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1.
Plant regeneration from the nodal explants of 1-month-old in vitro grown plants and cotyledonary node explants of 15-days-old seedlings of Sterculia urens is reported. Nodal explants were grown on MS medium supplemented with various growth regulators like BA, KIN and TDZ. For shoot induction 13.3 μM BA, 0.9 μM TDZ and 9.3 μM KIN were found optimum. Among the three growth regulators 0.90 μM TDZ was used for the growth of cotyledonary node explants. An average of 8.6 shoots per node and 11.2 shoots per cotyledonary node were observed in 4 to 5 weeks. These shoots were subsequently rooted in vitro on half strength MS medium containing various concentrations of auxins like IBA and NAA. The best concentrations for rooting of shoots were 19.7 μM IBA and 16.1 μM NAA. Plantlets were acclimatized to ex vitro conditions and established in the field.  相似文献   

2.
Regeneration of transgenic shoots was achieved from Hypericum perforatum L. hairy roots on hormone-free MS/B5 medium for a period of 4 weeks under a photoperiod of 16-h light. A control experiment was set up with root segments obtained from in vitro grown seedlings. Investigations have been made to study the production of phenolic compounds in non transgenic and transgenic shoot cultures. Six groups of phenolic compounds such as phenolic acids, flavonols, flavan-3-ols, naphtodianthrones, phloroglucinols, and xanthones were recorded in the transgenic shoots. Chlorogenic acid was found as the most representative phenolic acid in shoot extracts. With regard to the class of quercetin derivatives in transformed shoots, quercetin 6-C-glucoside usually dominated among the glycosides followed by quercitrin and hyperoside. The analysis of flavan-3-ols in transgenic shoots resulted in the identification of epicatechin and proanthocyanidin dimers. One of the main achievements in this study was considerably enhanced hypericin and pseudohypericin production in transgenic shoots. The concentration of identified naphtodianthrones was about 12-fold higher in transformed shoots compared to control. Chromatographic analysis of phloroglucinols in transgenic shoots resulted in the identification of hyperforin, while its homolog adhyperforin was detected in traces. A twofold higher content of hyperforin was observed in transgenic shoots compared to control. Although mangiferin was found as the main representative xanthone in shoot extracts, several other xanthones identified as γ-mangostin isomers, trihydroxy-1-methoxy-C-prenyl xanthone, garcinone E, and banaxathone E were de novo synthesized in transformed shoots. Therefore, H. perforatum transgenic shoots could be considered as a source for rapid and increased production of naphtodianthrones and other specific phenolic compounds.  相似文献   

3.
Iranian (Papaver bracteatum Lindl.) and opium poppy (P. somniferum L.) plantlets obtained from germinated seeds grown on a Murashige and Skoog basal medium (BM) readily manifest alkaloids. Temperature had a profound effect on growth and alkaloid production after 8 weeks in culture. Plantlets of poppy cultivars (cvs.) grew best at 18.5 and 20°C compared to 15 or 25°C. An alkaloid survey study with 24 Iranian and 21 opium poppy cvs. revealed that total morphinan alkaloids ranged from 0 to 6.55 mg/g dw. Prolific axillary branching was achieved from poppy cvs. by maintaining shoots on BM containing 1.0 mg/L N6‐benzyladenine and 0.01 mg/L α‐naphthalene acetic acid for an additional 16 weeks. The influence of vessel size on the growth response of established shoot clumps was determined by subculture in a variety of culture vessels for 8 weeks. The tested culture vessels included culture tubes (55 mm3 capacity (cap.)), babyfood jars (143 mm3 cap.), Magenta GA‐7 containers (365 mm3 cap.), and polycarbonate jars (1890 mm3 cap.) employing an in vitro hydroponics system (i.e. an automated plant culture system (APCS)). Highest growth rates occurred employing the APCS. The culture vessel capacity had a significant positive correlation on shoot length, fresh weight, number of leaves, and number of shoots. Shoot length, fresh weight, leaves, and shoots grown in the APCS exhibited increases of 1‐, 21.5‐, 7.8‐, and 8.3‐fold, respectively, compared to shoots grown in culture tubes. Higher culture growth rates that occurred in the larger‐size vessels were correlated with lower alkaloid production (mg alkaloids/g dw). However, the overall total alkaloids/vessel [(mg alkaloid/g dw)×g culture dw] increased because of greater biomass production per vessel. The alkaloid content was found to remain stable for shoots grown over a 6–month evaluation period.  相似文献   

