Genetic analyses of heading date of Japonica rice cultivars from Northeast China

Northeast regions of China (38–55°N latitude) play an important role in Japonica rice planting. Heading dates of 10 Japonica rice cultivars native to the Northeast of China were investigated and their sensitivities to photoperiod and temperature were analyzed. The results showed that these Japonica rice cultivars were insensitive to photoperiod but strongly sensitive to temperature, i.e. a high temperature can markedly shorten the heading date. Genetic analyses were conducted on these 10 cultivars using a set of heading date tester lines. All these Japonica cultivars carried a dominant early heading gene Ef-1, and most of these cultivars carried the photoperiod insensitivity allele e₁, and two types of alleles were presented at the Se-1 locus, including recessive photoperiod insensitivity gene Se-1^e and dominant strong photoperiod-sensitivity (PS) gene Se-1ⁿ. The PS of these cultivars carrying E₁ or Se-1ⁿ can be repressed or weakened by Ef-1 and the recessive allele hd2 they carried. These results provided a reasonable explanation to the adaptability of Japonica rice to the high latitude environment of the Northeast China, and could be useful for breeding new cultivars well adapted to the high latitude regions and expanding the rice cultivation range.     

Activities of essential oils from Asarum heterotropoides var. mandshuricum against five phytopathogens

Some secondary metabolites of plants function as antimicrobial products against phytopathogens and constitute an increasingly important class of pesticides. In the present study, the essential oil of Asarum heterotropoides var. mandshuricum was analyzed by GC/MS and its antimicrobial activity was evaluated against five phytopathogenic fungi. Major components of the oil were methyleugenol (59.42%), eucarvone (24.10%), 5-allyl-1,2,3-trimethoxybenzene (5.72%), and 3,7,7-trimethylbicyclo(4.1.0)hept-3-ene (4.93%). The essential oil and the most abundant component, methyleugenol, were separately assayed for inhibition of 5 pathogens: Alternaria humicola, Colletotrichum gloeosporioides, Rhizoctonia solani, Phytophthora cactorum and Fusarium solani. Both the oil and methyleugenol strongly inhibited the growth of the test pathogens (IC₅₀ values <0.42 μg ml⁻¹) except F. solani, with the best activity against P. cactorum (IC₅₀ values = 0.073 and 0.052 μg ml⁻¹, respectively). It is concluded that the essential oil of A. heterotropoides var. mandshuricum has a broad antiphytopathogenic spectrum, and that methyleugenol is largely responsible for the bioactivity of the oil. The mode of action of methyleugenol against P. cactorum is discussed based on changes in the mycelial ultrastructure.     

Highly efficient in vitro regeneration of the industrial oilseed crop Crambe abyssinica

A highly efficient regeneration protocol for oilseed crop Crambe abyssinica has been developed using hypocotyls as explants in this study. Crambe is a potential engineering oilseed crop for industrial purposes as it contains 55-60% erucic acid in its oil and, more importantly, it does not outcross with any food oil seed crops. However, the low regeneration frequency with the currently available protocols is still a limiting factor for genetic modification of Crambe. In this study, we investigated the effects of N-source, C-source, AgNO₃, cultural conditions as well as the concentration and combination of plant growth regulators (PGR) on the regeneration frequency of C. abyssinica. The results showed that all these factors, especially the N-source and PGR concentrations and combinations, played an important role in shoot regeneration. Among all the factors tested, the combination of using hypocotyls from C. abyssinica cv. galactica, the Lepiovre basal medium supplemented with 16 g l⁻¹ glucose, 0.5 g l⁻¹ AgNO₃, 2.2 mg l⁻¹ thidiazuron (TDZ), 0.5 mg l⁻¹ α-naphthaleneacetic acid (NAA), 2.5 g l⁻¹ Gelrite, seeds germinated in dark for 3 days and explants cultured in light, gave the best regeneration frequency (over 95%). The results also suggest that reducing the content of NH₄⁺ or keeping a suitable NO₃⁻/NH₄⁺ ratio in the regeneration medium would be crucial to Crambe shoot regeneration.     

