Underground system of Mandevilla atroviolacea (Stadelm.) Woodson (Apocynaceae,Apocynoideae) from the Brazilian high-altitude grassland

The underground systems of plants show the most varied structures adapted to survival in unfavorable environmental conditions. For instance, long-term droughts may induce the development of either water and reserve substances or vegetative propagation structures. Since Mandevilla atroviolacea is a species found at high altitudes on rocky outcrops, this study aimed to provide information on the morpho-anatomy of the underground system of this species that may assist in understanding the adaptive strategies at play. Samples from the underground system of three young and two mature plants collected in the field were sectioned and processed according to standard plant anatomical techniques. The upper area of the underground system corresponds to a xylopodium, while the lower is a tuberous stem organ. Tuberous roots account for the majority of the underground system. The increase in tuberous root diameter results from the action of a typical cambium. The high shoot-forming potential and storage of water and reserve substances observed in the underground system of this species are important strategies for it to survive in this habitat.     

Flavonoids from the flowers of Adenium obesum (Forssk.) Roem. & Schult., Mandevilla sanderi (Hemsl.) Woodson,and Nerium oleander L. (Apocynaceae)

Twenty-two ornamental flowers from different Adenium obesum, Mandevilla sanderi, and Nerium oleander cultivars/seedlings were analyzed for the presence of anthocyanins, flavonols, and chlorogenic acid using nuclear magnetic resonance (NMR) and mass spectrometry (MS). Cyanidin 3-O-[6-O-(rhamnosyl)-galactoside] and cyanidin 3-O-(galactoside) were identified as the major and minor anthocyanins, respectively, in three A. obesum seedlings that had red and red-purple flowers.Cyanidin 3-O-[2-O-(xylosyl)-galactoside] was identified as the major anthocyanin, whereas cyanidin 3-O-[6-O-(rhamnosyl)-galactoside] and cyanidin 3-O-(galactoside) were identified as the minor anthocyanins in 8 M. sanderi cultivars that had red and red-purple flowers. Cyanidin 3-O-[6-O-(rhamnosyl)-galactoside] and cyanidin 3-O-(galactoside) were identified as the major anthocyanins, whereas cyanidin 3-O-[2-O-(xylosyl)-galactoside] was identified as the minor anthocyanin in 8 N. oleander cultivars with red and red-purple flowers. Low levels of anthocyanins were detected in the N. oleander and M. sanderi cultivars that had white flowers, and there were no anthocyanins detected in the N. oleander cultivars with yellow flowers. Chlorogenic acid and four flavonols, quercetin 3-O-[6-O-(rhamnosyl)-galactoside], quercetin 3-O-[6-O-(rhamnosyl)-glucoside], kaempferol 3-O-(galactoside), and kaempferol 3-O-[6-O-(rhamnosyl)-galactoside], were identified in the flowers from all 22 cultivars/seedlings investigated.     

Micropropagation of Lippia junelliana (Mold.) Tronc.

A method for the micropropagation of Lippia junelliana (Mold.) Tronc. from shoot tips or nodal segments was developed. Proliferating microshoot cultures were obtained by placing shoot tips or nodal segments on full strength Murashige and Skoog medium (MS) supplemented with 4.4 μM benzyladenine (BA) or 0.04 μM indolebutyric acid- (IBA) plus 4.4 μM BA. The rooting of shoots was better on full-strength MS medium without growth regulators. Rooted plantlets were successfully acclimatized to soil. The shoot cultures showed a lower essential oil accumulation in comparison with parent plants. Essential oil accumulation is closely related with growth and shows a negative correlation with shoot proliferation. This revised version was published online in June 2006 with corrections to the Cover Date.     

仙人掌的微繁殖

成功建立了仙人掌离体快繁的实验体系 ,并且对影响微繁殖的一些因素 ,诸如激素组合、外植体的物理状态、大量元素的含量等进行了研究。结果表明 :BA对仙人掌芽增殖具明显作用 ,MS +BA 5 .0mg/L +IBA 0 .1mg/L为最适增殖培养基 ;接种方式实验表明劈接优于整棵。钙、镁离子浓度对试管苗生长没有影响 ,但影响生根数及根长 ;NAA抑制根的伸长 ,但一定浓度可促进生根。总体而言 ,最适的生根培养基为 1 /2MS。同时发现块接比单芽接具有优势。试管苗在形态上出现一些变异。实验结果对仙人掌科其它植物的快速繁殖具有参考意义     

Micropropagation of Alnus cordata (Loisel.) Loisel.

