毛地黄优良变异植株组织培养及无性系的建立

研究了毛地黄优良变异植株嫩茎愈伤组织的诱导、分化及无性系的建立。结果表明:MS+1mg/L BA+0.6 mg/L 2,4-D是毛地黄嫩茎愈伤组织诱导的理想培养基;MS+0.5 mg/L BA+0.1 mg/LNAA是愈伤组织分化的理想培养基;MS+0.2 mg/L BA+0.2 mg/LIAA是不定芽壮苗培养的理想培养基;1/3 MS+0.3 mg/LIAA是生根培养的理想培养基;炉灰渣、河沙是生根试管苗移栽的理想基质。     

Metabolism of [ring-2,6-14]parathion in plant cell suspension cultures of carrot (Daucus carota), purple foxglove (Digitalis purpurea), soybean,thorn apple (Datura stramonium) and wheat (Triticum aestivum)

By means of standardized procedures, the metabolism of [ring-2,6-¹⁴C]-parathion was investigated in carrot (Daucus carota L.), purple foxglove (Digitalis purpurea L.), soybean (Glycine max Merrill cv. ?Mandarin’?, and Glycine max Merrill cv. ?Harosoy 63’? cultivated on B5 and Miller media, respectively), thorn apple (Datura stramonium L.), and wheat (Triticum aestivum L.) cell suspension cultures. In the wheat and soybean (Mandarin) cells only 2.9 and 8.9%, respectively, of the applied parthion remained unmetabolized after 48 h of incubation, while 51.2, 57.9, 60.3, and 62.4% of the unchanged parent were detected in the D. purpurea, D. Stramonium, carrot and soybean (Harosoy) cultures, respectively. In all suspensions, paraoxon and 4-nitrophenol were found as phase I metabolites, thus demonstrating that plant tissues can catalyse oxidative desulfuration and dearylation of parathion. 4-Nitrophenol was also glycosylated with glucose and possibly galactose. Further, as yet unidentified, metabolites indicated that bio-transformations had also occurred at the aromatic moiety. Large amounts of non-extractable residues were detected in the wheat suspension (38.3%), while the other cultures showed a lower incorporation of ¹⁴C into insoluble cell material (0.9-9.4%). For a prospective ecotoxicological evaluation of the metabolic fate of pesticides and xenobiotics in plants in general, the differential metabolic capacity of plant cell cultures and plants should be taken into account.     

Antiproliferative steroidal glycosides from Digitalis ciliata

Two new compounds, a furostanol glycoside (1) and a pregnane glycoside (4), along with eight known compounds, belonging to the classes of spirostane (2,3), pregnane (5–7) and cardenolide (8–10) glycosides, were isolated from the seeds of Digitalis ciliata. Their structures were elucidated by 1D and 2D-NMR experiments as well as ESI-MS analysis. For the first time pregnane glycosides of the diginigenin series have been isolated from D. ciliata. The cytotoxic effects of compounds 1–10 on cell viability of several cancer cell lines, namely human breast cancer (MCF-7), human glioblastoma (T98G), human lung adenocarcinoma (A549), human colon carcinoma (HT-29), and human prostate cancer (PC-3) cell lines were evaluated. Compounds 1, 4, 7 and 8 showed antiproliferative effects against MCF-7, HT-29 and A549 cancer cells with IC₅₀ values ranging from 8.3 to 20 μM. The effects of compounds 1–10 on cell proliferation were evaluated on these three cancer cell lines by cell cycle analysis of DNA content using flow cytometry. Compounds 7, 8 and 10 induced significant changes in G₂/M cell cycle phase of all analyzed cells. The obtained results indicate that compounds 7, 8 and 10 are cytostatic compounds effective in reducing cell proliferation by inducing accumulation of the cells in the G₂/M phase of the cell cycle.     

Acaricidal effects of cardiac glycosides, azadirachtin and neem oil against the camel tick, Hyalomma dromedarii (Acari: Ixodidae)

The cardiac glycoside, digitoxin, from Digitalis purpurea L (Scrophulariaceae), a cardiac glycosidal (cardenolide) extract from Calotropis procera (Ait) R Br (Asclepiadaceae), azadirachtin and neem oil from Azadirachta indica A Juss (Meliaceae) were tested for their effects against larvae and adult stages of the camel tick, Hyalomma dromedarii Koch (Acari: Ixodidae). The contact LC50 values of the first three materials against adults were 4.08, 9.63 and >40.7 microg cm(-2), respectively, whereas the dipping LC50 values of the four materials were 409.9, 1096, >5000 and >5000 mg litre(-1), respectively. Contact and dipping LC50 values of the extract and azadirachtin against larvae were 6.16, >20.3 microg cm(-2) and 587.7 and >2500 mg litre(-1), respectively. Azadirachtin had no effects on egg production or feeding of adults up to 5000 mg litre(-1); however at 2500 mg litre(-1), it caused significant reduction in feeding activity of larve, prolonged the period for moulting to nymphal stage, and caused 60% reduction in moultability. Results of the two cardiac glycoside materials are comparable with those of several commercial acaricides. The risks and benefits associated with the use of cardiac glycosides are considered.     