4.
The alkaloid patterns of sea daffodil (Pancratium maritimum L.) shoot culture, cultivated in a temporary immersion cultivation system were investigated. The shoots accumulated maximal amounts of biomass (0.8 g dry biomass/L and Growth Index?=?1.6) at immersion frequency with 15 min flooding and 12 h stand-by periods. At this regime P. maritimum shoots achieved the highest degree of utilization of carbon source. Twenty-two alkaloids, belonging to narciclasine, galanthamine, haemanthamine, lycorine, montanine, tazettine, homolycorine and tyramine types were identified in intracellular and extracellular alkaloid extracts. The immersion frequency affected strongly the capacity of alkaloid biosynthesis in P. maritimum shoots and at the optimum conditions of cultivation, the total intracellular alkaloid content reached up to 3,469 μg/g dry biomass. The main biosynthesized alkaloids were haemanthamine (900.1 μg/g) and lycorine (799.9 μg/g). The obtained results proved that temporary immersion technology, as a cultivation approach, and P. maritimum shoots, as a biological system, are prospective for producing wide range bioactive alkaloids.  相似文献   

5.
The current study estimates the effect of different cytokinins on shoot proliferation and biosynthesis of caffeic acid derivatives in Dracocephalum forrestii in vitro culture. The shoots were grown on Murashige and Skoog (MS) agar medium with 1 µM indole-3-acetic acid (IAA) and different content of 6-benzyloaminopurine (BAP), zeatin, kinetin (1, 2, 4, 8, 18 µM) or thidiazuron (TDZ) (0.1, 0.2, 0.5, 1, 2 µM). The highest multiplication rate (about seven shoots and/or buds per explant) was obtained after 4 weeks of culture on MS medium with 1 µM IAA and 8 or 16 µM BAP. Optimal biomass of plant material was also received on the same media. The identity of the compounds present in the hydromethanolic extracts from D. forrestii shoots grown on cytokinin-supplemented media was confirmed using UPLC–PDA–ESI–MS method. The analysis revealed the presence of nine metabolites recognized as caffeic acid derivatives. The content of the predominant phenolic acids in the extracts, i.e. rosmarinic acid (RA) and salvianolic acid B (SAB), was determined with UHPLC. The highest yield of RA was found in shoots cultivated in the medium containing 1 µM IAA and 2 µM BAP (18.7 mg/g DW). The highest level of SAB (5.3–5.9 mg/g DW) was identified in multiple shoots grown in the presence of 1 µM IAA and 0.5–1 µM TDZ or 2 µM BAP.  相似文献   

6.
Ruta graveolens L. is a plant of agricultural interest due to its biological activity against phytopathogenic microorganism. This activity has been attributed to the presence of alkaloids, furanocoumarins, terpenes and fatty acids. The accumulation of these compounds is affected by environmental factors. The aim of this work was to analyze the seasonal variation the chemical profile of rue and to evaluate the biological activity against phytopathogenic fungi. Extracts were obtained in January, April, June and September 2017, chemically characterized by GC-MS. The antifungal activity was determined by absorbance, four concentrations were evaluated: 4, 8, 16 and 32 mg/mL. In extracts from the aerial parts, fatty acids were the most abundant group in the January sample (29.7%), and alkaloids in the April and June (27.7%) samples. In extracts from the roots, alkaloids were the most abundant group in the four collections. The highest percentage of growth inhibition on Fusarium and Stemphylium was obtained with the extracts of the January sample. Ruta graveolens can be considered an alternative for the management of phytopathogenic fungi of crops of economic importance.  相似文献   

