Ecballium elaterium: an in vitro source of cucurbitacins

Biomass and secondary metabolite accumulation were assayed on a wide range of auxins, cytokinins and their combinations using Ecballium elaterium callus tissue. The best combination was that consisting of alpha-naphthalene acetic acid (NAA) and benzylamino purine (BAP) (1 mg/l of medium, each). NAA and BAP were set up in a grid to determine concentrations that produced the best callus as regards fresh and dry weights and yield of cucurbitacins, particularly cucurbitacin E. The best combination for callus proliferation consisted of 5 mg/l NAA and 5 mg/l BAP, reaching a peak at week 5. For cucurbitacin production, an optimum yield was obtained at week 3 with 5 mg/l NAA.     

Immunomodulatory activity of cucurbitacin E isolated from Ecballium elaterium

The immunomodulatory effect of cucurbitacin E, extracted from Ecballium elaterium, was tested on peripheral human lymphocytes. These lymphocytes were co-cultured with cancer cells and an interesting lymphocyte-mediated cytotoxicity was observed.     

In vitro plantlet regeneration from Australian Red Cedar (Toona ciliata,Meliaceae)

In vitro regeneration of complete plants from nodal single-bud segments of 2-year-old Australian Cedar (Toona ciliata) trees were obtained under defined nutritional and environmental conditions. Explants were dissected from plants obtained by germination of seeds and growth in pots in a greenhouse. The best medium for shoot regeneration was that of Murashige and Skoog at 1/4 strength with 3% sucrose (1/4 MS), supplemented with 0.1 mg/l IBA and 0.5 mg/l BAP. Rooting of regenerated shoots was observed in MS medium with 0.1 mg/l IBA. Using mature tree material was more difficult. Forced flushing was used to induce shoot development on branches of a 10-year-old tree. Nodal segments of these epicormic shoots formed shoots in vitro on 1/4 MS + 0.01 mg/l IBA + 5 mg/l BAP, but rooting was never observed.     

Micropropagation of field tested superior Eucalyptus grandis hybrids

Three superior clones of Eucalyptus grandis hybrids were micropropagated through several steps. Five-year-old trees were girdled to induce juvenile sprouts. Cultures were attempted from mature branches and sprouts. Branches from mature trees were 100% contaminated while sprouts were only 40% contaminated. Pre-initiation hormone free medium and dark environment were used to screen for contaminants and to reduce production of phenolic compounds. Initiation of auxillary buds was achieved with modified MS plus 0.05 mg/l NAA and 0.5 mg/l BAR High multiplication rates were obtained on auxin-free medium with 0.6 mg/1 BAR Elongation of shoots was best on media with high auxin (2.5 mg/l of IBA) and cytokinin (1–1.5 mg/l of zeatin). Continual subculture on the multiplication medium improved rooting significantly. Up to 98% rooting was achieved on 1/4 MS with 2 mg/l IBA. Rooted propagules were successfully transferred to a mist greenhouse with 82% survival, and then to greenhouse conditions.     

Antitrypsin activity of extracts from Ecballium elaterium seeds.

The antitrypsin activity of extracts obtained from two different seed types of Ecballium elaterium, was tested in vitro. The presence of trypsin inhibitors in three chromatographed samples for each extract, showed a strong and specific antitrypsin activity.     

The aromatic and polyphenolic composition of lemon verbena tea

The chemical composition of the largely used tea from lemon verbena, Aloysia triphylla, was investigated. The qualitative and quantitative composition of the main aromatic and polyphenolic constituents of tea made by infusion from leaves were examined. The results showed that the relative proportions of the active constituents differ from those of the original leaves. The tea contained a large amount of polyphenolic compounds (mean value 675 mg/l; extraction yield 65%) including verbascoside (400 mg/l) and luteolin 7-diglucuronide (100 mg/l). It contained also 42 mg/l of essential oil (extraction yield 51%) with much more citral (77% of the essential oil) than the original leaves (41%).     

Direct shoot bud regeneration from shoot tip explants of <Emphasis Type="Italic">Enicostema axillare</Emphasis>: an important medicinal plant

An efficient protocol has been developed for in vitro propagation of Enicostema axillare using shoot tip explants. The shoot tip explants were cultured on MS medium supplemented with various combinations of (BAP, KIN) and (NAA/IAA & IBA) in different concentrations between 0.5 and 2.0 mg/l for multiple shoot bud induction. The highest percent of (98.51 %) was observed at 1.0 mg/l BAP in combination with 0.2 mg/l KIN while maximum number of shoot buds (8.41 shoots/explant) was noticed on MS medium containing 1.0 mg/l BAP and 0.2 mg/l KIN combination. The highest frequency (90.82 %) of multiple shoot bud regeneration was observed at 1.0 mg/l BAP and 0.5 mg/l IBA with 15.12 ± 2.12 shoots/explants. The regenerated multiple shoots were transferred to half-strength MS medium augmented with different concentration of 0.5–2.5 mg/l IBA for rooting. Among the different concentrations of IBA tested, maximum percentage of rooting (100 %) was observed in MS medium augmented with 1.5 mg/l IBA. The rooted plantlets were successfully transferred into plastic cups containing soil and sand in the ratio of 1:1. Subsequently established in the field conditions with 90 % of survival rate. The protocol developed can be utilized for both large scale plant production and conservation of germplasm of this species. The described method can be successfully employed for large-scale multiplication and in vitro conservation as well as production of secondary metabolites of E. axillare.     