Screening of three sweet potato (Ipomoea batatas L.) cultivars for resistance to different virulence groups of root-knot nematodes (Meloidogyne spp.) under controlled conditions

The susceptibility of three sweet potato cultivars (Ipomoea batatas L.) C4, TIS 3290 and TIS 9162 was evaluated against 156 isolates of Meloidogyne spp. with the aim to include resistant/tolerant sweet potato cultivars in a crop rotation scheme for the management of root-knot nematodes. The nematode isolates corresponded to races 1, 2 and 3 of Meloidogyne arenaria (n = 7), races 1, 2, 3 and 4 of M. incognita (n = 131) and Meloidogyne javanica (n = 18). Also, the isolates of M. incognita were differentiated in virulence groups: Pepper (n = 35), Pepper-Mi (n = 25), Tomato (n = 41) and Tomato-Mi (n = 30), depending on their ability to parasitize resistant pepper and tomato cultivars. The tested isolates of M. javanica parasitized C4 and TIS 3290, but not TIS 9162, whereas M. arenaria parasitized C4 and TIS 9162, but not TIS 3290, and M. incognita was able to parasitize the three sweet potato cultivars tested. C4 was the most susceptible cultivar to all nematode species tested, especially M. incognita, TIS 3290 was the most resistant and TIS 9162 was in between (7.2, 62.9 and 26.9% of resistant plants, respectively). Susceptibility of the sweet potato cultivars showed slight variations depending on the race or virulence group of M. incognita. The results suggest that sweet potato cultivars TIS 3290 and TIS 9162 may be used as rotation crops in fields where root-knot nematodes are present, their selection depending on the Meloidogyne isolates present. The use of resistant sweet potato cultivars would be preferably combined with other management practices to avoid virulence selection in nematode isolates.     

Source/sink ratio and the relationship between maximum water content,maximum volume,and final dry weight of maize kernels

Final kernel weight (KW) is closely related to maximum kernel volume (KV) and maximum kernel water content (KWC). It is not clear, however, how changes in the reproductive sink capacity, assimilate availability during grain filling and physical restriction to kernel expansion affect the relationship between KW and KWC or between KW and KV. Three experiments were conducted at Balcarce, Argentina and Ames, USA. Defoliation, thinning, plant density, restricted pollination and volume restriction treatments were imposed to manipulate KV, sink and source capacity. KW varied from 111 to 436 mg across all hybrid–treatments combinations and was related to the source/sink ratio during grain filling (r² = 0.85). Treatment variation in KW was related primarily to changes in kernel growth rate, except for the complete defoliated treatment, which also shortened the duration of grain filling. KW was correlated with maximum KWC (r² = 0.77, p < 0.001) and with maximum KV (r² = 0.91, p < 0.001). The developmental patterns for KW, KWC and KV during grain filling were not affected by the source/sink manipulations, except for severe defoliation. In the latter case, maximum KWC was not a good estimate of final KW. KV, however, was sensitive to reductions in carbohydrate supply during grain filling and was closely correlated to KW. Physical restriction to kernel expansion reduced kernel weight 13% relative to its control (p < 0.01). But restricting kernel expansion did not alter the general relationships between KW and KWC or between KW and KV, because kernel density was not affected.     

Flowering time in oat: Genotype characterization for photoperiod and vernalization response

Flowering time is an important component in the adaptation of oat to sub-tropical environments. Genotypes differ in their response to photoperiod and to vernalization. The objectives of this study were to identify photoperiod insensitive oat genotypes and then investigate the response of different oat genotypes to a period of vernalization (cold treatment), and to evaluate F₆ recombinant inbred lines from two oat crosses in two different environments (day-length increases and day-length decreases). The genotypes used in the studies were from the UFRGS Oat Breeding Program: UFRGS 8, UFRGS 881971 and UFRGS 930605 and from the University of Minnesota, USA: Amagalon/*Ogle, Coker 492/Starter-1 and PC68/5*Starter, and F₆ lines from the crosses UFRGS 8 × UFRGS 930605 and UFRGS 8 × PC68/5*Starter. UFRGS 8 showed no photoperiod sensitivity by flowering early regardless of the photoperiod length. Late flowering in the winter and early flowering in the summer characterize day-length-dependent genotypes like PC68/5*Starter. UFRGS 881971 responded to vernalization. Genotypes showed variability that can be used in the selection of lines/varieties that more effectively use the complete season.     

