Chemical composition of newborn larvae, muscle larvae and adult Trichinella spiralis

Chemical composition of newborn larvae, muscle larvae and adult Trichinella spiralis. International Journal for Parasitology16: 455–460. The chemical composition of newborn larvae (NBL), muscle larvae (ML) and 4-day-old and 7-day-old adult Trichinella spiralis were compared. Total protein constituted 44.1% of the dry wt of ML, 36.9% of NBL and 30.3% of 7-day-old adult worms. Glycogen content was 16.1% of worm dry wt in ML and 7.8% in NBL and 7.2% of worm dry wt in adults. Trehalose content was 5.2% in NBL, 8.3% in ML and 8.7% in 7-day-old adult worms. Significantly lower levels of trehalose and glycogen were found in 4-day-old worms than were present in 7-day-old worms. Free glucose amounted to 0.1% of dry wt in ML, 0.37% in NBL and 0.87% in 7-day-old adults. RNA accounted for 3.5% of the dry wt of ML, 3.2% of the dry wt of 7-day-old adults and 1.1% of the dry wt of NBL. The DNA content of NBL was 0.23% of worm dry wt, in ML 0.48% of worm dry wt and 0.51% of worm dry wt in 7-day-old adults. The three parasite stages examined agreed closely with regard to types of amino acids in the free pool. Exceptions were as follows: NBL lacked taurine which both adults and ML contained; adults lacked methionine which both ML and NBL contained; and, trace amounts of cysteine were present in ML but absent from the other two stages.     

Enhanced Alkaloid Production in Callus Cultures of Heliotropium indicum L and Regeneration of Plantlets

Heliotropium indicum L (Boraginaceae) contains the anticancer pyrrolizidine alkaloid, indicine-N-oxide (INO). To study the yield of INO as a function of plant development, plantlets were regenerated in vitro from nodal and hypocotyl explants and also from hypocotyl callus, on Murashige and Skoog’s (MS) medium supplemented with 1-naphthaleneacetic acid (NAA), 6-benzylaminopurine (BAP), asparagine (Asp) and glutamine (Glu). The regenerated plantlets were rooted on MS supplemented with Glu or gibberellic acid (GA₃). While 5-week-old seedlings showed a high amount of INO (0.12% dry wt) that depleted gradually as the plants attained maturity and flowered, fast growing callus produced a much higher yield of INO (0.32% dry wt). As the callus cultures differentiated into shoots and subsequently into plantlets, the INO content decreased to about 0.2% dry wt. It appears that INO is the primary product of pyrrolizidine alkaloid biosynthetic pathway in rapidly growing meristematic tissue of Heliotropium indicum, later reduced to its alkaloidal base and reallocated to other tissues. The article includes an efficient micropropagation method for Heliotropium indicum.     

High berberine-producing cultures of coptis japonica cells

The highest berberine content of unselected Coptis cells cultured under the best conditions for berberine production (darkness, high aeration, 3% sucrose and White's basal medium) was about 5% on a dry wt basis. Fluoromicroscopy showed that cultured Coptis cells had heterogeneous characters; therefore, selection was used to establish a high berberine-producing culture of Coptis cells. When small cell aggregates were cloned, high berberine-producing cell lines were produced. Repeated cloning, however, was needed to obtain stable cell lines that produced large amounts of berberine. The highest berberine production in a selected cell line was 13.2% on a dry wt basis (1.39 g/l. culture). The average production was 8.2% (0.90 g/l. culture).     

Production of thiophenes from callus cultures of Tagetes patula L. and its mosquito larvicidal activity

Calli were initiated from leaf tissues of T. patula, on Murashige and Skoog's medium supplemented with 2-4 dichlorophenoxacetic acid (2,4-D) and kinetin (Kn). The maximum accumulation of biomass was recorded on 6th week at 1.11 g. dry wt/culture. Maximum thiophene content was recorded on 4th week at 0.008% on dry wt basis in callus cultures of T. patula subjected to nutrient stress. Nitrogen stress induced 3-fold increase in thiophene production level in six weeks (0.024% on dry weight basis). The best hormonal supplementation required for thiophenes production was found to be 2,4-D (2.0 mg L(-1)) and kinetin (2.0 mg L(-1)). The thiophenes produced in callus cultures of T. patula showed larvicidal effect against mosquito larvae.     