7.
Axillary shoots from three selected white ash (Fraxinus americana L.) clones were harvested from in vitro shoot cultures. Roots were initiated by pulsing excised shoots for eight days in the dark in MS medium supplemented with 2% sucrose, 0.7% agar, 5 M NAA, and 1 M IBA. Pulsed shoots were transferred to a root elongation medium consisting of 25% MS macrosalts, full-strength microsalts and organics, 1% sucrose, 0.7% agar and no auxins. When roots were visible (6–10 days after transfer to root elongation medium), microplants were transferred to vessels containing the same minimal medium and tall fescue (Festuca elatior var. arundinacea (Schreb.) Wimm.) leaf extracts, leaf leachates, or soil leachates from plant boxes with and without tall fescue sod. After four weeks in vitro, primary adventitious and secondary root growth was reduced by extracts obtained from 5 and 10 g ground leaves per 100 ml of medium. Leachates obtained from 5 g soaked leaves per 100 ml of medium stimulated primary root growth. Soil leachates from bare soil also stimulated primary root growth. Variation was observed among the clones for root growth when plantlets were grown in extracts or leachates from tall fescue.  相似文献   

8.
Factors affecting in vitro propagation were evaluated for Ceropegia attenuata Hook., an endemic and endangered plant having ornamental potential but a limited reproductive capacity. Rapid shoot multiplication from nodal explants was established using varying concentrations of cytokinins and auxins either alone or in combinations. The highest frequency of shoot induction was achieved when nodal explants were inoculated on Murashige and Skoog (MS) medium supplemented with 13.31 μM 6-benzylaminopurine with a mean of 12.9?±?0.5 shoots per explant. High concentrations of TDZ (6.81–11.35 μM) and KN (6.78–11.61 μM) resulted in stunted and vitrified shoots. Factors implicated in the promotion of floral transition of the C. attenuata have been identified which are 4-amino-3, 5, 6-trichloropicolinic acid (picloram), 6-benzylaminopurine, sucrose and photoperiod. The highest frequency of flowering (100%) was obtained when axillary shoot explants were transferred to MS medium supplemented with picloram (4.14 μM) within 4 weeks of culture. Transfer of in vitro regenerated shoots to half strength MS medium with 2.46 μM indole-3-butyric acid (IBA) showed maximum root induction. The in vitro grown plantlets were successfully acclimatized in the glasshouse with 85% of survival and showed normal development. The developed protocol provided a simple, cost-effective approach for the conservation of endangered plant C. attenuata for replenishing its declining populations.  相似文献   

9.
Cultures of Ruta graveolens have been grown and maintained on Murashige and Skoog's medium supplemented with α-naphthalene acetic acid (1 mg/l.) and kinetin (0.1 mg/l.) or 2,4-dichlorophenoxyacetic acid (1 mg/l.), and glucose (30 g/l.). The production of dihydrofuroquinoline alkaloids has been investigated in these cultures. The present study showed that: (a) cultures of Ruta produced both differentiating and non-differentiating strains; (b) quaternary alkaloids (platydesminium, ribalinium and rutalinium) were present in all the strains and maximum contents were detected at 3-4 weeks growth; (c) high platydesminium and rutalinium content was associated with differentiation and callus formation, respectively; (d) changed concentrations of quaternary alkaloids were recorded in all the five strains grown on media supplemented with tryptophan and precursors (anthranilic acid, 5-methylanthranilic acid and 2,4-dihydroxyquinoline).  相似文献   

10.
Alcohol dehydrogenase and an inactivator from rice seedlings   总被引:6,自引:4,他引:2       下载免费PDF全文
Alcohol dehydrogenase (ADH) was measured in the various organs of rice seedlings (Oryza sativa) growing in air. In extracts from ungerminated seeds, the ADH is stable, but in extracts from seedlings more than 2 days old the enzyme initially present loses activity in a time- and temperature-dependent fashion, due to the presence of an inactivating component which increases with age in roots and shoots. The inactivation can be prevented completely by dithiothreitol, and when this is included in the extraction medium the apparent loss of total ADH in roots and shoots with age is not observed. In seedlings grown in N2, ADH levels in coleoptile extracts are higher than those in air, the enzyme is stable, and no inactivator can be detected. When seedlings grown for 5 days in air were transferred to N2 for 3 days, ADH levels increased and there was a decline in inactivator activity. Transfer back to air after 1 day in N2 led to loss of the accumulated ADH and increase in inactivator. These reciprocal changes and the fact that the inactivator is absent from coleoptiles of seedlings grown in N2 appear to suggest a regulatory role for the inactivator in vivo. However, it is clear that high levels of inactivator and ADH can exist in cells of seedlings grown in air for long periods without loss of enzyme activity, and it is argued that they must normally be separately compartmented.  相似文献   

