Analysis of artemisinin in Artemisia annua L. by LC-MS with selected ion monitoring

A liquid chromatography--mass spectrometry (LC-MS) method with selected ion monitoring (SIM) was developed and validated for the analysis and standardization of artemisinin in Artemisia annua L. This method is simple and accurate and requires only an 11 min per sample running time for the direct detection and quantification of artemisinin, a sesquiterpene lactone with antimalarial activity. To accurately analyze artemisinin, SIM was used to record the abundance of the [M - 18 + H]+ ion peak at m/z 265.3, with a scan range between m/z 250 and 270. Quantification was based on the LC-MS peak area of artemisinin, and the standard curve was used for calculation. This method was then validated and applied to several populations of A. annua to assess the population means and the diversity within a selected population. The distribution of artemisinin was found to vary by plant origin with population means ranging from 0.03 to 0.71% artemisinin dry leaf weight. Individual plants within the selected population that exhibited the highest artemisinin mean were then sampled, and the contents ranged from trace amounts to 1.5% artemisinin dry leaf weight basis, making these single plants accumulating artemisinin in concentrations >1.0% promising candidates for plant breeding and varietal development for high-yielding artemisinin.     

栽培密度和施肥水平对黄花蒿生长特性和青蒿素的影响

采用大田试验,研究不同密度和施肥水平对黄花蒿生长、生物量分配和青蒿素含量的影响。试验设3个密度水平: 高密度（111111株/hm2）、中密度（55555株/hm2）和低密度（27778株/hm2）。 各密度设3个施肥水平(复合肥,N-P2O5-K2O为15-15-15): 不施肥、低肥（60 kg/hm2）和高肥（120 kg/hm2）。结果表明,黄花蒿对密度和养分条件变化的适应性较强,其中密度是植株大小、生物量分配和产量有关参数的主要决定因子,而青蒿素含量由施肥水平决定。相同施肥水平下,黄花蒿的基径与分枝数均随密度的降低而显著增大,其单株生物量也随密度的降低而显著增大; 中、高密度黄花蒿的支持结构生物量分数均显著大于低密度,低密度黄花蒿的根生物量分数和根/冠比显著大于中、高密度。相同密度下,施肥水平对黄花蒿的单株生物量影响不显著,但显著影响其生物量分配。低密度下,黄花蒿根生物量分数和根/冠比随施肥水平的增高而显著降低; 高密度下,黄花蒿叶生物量分数随施肥水平的增高而显著增大。所有处理中,低密度低施肥水平黄花蒿的青蒿素含量最高,中密度低施肥水平的叶产量和青蒿素产量最大。本试验条件下,黄花蒿栽培以密度55555株/hm2、 施肥60 kg/hm2为宜。     

Synergistic effect of Glomus mosseae and nitrogen fixing Bacillus subtilis strain Daz26 on artemisinin content in Artemisia annua L.

Artemisia annua L. (Asteraceae) is an important medicinal plant whose secondary metabolite artemisinin is used for the treatment of cerebral malaria. A study was undertaken to determine the effect of arbuscular mycorrhizal (AM) fungi, Glomus mosseae, Glomus aggregatum, Glomus fasciculatum, Glomus intraradices and two free living nitrogen fixing bacteria (NFB) (identified as Bacillus subtilis and Stenotrophomonas spp. through 16S rRNA gene sequence analysis), inoculated alone or in combinations on the biomass, nutrient uptake, and content of artemisinin in A. annua under glass house conditions. Various parameters like plant height, total plant biomass, NPK content, leaf yield, mycorrhizal colonization, bacterial population in rhizosphere, phosphatase activity and artemisinin content were determined and found to vary with different treatments. Among all the treatments, plants inoculated with G. mosseae + B. subtilis performed better than any other treatment or uninoculated control plants. The results of the experiment clearly indicated the compatibility and synergy between G. mosseae and B. subtilis and suggested the use of this microbial consortium in A. annua for enhancing growth, biomass yield, and the content and yield of artemisinin.     