Environmental effects on seed yield determination of irrigated peanut crops: Links with radiation use efficiency and crop growth rate

In Argentina, delayed sowing causes a decrease in seed yield and in radiation use efficiency (RUE) of peanut crops (Arachis hypogaea L.), but it is not known if RUE reduction is mainly due to reduced temperature during late reproductive stages or to a sink limitation promoted by decreased seed number in these conditions. We analyzed seed yield determination and RUE dynamics of two cultivars (Florman and ASEM) in four irrigated field experiments (Exp_n) grown at three sites and five contrasting sowing dates (between 17 October and 21 December) in three growing seasons. An additional field experiment was performed with widely spaced plants (i.e. with no interference among them) to evaluate the effect of peg removal on RUE and leaf carbon exchange rate (CER). Seasonal dynamics of mean air temperature and irradiance, biomass production (total and pods), and intercepted photosynthetically active radiation (IPAR) were followed. Seed yield and seed yield components (pod number, seeds per pod, seed number and seed weight) were determined at final harvest. Crop growth rate (CGR) and pod growth rate (PGR) were computed for growth phases of interest. RUE values for crops sown until 14 November were 1.89–1.98 g MJ⁻¹ IPAR, within the usual range. RUE decreased significantly for cv. Florman in the late sowing of Exp₁ (29 November) and for both cultivars in Exp₃ (21 December sowing). Across experiments, seed yield (4.5-fold variation relative to minimum) was strongly associated (r² = 0.87, P < 0.0001) with variations in seed number (3.5-fold variation relative to minimum), and to a lesser extent (r² ≤ 0.54, P ≤ 0.001) to variations in seed weight (1.9-fold variation relative to minimum). Seed number was positively related (P < 0.01) to CGR (r² = 0.66) and to PGR (r² = 0.72) during the R₃–R_6.5 phase (seed number determination window), while crop growth during the grain-filling phase (i.e. between R_6.5 and final harvest) was positively associated with grain number (r² = 0.80, P < 0.001). No association was found between RUE and mean air temperature, neither for the whole cycle nor for the phase between R_6.5 and final harvest, which showed the largest temperature variation (16.4–22.4 °C) across experiments. Use of mean minimum temperature records (range between 13.8 and 18.5 °C) did no improve the relationship. However, grain-filling phase RUE showed a positive (r² = 0.69, P = 0.003) linear response to seed number across experiments. This apparent sink limitation of source activity was consistent with the reduced RUE (from 2.73 to 1.42 g MJ⁻¹ IPAR) and reduced leaf CER at high irradiance (from ca. 30 to 15 μmol m⁻² s⁻¹) for plants subjected to 75% peg removal.     

High level of resistance to Sclerotinia sclerotiorum in introgression lines derived from hybridization between wild crucifers and the crop Brassica species B. napus and B. juncea

