Influence of ventilation and sucrose on growth and leaf anatomy of micropropagated potato plantlets

Potato single nodes were cultured in vessels containing MS medium supplemented with 10, 20 and 30 g/l of sucrose. Vessels were closed with a clear polypropylene lid with or without 10 mm microporous polypropylene membrane. Sucrose concentration significantly increased plantlet height, shoot fresh weight and chlorophyll a content. Plantlets grown in ventilated vessels were significantly shorter, had lower shoot fresh weight and higher shoot dry weight than those in non-ventilated vessels. The highest leaf chlorophyll a content (21.83 mg/g fresh weight) was found in plantlets grown in ventilated vessels using MS medium with 20 g/l of sucrose, whereas those grown on medium with 10 g/l of sucrose had the highest chlorophyll b content (24.00 mg/g fresh weight). Total chlorophyll content was significantly higher when plantlets were grown in ventilated vessels containing medium with 10 or 30 g/l sucrose than in non-ventilated vessels. There was no significant difference in total chlorophyll content among plantlets grown in ventilated vessels with different concentrations of sucrose. Stomatal density was significantly lower when plants were grown under ventilated conditions. Leaf replica examination showed that stomata under non-ventilated condition were spherical with wide openings whereas, those in ventilated vessels were elliptical with narrow openings. Plantlets grown in non-ventilated vessels had thinner leaves and failed to build up a distinct defined upper epidermis, palisade parenchyma layer and spongy cells. On the other hand, leaves under ventilated conditions showed comparatively well organized layers with small intercellular space. The vascular system of leaves under the ventilated conditions demonstrated very well developed xylem unlike leaves under non-ventilated conditions. Thus, ventilated vessels with the 20 g/l of sucrose under ambient CO₂ in the growth room could successfully promote photomixotrophic culture and produce healthy plantlets.     

Effect of cutting date and position on rooting ability and fatty acid composition of Carignan (Vitis vinifera L.) shoot

The effects of cutting date and position (apical, basal and central) on rooting ability and fatty acid composition from Carignan (Vitis vinifera L.) shoot were determined. Root number and weight depended of the cutting date and position. Only in the case of the cutting in the basal position, there was a highly positive correlation between number and percentage of roots (r = 0.95) during sampling date. Concerning the influence of the cutting date, the root number by cutting oscillated in a saw tooth. The root weight and percentage showed a positive correlation for the three cutting positions and they increased with time. Cutting date and position had an irregularly effect on the contents of total lipid and different fatty acids. Independently of cutting position, the contents of oleic acid (C18:1) and palmitic acid (C16:0) correlated negatively with those of linoleic (C18:2) and linolenic acids (C18:3) during sampling date.     

Influence of medium composition and vessel ventilation on in vitro propagation of Phillyrea latifolia L.

Micropropagation of Phillyrea latifolia L. a wild species present in Mediterranean coastal areas having drought and salt tolerance was performed using explants from adult plants. Shoots were induced from nodal explants on the Rugini’s initial medium (IM). Then these were proliferated on either Rugini olive medium (OM) or Linsmaier and Skoog (LS) medium, each supplemented with 2.22 μM 6-benzylaminopurine (BA) or 4.56 μM zeatin (Z). Rooting (66.1±11%) was induced on shoots grown in perlite soaked with half-strength Rugini olive proliferation medium (OMr) containing 2.69 μM α-naphthaleneacetic acid (NAA) and 160 mg l⁻¹ putrescine. Both shoot multiplication and rooting were performed using Magenta^® GA-7 (Sigma) vessels either non-permeable or permeable to gas exchanges. Contamination (about 40%) was observed during the first five passages notwithstanding the addition of cefotaxime to the culture medium, but a high proliferation rate (90%) of explants provided enough healthy plant material. The highest shoot proliferation was observed on LS medium and zeatin whereas the presence of the ventilated filters reduced fresh weight of explants growing on LS media and did not affect shoot growth on OM media. During rooting, the use of ventilated vessels in comparison with the closed ones enhanced development of roots, and doubled the dry weight of plantlets. The vessel ventilation combined with the artificial substrate (perlite) was beneficial for in vitro acclimatization of rooted Phillyrea plantlets.     