Procedures were developed for micropropagation of Alnus cordata through in vitro axillary shoot multiplication of axillary bud explants cultured in Murashige & Skoog (MS) medium. Establishment of cultures from plants grown in the field was very difficult due to bacterial contamination and phenolic oxidation in explants causing severe browning. Explants were first cultured on an MS medium containing 4.4 M 6-benzyladenine and 87.6 mM sucrose (initiation medium) for 7 days and then transferred to an MS medium containing 1.1 M 6-benzyladenine and 333 mM glucose (multiplication medium) for a further 20–25 days. It was necessary to transfer cultures from initiation medium to multiplication medium after 7 days to minimize excessive callus growth, abnormally thick and brittle leaves, inhibition of shoot elongation, and senescence. Shoot multiplication comparable to the above method was achieved by culture of axillary bud explants in MS medium supplemented with 1.1–4.4 M 6-benzyladenine and 333 mM glucose 4–5 weeks after culture. Shoots rooted in MS medium (1/2 x macro-nutrients) supplemented with 1.2–4.9 M indolebutyric acid. Also, 98% rooting was achieved when cultures were treated with 625 mgl^-1 indolebutyric acid for 24 h at the end of the shoot production stage and rooted in vivo as mini-cuttings. Plantlets established well in soil.     

High specialisation in the pollination system of Mandevilla tenuifolia (J.C. Mikan) Woodson (Apocynaceae) drives the effectiveness of butterflies as pollinators

Butterfly pollination in the tropics is considered somewhat effective or solely effective in a few plant species. In the present study, we tested the hypothesis that Mandevilla tenuifolia (Apocynaceae), which has floral attributes associated with psychophily, has strategies adapted to pollination by butterflies, restricting other floral visitors and making these insects act as efficient pollinators. We analysed the floral and reproductive biology of M. tenuifolia, as well as the frequency and efficiency of its flower visitors. M. tenuifolia is an herb whose flowers have strong herkogamy and secondary pollen presentation on the style head, which corresponds to 60.4% of pollen on the anthers. Flower longevity and the long period of receptivity of the stigmatic region associated with the large amount of pollen removed in the first visits suggest that flowers remain functionally female during part of anthesis. Butterflies, mainly of the families Nymphalidae and Pieridae, are the only pollinators of M. tenuifolia. Despite being self‐compatible, M. tenuifolia depends on biotic vectors for fruit production. A non‐significant difference in fruit set between controlled treatments and natural conditions suggests that the pollinators are efficient. The inclination resulting from the landing of butterflies on flowers, together with flower morphology, guiding the insect proboscis inside the floral tube, as well as the frequency and efficiency of butterfly visits, are evidence of the close relationship between butterflies and M. tenuifolia, and also of the efficiency of these insects as pollinators.     

Micropropagation of Isoplexis canariensis (l.) G. Don

Isoplexis canariensis is a source of compounds related to the Digitalis cardenolides and anthraquinones. It is a protected endemic plant, but even so, it is rapidly disappearing from nature. An optimal micropropagation procedure was achieved using nodal segments with two axillary buds as explants. A concentration of 0.5 μM kinetin in Murashige and Skoog liquid basal medium was found to be optimal for micropropagation. Rooting in vitro was unnecessary for ex vitro survival. This revised version was published online in June 2006 with corrections to the Cover Date.     

Micropropagation of meadowfoam (Limnanthes spp.)

Summary A rapid micropropagation system was developed for meadowfoam (Limnanthes spp. Brown) using four genotypes of three species. Murashige and Skoog (MS) medium supplemented with N⁶ benzyladenine (BA) and indole-3-acetic acid (IAA) at 0, 0.1, 0.5, 1.0 and 2.0 mg/l was tested for multiplication, shoot elongation and rooting. Expiants were taken from pot-grown plants. The most useful level for shoot growth and multiplication of both floral induced and non-induced plants was 0.5 mg/l BA. IAA failed to affect shoot growth or multiplication. Expiants from non-induced plants multiplied at moderate to high rates on 0.5 mg/l BA, while those from induced plants multiplied slowly and tended to elongate and flower. Non-induced plants on 2 mg/l BA produced large numbers of tiny shoots; induced plants did not respond. Shoots of all genotypes rooted on MS medium without hormones and all plants grew normally after transplanting to soil. This system provides a new tool for the development of meadowfoam as a crop plant.Abbreviations (BA) N ⁶ -benzyladenine - (IAA) indole-3-acetic acid - (MS) Murashige and Skoog medium, 1962     

Micropropagation of sugarcane (Saccharum spp.)