低温处理对青蒿叶片脯氨酸、SOD和CAT的含量及蛋白质的影响

青蒿是一种重要的抗痢疾药物的原材料,低温是影响青蒿的青蒿素含量的重要因素。本试验以2个不同产地的青蒿为材料,研究低温处理对青蒿叶片脯氨酸、SOD和CAT的含量及蛋白质的影响。结果表明,低温处理条件下,青蒿叶片脯氨酸、超氧化物歧化酶活性(SOD)和过氧化氢酶活性(CAT)与室温相比均增加,且差异显著。SOD含量的增加,会增强青蒿植株抵抗低温的能力;CAT含量的增加,有利于缓解过量的过氧化物对青蒿叶片细胞的伤害;脯氨酸含量的增加,有利于调节细胞渗透压,促使细胞正常生长。进一步研究表明,在低温和室温条件下,青蒿叶片蛋白质电泳图谱存在强弱差异条带,说明低温处理对青蒿叶片中蛋白质产生影响。本文结果将为今后青蒿的栽培生产提供有价值的参考。     

黄蒿和大籽蒿与两个园林小菊砧穗组合对生长和开花的影响

为了探究黄蒿(Artemisia annua)和大籽蒿(Artemisia sieversianae)作为砧木嫁接菊花的效果,拓展菊花嫁接的砧穗组合,本研究以园林小菊钟山粉韵和钟山紫辰为接穗,黄蒿和大籽蒿为砧木,比较4种砧穗组合嫁接对嫁接株的成活和发病、生长特性(冠幅、冠丛周径、砧木茎粗、最大根长、接穗干重、根干重、总...     

模拟增温和增雨对冷蒿营养元素分配的影响

通过野外控制温度和降雨条件模拟气候变化,设计4种试验处理,研究对照（CK）、增温（＋2℃,IR）、增雨（＋ 120mm,W）和增温＋增雨（IRW）处理对冷蒿（Artemisia frigida Willd.）不同部位（生殖枝与营养枝）营养元素吸收和分配的影响.结果表明：增温、增雨及其耦合作用对冷蒿不同部位营养元素的吸收和分配存在显著差异（P＜0.05）;与对照相比,增温效果不显著,而增雨显著影响冷蒿对N、K、Mg、Zn、Fe、Mn、Ni元素的吸收和分配（P＜0.05）;冷蒿不同部位对N、K、Mn吸收与分配比例存在显著差异（P＜0.05）,但是增温、增雨与不同部位之间无明显耦合作用.说明长期增温与增雨通过改变土壤水分、温度、养分状况和植物内在生长节律,直接或间接影响植物对营养元素的吸收和分配过程.     

Nutrient deficiency and physiological disease of lowland rice in Ceylon

In Ceylon, rice crop is grown chiefly on alluvial as well as low-humic gley soils which are distributed throughout the island. Parent materials of these soils are derived mainly from the weathered products of Khondalite and Charnackite groups of rocks among which acidic schists and gneisses dominate (1). The lateritic weathering of rocks, the most typical lOil formation under tropical climates, results in a general supply of soil materials rich in iron and alumina but lacking in nutrient bases and silica. This is one of the possible reasons for the low fertility of tropical rice soils compared to those of temperate zones.     