Sclerotinia rot caused by the fungus Sclerotinia sclerotiorum is one of the most serious and damaging diseases of oilseed rape and there is keen worldwide interest to identify Brassica genotypes with resistance to this pathogen. Complete resistance against this pathogen has not been reported in the field, with only partial resistance being observed in some Brassica genotypes. Introgression lines were developed following hybridization of three wild crucifers (viz. Erucastrum cardaminoides, Diplotaxis tenuisiliqua and E. abyssinicum) with B. napus or B. juncea. Their resistance responses were characterized by using a stem inoculation test. Seed of 54 lines of B. napus and B. juncea obtained from Australia, India and China through an Australian Centre for International Agricultural Research (ACIAR) collaboration programme were used as susceptible check comparisons. Introgression lines derived from E. cardaminoides, D. tenuisiliqua and E. abyssinicum had much higher levels (P < 0.001) of resistance compared with the ACIAR germplasm. Median values of stem lesion length of introgression lines derived from the wild species were 1.2, 1.7 and 2.0 cm, respectively, as compared with the ACIAR germplasm where the median value for stem lesion length was 8.7 cm. This is the first report of high levels of resistance against S. sclerotiorum in introgression lines derived from E. cardaminoides, D. tenuisiliqua and E. abyssinicum. The novel sources of resistance identified in this study are a highly valuable resource that can be used in oilseed Brassica breeding programmes to enhance resistance in future B. napus and B. juncea cultivars against Sclerotinia stem rot.     

Incorporating vernalization response functions into an additive phenological model for reanalysis of the flowering data of annual pasture legumes

Vernalization is a physiological process by which plants acquire competence to flower by prolonged exposure to cooling temperatures. A non-linear vernalization module was incorporated into the thermal and photothermal additive models for modelling the time to flowering. The model was applied to published data of time to flowering for annual pasture legumes. Fourteen out of 15 cultivars were detected as vernalization-requiring genotypes. All vernalization-requiring cultivars fitted to the thermal model had R² of below 0.51. After incorporating a vernalization module, the model (vernalizing-thermal model) fitted to nine cultivars with R² of over 0.9. All cultivars were well fitted to the photothermal model with an average R² of 0.94. However, the accountability of variance in observed time to flowering dropped from 94% when the model was tested without cross validation to 68% when the model was tested with cross validation, indicating a poor predictability of the model. Further, the values of parameters for the photothermal model, such as base temperature and coefficients associated with the effect of temperature on the rate of development are unrealistic. The vernalizing-photothermal model not only provided a good fit to the data (averaged R² = 0.98), but it also was able to account for 94% of the variance in the observed data when the model was tested by cross validation. The vernalizing-photothermal model produced meaningful base temperature and the coefficients for the effects of temperature and photoperiod were positive, which are consistent with the suboptimum temperature promotion of flowering and quantitative long day plant behaviour. The vernalizing-thermal model and vernalizing-photothermal model gave an averaged root mean squared error (RMSE) of 3.6 and 2.3 days, compared with the averaged RMSE of 32.4 and 5.9 days for the two respective additive models without the vernalization component. It was concluded that if the vernalization occurs, the additive models without the vernalization module can confound the effects of vernalization that hastens flowering at low temperature with the effects of temperature that promotes flowering at a suboptimum temperature regime. The confounded effects may be forced to be expressed as the unrealistic effects of temperature and base temperature in the model parameters and the model overall has low predictability. The modified additive model that incorporating the vernalization module can be used to quantitatively measure the length of vernalization and is a more valid approach to modelling the time to flowering.     

Effects of temperature and photoperiod on flowering time of forage legumes in a Mediterranean environment

