commercial phagodeterrent. Rev. Biol. Trop. 66(1): 58-69. Epub 2018 March 01.

Advances in plant stem cell culture / 生物工程学报

Plant stem cells are the cells that are located in meristems and are kept in a state of undifferentiation. Plant stem cell possesses lower vacuolization, higher mitochondrial activity, more genetic stability and stronger self-renewal capacity compared with calli. Plant stem cell culture has a wide application in pharmaceutical, functional food as well as cosmetic industries. Here we describe the procedure of induction, isolation and identification of plant stem cells, to provide a reference for further research in this field.

The Effect of 4-hydroxybenzaldehyde on the γ-aminobutyric Acid Type A Receptor

in the mammalian cortex and hippocampus. It is expressed heterologously in the Xenopus laevis oocyte as a α1β2γ2S/L subtype for application as an in vitro model for the screening of compounds that modulate receptor activities. In fact, 4-hydroxybenzaldehyde (4-HB) has been identified as one of the major components in Dendrocalamus asper bamboo shoots in our previous study, and the current study showed that at 101.7 μM, 4-HB significantly reduced the GABA-induced chloride current of GABAA receptors expressed on Xenopus oocytes, indicating a possible GABAergic antagonistic effect at high concentrations.

Antioxidant capacity and acute testicular toxicity of Leucaena leucocephala aqueous shoot tips plus young leaves extracts / Capacidad antioxidante y toxicidad testicular aguda de brotes acuosos con extracto de hojas nuevas de Leucaena leucocephala

This study aimed to determine the antioxidant capacity of the Leucaena leucocephala aqueous shoot tips plus young leaves (LL-spl) extracts among three different fractions (LL-spl 10, 20, and 40 min) and to examine its acute toxicity on male reproductive parameters. The amount of the total phenolics in LL-spl extract was determined using a Folin-Ciocalteu reagent method and its antioxidant capacity was analyzed using 1, 1-diphenyl l-2-picrylhydrazyl radical scavenging and ferric reducing antioxidant powder methods. The LL-spl extract fraction with highest antioxidant capacity was used in animal treating. Male rats were divided into three groups (n= 5); control and groups treated with LL-spl 400 and 600 mg/Kg body weight for consecutive 40 days. The results showed that the LL-spl 40 min fraction possessed the highest antioxidant capacity. In addition, the LL-spl 400 and 600 groups showed no differences in weights of body, testis and epididymis, serum testosterone levels, and expression of testicular steroidogenic acute regulatory (StAR) protein. Significantly, LL-spl extract reduced the weight of seminal vesicle, sperm concentration, and seminiferous diameters compared with control. Moreover, LL-spl extract had adverse effect on testicular histology in inducing of seminiferous atrophy and degeneration including dilated blood vessels in interstitial tissue. It was concluded that although LL-spl extract possessing antioxidant capacity, in short term consumptions, it could be toxic to some male reproductive organs especially damaging testicular tissues.

El objetivo fue determinar la capacidad antioxidante del extracto de brotes acuosos con hojas nuevas de Leucaena leucocephala (LL-spl) en tres fracciones diferentes (LL-SPL 10, 20 y 40 min), además de examinar su toxicidad aguda sobre los parámetros reproductivos masculinos. Se determinó la cantidad de los fenoles totales en el extracto de LL-spl utilizando un método reactivo de Folin-Ciocalteu. La capacidad antioxidante se analizó por medio de 1-difenil-2-picrilhidracilo y/o métodos de reducción férrica de la capacidad antioxidante. La fracción de extracto LL-spl con mayor capacidad antioxidante fue utilizada en el tratamiento de los animales. Ratas macho fueron divididas en tres grupos (n= 5): el control y los grupos tratados con LL-spl 400 y 600 mg/kg peso corporal por 40 días consecutivos. El resultado mostró que la fracción LL-spl 40 min poseía la mayor capacidad antioxidante. Además, los grupos 400 y 600 LL-spl no mostraron diferencias según el peso corporal, testículos y epidídimo, niveles de testosterona y la expresión de proteínas testiculares. El extracto de LL-spl redujo de manera significativa el peso de la vesícula seminal, la concentración de espermatozoides y los diámetros de los túbulos seminíferos en comparación con el control. Por otra parte, el extracto de LL-spl tuvo un efecto adverso sobre la histología testicular por la inducción de atrofia y degeneración de los túbulos seminíferos, incluyendo a vasos sanguíneos dilatados en el tejido intersticial. Si bien el extracto LL-spl posee una capacidad antioxidante, ésta podría ser tóxica en el consumo a corto plazo para algunos órganos reproductores masculinos y especialmente dañino para los tejidos testiculares.

Influence of Benzyladenine and Thidiazuron on Shoot Regeneration from Leaf and Shoot tip Explants of Sedum sarmentosum Bunge

Effects of N6-benzyladenine (BA) or thidiazuron (TDZ) on adventitious shoot regeneration and axillary shoot multiplication of Sedum sarmentosum was investigated. Leaf and shoot tip explants obtained from in vitro-grown shoots of S. sarmentosum were cultured on Murashige and Skoog (MS) medium supplemented with 0, 2.0, 4.0 or 8.0 µM BA or TDZ. Of the two cytokinins studied, BA was found to be more responsive as compared to TDZ with respect to number of shoots produced per explant. High frequency of shoot regeneration (92.2%) with a mean of 12.3 shoots was produced when the leaf explants were cultured on MS medium supplemented with 8.0 µM BA. The highest number of shoots (25.4) was obtained when shoot tip explants were cultured on MS medium devoid of cytokinins after 35 days of culture. For root induction, regenerated shoots were cultured on MS medium supplemented with 0, 2.0, 4.0 or 8.0 µM indole-3-acetic acid (IAA), indole-3-butyric acid (IBA) or α-naphthaleneacetic acid (NAA). The highest number of (17.6) roots per shoot was obtained on MS medium supplemented with 2.0 µM IBA after 30 days of culture. Regenerated plantlets were successfully acclimatized in the greenhouse with 100% survival rate.