Nutrient deficiency symptoms and boron uptake mechanisms of rice

There is little agreement on the leaf and shoot boron (B) requirement of rice (Oryza saliva L.) and the optimum hydroponic solution B concentration for rice. Questions on the mechanism of B uptake active or passive are also unresolved. We grew rice in hydroponic solutions in a growth chamber for six weeks with B at 0, 0.05, 0.2, 1, 5, 10, 25, and 50 μM. Transpirational flow, dry matter yields and tissue B were determined. Results indicated that B deficiency occurred when there was <7.3 mg kg^‐1 B in the flag leaves, <3.6 mg kg^‐1 B in shoots, and <0.2 μM B in the nutrient solutions. Boron additions increased dry matter and plant height. Typical B‐deficiency symptoms were a light color or chlorosis on almost all of the youngest leaves and stems, an unthriftiness, leaf tip burn, and pale bands 2–3 mm wide on leaves, particularly on the moderately B‐deficient plants. Whitish and twisted new leaf tips occurred at >0.05 μM B. A mass balance analysis that compared the total mass of B in the plant versus B provided via transpirational flow showed that at high hydroponic B supply, passive uptake and active excretion of B, or, active blockage of B may have occurred, for > 10‐fold differences existed between the B mass in plant and B mass that could be provided via transpirational flow. Thus, B uptake was against a concentration gradient and high B supply, and as other evidence indicated, at low B supply.     

转青蒿反义鲨烯合酶基因对烟草鲨烯合酶基因表达的影响

利用反义技术，通过根癌农杆菌（Agrobacterium tumefaciens）介导，将青蒿（Artemisia annua）的反义鲨烯合酶基因导入烟草（Nicotiana tabacum）株系W38中，存转基因烟草中，鲨烯合酶的表达成功地得到抑制，其mRNA的水平降低。对鲨烯合酶的下游产物之一胆同醇的检测显示，转基因烟草的胆固醇含量低于对照，同时以法呢基焦磷酸为前体的二萜支路产物之一的GA，含量得到提高。实验结果表明，反义抑制烟草鲨烯合酶可以降低植物甾醇的生物合成，并促进类异戊二烯途径中以法呢基焦磷酸为前体的其它代谢支路的生物合成。     

Determination of the chemical composition and antioxidant activity of the essential oil of Artemisia dracunculus and of the antifungal and antibacterial activities of Turkish Artemisia absinthium, A. dracunculus, Artemisia santonicum, and Artemisia spicigera essential oils

The essential oil isolated from Turkish tarragon (Artemisia dracunculus) by hydrodistillation was analyzed by GC-MS. Thirty compounds representing 99.5% of total oil were identified. The predominant components in the oil were (Z)-anethole (81.0%), (Z)-beta-ocimene (6.5%), (E)-beta-ocimene (3.1%), limonene (3.1%), and methyleugenol (1.8%). The antibacterial and antifungal activities of the essential oils isolated from A. dracunculus, Artemisia absinthium, Artemisia santonicum, and Artemisia spicigera oils were also evaluated. In general, the oils exhibited potent antifungal activity at a wide spectrum on the growth of agricultural pathogenic fungi. Among the oils, the weakest antifungal activity was shown by the oil of A. dracunculus. In many cases, the oils of A. absinthium, A. santonicum, and A. spicigera completely inhibited the growth of some fungal species. As compared with antibacterial activities of all of tested oils, A. santonicum and A. spicigera oils showed antibacterial activities over a very wide spectrum. However, the essential oils tested showed lower inhibition zones than the inhibition zones of penicillin. In addition, antioxidant and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activities of tarragon oil were determined, and weak antioxidant and DPPH radical scavenging activities were found in comparison to butylated hydroxytoluene.     

Enhancement of acid phosphatase activity in Capsicum annuum L. plants in response to phosphate deficiency

This study examined acid phosphatase activity in the extracts of Capsicum annuum L. cv. ”︁Sweet Banana” seedlings grown axenically on water-agar medium or plants grown in a hydroponic system with or without phosphate. Initially, no elevated phosphatase activity was detected in the root surface, in root surface extractions or cell-free tissue extracts from plants that were not showing morphological symptoms of stress. Then elevated specific phosphatase activity was evident in all organ tissue extracts of the plants that showed signs of growth inhibition. The increase in specific activity in the cell free extracts of different organs appeared to be primarily the result of increased activity of the dominant isozyme present in all these extracts, instead of due to de novo synthesis of new isozymes. These and other experiments indicate the lack of phosphate starvation-inducible (psi) excreted phosphatase in Capsicum annuum L. cv. ”︁Sweet Banana”. Some limitations with the use of XP, 5-bromo-4-chloro-indolyl phosphate p-toluidine, particularly when incorporated into growth media and buffer with or without phosphate for the study of phosphate starvation-inducible excreted phosphatase activity were noted.     

