盐碱池塘养殖鲤肠道菌群的分子分析

为了了解盐碱池塘养殖鲤肠道细菌群落组成及多样性,提取鲤(Cyprinus carpio L.)肠道细菌基因组DNA,选用细菌通用引物对16S rRNA基因进行了PCR扩增,构建了细菌16S rRNA基因克隆文库。通过对阳性克隆子进行限制性片段长度多态性分析(RFLP),选出有代表性的克隆子进行测序、BLAST比对分析和构建系统发育树。本研究从16S rRNA基因文库中共筛选出176个阳性克隆,经RFLP分析得到28个不同分类操作单元(operational taxonomic unite,OTU)(GenBank登录号:JX262557~JX262584),文库覆盖度为88.6%。16S rRNA序列系统发育分析发现,盐碱塘养殖鲤肠道细菌归属于变形细菌门(Proteobacteria)(包含Alpha和Gamma亚群)、厚壁细菌门(Firmicutes)、拟杆菌门(Bacteroidetes)和梭杆菌门(Fusobacteria)4个门,分别占克隆总数的89.9%、7.9%、1.1%和1.1%。其中,变形细菌门的α-变形菌纲(占克隆总数的88.1%)为优势类群,气单胞菌属为优势菌属。本研究揭示了盐碱池塘养殖条件下健康鲤肠道细菌群落组成。     

Iron nutrition of sunflower(Helianthus annuus L.) as affected by various levels of p and ZN

The effects of various P and Zn levels on iron nutrition of sunflower (Helianthus annuus L.c.v. Record) were studied in two separate experiments in nutrient solution under greenhouse conditions.

In the first experiment, sunflower was grown in nutrient solutions containing four levels of P(1.5, 2.5, 3.5 and 4.5 mM/l) and three levels of Fe(0.25, 0.75, and 1.5 ppm) as FeCl₃ or FeEDDHA. In the second experiment (following the first experiment), the treatments were three P levels (0.75, 1.50 and 3.00 mM/l), three Fe levels (0.25, 0.75 and 1.5 ppm) as FeEDDHA and three Zn levels (0.1, 0.2 and 0.4 ppm).

The plants receiving Fe‐chelate, except for 0.25 ppm Fe, showed no symptoms of iron chlorosis. With inorganic Fe treatments, iron chlorosis appeared after 7–10 days depending on P level, but except for 0.25 ppm Fe which remained chlorotic, plants recovered completely within 3–4 days thereafter due to pH regulating mechanism of sunflower under iron stress condition. With both sources of Fe, chlorosis was associated with high P:Fe ratio.

Increased P and Fe levels in nutrient solution resulted in general increases in the dry weights of roots and shoots. The Fe concentration of shoots, except in few instances, was not affected by P levels, indicating that the sunflower cultivar used in this experiment could utilize inorganic Fe as well as Fe‐chelate under our experimental conditions.

Increasing P levels caused significant increases in Mn content of the shoots as 0.25 and 0.75 ppm inorganic Fe³⁺. Increased Fe levels increased shoot Mn content with inorganic Fe and decreased it with Fe‐chelate. The effects of P, Fe and Zn on sunflower indicated an antagonistic effect of Zn on 1.5 ppm Fe for all P levels. Increased Zn levels in nutrient solution generally increased Zn content of the shoots without having any marked effect on their Mn content.     

In vitro effect of malathion and endosulfan on the LH-induced oocyte maturation in the common CARP,Cyprinus carpio (L.)

Effect of malathion (organophosphorus insecticide) and endosulfan (organochlorine insecticide) on in vitro LH-induced oocyte maturation was investigated in the oocytes of common carp, Cyprinus carpio. In control incubation, LH at the concentration of 10 μg mL^?1 induced 41.2 ± 1.6% of germinal vesicle breakdown (GVBD). When the oocytes were incubated with malathion at the concentrations of 1000, 500, 100, and 50 ppb together with LH it could induce only 13.4 ± 0.4, 14.0 ± 1.0, 12.9 ± 3.5, and 18.1 ± 3.9% of GVBD, respectively. Similarly, when the oocytes were incubated with endosulfan at the concentrations of 0.5, 0.1, 0.05, and 0.01 ppb together with LH, it induced only 12.8 ± 1.6, 8.8 ± 1.2, 20.9 ± 2.1 and 26.0 ± 2.2% of GVBD, respectively. The significance of the result obtained were discussed on the basis of available literature.     