Distribution of hydroxamic acids in zea mays tissues

The hydroxamic acid content of leaves of cereals correlates well with resistance to aphids. In maize these compounds were absent from xylem exudates and guttation drops. Lateral veins of leaves of 7-day-old maize plants contained 8 mmol/kg fr. wt. while the entire leaf contained only 4.2 mmol/kg fr. wt. In leaves of 20-day-old plants, these amounts decreased by ca one-third. In mesocotyls, the cortex and central vascular cylinder contained 1.3 and 2.2 mmol/kg fr. wt, respectively. In 12-day-old wheat plants, the complete leaves and their veins contained 2.4 and 6.4 mmol/kg fr. wt respectively. Thus, the concentration of hydroxamic acid was always higher in the vascular bundles.     

Catechin production in cultured cells of Taxus cuspidata and Taxus baccata

The main polyphenols in callus and cell suspension cultures of Taxus cuspidata and T. baccata were (+)-catechin and (−)-epicatechin, while lignans, such as (+)-taxiresinol, (+)-isotaxiresinol, (+)-isolariciresinol and (−)-secoisolariciresinol, were present in trace amounts. T. cuspidata cells contained 1.7% (+)-catechin and 2.4% (−)-epicatechin on dry wt basis but when stimulated with methyl jasmonate produced 3.4% catechin and 5.2% epicatechin. These are the highest levels of these metabolites obtained in plant cell cultures.     

Comparative analysis of fatty acids in pollen and seed of rapeseed

Lipids accounted for 31.7% of the dry wt of mature, viable pollen of rapeseed (Brassica napus). External lipids of the pollen coat, specified by the diploid parental genome, and internal cytoplasmic lipids, presumably specified by the haploid pollen genome, were separated by differential extraction. External lipids and paraffins represented 9.8 % of pollen dry wt, and contained predominantly linolenic (18:3) with significant amounts of palmitic (16:0), stearic (18:0) and myristic (14:0) acids, as well as paraffins. Internal lipids accounted for 21.9% of pollen dry wt and comprised predominantly 18:3 and 16:0. Mature seeds of B. napus comprised 45.0 % lipid, with a fatty acid composition different from that of either pollen fraction. Decanoic (10:0), lauric (12:0) and hexadecatrienoic (16:3) acids were detected in pollen but not in seeds. These data are discussed in relation to pollen biochemistry and the potential for pollen selection in rapeseed breeding.     

Production of allantoin, rabdosiin and rosmarinic acid in callus cultures of the seacoastal plant Mertensia maritima (Boraginaceae)

A callus culture of the extreme halophyte seacoastal plant Mertensia maritima (Boraginaceae) was established from apical shoots of the plant using a modified Murashige and Skoog medium supplemented with 6-benzyladenine (0.5?mg/L) and ??-naphthaleneacetic acid (2.0?mg/L). Three main compounds, (?)-R-allantoin, (+)-rabdosiin and rosmarinic acid, were isolated from extracts of M. maritima calli by liquid chromatography and identified by ¹H and ¹³C NMR, UV, ECD and HPLC?CMS. Quantitative HPLC analysis showed that the calli produce (+)-rabdosiin (0.14% dry wt), rosmarinic acid (0.74% dry wt) and (?)-R-allantoin (3.7% dry wt). Allantoin was detected in plant cell cultures for the first time. All of these metabolites were also present in lower quantities in different parts of the plant. The presence of rabdosiin and rosmarinic acid, in combination with the skin-conditioning agent (?)-R-allantoin, represents a potentially useful novel composition for skin protection.     

A laboratory energy budget for the ormer Haliotis tuberculata L.

Each component of the energy budget (ingestion, egestion,somatic growth, reproductive investment, respiration, excretion, and mucus production) was measured for Haliotis tuberculata L. held at 15°C in a 12-h light: 12-h dark regime. Ulva lactuca L. was used as the food source throughout, and the budget was assessed over the whole size range of the animal. Ingestion rates ranged from 1.94 to 997.2 cal·animal⁻¹·day⁻¹ in 0.01 to 50 g dry wt (3.71 × 10⁻³ to 17.3 g dry flesh wt) animals, respectively. The major component of the energy budget was somatic growth (37.5% of I) in a 0.01-g dry wt animal while it was respiration (31.1% of I) in a 50-g dry wt animal. Mucus production formed a large part of the budget (from 23.3% of I in a 0.01-g dry wt animal to 29.l% of I in a 50-g dry wt animal). Scope for growth, I − (E + R + U + M), was calculated as ranging from 24.5% of ingestion in a 50-g dry wt animal to 36.8% in a 0.01-g dry wt animal.Each component was measured independently and allometric relationships with animal dry weight calculated. Exponents ranged from 0.60 (somatic growth) to 1.06 (reproductive investment).The calorific value of food was 3419 cal·g dry wt⁻¹ and for faeces was 2817 cal·g dry wt⁻¹. Absorption as a percentage of ingestion in terms of dry weight ranged from 78% for 95-mm length animals (50 g dry wt) to 81% in 6-mm animals (0.01-g dry wt⁻¹). Gross and net growth efficiencies (K₁ and K₂) were calculated on an energy basis and both were logarithmically related to animal dry weight.     