Nutrient deficiency diagnosis of corn (Zea Mays L.) plants from yield and composition responses to additions of N and S

Abstract

Corn (Zea mays L.) was grown in the greenhouse in a Laughlin (Ultic Haploxeroll) loam soil, with various amounts of N and S added in order to determine possible interactions of these nutrients with the relationship between plant composition and grain yields. Previous field experience and preliminary experiments had shown that this soil gave yield responses to N and S additions.

Regression equations were used to describe the relationship between composition of various plant parts and grain yields. The best correlation (R² = 0.943) was obtained using the total N concentration in leaf samples taken at the silking stage, but excluding data from plants which, based on their amide N concentrations (greater than 500 ppm), were considered S deficient. Calculations using the first derivative of the cubic polynomial indicated that a concentration of 2.5% N in the lower leaves was necessary in order to obtain maximum grain yields. The concentrations of total N in the upper leaves and the stalks at the silking stage also correlated well with the grain yields. The relationship of NO₃‐N in the stalks at silking to grain yields could be better described mathematically with an exponential function, but the correlation coefficient was low (R² = 0.58). The responses of two genotypes, one containing the opaque ‐ 2 gene, the other its normal counterpart, were similar.

The total N concentration in the leaves collected at the tassel stage did not correlate quite as well with grain yield as those collected later, but using the exponential model NO₃‐N concentrations in the stalks at the earlier stage showed a closer relationship to grain yield than for samples collected at silking. Excluding data for the plants showing S deficiency, a correlation coefficient of 0.90 was obtained. At both tasseling and silking stages, the S deficient plants were characterized by high N:S ratios, with values of 18 to 50 for the stalks, compared to values of less than 10 for the S adequate plants. The marked effect of inadequate S on grain formation was not evident in the amounts of leaves and stalks produced.

Field studies will be necessary to evaluate further the merit of the diagnostic procedure indicated by these experiments.     

Rise and fall of Stachys annua (L.) L. in the Carpathian Basin: a historical review and prospects for its revival

Genetic Resources and Crop Evolution - Stachys annua (L.) L., a melliferous archaeophyte plant became a dominant weed of the cereal stubbles of the Carpathian Basin in the medieval three-field...     

Improvement in Productivity,Water-Use Efficiency,and Soil Nutrient Dynamics of Summer Peanut (Arachis hypogaea L.) through Use of Polythene Mulch,Hydrogel, and Nutrient Management

Agronomic management through better use of inputs benefits farmers both by enhancing productivity and profitability. A field experiment was conducted in consecutive summer seasons (2011–2013) consisting of two mulching (no mulch, polythene mulch), three hydrogel (0, 2.5, 5.0 kg ha^?1), and three nutrient management treatments (organic, inorganic, and integrated) in a split–split plot design. Use of mulching and 2.5 kg hydrogel ha^?1 and integrated nutrient management enhanced pod, haulm, kernel and oil yields, and net economic returns. Partial factor productivity and water-use efficiency were higher under polythene mulch and 5.0 kg hydrogel ha^?1. Higher nutrient uptakes were obtained under both mulching and integrated nutrient management. Use of 2.5 kg hydrogel ha^?1 resulted in more removal of N; P and K uptakes were higher in 5.0 kg hydrogel ha^?1. Combination of three managements had a consequence of actual soil N loss, but gains in soil P and K after three cropping cycles.     