Acaricidal,repellent and oviposition deterrent activities of <Emphasis Type="Italic">Datura stramonium</Emphasis> L. against adult <Emphasis Type="Italic">Tetranychus urticae</Emphasis> (Koch)

The ethanol extracts obtained from both leaf and seed in the Thorn apple (Datura stramonium L.) (Solanaceae) were investigated for acaricidal, repellent and oviposition deterrent properties against adult two-spotted spider mites (Tetranychus urticae Koch) (Acari: Tetranychidae) under laboratory conditions. Leaf and seed extracts, which were applied in 167,250 and 145,750 mg/l concentrations, respectively (using a Petri leaf disc-spray tower method), caused 98 and 25% mortality among spider mite adults after 48 h. The simple logistic regression analysis showed that the independent variable, an increase in the dose of leaf extract was associated with a significant increase in the death rate of T. urticae females, but an increase in the dosage of seed extracts did not have a significant effect. Using probit analysis and estimating the parameters with a confidence limit of 95%, we determined the LC₅₀ values of leaf extract to be 70,593 mg/l. According to Pearson’s χ² test, mites showed the strongest run off to bean leaf surfaces sprayed with both leaf and seed extracts (in sub-lethal doses: <7,500 mg/l and <25,000 mg/l, respectively) and moved towards surfaces that had not been sprayed with extracts. Furthermore, repeated measures ANOVA indicated a significant difference between the number of eggs laid on unsprayed bean leaves compared to bean leaves that were sprayed with leaf and seed extracts at sub-lethal doses, 2,500 and 25,000 mg/l concentrations, respectively. These results suggest that D. stramonium extracts could be used to manage the two-spotted spider mite.     

Comparative performance of seventeen upperstorey tree species associated with crops in the highlands of Uganda

Trials were established at three sites in Uganda to test the suitability of multipurpose trees (MPTs) as upperstorey in crop lands to provide poles, small timber and fuelwood. The three sites were Kachwekano District Farm Institute (1°16 S, 29°57 E, 2000 m.a.s.l.) in Kabale District, Kabanyolo University Farm (0°28 N, 32°27 E, 1250 m.a.s.l.) in Mpigi District and Bushenyi District Farm Institute (0°34 S, 30°13 E, 1610 m.a.s.l.) in Bushenyi District. The MPTs were planted in single rows at intra spacing of 2 m and each plot contained seven or nine trees. On both sides of the tree row, crops were raised. Data on crop yields were collected every season, while data on the growth of the trees were collected four times each year.In terms of tree growth,Grevillea robusta, Casuarina cunninghamiana andAlnus acuminata performed well with height growth of 1.8–2.4 m per year at Kachwekano, while at BushenyiGrevillea robusta, Casuarina junghuhniana, Cupressus lusitanica andCedrela serrulata averaged 1.6–2.0 m height per year. At Kabanyolo,Melia azedarach, Cassia siamea, Jacaranda mimosifolia, Grevillea robusta andMaesopsis eminii had height increments ranging from 1.8–2.7 m annually. Crop yields were affected by the presence of the MPTs, withMaesopsis eminii being the most competitive (averaging 60% reduction, over five seasons). The crop rows nearest to the tree line were the most affected. OnlyAlnus acuminata seems to have had a positive effect on crop yields. The installation of a root mesh to reduce tree root competition for nutrients and water in four species increased yields in plots with MPTs by 5% (Melia azedarach) to 152% (Maesopsis eminii), but the control plot still had significantly higher bean yields, suggesting that shading could also be important. In the case of maize, suppression seems to be due mainly to root competition because after its elimination yields obtained thereafter did not differ significantly from those of the control except for the Maesopsis plots. The negative influence of the MPTs could, therefore, be minimized by periodically pruning the tree crowns and roots.     