A method for callus induction, adventitious bud regeneration, shoot multiplication and rooting of in vitro formed shoots of Helianthus annuus L. var. Argentario is described. Hypocotyl and cotyledon explants formed callus on medium containing 2 mgl^–1 naphthalene acetic acid and 0.5 mgl^–1 benzyladenine. Adventitious buds were formed on hypocotyl segments on medium containing 0.5–2 mgl^–1 benzyladenine. The optimal level of sucrose concentration for shoot regeneration from hypocotyls was 1.5%. Multiplication from shoot apices was promoted by kinetin (2 mgl^–1) plus gibberellic acid (5 mgl^–1), benzyladenine (2 mgl^–1) plus gibberellic acid (10 mgl^–1) or at lower frequency by benzyladenine (1 mgl^–1). A general feature of the plantlets formed in vitro was the precocious flowering.     

Micropropagation of Rollinia mucosa (JACQ.) baill

Summary A protocol for in vitro propagation of Rollinia mucosa, an important medicinal plant, was developed. The presence of 500 mg l⁻¹ polyvinylpyrrolidone (PVP) during explant excision was important to avoid browning. Axillary buds, adventitious buds, and shoot cluster proliferation were achieved from epicotyl and hypocotyl explants from nursery-grown seedlings. The highest direct organogenesis percentage from hypocotyl explants was obtained upon culture of explants on Murashige and Skoog medium supplemented with 2.2 μM benzyladenine (BA) plus 2.32 μM kinetin. Epicotyl explants display highest regeneration frequency on a medium containing 8.8 μM BA and 0.54 μM naphthaleneacetic acid. Gibberellic acid was necessary for shoot elongation. Root induction was observed when shoots were pretreated with activated charcoal for 7 d in the dark before culture on Woody Plant Medium supplemented with 49.21 μM indolebutyric acid for 10 d. Root development was observed when 20 g l⁻¹ sucrose was used. Rooted plantlets were acclimatized and grown in the greenhouse.     

Role of Achromobacter xylosoxidans AUM54 in Micropropagation of Endangered Medicinal Plant Naravelia zeylanica (L.) DC

This study was carried out to evaluate the inoculation effects of Achromobacter xylosoxidans AUM54 and Indole-3-butyric acid (IBA) on the growth of the medicinal plant Naravelia zeylanica (L.) DC under micropropagation conditions. Results revealed that the micropropagated shoots treated with the combination of endophytic bacterium and IBA promoted shoot growth, root length, number of roots, chlorophyll content, nitrogen content, antioxidant enzymes, and stress tolerance compared with the control plants. A significant increase in shoot fresh and dry weights (64.65 and 8.85 %), root fresh and dry weights (61.65 and 3.91 %), shoot length (30.17 %), root length (28.57 %) and number of roots (276.9 %) was observed in treated plants over controls. Total chlorophyll and nitrogen content of bacterized plants also treated with IBA showed a 48.39 and 116.66 % increase, respectively, compared with controls. A significant increase in peroxidase (22.52 %) and superoxide dismutase levels (48.38 %) and fewer changes in the polyphenol oxidase level were observed in plants treated with A. xylosoxidans AUM54 and IBA. Moreover, stress ethylene levels were reduced by 21.4 and 14.5 % due to bacterization with A. xylosoxidans AUM54 and IBA treatment during postacclimatization and acclimatization stages, respectively. The shoot primordial with application of A. xylosoxidans AUM54 and IBA (1 mg l^?1) had increased survivability of N. zeylanica plants by 30 % during the acclimatization stage under greenhouse conditions. From the present study it could be inferred that the association of endophytic bacterium A. xylosoxidans AUM54 and IBA with in vitro shoots of N. zeylanica improved root initiation, promoted plant growth and development under micropropagation conditions, reduced stress ethylene levels, and increased survivability during the postacclimatization stage. Therefore, A. xylosoxidans AUM54 along with IBA treatment can be used as a valuable tool for micropropagation of N. zeylanica and other endangered plants.     

