In vitro production of solasodine from Solanum trilobatum

Sucrose concentration in the culture medium affected chlorophyll content, trichome development and amount of solasodine in regenerated plantlets of Solanum trilobatum. High chlorophyll content and glandular trichomes were observed in the plants grown on Murashige and Skoog basal medium supplemented with 131.85 mM sucrose. The solasodine was quantified using reverse phase high performance liquid chromatography. The plantlets cultivated on this medium yielded 35.97 mg g⁻¹ (d.m.) solasodine whereas the field plants used as control yielded only 2.32 mg g⁻¹ (d.m.) of solasodine.     

In vitro regeneration of Solanum nigrum with enhanced solasodine production

High frequency of shoot formation was achieved from Solanum nigrum L. leaves on Murashige and Skoog (MS) medium without any callusing stage. Shoot forming ability was more pronounced on leaves positioned dorsally. For shoot induction, 2.0 mg dm⁻³ benzylaminopurine and 1.5 mg dm⁻³ kinetin were observed to be the most effective plant growth regulators (PGRs). The present paper also describes first successful induction of in vitro flowering in S. nigrum. The leaf derived shoots were excised and treated with various root promoting PGRs and 0.25 mg dm⁻³ indole-3-butyric acid produced maximum number of roots (15.2 per plant). Plants were later transplanted in field with 100 % survival. Solasodine content was higher in in vitro raised shoots and leaf derived callus, compared to ex vitro grown shoots.     

Exogenously applied ascorbic acid ameliorates detrimental effects of NaCl and mannitol stress in Vicia faba seedlings

The adverse effects of either NaCl or mannitol on growth, nitrogen content, and antioxidant system in Vicia faba seedlings were investigated. The role of exogenous ascorbic acid in increasing resistance to these stressors was also evaluated. Thus, with an increase in concentration of either NaCl or mannitol in culture media, a progressively greater significant decrease in percentage germination, in growth parameters, and in nitrogen constituents of the germinating beans, was observed. On the other hand, amide-, nitrate-, and total soluble-N contents appeared to show a progressive significant increase. Exogenous addition of ascorbic acid (4 mM) to the stressful media induced a pronounced significantly increased percentage germination and the growth attributes, whereas nitrogen constituents were variably changed in relation to values maintained in beans treated with either NaCl or mannitol. Furthermore, exogenous addition of ascorbic acid to NaCl or mannitol media induced a significant increase in the contents of ascorbate and glutathione and enzymatic antioxidant activities, in particular, in beans treated with the three highest concentrations of NaCl or mannitol, throughout the period of the experiments (12 days). Thus, ascorbic acid ameliorates the adverse effects of the stressful media; the magnitude of amelioration being a function of the type and the concentration of the stressful agent as well as of the duration of treatment. The importance of the above-mentioned changes in growth and metabolism to stress tolerance in broad bean is discussed.     

Glycinebetaine application ameliorates negative effects of drought stress in tobacco

Two tobacco (Nicotiana tabacum L.) cultivars differing in drought tolerance were used to study the effects of foliar-applied glycinebetaine (GB, 80 mM) under well-watered and water-deficit conditions. The latter affected shoot biomass and height, with a more significant decrease observed in drought-sensitive cultivar than in drought-resistant cultivar. Foliar-applied GB was absorbed, accumulated by tobacco leaves and improved growth of plants subjected to water deficit. GB-treated plants maintained leaf water status apparently due to the improved osmotic adjustment. GB application enhanced the photosynthesis in water-deficit experiencing plants, mostly due to a greater stomatal conductance and carboxylation efficiency of CO₂ assimilation. photosystem II (PSII) activity in GB-treated plants was higher, as suggested by higher actual efficiency of PSII (ΦPSII). GB increased anti-oxidative enzyme activities under water deficit. All these effects resulted in an improved shoot biomass and height. Therefore, foliar GB application at the rapid growth stage favors plant growth in drought-stressed plants, mainly by improving water status and increasing PSII activity. Published in Russian in Fiziologiya Rastenii, 2007, Vol. 54, No. 4, pp. 534–541. The text was submitted by the authors in English.     