Effect of three Greek arbuscular mycorrhizal fungi in improving the growth,nutrient concentration,and production of essential oils of oregano and mint plants

Arbuscular mycorrhizal fungi have been widely used in agriculture to improve the cultivation of many crops. One of the aims of this study was the isolation and molecular identification of arbuscular mycorrhizal fungi isolated from mountain areas of Northern Greece (Ritini Pieria, Elatochori Pieria, Ambelakia Ossa). Only three isolates were obtained; two of Glomus etunicatum and one of G. lamellosum. The second objective of this study was to investigate the effect of these arbuscular mycorrhizal fungi on the concentration of macro- and micronutrients in tissues, the quantity and quality of essential oils and the growth of oregano and mint plants (two widely used aromatic plants in Greece). It was found that mycorrhizal oregano and mint plants had a higher content of essential oils and nutrient elements, and grew better than non-mycorrhizal plants. In addition, the composition of the essential oil in mycorrhizal plants differed from the oil of non-mycorrhizal plants. These results suggest that the use of mycorrhizal fungi may allow plant growth in low fertility soils, reduce fertilizer inputs and increase aromatic plant production of essential oils, They also indicate that it may be possible to use mycorrhizae to affect the quality of the essential oil produced.     

Effect of cold cathode fluorescent lamps on growth of Gerbera jamesonii plantlets in vitro

The goal of this study was to evaluate the effect of cold cathode fluorescent lamps (CCFLs) on the growth of Gerbera jamesonii var. ‘Rui Kou’ plantlets in vitro in six different light quality ratios: 100% red CCFL (R), 80% R + 20% blue CCFL (B), 70% R + 30% B, 60% R + 40% B, 100% B and white CCFLs (W). Control radiation was provided by conventional heat-generating plant growth fluorescent lamps (PGFLs). Plantlets under CCFLs showed better plantlet height, SPAD value (i.e., leaf chlorophyll content) and root activity (as assessed by root dehydrogenase activity) than those growing under PGFLs while all other growth parameters were comparable with plants under conventional lighting systems.     

Effect of phosphite fertilization on growth,yield and fruit composition of strawberries

Traditionally, phosphates (Pi, salts of phosphoric acid, H₃PO₄) have been used for plant fertilization, and phosphites (Phi, salts of phosphorous acid, H₃PO₃) have been used as fungicides. Nowadays several Phi fertilizers are available in the EU market despite the fact that in research trials Phi has often had a negative influence on plant growth. The objective of this study was to elucidate the effect of a Phi fertilizer on plant growth, yield and fruit composition of strawberries (Fragaria × ananassa Duch.). Experiments were carried out with ‘Polka’ frigo plants in South Estonia in 2005 and 2006. The number of leaves per plant, total and marketable yields, fruit size, fruit ascorbic acid content (AAC), soluble solids content (SSC), titratable acidity (TA), anthocyanins (ACY) and total antioxidant activity (TAA) were recorded.     

Effect of genotype,explant size,position, and culture medium on shoot generation of Gerbera jamesonii by receptacle transverse thin cell layer culture

An unique procedure for the mass shoot propagation of Gerbera using receptacle transverse thin cell layer (tTCL) culture procedure was developed. Genotype, flower bud age, explant size, position of receptacle tTCLs and culture media were found to affect the success of culture. Ten interspecific crosses of Gerbera showed different shoot regeneration rates and callus induction via receptacle tTCL culture, all of which had shoot regeneration rates higher than 57%. Flower buds collected on the 10th day resulted in 91% shoot regeneration after 6 weeks of culture on basal MS medium [Murashige, T., Skoog, F., 1962. A revised medium for rapid growth and bioassay with tobacco tissue cultures. Physiol. Plant. 15, 475–497] supplemented with 0.02 mg l⁻¹ thidiazuron (TDZ), 0.8 mg l⁻¹ adenine and 10% (v/v) coconut water (CW). This was significantly higher than those from flower buds on the 7th and 14th days (22% and 54%), respectively. Shoot regeneration rate was the highest (94–100%) in the middle layers of the receptacle. For mass shoot propagation, shoot clusters were subcultured on half-strength MS medium supplemented with 0.5 mg l⁻¹ indole-3-butyric acid (IBA), 0.5 mg l⁻¹ 6-benzyladenine (BA) and 2.0 mg l⁻¹ kinetin after every 4 weeks. Plantlets formed when single shoots were cultured on half-strength MS medium containing 1 mg l⁻¹ IBA. All plantlets acclimatized well in the greenhouse.     