Nutrient dynamics,dehydrogenase activity,and response of the rice plant to fertilization sources in an acid lateritic soil

Abstract

Rice variety IR 36, grown under flooding, was studied in 1998 to determine the effects of fly ash, organic, and inorganic fertilizers on changes in pH and organic carbon, release of nutrients (NH₄ ⁺-N, Bray's P, and NH₄OAc K), and dehydrogenase activity in an acid lateritic soil at 15-day intervals. Application of fly ash at 10?t?ha^?1 alone did not improve the availability of NH₄ ⁺-N, or P, as well as the rice grain yield. Availability of NH₄ ⁺-N (35.3–36.9?mg?kg^?1), and P (12.3–14.6?mg?kg^?1) at 15 days after transplanting, and rice grain yields (48.0–51.7?g per pot) were similar under the various fertilization sources such as inorganic fertilizer alone, inorganic fertilizer?+?fly ash or inorganic fertilizer?+?green manure?+?fly ash. Mean dehydrogenase activity was the highest (8.47?µg triphenyl formazon g^?1 24?h^?1) under the mixed fertilization treatments with green manure. At the end of the cropping season (75 days after transplanting), pH, organic carbon, and dehydrogenase activity were higher under the mixed fertilization treatments involving green manure by 3, 15 and 154%, respectively, compared with the inorganic fertilizer alone.     

Nutrient management in low input grazing-based systems of meat production

Abstract. Low input grazing-based meat production is often practised on land where alternative land use is restricted and where high biodiversity warrants specific environmental payments. Although these systems are characterized by low external nutrient inputs, significant nutrient losses may occur per unit of produce, even though losses per unit area are low. If the aim is to have animal production without further eutrophication of watercourses, these losses should be minimized,. However, if nutrient-poor grasslands characterized by high biodiversity are the aim, some losses must be accepted to balance nutrient inputs by atmospheric deposition. The impact of grazing animals on nutrient dynamics in grasslands is reviewed in this paper, and options to further improve nutrient management are discussed. It is concluded that nutrient management should largely be addressed indirectly through management of animal behaviour in relation to grazing and supplemental feeding, as well as through optimization of the seasonal distribution of livestock size. A case study indicated that N losses from a cattle farm could be reduced considerably by these means, while animal production could actually be improved. Sustainability issues and trends in legislative and social pressures on low input grazing-based systems of animal production are discussed in relation to nutrient management and practical recommendations.     

Nutrient and carbon balances in organic vegetable production on an irrigated,sandy soil in northern Oman

Our understanding of nutrient and carbon (C) fluxes in irrigated organic cropping systems of subtropical regions is limited. Therefore, leaching of mineral nitrogen (N) and phosphorus (P), gaseous emissions of NH₃, N₂O, CO₂, and CH₄, and total matter balances were measured over 24 months comprising a total cropping period of 260 d in an organic‐cropping‐systems experiment near Sohar (Oman). The experiment on an irrigated sandy soil with four replications comprised two manure types (ORG1 and ORG2) characterized by respective C : N ratios of 19 and 25 and neutral detergent fiber (NDF)‐to‐soluble carbohydrates (SC) ratios of 17 and 108. A mineral‐fertilizer (MIN) treatment with equivalent levels of mineral N, P, and potassium (K) served as a control. The three treatments were factorially combined with a cropping sequence comprising radish (Raphanus sativus L.) followed by cauliflower (Brassica oleracea L. var. botrytis) or carrot (Daucus carota subsp. sativus). Over the 24‐months experimental period gaseous N emissions averaged 45 kg ha^–1 (59% NH₃‐N, 41%N₂O‐N) for MIN, 55 kg N ha^–1 (69% NH₃‐N, 31%N₂O‐N) for ORG1, and 49 kg N ha^–1 (59% NH₃‐N, 41% N₂O‐N) for ORG2. Carbon losses were 6.2 t ha^–1 (98% CO₂‐C, 2% CH₄‐C) for MIN, 9.7 t C ha^–1 (99% CO₂‐C, 1% CH₄‐C) for ORG1, and 10.6 t ha^–1 (98% CO₂‐C, 2% CH₄‐C) for ORG2. Exchange resin–based cumulative leaching of mineral N amounted to 30 kg ha^–1 for MIN, 10 kg ha^–1 for ORG1, and 56 kg ha^–1 for ORG2. Apparent surpluses of 361 kg N ha^–1 and 196 kg P ha^–1 for radish‐carrot and 299 kg N ha^–1 and 184 kg P ha^–1 for radish‐cauliflower were accompanied by K deficits of –59 kg ha^–1 and –73 kg ha^–1, respectively, for both cropping systems. Net C balances for MIN, ORG1, and ORG2 plots were –7.3, –3.1, and 1.5 t C ha^–1 for radish‐carrot and –5.0, 1.3, and 4.6 t C ha^–1 for radish‐cauliflower. The results underline the difficulty to maintain soil C levels in intensively cultivated, irrigated subtropical soils.     