百两金的化学成分

从百两金(Ardisia crispa(Thunb.)A.DC.)的根及茎乙醇提取物中分离得到了8个化合物,通过MS和NMR等方法鉴定为(7S,7′R)-双(3,4-亚甲二氧苯基)-rel-(8R,8′R)-二甲基四氢呋喃(1),(-)-襄五脂素(2),(7S,8S,7′R,8′R)-3,4-亚甲二氧基-3′,4′-二甲氧基-7,7′-环氧脂素(3),异安五脂素(4),α-菠甾醇(5),26-羟基二十六烷酸甘油酸酯(6),百两金皂苷B(7),岩白菜素(8)。其中有化合物1~6为首次从该种植物中分离得到。     

Micropropagation of Hohenbergia penduliflora (A. Rich.) Mez. for sustainable production of plant proteases

Hohenbergia penduliflora (A. Rich.) Mez. inhabits the protected ecological area of Cunagua Highland, Ciego de Ávila, Cuba. The availability of this plant for experimental purposes is exceedingly limited. Tissue cultured plants of this specie, would be useful for propagation purposes. Experiments were carried out to optimize the micropropagation process from the disinfection of fruits to ex vitro hardening of regenerated plantlets. The best results were obtained when seeds were disinfected with 2 % (v:v) sodium hypochlorite for 20 min and placed in vitro for 45 days for seed germination. Tissue cultured shoots (1 cm) with vertical wounds in the basal region (5 mm long) were placed in a medium containing Murashige and Skoog (MS) salts, 100 mg l^?1 myo-inositol, 0.1 mg l^?1 thiamine-HCl, 30 g l^?1 sucrose, 8.8 μM 6-benzyladenine (BA) and 1.6 μM naphthalene acetic acid (NAA). Shoots were proliferated for 45 days to obtain 8.21 new shoots per explant; they were subsequently divided and rooted on a medium containing 1.6 μM NAA for 30 days. For ex vitro hardening, plastic trays containing 82 cm³ of filter-cake-sugarcane ashes were used; 100 % survival rate was recorded. After 6 months of hardening, plants were established ex vitro and ready for protease extraction. Comparisons between protein contents, proteolytic activities and specific proteolytic activities of extracts from stems of macro- and micropropagated plants were acquired. Tissue cultured stems showed statistically lower figures which is why Ethrel was tested here to increase proteolytic activity in micropropagated plant stems. After Ethrel applications, protein contents, proteolytic activities and specific proteolytic activities of extracts from stems were the three main indicators recorded. However, other biochemical effects of Ethrel were also evaluated, such as, levels of chlorophyll pigments, malondialdehyde and other aldehydes; and superoxide dismutase, and guaiacol peroxidase activities. Rising concentrations of Ethrel (0, 1.5, 3.0, 4.5 and 6.0 mg l^?1) decreased protein contents at 72 h but increased proteolytic and specific proteolytic activities of stem extracts. Ethrel was effective in increasing proteolytic activity in in vitro culture-derived plant stems, at a level higher than in field-grown plant stems. Moreover, Ethrel increased superoxide dismutase and guaiacol peroxidase-specific activities in leaves; and decreased chlorophyll pigments. Ethrel did not affect levels of malondialdehyde and other aldehydes.     

Micropropagation in Peanut (Arachis hypogaea L.)

Multiple shoots in Arachis hypogaea L. could be induced from the de-embryonated cotyledons (DC), embryo-axes (EA) and mature whole seeds (MWS) in MS medium supplemented with different levels of benzylaminopurine (BAP). DC was the most suitable explant with 57.9 % induction and more than 40 shoots per explant in 31.6 % of cases. Though EA and MWS had high percent induction at or above 30 mg dm^–3 BAP, only 10 – 14 shoots per explant were observed. In DC, multiple shoots were confined to the proximal end and in EA they originated from the axillary bud region. Histological studies on DC confirmed the origin of shoots from the region of attachment with the embryo. Shoots could be rooted in MS medium containing 2 g dm^–3 charcoal and 200 mg dm^–3 casein hydrolysate. Sixty percent of the rooted plantlets could be established in the field.     

A New Isoflavonoid from Belamcanda chinensis （L.） DC.

