Micropropagation of juvenile and adult Annona squamosa

A micropropagation system for Annona squamosa L. (Sugar Apple) using hypocotyls of seedlings and nodal cuttings from 3-year-old plants was developed. Shoot proliferation was achieved with Woody Plant Medium supplemented with BA. Silver thiosulphate was added at 0.5 mg l^–1 to control leaf abscission. Rooting was obtained when subcultured shoots were preconditioned for 2 weeks in medium with 10 g l^–1 activated charcoal before treatment with 43 µm NAA or 39 µm IBA. Rooting was improved when galactose was used instead of sucrose in the rooting medium. The rooted plantlets were acclimatised successfully.Abbreviations NAA naphthaleneacetic acid - IBA indolebutyric acid - MS Murashige & Skoog Medium - WPM Woody Plant Medium - NN Nitsch Medium - Juv juvenile explant - Adu adult explant     

A Humidity-induced Convective Throughflow Ventilation System Benefits Annona squamosa L. Explants and Coconut Calloid

A simple apparatus is described for generating pressurized throughflowventilation in plant tissue culture vessels. No pumps or gas-cylindersare required and the flow is driven by humidity-induced diffusionacross microporous membranes. In the experiments described,pressurized flows of sterile humidified air were supplied atrates of up to 1 ml min^-1and these had beneficial effects onleaf survival and production inAnnona cuttings and on calloidform in coconut. Ethylene (ethene) was removed more quicklyfrom the pressure-flow ventilated culture vessels (t ₅₀, 0.4–0.7h) than from those aerated by diffusion through conventionalpolypropylene membranes (t ₅₀, 1.6–2.4 h). InAnnona cuttings leaf production was greatly increased andethylene-induced leaf fall considerably delayed when culturedwith the forced as opposed to diffusion-based ventilation ofthe vessels. With throughflow ventilation, coconut calloid was more convolutedthan under wholly diffusive aeration and had a smooth distinctepidermal surface and clearly defined sub-epidermal meristematicnodules. It resembled freshly initiated calloid from which regenerationof plantlets via somatic embryogenesis can be obtained. Underwholly diffusive aeration, calloid developed a rough, relativelyundifferentiated surface, more haustorial (i.e. cotyledonary)in appearance, and characteristic of cultures where regenerationpotential has been lost. It is suggested that other benefits of the pressurized throughflowventilation may be the removal of volatiles such as ethanoland acetaldehyde, the removal of excess carbon dioxide at nightand its improved supply during the day, and sustained oxygenconcentrations at levels close to atmospheric both night andday. Abscission; callus; ethylene; explants; tissue-culture; growth; ventilation     

Monotetrahydrofuran annonaceous acetogenins from the seeds of Annona squamosa

Three new monotetrahydrofuran annonaceous acetogenins, Annosquamins A–C (1–3) were isolated from the 95% EtOH extract of Annona squamosa seeds, in addition to the known compounds, solamin, annotemoyin-1, uvariamicin-II, uvariamicin-III. Their structures were elucidated by spectroscopic analysis and the relative configurations of bearing tetrahydrofuran rings were established by comparing NMR data with model compounds.     

Pollen--stigma Interactions: Development and Cytochemistry of Stigma Papillae and their Secretions in Annona squamosa L. (Annonaceae)

The development and cytochemical features of the stigma andstyle have been investigated in Sugar apple, Annona squamosaL., using light and electron microscopy. The pistil is a syncarpwith an open stylar canal. Papillae of epidermal origin lineboth the surface of the stigma and the inner face of the stylarcanal. The papillae contain organelles characteristic of secretorycells with a highly thickened cellulosic wall. The wall is multi-layered,the zones differing in their microfibrillar stacking and orientation.The stigma is of the ‘wet’ type and the surfaceexudate is heterogeneous in microscopic appearance and reactscytochemically for proteins, carbohydrates and lipids. The surfacecuticle undergoes dissolution prior to anthesis. A secretionalso appears in the thickened middle lamella of the sub-epidermalcell layer which reacts cytochemically for pectinaceous acidicpolysaccharides. Esterase activity of papillae is indicative of the receptiveareas, and it is also related to the onset of receptivity. Acidphosphatase activity is intense in the sub-epidermal cell layerswhich probably reflects their secretory activity. Pollinationtriggers a copious flow of secretion onto the stigma surfacewhich engulfs the pollen grains. It appears that most of theacidic polysaccharides of this secretion come from the middlelamella of the sub-epidermal cell layer. Compatible pollen tubes have no apparent barriers to overcomeon their route to the embryo sac and the inherent protogynousdichogamy seems to control the acceptance or rejection of compatiblepollen. Annona squamosa L., sugar apple, stigma, style, secretions     