High Rabdosiin and Rosmarinic Acid Production in Eritrichium sericeum Callus Cultures and the Effect of the Calli on Masugi-Nephritis in Rats

During an investigation of plant cell cultures that might be useful in the treatment of renal disorders, we established a vigorously-growing E-4 callus culture of Eritrichium sericeum that produced large amounts of caffeic acid metabolites, (?)-rabdosiin (1.8% dry wt) and rosmarinic acid (4.6% dry wt). Elicitation of the calli by methyl jasmonate induced a 38% increase in total polyphenol production. The most efficient method of eliciting (?)-rabdosiin biosynthesis was through the treatment of E-4 calli with cuprum glycerate, which induced an increase in (?)-rabdosiin production of as much as 4.1% dry wt. Oral administration of E-4 callus biomass (100 mg/kg/d for 30 d) to rats with induced Masugi-nephritis caused an increase in diuresis and lowered creatinine excretion and proteinuria levels as compared with Masugi-nephritis untreated rats. While all of the Masugi-nephritis untreated rats began to suffer, near a quarter of the E-4 treated rats remained in good health. This result indicates that the E-4 culture has the potential to alleviate the symptoms associated with nephritis.     

Effects of temperature and gibberellic acid on 4-desmethylsterol composition of Avena sativa stem segments

Of the three morphological subunits of Avena sativa stem segments (node, leaf-sheath and internode) examined, internodes constituted the richest source of phospholipids and sterols, yielding almost double the concentration of lipid found in the leaf-sheath. The phospholipid compositions of nodes and internodes were similar, comprising mostly phosphatidylcholine (PC) and phosphatidylethanolamine (PE), with linoleic, linolenic and palmitic acids as the predominant component fatty acids. Leaf-sheath tissue contained mainly PE, with equally high amounts of palmitic, linoleic and linolenic acids. β-Sitosterol and cholesterol were the major 4-desmethylsterols of the internode, while β-sitosterol was predominant in the node and leaf-sheath tissues. The growth temperature of segments prior to isolation produced its major effect on the concentration of stigmasterol, which decreased markedly with temperature. The sitosterol/stigmasterol ratio increased significantly as temperature decreased. Stem segments isolated from plants treated with gibberellic acid (GA₃) for 3 weeks, showed a significant reduction in the amounts of 4-desmethyl sterols on a dry wt basis when compared with control segments. However, when stem segments were incubated with GA₃ for 20 hr, no change in 4-desmethylsterol composition or concentration was observed, even though significant growth in response to GA₃ occurred.     

High rabdosiin and rosmarinic acid production in Eritrichium sericeum callus cultures and the effect of the calli on masugi-nephritis in rats

During an investigation of plant cell cultures that might be useful in the treatment of renal disorders, we established a vigorously-growing E-4 callus culture of Eritrichium sericeum that produced large amounts of caffeic acid metabolites, (-)-rabdosiin (1.8% dry wt) and rosmarinic acid (4.6% dry wt). Elicitation of the calli by methyl jasmonate induced a 38% increase in total polyphenol production. The most efficient method of eliciting (-)-rabdosiin biosynthesis was through the treatment of E-4 calli with cuprum glycerate, which induced an increase in (-)-rabdosiin production of as much as 4.1% dry wt. Oral administration of E-4 callus biomass (100 mg/kg/d for 30 d) to rats with induced Masugi-nephritis caused an increase in diuresis and lowered creatinine excretion and proteinuria levels as compared with Masugi-nephritis untreated rats. While all of the Masugi-nephritis untreated rats began to suffer, near a quarter of the E-4 treated rats remained in good health. This result indicates that the E-4 culture has the potential to alleviate the symptoms associated with nephritis.     

Soybean protoplast culture and direct gene uptake and expression by cultured soybean protoplasts

A method was developed for culturing protoplasts freshly isolated from developing soybean (Glycine max L.) cotyledons. First cell divisions were observed within 5 days after protoplast isolation and microcalli, consisting of about 20 cells, were formed within 10 days. Thirty days after protoplast isolation, callus tissues were observed without the aid of a microscope. A 30 to 50% plating efficiency was consistently obtained. Using a polyethylene glycol-electroporation technique, DNA was introduced into these protoplasts. The protoplasts were then cultured to form callus. Chloramphenicol acetyltransferase (CAT) activity was detected in protoplast cultures 6 hours after introduction of a 35S-CAT-nopaline synthase 3′ chimeric gene. The highest CAT activity was detected in 3-day-old electroporated protoplast cultures, indicating transient expression of the introduced gene. Some CAT activity was detected in 40-day-old callus cultures and in geneticin (G418) selected callus tissues which also received a chimeric neomycin phosphotransferase II gene, indicating the presence of stable transformants. A control chimeric gene with an inverted 35S promoter failed to produce any CAT activity in this system.     