Purification and characterization of gelatinase-like proteinases from the dark muscle of common carp (Cyprinus carpio)

Gelatinolytic proteinases from common carp dark muscle were purified by 30-60% ammonium sulfate fractionation and a combination of chromatographic steps including ion exchange on DEAE-Sephacel, gel filtration on Sephacryl S-200, ion exchange on High-Q, and affinity on gelatin-Sepharose. The molecular masses of these proteinases as estimated by SDS-PAGE were 75, 67, and 64 kDa under nonreducing conditions. The enzymes revealed high activity at a slightly alkaline pH range, and their activities were investigated using gelatin as substrate. Metalloproteinase inhibitors, EDTA, EGTA, and 1,10-phenanthroline, almost completely suppressed the gelatinolytic activity, whereas other proteinase inhibitors did not show any inhibitory effect. Divalent metal ion Ca (2+) is essential for the gelatinolytic activity. Furthermore, these gelatinolytic proteinases hydrolyze native type I collagen effectively even at 4 degrees C, strongly suggesting their involvement in the texture softening of fish muscle during the post-mortem stage.     

Expression of soluble form carp (Cyprinus carpio) ovarian cystatin in Escherichia coli and its purification

A DNA encoding thioredoxin-mature carp ovarian cystatin (trx-cystatin) fusion protein was ligated into a pET-23a(+) expression vector and then transformed into Escherichia coli AD494(DE3) expression host. After induction by isopropyl beta-D-thiogalactopyranoside, a high level of the soluble form of recombinant trx-cystatin was expressed in the cytoplasm of E. coli. The recombinant trx-cystatin could be purified by Ni(2+)-NTA agarose affinity chromatography. The molecular mass (M) of the recombinant trx-cystatin was approximately 28 kDa composed of recombinant thioredoxin (16 kDa) and recombinant mature carp ovarian cystatin (12 kDa). Both recombinant trx-fused and mature carp ovarian cystatins were stable at pH 6-11. No obvious decrease in activity was observed even after 5 min of incubation at 60 degrees C. They exhibited papain-like protease inhibition activity comparable to that of the mature carp ovarian cystatin, which could inhibit papain and mackerel cathepsins L and L-like, but not cathepsin B.     

Formation of odorants in Maillard model systems based on l-proline as affected by pH

Formation of the odorants acetic acid, 4-hydroxy-2,5-dimethyl-3-(2H)-furanone (HDMF), 6-acetyl-1,2,3,4-tetrahydropyridine (ATHP), and 2-acetyl-1-pyrroline (AP) was monitored by isotope dilution assays at pH 6, 7, and 8 in Maillard model reactions containing glucose and proline (Glc/Pro) or the corresponding Amadori compound fructosyl-proline (Fru-Pro). In general, higher yields were obtained at pH 7 and 8. Acetic acid was the major odorant with up to 40 mg/mmol precursor followed by HDMF (up to 0.25 mg/mmol), the formation of which was favored in the Fru-Pro reaction systems. On the contrary, ATHP (up to 50 microg/mmol) and AP (up to 5 microg/mmol) were more abundant in Glc/Pro. However, the sensory relevance of the two N-heterocycles was more pronounced on the basis of odor activity values, confirming their contribution to the overall roasty note of the reaction samples. It was also found that formation and decomposition of Fru-Pro were faster at pH 7 as compared to pH 6, explaining in part the preferred formation of the four odorants studied under neutral and slightly alkaline conditions. After 4 h of reaction at pH 7 in the presence of proline, about one-fourth of the glucose was consumed leading to acetic acid with a transformation yield of almost 40 mol %.     