In vitro flowering of green and albino Dendrocalamus latiflorus

To propagate Dendrocalamus latiflorus, we used in vivo inflorescences to produce calli on Murashige and Skoog basal (MS) medium supplemented with 3 mg/l 2,4-dichlorophenoxyacetic acid (2,4-D), 2 mg/l kinetin, 250 mg/l polyvinyl pyrrolidone (PVP), and 1% coconut milk. Multiple shoots were generated on MS medium supplemented with 0.1 mg/l thidiazuron (TDZ). The green plantlets were successfully transferred to soil. Multiple albino shoots also regenerated and were able to proliferate on medium containing cytokinins, especially TDZ. Albino multiple shoots rooted in medium containing α-naphthaleneacetic acid (NAA), and callus formation was observed in the presence of 2,4-D and picloram. Green and albino regenerates flowered after 8 months of subculture. The flowering ratio increased to 44% after three treatments in medium containing 1 mg/l TDZ. Morphological observations revealed that the in vitro green and albino flower organs were normal. However, pollen derived from the in vitro flowers of both the green and albino plants were sterile.     

石蒜组培繁殖技术的研究

以石蒜的鳞茎为材料，采用不同的激素配对离体小鳞茎的分化和增殖效果进行了试验研究，结果表明，添加30g/l蔗糖。0.7%琼脂的MS BA（3mg/l) NAA(3mg/l)培养基对小鳞茎的分化效果最佳，而BA（3mg/l) NAA（1mg/l)的激素配比更有利于小鳞茎的增殖，试验中发现，蔗糖的浓度对小鳞茎的增殖与生长影响显著，以60g/l的浓度为最佳，将组织培养获得的小鳞茎切分成块后培养，可获得更多的小鳞茎，BA（0.5-1) NAA(0.5)的激素配比最有利于鳞茎切块增殖；当蔗糖浓度为40g/l时，鳞茎切块结成鳞茎的平均直径达到最大。     

枣胚轴组织培养的初步研究

以枣胚轴为外植体，用基本培养基MS和各种激素不同组合及其配比的6种培养基进行组织培养，结果表明：在MS（1／2MO3^-) 1.75mg/lZT 0.015mg/lNAA 4mg/lKT 2mg/lAgNO3和MS＋0．5mg/l IBA两种培养基上成苗率都达40％，但后一种培养基更有利于诱导生根；在用MS（1／2NO3^-)作基本培养基，添加0．2mg/l BA和3mg/l NAA的培养基上，愈伤率最高，达73％,由愈伤组织分化出来不定芽的分化率为27％。     

Regeneration of Psychotria umbellata and production of the analgesic indole alkaloid umbellatine

Psychotria umbellata Vell. (Rubiaceae), a Brazilian coastal woody species, produces umbellatine (also known as psychollatine), an analgesic indole alkaloid. An in vitro embryogenic regeneration protocol capable of yielding alkaloid-accumulating plants was developed. Rhizogenic calli, which were obtained from stem segments derived from rooted apical cuttings, were cultured on Murashige and Skoog's (MS) medium containing either 1 mg l(-1) NAA (naphthalene acetic acid) and no kinetin, or 5 mg l(-1) NAA + 1 mg l(-1) kinetin. Calli did not accumulate umbellatine. Segments of rhizogenic callus were cultured on complete MS medium with various concentrations of kinetin and sucrose. Plant regeneration was best in the light with 0.25 mg l(-1) of kinetin and 1.5% sucrose. After 3 months of acclimatization in soil mixture, plant survival was 81%. Leaves of 10-month-old regenerated plants yielded umbellatine concentrations equivalent to those of adult forest-grown plants.     

红木硬枝扦插试验研究

用浓度５０ｍｇ／ｌ、１００ｍｇ／ｌ、１５０ｍｇ／ｌ的ＡＢＴ生根粉１号对红木三、四年生枝条进行浸泡处理,将处理好的枝条分别插于黄心土和珍珠岩基质中,６０ｄ后检查成活率,结果表明：以１００ｍｇ／ｌ的ＡＢＴ生根粉１号处理,插于黄心土基质中的红木枝条成活率最高,达５４７％;以１５０ｍｇ／ｌ的ＡＢＴ生根粉１号处理,插于珍珠岩基质中的红木枝条成活率最低,为２９４％;所有处理与对均有显著差异。     

Plantlet regeneration from leaf protoplasts ofPopulus euphratica

Protoplasts were isolated from the leaves of sterile plants ofPopulus euphratica Oliv. by using 1% Cellulase “Onozuka” RS and 0.25% Pectolyase Y-23 in 0.6m of mannitol solution. Protoplasts were cultured in modified Murashige and Skoog's (MS) medium which contained no ammonium ions but was supplemented with BAP (6-benzylaminopurine), 2,4-D (2,4- dichlorophenoxy-acetic acid), and 1% sucrose at the cell density of 9×10⁴/ml. Cell divisions occurred in every culture medium, especially in the medium containing 0.5 mg/l of BAP and 0.1 mg/l of 2,4-D, in which callus was successfully induced by successive culture through cell cluster formation. Shoots were regenerated from the callus, and their growth was enhanced on 1/2 MS medium containing 0.8 mg/l of BAP. Finally, shoots were rooted and plantlets were regenerated on 1/2 MS medium without a hormone. A part of this paper was presented at the 106th Annual Meeting of the Jpn. For. Soc. (1995).     