Influence of medium composition on callus induction and camptothecin(s) accumulation in Nothapodytes foetida

Camptothecin(s) production was examined in callus cultures derived from cotyledons of Nothapodytes foetida (Weigh) Sleumer. The calluses were grown on various combinations of Murashige and Skoog's basal media supplemented with auxins 2,4-dichlorophenoxyacetic acid (2,4-D), 2,4,5-trichlorophenoxyacetic acid (2,4,5-T), a-napthalene acetic acid (NAA) and indole-3-acetic acid (IAA) with 6-benzyl aminopurine (BA)/kinetin in different concentrations. The presence of camptothecin (CPT) and 9-methoxycamptothecin (9-OMeCPT) were analyzed by HPLC in relation to the media composition. Hyper production of CPT(1.306% on dry wt. basis) was observed with a combination of 2,4-D with BA and 2,4,5-T and NAA in 1-month-old callus.     

The Sterol,Fatty Acid,and Hydrocarbon Composition of Globodera solanacearum

Globodera solanacearum females were found to have less than 0.01% of dry wt as sterols. Seven sterols were detected in the nematode, with stanols (campestanol and stigmastanol) making tip more than 50% of the total sterols present. Lipid amounted to 29.4% of the dry weight of the nematode. Triglyceride, free fatty acid, and phospholipid classes were composed predominantly of 20:4, 20:1, and 18:1 fatty acids. Of the total weight of fatty acids found in G. solanacearum females, the greatest portion occurred in the triglyceride fraction, followed by the free fatty acid fraction then the phospholipid fraction. Several unidentified hydrocarbons were detected in the nematode. Paraffinic hydrocarbons detected ranged in carbon length from C15 to C29. Total concentration of hydrocarbon composed 0.20% of the dry wt.     

Occurrence of adenosine 3′:5′-cyclic monophosphate in plant tissues

A procedure is described which unequivocally demonstrates the presence of adenosine 3′:5′-cyclic monophosphate in Phaseolus vulgaris. Its concentration was determined spectrophotometrically at 2·6–9·2 nmol g^?1 of tissue (dry wt) for 6-day-old seedlings and about one-tenth of this in 13-day-old plants.     

Selection of high nicotine-producing cell lines of tobacco callus by single-cell cloning

Single-cell clones of Nicotiana tabacum callus showed wide variation in the production of nicotine. An efficient screening of numerous clones was made possible by the adoption of the ‘cell squash method’ devised for estimating the approximate alkaloid content of a small piece of callus. From these clones, several cell lines with higher yield of nicotine (1.0–3.4% of dry wt) have been isolated by repeated clonal selection.     

Verbascoside production in callus and suspension cultures of Hygrophila erecta

Cell suspension cultures of Hygrophila erecta obtained from hypocotyl explants accumulate up to 2.6% of their dry wt as only one caffeic glycoside ester, verbascoside. On the contrary, different callus lines initiated from stem explants contain not more than 1 % of verbascoside and a few of them produced other caffeoyl glycosidic esters.     

咖啡酸代谢物的抗肾炎活性分析

在研究治疗肾功能紊乱植物的细胞培养时, 建立了一个Eritrichium sericeum的E-4愈伤组织株系, 发现此株系可产生大量的咖啡酸代谢物、(–)-rabdosiin (1.8%干重)和迷迭香酸(4.6%干重), 通过诱导(–)-rabdosiin的含量提高至4.1% (干重)。将E-4愈伤组织喂服Masugi肾炎大鼠, 结果发现, 与对照组(未喂服E-4愈伤组织的Masugi肾炎大鼠)相比, 处理组(喂服E-4愈伤组织)中的大鼠出现如下症状: 尿多、排泄物中肌氨酸酐降低、尿蛋白水平降低; 当对照组中所有大鼠都出现疼痛症状时, 处理组中仍有约/4的大鼠表现出健康状况良好。以上结果表明, E-4株系具有缓解肾炎症状的潜在功能。此外, 利用富含多酚的rolC转基因的细胞株系, 研究了愈伤组织中咖啡酸代谢物的诱导合成机制。结果发现,在rloC转基因的E. sericeum愈伤组织中,咖啡酸代谢物的高产与迷迭香酸生物合成中的关键基因CYP98A3的高表达有关。