In vitro responses of grape rootstocks to NaCl

An investigation was undertaken to adjudge the in vitro salt tolerance and biochemical changes due to NaCl on four grape rootstocks (Dogridge, SO4, H-144 and 3309C). The in vitro two-node micro-cuttings from the established cultures were sub-cultured on rooting medium comprising Murashige and Skoog (MS) basal medium supplemented with 0.5 μM indolebutyric acid (IBA), 200 mg dm⁻³ activated charcoal and different NaCl concentrations ranging from 0 to 125 mM. The surviving and proliferated cultures were further sub-cultured four times on respective media. Dogridge and H-144 tolerated 125 and 100 mM NaCl, respectively, while SO4 and 3309C survived only up to 75 mM NaCl. Contents of proteins, proline, K⁺ and Na⁺ in tissue increased in all genotypes due to NaCl supplementation, while contents of chlorophyll and total soluble sugars declined. Higher K⁺/Na⁺ ratio was registered in Dogridge and H-144 than in SO4 and 3309C. The relative NaCl tolerance for different grape rootstocks under study could be ranked as Dogridge > H-144 > SO4 and 3309C.     

Nitric oxide alleviates Fe deficiency-induced stress in Solanum nigrum

The possible involvement of nitric oxide in response of Solanum nigrum seedlings to Fe deficiency was investigated. Iron deficiency resulted in decreased shoot height and chlorophyll content and increased proliferation of root hairs and H₂O₂, K⁺ and Ca²⁺ content. NO donor S-nitrosoglutathione (GSNO) was effective in preventing Fe deficiency-induced increase in content of H₂O₂ and the ion uptake. The protective effects of GSNO were reversed by cPTIO, an NO scavenger, and tungstate, a nitrate reductase (NR) inhibitor.     

少花龙葵与黄果龙葵染色体核型分析

利用DAPI显带技术对少花龙葵和黄果龙葵染色体进行了核型分析。结果表明,少花龙葵染色体数目为24,为二倍体,核型公式为K(2n)=24=6sm+18m;黄果龙葵染色体数目为48,为四倍体,核型公式为K(4n)=48=48m,并进一步构建了这两个种的核型模式图。此外,还初步探讨了染色体的DAPI带型及其异染色质的分布。为中国龙葵的系统分类和进化趋向研究,以及育种和资源的开发利用提供帮助。     

黄果龙葵幼苗对镉胁迫的生理生长响应

黄果龙葵（Solanum diphyllum）是茄科的小灌木,已有的研究对黄果龙葵的镉（Cd）响应知之甚少。该研究采用水培控制试验,在160μmol·L^-1 Cd胁迫下,测定了黄果龙葵幼苗在不同胁迫时间点（0、0.5、1、3、7、19 d）的生长、耐性系数（TI）、Cd富集特性、超氧化物歧化酶（SOD）活性、渗透调节物质和光合色素等指标。结果表明：（1）在胁迫0～3 d时Cd对黄果龙葵幼苗的生长无显著影响。胁迫3～19 d时表现为明显抑制,且毒害随胁迫时间延长而加重。与对照组相比,Cd胁迫使黄果龙葵幼苗的生物量降低13.28%～62.40%。（2）黄果龙葵幼苗的最大Cd积累量为60.14μg·plant^-1,其地上部份的Cd积累量占植株的15.46%～35.24%。（3）丙二醛（MDA）含量呈上升趋势,根的MDA含量最大增幅为对照组的5.25倍。SOD活性、游离脯氨酸、可溶性糖、叶绿素a、叶绿素b和类胡萝卜素的含量均呈先升后降的趋势。（4）黄果龙葵幼苗的耐性系数TI随胁迫时间延长而降低,最小值为0.64。综上结果认为,黄果龙葵是一种潜在的Cd...     

Physiological effects of exogenous nitric oxide on Brassica juncea seedlings under NaCl stress

The study was conducted to investigate the physiological effects of exogenous NO on potherb mustard (Brassica juncea Coss.) seedlings under salt stress. The plants were grown in Hogland nutrient solution for 15 d and treated with 150 mM NaCl, NO donor sodium nitropruside (SNP) and NO scavenger methylene blue (MB-1) for 4 d. The NaCl stress increased superoxide dismutase, peroxidase and ascorbate peroxidase activities and malondialdehyde (MDA) and free proline contents, and decreased soluble protein content. However, the application of exogenous NO limited the production of MDA and free proline, while markedly promoted SOD, POD and APX activity.     