Integrated Nutrient Management Affects Fruit Yield of Sapota (Achras zapota L.) and Nutrient Availability in a Vertisol

ABSTRACT

We studied the effect of integrated nutrient management (INM) combinations on supplement of plant nutrient for quantitative and qualitative fruit production in sapota. Thus, 17 combinations of INM practices were evaluated on fruit yield of sapota and nutrient availability in a Vertisol of Chambal region, India. The results demonstrated that almost all treatment combinations comprised of recommended dose of fertilizer (RDF), i.e. 1,000:500:500 g NPK plant^?1 with application of organic and inorganic sources of nutrients had a significant effect on the fruit yield of sapota, soil microbial biomass, NPK content of leaf, fruit and soil over control (T₁). Among different treatments, application of 2/3rd part of RDF + 50 kg FYM + 250 g Azospirillum + 250 g Azotobacter plant^?1 (T₁₁) significantly enhanced the number of fruits plant^?1 (327.88), yield plant^?1 (29.03 kg) and yield ha^?1 (4.52 t). However, the soil microbial count of fungi (8.89 cfu g^?1 soil), bacteria (11.19 cfu g^?1 soil) and actinomycetes (5.60 cfu g^?1 soil) at fruit harvest was higher under the 2/3 of RDF +10 kg vermicompost + 250 g Azospirillum + 250 g Azotobacter plant^?1 (T₁₅). The leaf nitrogen content (N, 2.03%) was higher in T_15, while phosphorus (P, 0.28%) and potassium (K, 1.80%) content were higher in T₁₁. It is evident that treatment T₁₁ increased fruit yield by 32% in Sapota cv. Kalipatti compared to control. Therefore, combined application of nutrient sources proved not only beneficial for enhancing fruit yield of sapota but also sustaining soil health in Chambal region of south-eastern Rajasthan.     

Nutrient solution concentration and collection time in phytomass production,content, yield and chemical composition of essential oil of rosemary

Rosmarinus officinalis L. (rosemary), is a medicinal and aromatic species used in the cosmetics and food processing industries. The present study aimed to evaluate the biomass production, content, yield and chemical composition of the essential oil of rosemary in cultivation out of soil with different concentrations of nutrient solution in both harvests. The cultivation of rosemary was conducted in a protected system off the ground, with concentrations of nutrient solutions of 1, 2, 3, 4 and 5 dS.m^?1 for fertigation with collections of leaves 100 and 160 days after planting. Nutrient solution concentration influences phytomass production and the content and yield of essential oil of rosemary, though not its chemical composition. The collection time does not interfere in the content of essential oil; however, does interfere in the chemical composition; the production of phytomass and the yield of oil per plant are higher in collections at 160 days after planting.     

Antifungal constituents of the essential oil fraction of Artemisia dracunculus L. Var. dracunculus

The isolation and structure elucidation of antifungal constituents of the steam-distilled essential oil fraction of Artemisia dracunculus are described. Antifungal activities of 5-phenyl-1,3-pentadiyne and capillarin against Colletrotichum fragariae, Colletrotichum gloeosporioides, and Colletrotichum acutatum are reported for the first time. The relative abundance of 5-phenyl-1,3-pentadiyne is about 11% of the steam-distilled oil, as determined by GC-MS. Methyleugenol was also isolated and identified as an antifungal constituent of the oil.