In vitro antioxidant studies of Annona squamosa Linn. leaves

The free radical scavenging potential of the leaves of A. squamosa was studied by using different antioxidant models of screening. The ethanolic extract at 1000 microg/ml showed maximum scavenging of the radical cation, 2,2-azinobis- (3-ethylbenzothiazoline-6-sulphonate) (ABTS) observed upto 99.07% followed by the scavenging of the stable radical 1,1-diphenyl, 2-picryl hydrazyl (DPPH) (89.77 %) and nitric oxide radical (73.64%) at the same concentration. However, the extract showed only moderate scavenging activity of superroxide radicals and antilipid peroxidation potential, which was performed using rat- brain homogenate. The findings justify the therapeutic applications of the plant in the indigenous system of medicine, augmenting its therapeutic value.     

An improved method for DNA isolation from polyphenols rich leaf samples of Annona squamosa L.

A reliable and efficient method for isolating Annona squamosa L. genomic DNA, free from polyphenols and polysaccharides has been developed. Different methods involving use of CTAB and SDS with or without modifications were used. A CTAB based extraction method which uses diatomite to remove polyphenols and polysaccharides proved to be the best. This method allowed recovery of good quality DNA in sufficient quantity suitable for complete digestion by restriction endonucleases and amplifiable in polymerase chain reaction as compared to other methods.     

番荔枝内生真菌Hypoxylon sp.B38的代谢产物研究

从番荔枝内生真菌Hypoxylon sp.B38的固体平板发酵提取物中分离到5个化合物,通过NMR和MS等方法把它们分别被鉴定为Lachnellin D(1)、D ienetriol 9(2)、Lachnellin C(3)、Lachnellin B(4)和Benzo[d][1,3]d ioxol-5-ylm ethanol(5)。     

Effects of Annona squamosa extract on early pregnancy in rats.

Annona squamosa Linn., family Annonaceae, is said to show varied medicinal effects, including insecticide, antiovulatory and abortifacient. The purpose of present study was to investigate if A. squamosa seed aqueous extract, in doses higher than that popularly used to provoke abortion, interferes with reproductive performance, and to correlate the ingestion of this extract with possible alterations in rat embryonic implantation. Doses of 300 mg/kg (Treated Group I, n = 17) and 600 mg/kg (Treated Group II, n = 12) body wt. were administered by gavage, during days 1 to 5 of pregnancy (preimplantation period). The control group (n = 13) received water in the same manner, during the same period for comparison with experimental groups. The animals were euthanized on day 10 of pregnancy. Treatment of dams during the preimplantation period showed no signs of toxicity, and no alteration in the corpora lutea, implantations and embryo in terms of development numbers. The percentage of preimplantation and postimplantation losses in treated groups I and II did not differ from those of control. Treatment with aqueous extract of A. squamosa seeds caused no morphological change in the endometrium. The absence of morphological alterations in uterine epithelial cells in treated groups I and II permitted a viable embryonic implantation, as verified by the number of embryos in development at day 10 of pregnancy. Thus, A. squamosa seed aqueous extract did not interfere with the reproductive performance of pregnant rats.     