黄果龙葵和龙葵染色体制片优化及核型分析

该实验以黄果龙葵和龙葵的根尖为实验材料,进行不同的预处理、固定和解离,确定出各种材料适合于核型分析的制片方法。结果表明:龙葵于15℃条件下经0.05%秋水仙素预处理2.5h,固定后用1mol/L HCl酸解后,染色观察,得到的染色体分散,易于染色体计数和形态观察。用此方法对黄果龙葵和龙葵进行核型分析,结果发现:黄果龙葵和龙葵都属于小型染色体,黄果龙葵为四倍体,核型公式为K(4n)=48=4sm+44m,核型不对称系数为56.22%,属于2B核型。龙葵为六倍体,核型公式为K(6n)=72=72m,核型不对称系数为55.89%,属于1B核型。     

In vitro effects of propolis on Giardia duodenalis trophozoites

In order to improve the current chemotherapy of Giardia infection, potential antigiardial agents have been screened, including natural products. Propolis, a resinous hive product collected by bees, has attracted attention as a useful and popular substance with several therapeutic activities. The present study was carried out aiming to evaluate the in vitro effects of an ethanolic extract of propolis on the growth and adherence of Giardia duodenalis trophozoites. Propolis inhibited the growth of trophozoites and the level of inhibition varied according to the extract concentration and incubation times. The highest reduction of parasite growth was observed in cultures exposed to 125, 250 and 500 microg/ml of propolis, in all incubation periods (24, 48, 72 and 96 h). Growth reduction by 50% was observed in 125 microg/ml propolis-treated cultures, while the concentrations of 250 and 500 microg/ml were able to inhibit growth by more than 60%. Propolis also inhibited parasite adherence and all assayed propolis concentrations promoted the detachment of trophozoites. Light microscope observations revealed changes of the pear-shaped aspect of the cell and reduction of flagellar beating frequency in the great part of the trophozoites. Our results hold the perspective for the utilization of propolis as an antigiardial agent.     

Moderate NaCl alleviates osmotic stress in Lycium ruthenicum

Plant Growth Regulation - Lycium ruthenicum is a salt-accumulating xerophytic species with excellent adaptability to adverse environments. Previous studies showed that a certain amount of NaCl...     

Salt-induced osmotic stress for glutathione overproduction in Candida utilis

The effect of NaCl-induced osmotic stress on glutathione (GSH) production was investigated in Candida utilis. Based on the fact that NaCl stress can enhance GSH production but inhibit cells growth simultaneously, the novel strategies of multiple osmotic stresses with different NaCl additions (0.2 mol/l at 4 h, 0.4 mol/l at 8 h, and 0.6 mol/l at 12 and 16 h) were developed for GSH overproduction. After 30 h cultivation, GSH yield reached 238 mg/l and intracellular GSH content was 2.34%, increased by 66.4% and 70.7% respectively compared to the control. Further applying the strategies to 7 l fermentor, GSH yield of 356 mg/l was achieved at 30 h, which was 65.6% higher than the control. Moreover, NaCl stress led to an increase in intracellular cysteine content and activities of γ-glutamylcysteine synthetase, GSH synthetase and GSH reductase, explaining the mechanism involved in inducing cellular GSH accumulation.     

The foliar micromorphology of Solanum pseudocapsicum

Foliar micromorphology of Solanum pseudocapsicum was investigated by electron microscopical examination. The leaves are characterized by anisocytic stomata which are more abundant on the abaxial surfaces. The leaves have short multicellular glandular trichomes sparsely distributed over the entire leaf surfaces. Crystal deposits were also observed on the surfaces above the stomata. Energy dispersive X-ray spectroscopy-SEM showed that Al, K, Na and Si were the major constituents of the crystals analyzed. The presence of glandular trichomes in this plant could be the source of poisonous compounds that are characteristic of this species.     