Flowering time plasticity is a commonly occurring adaptive characteristic of fodder crops, including legumes, in arid and semiarid environments of the Mediterranean regions. Time of flowering is mainly influenced by genotype, temperature and photoperiod. Field experiments were carried out at Foggia (southern Italy) during successive growing seasons (from 8 to 16 growing cycles according to species) to study the relation among air temperature, photoperiod and duration of the morphological development of flowering in eight forage legume species: sulla (Hedysarum coronarium L.), sainfoin (Onobrychis viciifolia Scop.), pea (Pisum sativun L.), berseem clover (Trifolium alexandrinum L.), Persian clover (Trifolium resupinatum L.), faba bean (Vicia faba L.), common vetch (Vicia sativa L.) and hairy vetch (Vicia villosa Roth). Time to reach 10% flowering (EF) and 100% flowering (FF) were recorded. Rate of progress to flowering, defined as the inverse of time from sowing to EF and FF, was related to mean daily temperature, or to both mean daily temperature and mean photoperiod. Using the linear equations, the thermal time requirements (T_t) and the base temperature (T_b) expressed as heat units were determined by the x-intercept method for both EF and FF stages. Evaluation of flowering time was also based on days after planting (DAP), day of year (DOY) and on a photothermal index (PTI). For all species, a significant negative correlation (P ≥ 0.01) was found between planting date (PD) and DAP whereas PTI showed a significant negative relationship (P ≥ 0.05) only for faba bean, pea, berseem clover and common vetch. In sainfoin, sulla and berseem clover, the rate of progress to flowering was affected significantly (P ≥ 0.05) by both mean temperature and photoperiod. The T_t requirements to reach the EF and the FF stage ranged from 871 to 1665 °C day and from 1043 to 1616 °C day, respectively, for the studied species. Both phenological stages considered depended upon accumulated thermal time above a species-specific base temperature. Furthermore, in all legumes the onset of flowering only occurred when dual thresholds of a minimum T_t and a minimum photoperiod were reached, which were specific to each species.     

Antioxidant, antibacterial, and antiviral effects of Lactuca sativa extracts

Antioxidant, antibacterial and antiviral effects of aqueous and methanol extracts of Lactuca sativa var longifolia leaves were investigated. The antioxidant activity was evaluated using the DPPH assay. The effect of the extracts against 5 Gram-positive and 6 Gram-negative bacteria was tested. The antiviral activity was determined against human cytomegalovirus (HCMV) strain AD-169 (ATCC Ref. VR 538) and coxsackie B virus type 3 (CoxB-3) using a cytopathic effect (CPE) reduction assay. The methanol extract had the highest total phenolic contents (235.31 mg CE/g extract). It exhibited a significantly (p < 0.05) greater hydroxyl radical-scavenging activity (IC₅₀ = 3.5 μg/ml) than the aqueous extract (4.1 μg/ml). It was also the most effective extract with the lowest MIC (2.5 mg/ml) against all Gram negative and Gram positive bacteria. Methanol and aqueous extracts exhibited antiviral activity against HCMV and Cox-B3 viruses with IC₅₀ of 200 μg/ml.     

Comparative assessment of antioxidant and cholinesterase inhibitory properties of the marigold extracts from Calendula arvensis L. and Calendula officinalis L.

Calendula sp. is an important medicinal and industrial plant with various bioactivities. In this study, we examined enzyme inhibitory effects of the n-hexane, dichloromethane, acetone, ethyl acetate, methanol, and water extracts of the leaf and flowers of Calendula arvensis L. and C. officinalis L. against acetylcholinesterase (AChE) and butyrylcholinesterase (BChE). The extracts were screened for their antioxidant activity using 2,2-diphenyl-1-picrylhydrazyl (DPPH), ferric ion-chelating capacity and ferric-reducing antioxidant power (FRAP) assays at 250, 500, and 1000 μg mL⁻¹. Total phenol and flavonoid quantification of the extracts was achieved using Folin-Ciocalteau and AlCl₃ reagents, respectively. The ethyl acetate extract of C. arvensis flowers was the most active in AChE inhibition assay (31.24 ± 1.29%), while the n-hexane extract of C. officinalis leaves exerted the highest ferric ion-chelating capacity (74.27 ± 2.25%). Thin layer chromatographic analysis indicated presence of flavonoid and triterpene derivatives mainly in the extracts.     