11.
The in vitro response of kiwifruit (Actinidia deliciosa) to increasing concentrations of boron (B) and NaCl in the culture medium was studied. Kiwifruit shoot cultures were grown in vitro for 12 weeks on an MS medium containing two B concentrations (0.1 and 2 mM) combined with five NaCl concentrations (0, 10, 20, 40 and 80 mM). Kiwifruit produced the longest shoots with 2 mM B when NaCl concentration was 0--20 mM. More shoots were produced with 2 mM B for all NaCl treatments. More shoots were produced with 2 mM B and 10 and 20 mM NaCl. High B concentrations in the culture medium significantly increased shoot proliferation. Explants exhibited a moderate chlorotic appearance with 40 mM NaCl and shoots died with 80 mM NaCl. With 2 mM B, the B concentration of explants was 5--9X greater for the various NaCl treatments compared to the control. Increasing the NaCl concentration from 10 to 80 mM, resulted in higher Na and Cl concentrations in explants for all B treatments, while K and Ca concentrations decreased. Phosphorus concentration in the explants was significantly increased by increasing the NaCl concentration reaching a maximum value at 80 mM NaCl for the two B concentrations.  相似文献   

12.
The effects of Cd have been investigated in tomato (Lycopersicon esculentum) plants grown in a controlled environment in hydroponics, using Cd concentrations of 10 and 100 μM. Cadmium treatment led to major effects in shoots and roots of tomato. Plant growth was reduced in both Cd treatments, leaves showed chlorosis symptoms when grown at 10 μM Cd and necrotic spots when grown at 100 μM Cd, and root browning was observed in both treatments. An increase in the activity of phosphoenolpyruvate carboxylase, involved in anaplerotic fixation of CO2 into organic acids, was measured in root extracts of Cd-exposed plants. Also, significant increases in the activities of several enzymes from the Krebs cycle were measured in root extracts of tomato plants grown with Cd. In leaf extracts, significant increases in citrate synthase, isocitrate dehydrogenase and malate dehydrogenase activities were also found at 100 μM Cd, whereas fumarase activity decreased. These data suggest that at low Cd supply (10 μM) tomato plants accumulate Cd in roots and this mechanism may be associated to an increased activity in the PEPC–MDH–CS metabolic pathway involved in citric acid synthesis in roots. Also, at low Cd supply some symptoms associated with a moderate Fe deficiency could be observed, whereas at high Cd supply (100 μM) effects on growth overrule any nutrient interaction caused by excess Cd. Cadmium excess also caused alterations on photosynthetic rates, photosynthetic pigment concentrations and chlorophyll fluorescence, as well as in nutrient homeostasis.  相似文献   

13.
Tissue cultures of Nicotiana labacum consisting of green, albino and habituated (normal origin) and teratoma (tomorous origin) were grown under asceptic conditions for 6 to 8 weeks and their extracts were analyzed for phosphatase activity. Comparative enzyme analyses were also made on crude stem extracts of greenhouse-grown normal and tumor tissues of Nicotiana tabacum (var. Wisconsin) and a hybrid (N. glauca × N. langsdorffii).

All the crude extracts showed acid phosphatase activity with a pH optimum at 5.8 to 6.0. The total protein content and enzyme acivity of teratoma tissue (tumor) was higher than that of green, albino or habituated tissue (normal). Similar increased levels were seen in tumor tissue grown in greenhouse in comparison with greenhouse-grown normal tissues. The crude extracts of each of the tissues did not exhibit any qualitative difference in specificity with the 5 different substrates tested; however, differences in the level of activity was observed.

The effect of 4 different culture media was tested on the growth, protein content and acid phosphatase activity of habituated tobacco in tissue culture. Tissues growing in medium containing high salt concentrations showed higher activities than tissues grown in a basal control medium. From the results, it is suggested that although many factors like auxin and other growth factors can influence growth of habituated tobacco tissue, they need not necessarily affect this specific enzyme activity.