Annual evolution of fatty acid profile from muscle lipids of the common carp (Cyprinus carpio) in Madagascar inland waters

Annual evolution of muscle lipids fatty acid (FA) from common carp (Cyprinus carpio) has been determined in 2001 through monthly samplings in the reserve pond of Sisaony (SIS series) and Itasy Lake (ITA series) of the Madagascar highlands. Total lipids from muscle were extracted and quantified according to the Bligh and Dyer method. FA identification was performed by GC-MS of FA methyl esters and FA pyrrolidides and led to the identification of 41 FA; routine analyses of FA were made by capillary GC. Principal component analysis (PCA) was performed on the data set to compare FA profiles. Lipid content is low, ranging from 0.91 to 1.73% of wet muscle, with a low stage during the hot season (January-April) and a higher stage during the cold season (July-October). Three FA dominated the FA composition: oleic acid (17.0-21.5%), palmitic acid (13.1-16.1%), and linoleic acid (9.6-13.2%). Polyunsaturated fatty acids (PUFA) were present in appreciable amounts: arachidonic acid (AA; 2.9-5.9%), docosahexaenoic acid (DHA; 2.9-6.7%), eicosapentaenoic acid (EPA; 1.9-3.4%), and docosapentaenoic acid (DPA; 1.9-4.3%). Two opposite evolution schemes appear within two groups of FA; on the one hand PUFA (both n-3 and n-6 series) show a maximum in August-October and a minimum in January-April, and, on the other hand, oleic, palmitic, and linoleic acids show the opposite maxima and minima. PCA results give confirmation of these evolution schemes, the two groups of FA giving opposite high factor loadings on axis 1. The SIS and ITA series are differentiated by axis 2 by mean of minor FA, mostly odd- and branched-chain. Results indicate that common carp, the second most abundant freshwater fish in Madagascar highlands waters, may be an interesting source of dietary PUFA.     

Compositional changes in 'Bartlett' pear ( Pyrus communis L.) cell wall polysaccharides as affected by sunlight conditions

Preharvest conditions can have a great impact on fruit quality attributes and postharvest responses. Firmness is an important quality attribute in pear, and excessive softening increases susceptibility to bruising and decay, thus limiting fruit postharvest life. Textural characteristics of fruits are determined at least in part by cell wall structure and disassembly. Few studies have analyzed the influence of fruit preharvest environment in softening, cell wall composition, and degradation. In the current work 'Bartlett' pears grown either facing the sun (S) or in the shade (H) were harvested and stored for 13 days at 20 °C. An evaluation of fruit soluble solids, acidity, color, starch degradation, firmness, cell wall yield, pectin and matrix glycan solubilization, depolymerization, and monosaccharide composition was carried out. Sun-exposed pears showed more advanced color development and similar levels of starch degradation, sugars, and acids than shaded fruit. Sunlight-grown pears were at harvest firmer than shade-grown pears. Both fruit groups softened during storage at 20 °C, but even after ripening, sun-exposed pears remained firmer. Sunlight exposure did not have a great impact on pectin molecular weight. Instead, at harvest a higher proportion of water-solubilized uronic acids and alkali-solubilized neutral sugars and a larger mean molecular size of tightly bound glycans was found in sun-exposed pears. During ripening cell wall catabolism took place in both sun- and shade-grown pears, but pectin solubilization was clearly delayed in sun-exposed fruit. This was associated with decreased removal of RG I-arabinan side chains rather than with reduced depolymerization.     

Maize (Zea mays L.) yield and soil properties as affected by no tillage in the black soils of China

A four-year tillage experiment on maize was conducted in the high latitude region of Northeast of China. The main objective of this study was to investigate the effects of different field tillage practices on maize grain yield, soil physical properties, and soil water and temperature dynamics in Northeast of China. The tillage practices included moldboard plow tillage (MOT), rotary tillage (ROT), reduced tillage (RET), combined tillage (COT), and no tillage (NOT). The surface soil water content at depths of 0–20 cm was higher under NOT compared with other tillage practices, but not different at the deeper soil depths in 2011. The soil temperatures under NOT and RET were lower than those under moldboard plow tillage and ROT at depths of 5 and 15 cm, respectively, measured at 9:00 am in 2005. From the hourly dynamics of soil temperature, the differences among the tillage practices mainly appeared during the daytime (from 8:00 am to 7:00 pm). Among all the practices, the average daily soil temperature under ROT was the highest, while that under NOT was the lowest. MOT, ROT, and RET had higher soil accumulative infiltration compared with NOT and COT. The surface soil bulk density under NOT was higher than or equal to that under the other four tillage practices. The maize yields under NOT were the lowest among all the tillage practices for three years straight. Meanwhile, the yields under MOT were the highest, which were about 47%, 61%, and 38% higher than those under NOT. NOT practice is not recommended for use in spring-planted maize under the high latitude humid cool climate Mollisol region in Northeast of China.     