The effect of different extraction techniques on extraction yield, total phenolic, and anti-radical capacity of extracts from Pinus radiata Bark

The performance of four techniques, conventional maceration, Soxhlet extraction, microwave assisted extraction (MAE), and ultrasound assisted extraction (UAE), for extraction of Pinus radiata bark, in one and several stages, were evaluated. For each technique, the mass extracted (g extract/g bark), total phenols (by Folin-Cicalteau), and tannin (by precipitation) concentration and anti-radical capacity (diphenyl-picrylhydrazyl, DPPH) were quantified. In one stage, the extracted mass increased in the following order: maceration < UAE < MAE < Soxhlet (p < 0.05). The total phenols and tannin levels were also higher with the Soxhlet technique. With additional extraction stages, only the samples produced with MAE and UAE techniques improved their parameters. Additionally, MAE extracts presented a higher anti-radical capacity than does Soxhlet and Pycnogenol^® extracts. Therefore, MAE was a simple and rapid method that was useful for extraction of P. radiata bark. Scanning electron micrographs (SEM) provided evidence of the mechanical effects on cell walls, mainly evidenced by cell destruction produced by Soxhlet, MAE, and UAE on the bark. In contrast, maceration only results in slightly ruptured cell pores, which could explain its low extraction yield.     

Genetic structure of natural Tunisian Hypericum humifusum L. (Hypericacae) populations as assessed by allozymes and RAPDs

The genetic diversity within and among seven Tunisian natural populations of Hypericum humifusum L., from different geographic regions and bioclimates, was assessed using 11 isozymic polymorphic loci, and 166 RAPD markers amplified by 8 primers. The genetic diversity within populations based on allozymes was higher (P = 72.46%, Ap = 2.01 and H_e = 0.29), than that revealed by RAPDs (29.52 < P < 39.16% and 0.150 < H < 0.200). Both markers yielded high estimates of genetic differentiation and low gene flow (Nm = 0.257 and 0.508 for RAPD and allozymes, respectively) among populations at all space scales. However, the level of differentiation revealed by RAPDs (Φ_ST = 0.494; G_ST = 0.561) was higher than that based on allozymes (F_ST = 0.117). No correlation (Mantel test) among genetic (F_ST and Φ_ST matrices) and geographical distance matrices was observed indicating no isolation by distance. Cluster analyses from allozyme and RAPD loci did not completely agree. The dendrogram based on allozymes yielded higher separation among most populations, while that from RAPDs separated populations into three distinct subclusters. Groupings of populations, in both dendrograms, did not reflect spatial geographic or bioclimatic patterns, indicating specific adaptation of populations to local environments. The correlation between matrices of allozyme and RAPD band frequencies was not significant (Mantel test). The dendrogram obtained from combined data yielded similar population groupings to that probed by RAPDs suggesting higher accuracy of these markers. Given the high differentiation among all populations even at a low geographic distance, ex situ conservation should involve extensive seed collection from all populations from all bioclimatic zones. The populations from the upper semi-arid bioclimate exhibiting relatively high level of genetic diversity should be first protected.     

Impact of organic manure and chemical fertilizers on artemisinin content and yield in Artemisia annua L

Artemisinin isolated from the aerial parts of Artemisia annua L. is a promising and potent antimalarial drug. It posses remarkable activity against both chloroquinine resistant as well as chloroquinine sensitive strains of Plasmodium falciparum. It is also useful in the treatment of cerebral malaria. The relatively low content of artemisinin in A. annua and unavailability of cost effective and viable synthetic protocol however, are major obstacles to the commercial production of the drug. The enhanced production of artemisinin is hence, highly desirable, which can be achieved by adequate and judicious supply of plant nutrients. The present experiment was therefore, designed to study the effect of organic manure (15 tonnes ha⁻¹) and chemical fertilizers (N₄₀₊₄₀, P₄₀, K₄₀, S₁₅₊₁₅ kg ha⁻¹; nitrogen, phosphorus, potassium and sulphur) on the accumulation of artemisinin and biomass in various plant parts through the developmental stages of A. annua L. Artemisinin yield (kg ha⁻¹) was also determined through the developmental stages of A. annua L. Artemisinin content and artemisinin yield of dried leaves were increased significantly at pre-flowering stage in the plants treated with NPKS (27.3% and 53.6%) and NPK (18.2% and 33.5%), respectively, when compared with control. Maximum dry yield of leaf ranging from 2596 to 3141 kg ha⁻¹ was observed at pre-flowering stage with various treatments.     