我国油橄榄中试园土壤条件及其营养指标的研究

1983—1987年对湖北省武昌、宜昌、当阳县和陕西省城固县油橄榄中试园土壤与营养状况进行了研究。结果表明:油橄榄生长较好的(一类树下)土壤条件是pH7—8,CaCO_3>5％,有机质>1.8％,速效N>74.4—87.4mg/kg土,速效P>40mg/kg土,速效K>200mg/kg土,交换性Ca103—181mmol/kg土,交换性Mg24—34mmol/kg土。生长结实好的一类树叶片营养指标为N2.0—2.5％,P_2O_5 0.14％,K_2O 1.5％,CaO 4.0％,MgO 0.8％。通过分析建立了预测产量和施肥控制的回归方程。     

Effects of different neem preparations in comparison to synthetic insecticides on the whitefly parasitoid Encarsia sophia (Hymenoptera: Aphelinidae) and the predator Chrysoperla carnea (Neuroptera: Chrysopidae) on cotton under laboratory conditions

The contact and feeding toxicity of NeemAzal T/S (EID Parry, Chennai, India) to the parasitoid Encarsia sophia (Girault & Dodd) was compared with three synthetic insecticides (chlorpyriphos, endosulfan and triazophos) against immatures and adult emergence. NeemAzal T/S (1.0%) at lower dose (200 mg/l) did not cause any effect on the emergence of E. sophia adults, but there was a significant reduction in emergence at higher doses (800 mg/l). Also, it did not show contact toxicity to adults of E. sophia, but there was a significantly high feeding mortality in a dosage dependant manner, whereas chlorpyriphos, endosulfan and triazophos showed high toxicity both by contact and feeding method. Three neem preparations registered in India, NeemAzal T/S, Nimbecidine (T. Stains, Coimbatore) and Godrej Achook (Godrej Agrovet, Mumbai) were tested against egg and larval stages of Chrysoperla carnea (Stephens) at three dosages, using a synthetic insecticide, triazophos 40 EC (200 mg/l), for comparison under laboratory conditions. The three biorationals did not induce any adverse effect on the hatchability of C. carnea eggs. The mortality of the first instars of C. carnea was not affected by any of the azadirachtin enriched formulations; however, at higher dosage of 800 mg/l these neem based products resulted in increased mortality of the first and second instar larvae of C. carnea relative to the untreated controls. Whereas, triazophos induced very high mortality rates (85.0, 89.0 and 81.5%) of all the three larval instars. The neem based insecticides showed a dosage-dependant effect on the larval instars of C. carnea. Thus, these biorationals show that there is a potential to use them in an IPM system, being safe for natural enemies of B. tabaci in cotton.     

Breaking Seed Dormancy of Laurel (Laurus nobilis L.)

Laurel (Laurus nobilis L.) seeds were collected from the west part of Turkey in the fall of 2002. Seeds with pericarp (+) or without pericarp (−) were treated with 1000 mg/l, 2000 mg/l and 3000 mg/l GA₃ concentrations, cold stratified at +4 ± 1°C for 25 and 50 days, punctured and seed coat removed to overcome and assess the mechanism of laurel seed dormancy. A period of 50 days cold stratification and removing seed coat significantly increased germination rate to 55% ± 1.91 (Mean ± SE) and 85% ± 3.00, respectively. None of the seeds with pericarp germinated regardless of treatment they underwent. Results suggested that seed dormancy was mainly due to pericarp and perhaps inhibitors linked to seed coat.     

Efficacy of aerial application ofBacillus thuringiensis to controlSpodoptera littoralis (Lepidoptera, Noctuidae) infesting soybean cultivations in Egypt

The calculated percentage of droplets deposition ofBacillus thuringiensis var.kurstaki HD-1 Dipel 2X on the foliage of soybean plants in relation to the sensitive “Ciba Geigy” papers showed to be the highest (92.4%) after ground spray application, compared to an average of 67.4, 85.8 and 89.3% following aerial application of the same preparation in spray volumes of 10, 20 and 30 l/F, respectively. Aerial application ofB. t. showed to be more effective during the first 3 days after spraying soybean cultivations and the mortality ofSpodoptera littoralis larvae varied between 36.9–67.2% in correlation to the spray volume. The volume achieving the highest and significant suppression of infestation was 20 l/F. On the other hand, the yield of soybeans obtained from plots receiving a spray volume of 30 l/F was higher compared to plots sprayed with a volume of 20 or 10 l/F, but with no significant difference when compared to those plots receiving ground spray application.