  相似文献   

14.
Different explants of Rauwolfia serpentina were tested for their capacity and root differentiation. MS sedium containing 2.0 mg I-1 BAP or 1.5 mg I-1 BAP with 0.5 mg I-* NAA gave the best shoot proliferation from shoot apices (84.12%). Multiple shoots subcultured on the same media gave higher number of shoots (6–9). Callus formed at the cut ends of explants produced shoots when subcultured on the above media. Rooting was achieved after transfer of shoots either to hormone free or half strength or full strength MS medium with different concentrations of IAA (0.5,1.0 mg I-1 and IBA (0.5,1.0 mg I-1).The complete regenerated plantlets have been successfully transferred to earthen pots in green house. Maximum root differentiation from leaves, stem and nodal cuttings derived calli was obtained on MS medium supplemented with NAA (2.0 mg I-1) and BAP (1.5 mg I-1). Identification of reserpine and other alkaloids from differentiated roots holds an interesting alternative for controlled production of alkaloids from this endangered, overexploited medicinal plant species.  相似文献   

15.
A protocol was developed for micropropagation of Mandevilla moricandiana (A.DC.) Woodson, a native plant from Brazil. Shoots, obtained from in vitro plantlets were used as source of nodal segments for shoot production from axillary buds. The nodal segments were grown on Murashige and Skoog medium supplemented with different concentrations of 6-benzyladenine and/or indole-3-acetic acid to induce axillary bud elongation. After a 2-mo culture period, the medium supplemented with 1.0 mg?L?1 6-benzyladenine gave the largest number of nodal segments per explant. The nodal segments obtained from plants developed under these conditions were grown on medium supplemented with different concentrations indole-3-acetic acid, ??-naphthaleneacetic acid, and indole-3-butyric acid. The use of the medium supplemented with indole-3-acetic acid and indole-3-buryric induced shoot elongation and shoot development, formation of basal callus, and/or indirect organogenesis of roots. Following transfer of shoots to soil, the plants with only basal callus showed 10% survival and developed roots from callus, while in vitro-rooted plants had a maximum 40% survival rate ex vitro. Regardless of the auxin added to the rooting medium, the acclimatization period allowed the plants rooted in vitro to develop their shoots fully. The protocol developed here is suitable for the production of shoots and rooted plantlets of M. moricandiana.  相似文献   

16.
In Vitro Regeneration of Stevia rebaudiana (Bert) from the Nodal Explant   总被引:1,自引:0,他引:1  
Procedure for micropropagation of Stevia rebaudiana Bertoni, containing stevioside, a natural noncaloric sweetner, has been developed using nodal segments as explant. Higher proliferation of shoots and multiplication was obtained on Murashige and Skoog basal medium (MS) supplemented with 1.0 mg l-1 indoleacetic acid (IAA) plus 10.0 mg l-1 kinetin and 30.0 mg l-1 adenine sulphate. Sprouting of 90% of the axillary buds was observed within 4 weeks of inoculation, producing >10.0 shoots per explant within 12 weeks. Profuse roots were induced from 90% of the regenerated shoots within 4 weeks of inoculation on half strength MS solid medium supplemented with 1.0 mg l-1 IAA. High survival rate, > 60%, was obtained when the plantlets were transferred to field conditions. The survival rate of taller plants was always higher. The in vitro regenerated plants were morphologically indistinguishable from the donor plants and leaves were of intense sweet taste upon chewing. The heterogenic nature of S. rebaudiana necessitates establishment of protocol for every genotype independently.  相似文献   

17.
[3H]Indole-3-acetyl-myo-inositol was hydrolyzed by buffered extracts of acetone powders prepared from 4 day shoots of dark grown Zea mays L. seedlings. The hydrolytic activity was proportional to the amount of extract added and was linear for up to 6 hours at 37°C. Boiled or alcohol denatured extracts were inactive. Analysis of reaction mixtures by high performance liquid chromatography demonstrated that not all isomers of indole-3-acetyl-myo-inositol were hydrolyzed at the same rate. Buffered extracts of acetone powders were prepared from coleoptiles and mesocotyls. The rates of hydrolysis observed with coleoptile extracts were greater than those observed with mesocotyl extracts. Active extracts also catalyzed the hydrolysis of esterase substrates such as α-naphthyl acetate and the methyl esters of indoleacetic acid and naphthyleneacetic acid. Attempts to purify the indole-3-acetyl-myo-inositol hydrolyzing activity by chromatographic procedures resulted in only slight purification with large losses of activity. Chromatography over hydroxylapatite allowed separation of two enzymically active fractions, one of which catalyzed the hydrolysis of both indole-3-acetyl-myo-inositol and esterase substrates. With the other fraction enzymic hydrolysis of esterase substrates was readily demonstrated, but no hydrolysis of indole-3-acetyl-myo-inositol was ever detected.  相似文献   