Jasmonic acid inhibition of in vitro shoot organogenesis in Pinus radiata cotyledons

The effect of jasmonic acid (JA) onde novo shoot formation in excised cotyledons of radiata pine (Pinus radiata D. Don), was examined. JA had no effect on shoot-forming (SF) tissues at concentrations up to 10⁻⁶ mol · L⁻¹. At concentrations greater than that, JA caused a reduction in the number of shoots formed, as well as the lengths of the shoots and fresh and dry masses of the tissues. Reciprocal transfer of excised radiata pine cotyledons from a SF medium with 10⁻⁵ mol · L⁻¹ JA to a SF medium without JA and vice versa showed that any exposure of the cotyledons to JA either during the shoot induction phase (days 0-21) or the shoot development phase (beyond day 21) led to a reduction in shoot formation. However, the JA effect was significantly less if the cotyledons were not exposed to JA during the initial 10 days in culture; indicating that the JA effect was mainly during shoot primordia formation and the subsequent development into shoots.     

Transposition of the maize transposable element Ac in Solanum tuberosum

Summary The maize transposable element Ac has been introduced into potato via the T-DNA (transferred DNA) of Agrobacterium tumefaciens. Ac was inserted within the untranslated leader region of a neomycin phosphotransferase II (NPT-II) gene such that excision restored NPT-II activity. Two approaches to monitor Ac excision were used. (i) Using an Agrobacterium strain harbouring plasmid pGV3850::pKU3, leaf discs were selected on kanamycin (Km) after exposure to Agrobacterium. (ii) Using a strain containing plasmid pGV3850HPT::pKU3, the leaf discs were selected on hygromycin (Hm) and the resulting shoots were checked for NPT-II expression. Thirteen kanamycin resistant shoots transformed with pGV3850::pKU3 were isolated, suggesting that Ac had excised from the NPT-II gene. Out of 43 hygromycin resistant shoots transformed with pGV3850HPT::pKU3, 22 expressed the NPT-II gene, indicating that Ac had undergone excision in approximately 50% of the hygromycin resistant shoots. Southern analysis revealed that all kanamycin resistant plants contained the DNA restriction fragments expected when Ac excises from the NPT-II gene. The presence of Ac at new locations within the genomic DNA of several transformants was also detected.     

马铃薯萜类合酶基因StHcS的功能鉴定

为了研究马铃薯萜类代谢物的生物合成机制,从马铃薯基因组数据中筛选到一个萜类合酶基因。通过RT-PCR方法,从致病疫霉侵染后的马铃薯品种‘费乌瑞它’中成功克隆到该基因,命名为StHcS,并对其进行生物信息学分析、生化功能鉴定及表达模式分析。结果表明:(1)序列分析显示StHcS编码区序列长1 497 bp,编码498个氨基酸,分子质量为74.78 kD。(2)StHcS基因编码的蛋白序列含有DDXXD催化功能域,与短柱茶(Camellia maliflora)中的四甲基环癸二烯甲醇合酶相似度最高。(3)蛋白体外催化实验和大肠杆菌代谢工程分析表明,StHcS可以催化生成倍半萜化合物四甲基环癸二烯甲醇(hedycaryol)。(4)基因表达分析显示,StHcS可以被致病疫霉侵染诱导表达反式法尼烯焦磷酸(FPP),且在侵染后72 h表达最高;GC-MS分析显示,在受侵染的马铃薯块茎中检测到四甲基环癸二烯甲醇。StHcS生化功能的鉴定为倍半萜合酶的研究提供了多样性,也是首次在马铃薯中发现的四甲基环癸二烯甲醇合酶,为马铃薯萜类代谢途径解析提供了参考。     

In vitro effects of levamisole and ivermectin against Echinococcus granulosus protoscoleces

The in vitro effects of levamisole and ivermectin against Echinococcus granulosus protoscoleces were studied by means of light and electron microscopy. Both drugs had a protoscolicidal activity that increased proportionally with increasing concentrations of the drugs. Ivermectin showed the more rapid effects and caused contraction and paralysis of protoscoleces. A paralyzing effect was also observed with levamisole, followed by irreversible tissue vacuolation leading to death.     

Inhibitory effects of phospholipase D on chondrogenesis in vitro

Mesenchymal cells from the wing buds of stage 24 chick embryos undergo differentiation to cartilage when plated at high density. Treatment of these cultures with phospholipase D resulted in inhibition of chondrogenesis. Phospholipase D treatment (which produces phosphatidic acid from membrane phospholipids) was found to affect cell proliferation and to dramatically increase intracellular free calcium levels and inositol phosphate production. Intracellular free Ca²⁺, mobilized as a result of phosphatidylinositol phosphate hydrolysis, may therefore inhibit chondrogenesis in embryonic mesenchymal cells.