Three cytotoxic Annonaceous acetogenins from the seeds of Annona squamosa

Three new Annonaceous acetogenins, squamocin-IV (1), squamocin-V (2) and squamoxinone-E (3), together with seven other known Annonaceous acetogenins (4–10) were isolated from the seeds of Annona squamosa. The structures of all of the isolates were established and characterized through spectral and chemical methods. The new Annonaceous acetogenins 1–3 were evaluated for their cytotoxicity against the Hep-G2, SMMC-7721, BEL-7402, BGC-803 and H460 human cancer cell lines. Compound 3 exhibited the best cytotoxic activity, with IC₅₀ values of 0.103, 0.687, 4.19, 0.43 and 6.56 μg/mL, respectively for the cell lines above; meanwhile, Compound 1 showed selective cytotoxic activity against H460, with an IC₅₀ value of 0.049 μg/mL. The first mono-ACG (2) composed of 38 carbons was discovered.     

Studies on endosperm culture of Annona squamosa Linn

Mature endosperm tissue excised from germinated seeds (2–4 days after radicle emergence) of Annona squamosa grew and proliferated on White's basal medium supplemented with two cytokinins, an auxin and gibberellic acid. The callus obtained could be periodically subcultured. Shoot differentiation and root induction were obtained from callus on media of different compositions. Analyses of the root and young leaf tips showed triploid number of chromosomes (3n=21).Abbreviations Kin Kinetin - BAP 6-Benzylaminopurine - IAA Indole-3-acetic acid - IBA Indole-3-butyric acid - NAA naphthalene abetic acid - GA₃ Gibberellic acid Communication No. 3885     

Annonaceous acetogenins: Precursors from the seeds of Annona squamosa

From the less polar fractions of an Annona squamosa (Annonaceae) seeds extract, four new annonaceous acetogenins, dieposabadelin (1), squamocenin (2), lepirenin (3) and dotistenin (4), were isolated, along with sixteen known acetogenins: corepoxylone, diepomuricanins A and B, dieporeticenine, tripoxyrollin, bullatencin, glabrencin B, reticulatains-1, -2, uvariamicins I, II, III, erythrosolamin, annotemoyin-1 and -2, and solamin. Fourteen are reported for the first time in this species.     

番荔枝化学成分研究(7)

从番荔枝（Ａｎｎｏｎａ ｓｑｕａｍｏｓａ Ｌ．）种子中得到化合物１、２和４,化合物２和４分别为已知化合物ｍｏｔｒｉｌｉｎ（莫垂林,２）和ｂｕｌｌａｔａｎｏｃｉｎ（布拉他诺辛,４）。化合物１的化学结构经ＩＲ、ＭＳ、＾１Ｈ ＮＭＲ及＾１３Ｃ ＮＭＲ谱分析及其乙酰化物（１ａ）的ＭＳ、＾１Ｈ ＮＭＲ谱分析推定如（１）式并命名为２２－表－莫维查灵（２２－ｅｐｉ－ｍｏｌｖｉｚａｒｉｎ）。     

In vitro organogenesis from leaf explants of Annona squamosa Linn

Multiple shoot formation was induced from excised leaf explants of Annona squamosa Linn. (custard apple) seedlings on a Murashige and Skoog basal medium containing benzylaminopurine and kinetin. Various auxins in combination with the above medium produced callusing of the explants. In an investigation of environmental factors affecting shoot induction it was seen that the maximum number of shoots were obtained using the leaf base with petiole at a temperature of 27°C and a light intensity of 1000 lux. Roots were initiated erratically when individual shoots were treated with an auxin and then transferred to an auxin free medium. The process of the development of adventitious buds in leaf culture was analysed histologically.NCL Communication No. 3104.     

番荔枝化学成分研究(6)

自番荔枝（ ＡｎｎｏｎａｓｑｕａｍｏｓａＬ．） 种子中得到化合物１ 和２ , 化合物２ 是已知的ｂｕｌｌａｔａｃｉｎｏｎｅ （２ , ４－ 顺式和反式－ ｂｕｌｌａｔａｃｉｎｏｎｅ 的混晶） , １ 是新化合物, 命名为番荔枝塔亭丁（ｓｑｕａｍｏｓｔａｔｉｎ Ｄ） , 其结构经ＩＲ、ＭＳ、１Ｈ－ ＮＭＲ和１３ＣＮＭＲ谱解析推定如（１） 式。     

Studies on the Chemical Constituents of Annona squamosa(5)

From the seeds of Annona squamosa L.,three compounds were obtained,in which compounds i and 2 are known compounds,i.e. bullatacin and annonin I,respectively.Compounds 3 is a new non -adjacent bis -tetrahydrofuranyl annonaceous acetogenin,named squamostatin C.Based on the analysis of IR MS,1H NMR and 13 C NMR spectra,the structure of compound 3 was elucidated as (3).     