18.
Catharanthus roseus (L.) G. Don is an economically and medicinally important plant since its leaves and flowers contain terpenoid indole alkaloids. The present study, for the first time, encompasses the influence of silver nitrate (AgNO3), in consort with cytokinins like N 6-benzyladenine (BA) and 6-furfurylaminopurine (kinetin), to regenerate multiple shoots from nodal segments explants and to induce high-frequency precocious flowering of C. roseus under in vitro condition. Synergistic effect of equal concentrations of BA and kinetin was enhanced following the amalgamation of AgNO3. As high as 98% explants responded to multiple shoot initiation and proliferation in Murashige and Skoog medium supplemented with 3 µM BA, 3 µM kinetin and 0.1 µM AgNO3. As many as 7 shoots were developed per explant following 12 days of inoculation. Continuous culture in the same medium for 21 days induced precocious flowering from 75% shoots, wherein a maximum of ~?6 (5.67?±?0.88) flowers was observed per in vitro shoot. On the other hand, in the combinations of BA and kinetin excluding AgNO3, a maximum of 6.67% explants responded and initiated merely 3.33 shoots per explant. Nevertheless, no induction of flower was observed in the media devoid of AgNO3. Our results on the induction and proliferation of multiple shoots with simultaneous flowering would help the global pharmaceutical industry to produce in vitro shoots and flowers in bulk, as an alternative source of alkaloids.  相似文献   

19.
张科  田长彦  李春俭 《生态学报》2012,32(10):3069-3076
选用古尔班通古特沙漠南缘荒漠-绿洲交错带常见的一年生盐生植物盐角草、刺毛碱蓬、叉毛蓬、猪毛菜和碱地肤为材料,比较了它们在原状盐土和沙土中的生长及体内矿质元素组成的差异。结果表明:① 原状盐土0—100 cm各土层的pH值低于沙土,但电导率和含水量明显高于沙土;② 原状盐土中生长的植物干重是沙土中生长植物干重的7—118倍,后者的根冠比是前者的2—6倍。③ 体现肉质化程度的地上部含水率为52%—81%,中低耐盐植物含水率在两种土壤中差异显著,强耐盐植物差异不显著;④ 5种一年生盐生植物地上部氮浓度为11—34 g/kg,在有效氮含量高的盐土上植物氮浓度也高;磷浓度为1—4 g/kg,在有效磷高的盐土上植物磷浓度也较高(盐角草除外);但沙土中的植物地上部钾浓度明显高于盐土中植株的地上部钾浓度,这与两种土壤在0—60 cm土层中的钾浓度差异相反;⑤ 尽管原状盐土0—100 cm土层中的水溶性钙、镁、钠、氯、硫浓度显著高于沙土,盐土与沙土中生长的植物地上部钠、水溶性氯和硫的浓度比值远远低于土壤中的相应元素浓度的比值,甚至盐土中的植株钙、镁浓度等于或显著低于沙土中生长的植物。表明盐土不仅影响一年生盐生植物的生长,也显著影响这些植物对矿质元素的吸收和累积。一年生盐生植物能够选择性吸收不同生境中的矿质元素。本研究期望为进一步深入研究盐生植物耐盐的适应机制提供依据,也可为植物修复盐碱土的品种选择提供参考。  相似文献   

20.
Shoot formation was observed in callus from apical cells of pea (Pisum sativum L. cv. Century). Shoot apices from 4-day-old plants were macerated and the resulting cell masses grown on agar media. The callus formation and shoot production occurred within 4 to 6 weeks in defined media containing 0.2 to 5.0 μM benzyladenine and 1 μM naphthaleneacetic acid. While most callus produced one or more shoots at high frequency, root formation did not occur regularly. Plants obtained by these procedures were grown to maturity producing flowers and pods.  相似文献   

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