The effect of nitrogen source and concentration,medium pH and buffering on in vitro shoot growth and rooting in Eucalyptus marginata

This work examined the effect of nitrogen source and medium buffering on the micropropagation of Eucalyptus marginata Donn ex Sm. The number of shoots was increased when media contained 2-(N-morpholino) ethanesulfonic acid (MES) but this increase was minor and only applied to one of the two clones tested. Highest root production was obtained when the medium contained 7.5 mM nitrogen in a ratio of 2NO₃⁻:1NH₄⁺ and was buffered with 10 mM MES. In the rooting medium the pH was influenced most significantly by the nitrogen source, and then whether the medium was buffered. The media pH remained relatively constant when nitrate was the sole nitrogen source and this was assisted by the addition of 10 mM MES. Lower concentrations (<10 mM) of MES were less effective in buffering media over a four-week culture period in both shoot multiplication and rooting medium.     

Eggplant relatives as sources of variation for developing new rootstocks: Effects of grafting on eggplant yield and fruit apparent quality and composition

We propose the utilization of eggplant (Solanum melongena L.) interspecific hybrids derived from crosses with closely related species as an approach for developing new improved rootstocks for eggplant. Here we investigate rootstock effects on fruit yield, apparent quality and proximate and mineral composition of S. melongena ‘Black Beauty’ (BB) scions grafted on interspecific hybrid rootstocks developed from crosses of S. melongena with Solanum incanum L. (SI × SM) and Solanum aethiopicum L. (SM × SA). The results are compared with non-grafted (BB control) and self-grafted (BB/BB) controls and with S. melongena ‘Black Beauty’ scions grafted onto Solanum torvum Sw. (STO) and Solanum macrocarpon L. (SMA) rootstocks. All treatments were grown in a soil naturally infested with root-knot nematodes (mostly Meloidogyne incognita (Kofoid and White) Chitwood). SI × SM and SM × SA interspecific hybrids had high germination (≥90%) and total graft success (100%). Contrary to what occurred with all other treatments, no plants from scions grafted onto these hybrid rootstocks died during the experiment. In particular, the SI × SM hybrid rootstock conferred the highest vigour to the scion, which resulted in the highest values for fruit earliness and early and total yield. Little difference was observed among treatments for apparent fruit quality traits, except for a greater fruit calyx length and prickliness of fruit grafted onto SMA rootstocks. A similar result was obtained for fruit composition where phenolics content was higher in fruit from plants grafted onto SMA rootstocks. Grafting eggplant onto interspecific eggplant hybrids, especially on the SI × SM hybrid, has proved advantageous for eggplant production, as the high vigour and good compatibility of the rootstock with scion results in improved early and total yield without negative effects on apparent fruit quality or composition. Interspecific hybrids represent an alternative to the commonly used STO rootstock, which is a wild species with irregular germination.     

Effect of fruit-load on the growth,absorption, and partitioning of inorganic nutrients in young ‘Fuyu’ persimmon trees

Tree growth and the increase in inorganic nutrients during a growing season were studied in 3-year-old, container-grown ‘Fuyu’ persimmon trees with different levels of fruit-load. Fruit-load was varied by adjusting the leaf–fruit ratio to 10, 20, 30, and 60 on July 5. Some trees were completely defruited. The whole trees were harvested on November 5 of that year, and increases in dry weight and inorganic elements in different tree parts during the season were measured. A lower leaf–fruit ratio resulted in a higher yield but also decreased fruit weight, soluble solids, and fruit color. Dry weight of the aerial woods and root increased significantly with a greater ratio. However, total dry weight including the fruits was higher in the lower ratio trees than in the higher ratio trees; the weight of a tree that maintained the ratio of 10 was 35% higher than a tree without fruit. Fruits accounted for 62% of total dry weight when the ratio was 10 and 27% when the ratio was 60. The leaf–fruit ratio did not significantly alter the total increase in P, K, Ca, and Mg content. N tended to decrease with an increasing leaf–fruit ratio. Of the total increases of N, P, and K during the season, the fruits accounted for 30%, 50%, and 54% at the ratio of 20, respectively, and the percentages increased to 44, 81, and 74 at the ratio of 10. The inorganic element content in the aerial woods and root tended to increase with greater ratios, especially in the root. When trees were defruited, N and K increases in the root accounted for 44% and 35% of the total increase, respectively. These results indicated that persimmon orchard fertilization programs with different fruit-load levels should accommodate the proportion of nutrients partitioned to fruits that are removed permanently from the tree.     