番荔枝化学成分研究(5)

自番荔枝(AnnonasquamosaL)种子中分离到3个番荔枝内酯(1～3)。化合物1和2分别是已知化合物bullatacin和annoninⅠ,化合物3系新的间双四氢呋喃环型番荔枝内酯,命名为番荔枝塔亭丙(squamostatinC),经IR、MS、1HNMR及13CNMR谱分析,其结构推定如(3)式。     

In vitro organogenesis from leaf explants of Annona squamosa Linn

Multiple shoot formation was induced from excised leaf explants of Annona squamosa Linn. (custard apple) seedlings on a Murashige and Skoog basal medium containing benzylaminopurine and kinetin. Various auxins in combination with the above medium produced callusing of the explants. In an investigation of environmental factors affecting shoot induction it was seen that the maximum number of shoots were obtained using the leaf base with petiole at a temperature of 27°C and a light intensity of 1000 lux. Roots were initiated erratically when individual shoots were treated with an auxin and then transferred to an auxin free medium. The process of the development of adventitious buds in leaf culture was analysed histologically.     

Biosynthesis, characterization and antibacterial activity of silver nanoparticles by aqueous Annona squamosa L. leaf extract at room temperature

Silver nanoparticles (AgNPs) have gained great interest in nanotechnology, biotechnology and medicine. The green synthesis of nanoparticles has received an increasing attention because of it’s maximize efficiency and minimize health and environmental hazards as compared to other conventional chemical synthesis. In this study, we reported biosynthesis of AgNPs by aqueous Annona squamosa L. leaf extract and its characterization by UV-visible spectroscopy (UV–vis), Field emission gun scanning electron microscopy (FEG-SEM), X-ray energy dispersive spectroscopy (EDX), Transmission electron microscopy (TEM), Selected-area electron diffraction (SAED) and Fourier transform infra-red spectroscopy (FTIR). The results indicated that AgNPs formed were spherical in shape with size ranging from 14 to 40 nm with an average diameter 28.47 nm. Furthermore, it was observed that the AgNPs exhibited an antibacterial activity against different Gram positive and Gram negative microorganisms. Our report confirmed that the ALE is a very good eco-friendly and nontoxic bioreductant for the synthesis of AgNPs and opens up further opportunities for fabrication of antibacterial drugs, medical devices and wound dressings.     

Ultrastructure and Development of Oil Idioblasts in Annona muricata L.

The ultrastructure and development of oil idioblasts in theshoot apex and leaves in Annona muricata L. are described, andthree arbitrary developmental stages are distinguished: cellsin which no additional cell wall layers have been depositedagainst the initial primary cell wall, possessing an electron-translucentcytoplasm and distinct plastids which lack thylakoids (stage1); cells in which a suberized layer has been deposited againstthe primary wall (stage 2, the cytoplasm resembles that of thepreceding stage), and cells in which an additional inner walllayer has been deposited against the suberized layer, whichincreases in thickness with development (stage 3). In this stagean oil cavity is formed, surrounded by the plasmalemma, andattached to a bell-like protrusion of the inner wall layer,the cupule. A complex membranous structure occurs next to thecupule. Smooth tubular endoplasmic reticulum (ER), appearingas linearly arranged tubules, and groups of crystalline bodieswith an almost hexagonal outline are present. The final stagewas further subdivided into three subgroups (a, b, c) basedon the extent of the oil cavity, its contents, and the compositionof the cytoplasm, and increasing thickness of the inner walllayer. The oil is probably synthesized in the plastids, releasedinto the cytoplasm, and then passed through the plasmalemmasurrounding the oil cavity. Oil idioblasts, Annona muricata L., suberized layer, inner wall layer, oil cavity, cupule, smooth tubular ER, crystalline bodies