Identification and antimicrobial activity of two alkaloids from traditional Chinese medicinal plant Tinospora capillipes

Two antimicrobial alkaloids, palmatine and jatrorrhizine, were isolated from tubers of traditional Chinese medicinal plant Tinospora capillipes using activity-guided isolation method and chromatography. Their antimicrobial activity was determined in vitro. The results showed that palmatine and jatrorrhizine had inhibitory activity against plant pathogens Colletotrichum gloeosporioides, Fusarium oxysporum f. sp. niveum, Mycosphaerella sentina, Pestalotia mangiferae, Cercospora kaki, Gymnosporangium haraeanum, Rhizoctonia solani and Colletotrichum graminicola, with the EC₅₀ values of 0.0348-0.8356 g L⁻¹ and 0.0240-0.8649 g L⁻¹, respectively. Palmatine and jatrorrhizine also exhibited inhibition against animal pathogens Bacillus cereus, Bacillus megaterium, Bacillus subtilis, Staphyloccocus aureus, Staphylococcus epidermidi, Micrococcus lysodeikticus, Proteus vulgaris, Salmonella typhi and Escherichia coli, with the MIC values of 0.1-0.8 g L⁻¹ and 0.1-0.6 g L⁻¹, respectively. These results suggested that palmatine and jatrorrhizine showed relatively broad spectrum antimicrobial activity against plant and animal pathogens.     

Effects of leaf scales of different pineapple cultivars on the epiphytic stage of Fusarium guttiforme

The fungus Fusarium guttiforme (Syn. F. subglutinans f. sp. ananas) is responsible for fusariosis, one of the main phytosanitary threats to pineapple (Ananas comosus var. comosus). A structural study comparing epidermal differences in pineapple cultivars resistant and susceptible to fusariosis was performed, relating properties of the epidermis to known susceptibility to the disease. The basal, non-chlorophylled, portions of mature leaves of pineapple plants were analyzed by light and electron microscopy. All cultivars showed common morpho-anatomic aspects characteristic of Bromeliaceae, such as scutiform scales and unstratified epidermis. However, cultivar Vitoria (resistant) had less scales than cultivars Smooth Cayenne (susceptible, intermediate severity) and Perola (susceptible, with extreme severity of fusariosis symptoms). Inoculation of conidia suspension (10⁵ conidia ml⁻¹) of the fungus F. guttiforme to leaves and harvesting 24 h later yielded numbers of viable colonies related to the density of leaf scales. This suggests that scales can act as havens for fungal conidia and favour the epiphytic stage of the fungus on pineapple plants, and are involved in the interaction of plant and pathogen. A reduction in scale numbers was related to lower infection levels and is relevant to the future breeding programme for development of new pineapple cultivars resistant to fusariosis and their involvement in integrated control strategies.     

Foliar application of chitosan activates artemisinin biosynthesis in Artemisia annua L.

There has been much interest in artemisinin owing to its excellent activity against malaria, an infectious disease threatening the tropical world. However, the low artemisinin content (0.01-0.8%, DW) in Artemisia annua, which is the only commercial source of artemisinin, makes artemisinin expensive to produce and not yet available on a global scale. Here we show that foliar application of 100 mg l⁻¹ chitosan improved artemisinin biosynthesis in A. annua. The content of dihydroartemisinic acid and artemisinin in chitosan-treated leaves increased by 72% and 53% compared with control values, respectively. Chitosan induced the expression of ADS and DBR2, which could explain the increase in level of artemisinic metabolites. After chitosan treatment, the amounts of hydrogen peroxide (H₂O₂) and superoxide anion (O₂⁻) in leaves of A. annua were 1.4 and 3.0 times higher than those of the control, respectively. Accumulation of reactive oxygen species (ROS) probably accelerated the conversion of dihydroartemisinic acid to artemisinin. Foliar application of 100 mg l⁻¹ chitosan had no harmful effect on A. annua growth. The simple method described here could be an effective method to improve artemisinin production in A. annua field cultivation.     