A new isoflavonoid, 5, 6, 7, 3＇-terahydroxy-8, 4＇, 5＇-trimethoxyisoflavone （1）, along with 10 known isoflavonoids, namely 5, 6, 7, 4＇-tetrahydroxy-8-methoxyisoflavone （2）, irilone （3）, genistein （4）, tectorigenin （5）, irigenin （6）, irisflorentin （7）, dichotomitin （8）, dimethyltectorigenin （9）, iridin （10）, and tectoridin （11）, was isolated from the alcohol extract of the rhizomes of Belamcanda chinensis （L.） DC. The structures of these compounds were elucidated on the basis of results of spectroscopic analysis.     

A New Isofiavonoid from Belamcanda chinensis (L.) DC.

A new isoflavonoid, 5, 6, 7, 3'-terahydroxy-8, 4', 5'-trimethoxyisoflavone (1), along with 10 known isoflavonoids, namely 5, 6, 7, 4'-tetrahydroxy-8-methoxyisoflavone (2), irilone (3), genistein (4), tectorigenin (5), irigenin (6), irisflorentin (7), dichotomitin (8), dimethyltectorigenin (9), iridin (10), and tectoridin (11), was isolated from the alcohol extract of the rhizomes of Belamcanda chinensis (L.) DC. The structures of these compounds were elucidated on the basis of results of spectroscopic analysis.     

桔梗一新栽培变种

桔梗一新栽培变种,即重瓣桔梗Plantycodon grandiflorus(Jacq) A.DC.cv.plenus X.S.Wen。     

Micropropagation of tea (Camellia sinensis (L.) O. Kuntze) using Thidiazuron

The effect of thidiazuron (TDZ) on the micropropagation of Camellia sinensis (China hybrid) was compared with that of benzylaminopurine (BAP) using nodal segments from in vitro raised seedlings. Extremely low concentrations of TDZ (1pM–100nM) alone were effective in inducing shoot bud proliferation and maintaining high rates of shoot multiplication on hormone-free media. On the other hand, higher concentrations of BAP (1–10M) and its continued presence were required to initiate and sustain shoot proliferation. While wider ranges of BAP combined favourably with auxins like NAA or IBA, only specific combinations of TDZ and NAA were effective for shoot proliferation. TDZ treated explants yielded healthy shoots, with sturdy leaves, even during the initial stages of growth, whereas, the effect of BAP was cumulative over subcultures in attaining a high proliferative rate.     

Micropropagation of hornbeam (Carpinus betulus L.) and ash (Fraxinus excelsior L.)

Shoot multiplication of hornbeam was stimulated onWPM, QL andDKW medium supplemented with a low concentration of BAP or BPA (0.1-0.2 mg I -1) andIBA (0.1 mg I -1). Low concentration of thidiazuron promoted axillary bud formation, higher concentration inhibited shoot elongation. Microshoots were rooted onWPM supplemented with a low auxin concentration (IBA or NAA 0.2-0.5 mg I -1). High rooting percentages were obtained. Shoot proliferation of ash was stimulated on MS andDKW medium supplemented withBAP orBPA (2.0-5.0 mg I -1) andIBA (0.1 mg I -1). Root formation was promoted onWPM containing a low auxin concentration. Rooted plantlets were transplanted into soil and after hardening off the micropropagated trees were planted in the field. The planted trees grew normally without showing signs of abnormality.     

Micropropagation of adult Stone Pine (Pinus pinea L.)

This paper describes a micropropagation protocol for in vitro propagation of mature Stone Pine trees. Axillary bud development was achieved by culturing bud explants in media containing various cytokinins. Experiments were conducted to test the effect of asepsis conditions, type and concentration of cytokinin and rooting protocol. Four cytokinins were tested, namely, benzyladenine, meta-topolin, N-benzyl-9-(2-tetrahydropyranyl)-adenine and thidiazuron (TDZ) of which TDZ gave the best results, as 59% shoot development was obtained following the application of 1 μM TDZ to the culture medium. The shoot development was significantly influenced by the genotype of the tree, but was effective in explants from all 20 genotypes used in the trial. In vitro rooting was, however, difficult to achieve and could only be induced at low rates. This protocol represents the first successful biotechnological approach to the micropropagation of adult Pinus pinea trees. Paloma Moncaleán and Ricardo Javier Ordás contributed equally.