Yield,dry matter and polysaccharides content of the mushroom Agaricus blazei produced on asparagus straw substrate

The aim of this work is to evaluate the potential of asparagus (Asparagus officinalis L.) straw as a raw material for cultivating Agaricus blazei Murrill (ABM). On non-supplemented asparagus straw substrate, the yield and biological efficiency (BE) of ABM were respectively 6.7 kg/m² and 30.2%. Addition of appropriate amounts of cottonseed hull or cow manure to the substrate increased the mushroom yield significantly. The mushroom yield on asparagus straw + cottonseed hull substrate was higher than that on asparagus straw + cow manure substrate. Maximum mushroom yield (9.8 kg/m²) and BE (44.1%) were obtained on the substrate consisting of asparagus straw (600 kg) and cottonseed hull (300 kg). No significant differences were found in either the dry matter contents or the polysaccharides contents of fruit bodies among the treatments.     

Effect of increasing climatic water deficit on some leaf and stomatal parameters of wild and cultivated almonds under Mediterranean conditions

The prevailing environmental conditions, temperature in particular, drive seasonal changes both in leaf development and stomatal characteristics. In order to ascertain the effect of increases in climatic water deficit on some leaf and stomatal parameters under field conditions, a study was carried out on two sets of leaves (spring and summer) on a large sample of Amygdalus communis L. cultivars in comparison with several Amygdalus webbii Spach seedlings, a species more adapted to arid environments and probable ancestor of cultivated almonds. Observations were performed between spring and summer of a particularly hot season. The results showed a significant and general reduction of both leaf area and stomatal frequency and an increase in stomatal size. Nevertheless, there were evident differences between cultivated and wild almonds. A stronger reduction of leaf area was observed in A. webbii (−31%) with respect to A. communis (−14%); on the contrary, the latter reduced stomatal frequency more than the former (−25% and −19%, respectively). The examined cultivated almonds, in response to the increase in climatic water deficit, tended to arrange their stomatal structures like those of wild almonds. Finally, increasing the climatic water deficit, the slope of the linear regressions between stomatal frequency and size did not change in either species, leading to a better understanding of the mechanisms of almond acclimation to environmental stresses.     

Chloride and sodium excluding capacities of citrus rootstock germplasm introduced to Australia from the People's Republic of China

The responses to root zone salinity (0, 25 and 50 mM NaCl) by 40 citrus rootstock genotypes introduced from the People's Republic of China, measured as shoot chloride (Cl⁻) and sodium (Na⁺) ion accumulation, growth and dry matter accumulation, were investigated under glasshouse conditions. Two experiments, one using aerated nutrient solutions and the other irrigated sand cultures, were conducted with plants grown from rooted cuttings taken from representative trees of different mandarin (Citrus reticulata Blanco and C. erythrosa Hort. ex Tan.), yuzu (C. junos Sieb. ex Tan.), Ichang papeda (C. ichangensis Swing.), sour orange (C. aurantium L.) and trifoliate orange (Poncirus trifoliata (L.) Raf.) genotypes. Two standard genotypes, viz. Rangpur lime (Citrus x limonia Osbeck.) and William's trifoliate orange were included in the experiments as reference controls.     

Pome fruit viruses at the Canadian Clonal Genebank and molecular characterization of Apple chlorotic leaf spot virus isolates