Antifungal and antiaflatoxigenic efficacy of Caesulia axillaris Roxb. essential oil against fungi deteriorating some herbal raw materials, and its antioxidant activity

The study deals with evaluation of antifungal and antiaflatoxigenic Caesulia axillaris Roxb. essential oil (EO) against herbal raw materials deteriorating fungi and its free radical scavenging activity. During mycoflora analysis these herbal raw materials were found to be severely contaminated by different fungi and aflatoxins. A total of nine different fungal species were isolated from three herbal raw materials. Aspergillus flavus LHPtc was recorded as the highest aflatoxin B1 producing strain. EOs of some plants were tested for their fungitoxicity against the toxigenic strain A. flavus LHPtc, and C. axillaris EO was found as potent fungitoxicant. C. axillaris EO was chemically characterized through GC-MS analysis which depicted the presence of 18 compounds, dl-limonene and Euasarone being the major components. The EO exhibited broad spectrum of fungitoxicity against fungi causing postharvest deterioration of herbal raw materials. At 1.0 μl ml⁻¹ the oil showed complete inhibition of fungal growth and aflatoxin B₁ production was inhibited at 0.8 μl ml⁻¹. Free radical scavenging activity of the oil was also recorded by 2,2-diphenyl-1-picrylhydrazyl assay, and its IC₅₀ value was found 18 μl ml⁻¹. The safety limit of the EO was determined in terms of LD₅₀ on mice, which was 9166.6 μl kg⁻¹, suggesting its non mammalian toxicity. The EO of C. axillaris may be recommended as a plant based preservative in enhancement of shelf life of herbal raw materials by preventing their lipid peroxidation as well as biodeterioration due to fungal and aflatoxin contamination.     

Uptake and use efficiency of N, P, K, Ca and Al by Al-sensitive and Al-tolerant cultivars of wheat under a wide range of soil Al concentrations

The impacts of acidic soils and Al toxicity on wheat nutrient economy have been scarcely researched under field conditions even though these soils are widely spread in wheat production areas around the world. The main objective of this study was to quantitatively evaluate the element (N, P, K, Ca and Al) economy of an Al-sensitive and an Al-tolerant wheat cultivar growing under different soil Al concentrations at field conditions. To reach this objective, two field experiments were conducted in an Andisol in Valdivia (39°47′18″S, 73°14′05″W), Chile. Treatments were a factorial arrangement of: (i) two spring wheat cultivars (Al-sensitive, Domo.INIA and Al-tolerant, Dalcahue.INIA) and (ii) five exchangeable Al levels (0-2.7 cmol₍₊₎ kg⁻¹) with three replicates. At harvest, plant biomass was sampled and divided into 5 organ categories: ears, grains, blade leaves, stems plus sheath leaves and roots. The element content (N, P, K, Ca and Al) in each organ was measured to quantify element uptake and concentration, nutrient uptake efficiency (UPE) and nutrient utilization efficiency (UTE). Element uptake (N, P, K, Ca, and Al) was negatively affected by the increased soil Al concentration in above-ground and root biomass in both cultivars (R² = 0.61-0.98, p < 0.01), although clear differences were found between cultivars. On the contrary, the impact of soil exchangeable Al on the plant element concentration was minor, showing weak associations with soil Al levels. However, the Al concentration in above-ground tissues of the Al-sensitive cultivar was an exception because it increased exponentially with the Al soil concentration (R² = 0.96-0.99, p < 0.001). Nutrient uptake efficiencies, UPEs (N, P, K and Ca), were negatively affected by soil Al concentrations and were well described by linear equations in both cultivars (R² = 0.58-0.98, p < 0.05), with notable differences between them. Both nutrient uptake (capture) and UPE were the traits that best explained above-ground biomass production (R² = 0.82-0.99, p < 0.001, n = 20). Nutrient utilization efficiency, UTEs (N, P, K and Ca) responded more conservatively to the soil Al concentration, except for the Al sensitive cultivar under very high soil Al levels.