Effect of Whole Season CO2 Enrichment on the Cultivation of a Medicinal Plant, Digitalis lanata

A versatile method was developed for the application of 1000 ppm CO₂ during the whole growth period of plants. Temperature controlled water cooling and ventilation of the greenhouse resulted in a monthly CO₂ enrichment time of 60 to 90 % of the total light period. Digitalis lanata , grown in greenhouses with CO₂ enrichment during the whole growth phase from April to November, produced twice as much biomass as field cultivated plants.
The relative yield of the glycoside digoxin per gram Digitalis drug dry weight was 0.4% in field grown and 0.7% in greenhouse cultivated plants. The production of digoxin per hectare in the greenhouse at 1000 ppm CO₂ was almost 3.5-fold that by field cultivation. Drug yield and secondary metabolite production in D. lanata were remarkably influenced by increased temperature and elevated CO₂ partial pressure in the greenhouse.     

Cytology and digoxin production in hybrids between Digitalis lanata and D. grandiflora

Summary Digitalis lanata is the only commercial source of the cardiac glycoside digoxin. It is an outbreeder and displays considerable variation. An attempt to increase uniformity was made by making D. lanata × D. grandiflora diploid hybrids and from these produce allotetraploid hybrids using colchicine treatment. Such tetraploid plants would be homozygous. Tetraploid hybrids were shown to contain digoxin but at lower levels than in D. lanata. Progeny of selfed tetraploid hybrids were significantly more uniform than D. lanata grown from commercial seed, but were considerably less vigorous.     

毛地黄叶组织培养中光质对大分子物质含量及过氧化物酶同工酶酶谱的影响

光质不仅对毛地黄(Digitalis purpurea)叶愈伤组织的生长及器官发生有影响,同时也显著地影响了组织培养物中多糖类、蛋白质、DNA、叶绿素等生物大分子物质的含量。白光中蛋白质和叶绿素含量均较高,而蓝光、黑暗却抑制蛋白质的合成。生长值较高的绿光中含糖量较少,而生长值较低的黑暗中却有较多的糖。由此设想,光质对组织培养的影响与光合作用并无直接关系,光质可影响糖的转化过程,从而影响光形态建成。DNA 含量随光质的不同也有很大差异,并与生长值之间存在着显著的负相关。过氧化物酶活力测定表明,其比活力与形态发生有关,正处于器官发生高峰期的白光、黑暗、红光处理中,酶的比活力较高,而已有大量芽产生的黄、绿光处理组,其酶比活力较低。同工酶酶谱分析结果表明,黑暗中比有光条件下多一条 Px4谱带,各种光质既可影响到 Px1—3酶谱带的出现,也可影响各条带的宽窄和颜色深浅。这说明,光质对组织培养的影响,不但反映在基因的顺序表达水平上,而且也可作用在转录后的过程中。     

Seasonal Variation in Nutrient Status of Foxglove Leaves

The seasonal variation of mineral elements and the relationships among them were studied in natural populations of foxglove (Digitalis obscura). Young and mature leaves were collected in 10 different populations and on four sample dates (May, July, October, and February). Leaf mineral elements [nitrogen (N), phosphorus (P), potassium (K), calcium (Ca), magnesium (Mg), iron (Fe), manganese (Mn), zinc (Zn), and copper (Cu)] were determined. The highest concentrations of N, P, and K in young leaf were recorded in May, followed by a decrease in the other months, while in contrast Ca and Fe showed the lowest concentration in May. Mature leaves showed differential seasonal behavior. Besides seasonal variations, significant fluctuations of N/P and Ca/Mg ratios were observed in young leaves. Strong positive correlations existed among N, P and K, while negative correlations were found between Ca and N, P, or K.     

毛地黄甙对钉螺生理生化的影响

本实验研究了毛地黄强心甙对钉螺糖原和总蛋白含量的影响:大于0.8g/L毛地黄强心甙能够显著降低钉螺体内的糖原含量,减少幅度从10.69%到74.94%;毛地黄强心甙也能够在一定程度上降低钉螺体内的蛋白质含量(从5.79%到14.22%),但减少量不及糖原显著;灭螺植物对钉螺的致死作用可能更多与钉螺组织细胞中的能量代谢有关.     

Relationships among soil characteristics,plant macronutrients,and cardenolide accumulation in natural populations of Digitalis obscura

In the present study, we have investigated relationships among several soil parameters (pH, organic matter, total carbonate, macronutrients, electrical conductivity, cation‐exchange capacity) and macronutrient and cardenolide contents in leaves of wild Digitalis obscura plants. Young and mature leaves and soil samples were collected in ten different areas, corresponding to three Mediterranean bioclimatic belts (thermo‐, meso‐, and supramediterranean belts). Soil and leaf macronutrient (N, P, K, Ca, and Mg) contents and leaf cardenolide contents were determined. Bioclimatic conditions influenced the development of D. obscura, biomass being lowest in plant populations of the supramediterranean belt, but they did not exert any relevant effect on the contents of macronutrients in soils and plants. Magnesium (total and EDTA‐extracted) was the only soil macronutrient significantly correlated with its content in the plant. Cardenolide contents were negatively correlated with the N, P, and K contents in young leaves, whereas such correlations were highly significant and positive for Mg.