A survey for apple and pear viruses was carried out at the Canadian Clonal Genebank (CCG), Harrow, Ontario, Canada, during the fall/winter of 2007 and spring of 2008. Leaves and/or dormant cuttings were randomly collected from 438 to 122 accessions of apple and pear, respectively. Samples were tested by Double Antibody Sandwich-Enzyme Linked Immunosorbent Assay (DAS-ELISA) for the presence of Apple chlorotic leaf spot virus (ACLSV), Apple stem pitting virus (ASPV), Apple stem grooving virus (ASGV) and Apple mosaic virus (ApMV). Infection rates for apples were ACLSV (48.1%), ASGV (10%), ASPV (6.6%) and ApMV (7.1%), and for pears ACLSV (42.6%). ACLSV was detected and characterization by multiplex RT-PCR with primers targeting a fragment of 677 bp corresponding to the partial coat protein (CP), movement protein (MP) and untranslated (3′UTR) region in 22 accessions of apple and pear. Multiplex RT-PCR showed a higher sensitivity over the ELISA test. The nucleotide and amino acid deduced partial CP identities ranged from 82.6–100% to 91–100%, respectively, while partial MP identities was 62.5–100% at aa level based on the amplified fragment appropriate for partial MP using a frame shift, among 22 ACLSV isolates. Phylogenetic analyses based on the partial CP region clustered CCG ACLSV isolates in two different groups, while those based on the partial MP region embraced CCG ACLSV isolates in two sub-clusters within the same group. This is the first report on the detection of ACLSV, ASPV, ASGV and ApMV at CCG, and the molecular characterization of ACLSV isolates in apple and pear plants from worldwide countries to deduce possible heterogeneity and evolution.     

Phenolic composition of hazelnut leaves: Influence of cultivar,geographical origin and ripening stage

Nowadays, hazelnut tree leaf, a byproduct of hazelnut harvesting, is seen as a potential source of natural antioxidants. In this study, the phenolic composition of 93 samples (19 cultivars collected along 3 crop years in 2 geographical locations) was determined to evaluate the influence of genetic and edapho-climatic factors. Besides, a seasonal pattern variation study was performed, comprising the screening (from May to September) of the phenolic composition of four cultivars under the same agricultural, geographical and climatic conditions. Hazelnut leaf showed to be a good source of total phenolics since its content ranged from 2.5 to 30.5 g/kg. A seasonal pattern was observed consisting of an increase of the total phenolic content from May to July, a considerable decrease in August and a new increase in September. In all cultivars, the highest content of phenolics was achieved in July. Multivariated statistical analysis suggests that phenolics content in hazelnut leaves is mainly affected by the factor “crop year”, although genetic factors can also exert a moderate influence.     

Effect of co-cultivation and crop rotation on corm rot disease of Gladiolus

Field and pot experiments were conducted to evaluate the effect of co-cultivation and crop rotation on the growth and corm rot disease of gladiolus (Gladiolus grandiflorus sect. Blandus) cv. Aarti caused by Fusarium oxysporum f.sp. gladioli (Massey) Snyd. and Hans. In the field experiment, gladiolus was co-cultivated with 10 agricultural/horticultural crops viz. Allium cepa L., Brassica campestris L., Capsicum annuum L., Eruca sativa Mill., Helianthus annuus L., Tagetes erectus L., Zea mays L., Vinca rosea L. and Rosa indica L., in a soil infested with F. oxysporum. All the crops except V. rosea and R. indica reduced disease incidence. The effect of H. annuus and T. erectus was significant and more pronounced than other co-cultivated crops. In general, root and shoot dry biomass, corm fresh weight, number of cormlets and number of flowers per spike decreased as compared to the un-inoculated monoculture gladiolus treatment (negative control) but these parameters enhanced as compared to the F. oxysporum inoculated monoculture gladiolus treatment (positive control). In a pot experiment, all the crops of the field experiment except V. rosea and R. indica were sown in rotation with gladiolus. Pot grown plants of different species were harvested at maturity and the soil was inoculated with F. oxysporum. Gladiolus was cultivated 1 week after inoculation. Disease incidence was significantly suppressed in all the treatments ranging from 29% to 53%. The highest suppression of disease incidence was recorded in T. erectus (53%) followed by B. campestris (49%). The effect of preceding crops on various vegetative parameters was similar in the pot experiment to that of the field experiment. The present study suggests that corm rot disease of gladiolus can be managed by mixed cropping of H. annuus and T. erectus or cultivation of T. erectus and B. campestris in rotation.     

Influence of sulfur supply,ammonium nitrate ratio,and arbuscular mycorrhizal colonization on growth and composition of Chinese chive

Tissue concentrations of phytochemicals formed by Allium species may be influenced by plant nutrient supply or rhizosphere microorganism activity. To test these relations, three plant nutritional factors were varied in two experiments with Chinese chive [Allium tuberosum]: (a) increasing sulfur concentrations in the substrate, (b) two ratios of ammonium to nitrate in supply, and (c) inoculation with arbuscular mycorrhizal (AM) fungi. Shoot dry weight, nutrient composition (nitrate, N, P, S), and organosulfur compounds (measured indirectly as pyruvic acid) were determined.     

Effect of explants source on shoot proliferation and adventitious regeneration in 10 Buddleia cultivars

Viable shoot cultures and weaned plants were obtained from cultured apical meristems with 10 Buddleia cultivars giving viabilities of 32–72%. The number of shoots produced, the micropropagation rate and the root number produced in vitro was higher in meristem derived shoots compared to those derived from shoot-tips. The subsequent growth rate of meristem derived plants, in the greenhouse, was also greater. The number of roots produced by conventional cuttings collected from meristem derived plants was significantly higher than in cuttings which were collected from plants derived from shoot-tips or from the original stock plants.Endogenous bacteria were not detected in either shoot cultures derived from meristems or in 10-week-old weaned plants derived from meristems whereas those derived from shoot-tips showed the presence of endogenous bacteria when sterilized explants were cultured on nutrient agar or on tryptic soy broth.Factors affecting adventitious bud and shoot production in leaf and internode explants was determined for ‘Lochinch’, ‘Border Beauty’, ‘Ile de France’ and ‘Pink Delight’ using meristem derived shoot cultures. Adventitious shoots appeared after 4 weeks of culture, in both types of explant when cultured on MS supplemented with 0.5–5.0 μM TDZ. The highest percentage regeneration was achieved from bisected internode explants cultured on 0.5 μM TDZ, with 93–100% regeneration among the cultivars whereas BA was less effective. The best response was obtained using 5.0 μM TDZ which gave over 10–11 shoots per explant in all bisected explants for all cultivars.     

Effect of shade regime on flower development,yield and quality in lisianthus

The effects of shading on lisianthus (Eustoma grandiflorum) floral transition, plant development, flower yield and quality, and content of starch and soluble sugars were assessed in three cultivars, over two consecutive years. Shading nets affording 67% or 88% reduction in light intensity, were fitted at planting in the greenhouse for periods ranging from 3 to 8 weeks. Meristem morphology at floral transition was characterized by apical meristem widening and the appearance of two bract primordia. Floral transition time was affected by cultivars, but in general, longer and heavier shade treatments delayed floral transition; the longest delay (6 weeks) being recorded in Mariachi White under 88% shade for 7 weeks or under a combined shade treatment of 88% for 3 weeks followed by 67% for 5 weeks. Despite interactions between cultivar and shade treatment, consistent trends were discerned: the heaviest and most prolonged shading reduced yield (up to 40%), cut stem length (up to 15%), and number of flower buds/stem (up to 26%), within cultivar. Total carbohydrates levels were very low, and it is questionable whether changes observed in carbohydrate quantity following shade treatments had any effect on plant growth or flower yield. Rather, it appears that lisianthus is very dependent on current photosynthesis, so that even a brief shading interlude could reduce branching and flower quality. It may be concluded that the intensive shading usually applied is detrimental for lisianthus.     

The genotypic effects on the chemical composition and antioxidant activity of sea buckthorn (Hippophae rhamnoides L.) berries grown in Turkey

In this study, chemical composition of berries of 10 sea buckthorn (Hippophae rhamnoides L.) genotypes in Turkey was investigated. The total phenolic content of the berries ranged from 21.31 mg gallic acid equivalents (GAE) per g dry weight basis to 55.38 mg GAE per g. The highest antioxidant activity was 93.54% (similar to the standard BHT at 200 mg/L) and the lowest was 80.38%. There was no correlation (R = 0.688) between the total phenolic content and the antioxidant activity. The major fatty acids in berries were palmitoleic acid (35.48%), followed by palmitic acid (28.13%), oleic acid (22.89%) and linoleic acid (3.96%). Total soluble solid content of sea buckthorn genotypes varied from 10.15 to 14.80%, titratable acidity varied from 2.64 to 4.54%, the pH varied from 2.63 to 2.98 and Vitamin C varied from 19 to 121 mg/100 mL. The average content of minerals in the sea buckthorn berries of different genotypes was 20,800 ppm N, 7100 ppm P, 7260 ppm K, 1960 ppm Ca, 1465 ppm Mg, 32 ppm Zn, 24 ppm Cu, 22 ppm Mn and 7 ppm Fe.