Differences in key odorants of handmade juice of yellow-flesh peaches (Prunus persica L.) induced by the workup procedure.

Application of aroma extract dilution analysis (AEDA) on a flavor extract isolated from a freshly prepared, enzyme-inactivated peach juice using solvent extraction and high-vacuum distillation (extract I) revealed 24 odor-active regions in the gas chromatogram. Flavor dilution (FD) factors ranged from 4 to 512. The highest FD factors were determined for beta-damascenone (cooked-apple-like) and gamma-decalactone (peach-like). Cooking of peaches for 2 h in an apparatus equipped for simultaneous steam distillation/extraction (extract II) yielded an overall more intense aroma extract (extract II). By AEDA, 30 odorants were detected in the FD-factor region of 4-16384 and were subsequently identified. The results revealed that in extract II, besides the two above-mentioned aroma compounds, both had FD factors of 16 384; delta-decalactone, gamma-dodecalactone additionally, and 6-dodeceno-gamma-lactone contributed with very high FD factors (FD 8192) to the overall aroma. In general, the thermal treatment led to the formation of 15 new odorants which were not detected in I. Furthermore, the lactones and beta-damascenone were significantly increased in II, thereby indicating their generation from precursors in the fresh juice.     

Biological nitrogen fixation in faba bean (Vicia faba L.) in the Ethiopian highlands as affected by P fertilization and inoculation

 N₂ fixation by leguminous crops is a relatively low-cost alternative to N fertilizer for small-holder farmers in developing countries. N₂ fixation in faba bean (Vicia faba L.) as affected by P fertilization (0 and 20 kg P ha^–1) and inoculation (uninoculated and inoculated) with Rhizobium leguminosarium biovar viciae (strain S-18) was studied using the ¹⁵N isotope dilution method in the southeastern Ethiopian highlands at three sites differing in soil conditions and length of growing period. Nodulation at the late flowering stage was significantly influenced by P and inoculation only at the location exhibiting the lowest soil P and pH levels. The percentage of N derived from the atmosphere ranged from 66 to 74%, 58 to 74% and 62 to 73% with a corresponding total amount of N₂ fixed ranging from 169 to 210 kg N ha^–1, 139 to 184 kg N ha^–1 and 147 to 174 kg N ha^–1 at Bekoji, Kulumsa and Asasa, respectively. The total N₂ fixed was not significantly affected by P fertilizer or inoculation across all locations, and there was no interaction between the factors. However, at all three locations, N₂ fixation was highly positively correlated with the dry matter production and total N yield of faba bean. Soil N balances after faba bean were positive (12–58 kg N ha^–1) relative to the highly negative N balances (–9–44 kg N ha^–1) following wheat (Triticum aestivum L.), highlighting the importance of rotation with faba bean in the cereal-based cropping systems of Ethiopia. Received: 13 January 2000     

Soil temperature,growth and yield of maize (Zea mays L.) as affected by wheat straw mulch

Maize crop is grown mostly in tropical/subtropical environments where drought adversely affects its production. A field experiment was conducted on sandy loam soil for four years (1999 – 2002) to study the effect of wheat straw mulch (0 and 6 t ha^?1) and planting methods (flat and channel) on maize sown on different dates. Maximum soil temperature without mulch ranged from 32.2 – 44.4°C in channel and 31.6 – 46.4°C in flat planting method. Mulching, however, lowered soil temperature by 0.8 – 7.0°C in channel and 0 – 9.8°C in flat planting. Mulching, on an average, improved leaf area index by 0.42, plant height by 14 cm, grain yield by 0.24 t ha^?1 and biomass by 1.57 t ha^?1, respectively. Mulching improved grain yield only in flat sowing. Interaction between sowing date and planting method was significant. Seasonal variation in biomass were significantly correlated (p = 0.05) with mean air temperature during 0 – 45 days after planting (DAP) (r = ?0.95), pan evaporation during 0 – 15 DAP (r = 0.79) and negative correlation with rainfall in entire cropping season (r = ?0.89), whereas biomass increase with mulch in different cropping seasons had negative relation (r = ?0.74) with amount of rain during 0 – 15 DAP.     

Antioxidant activity of commercial soft and hard wheat (Triticum aestivum L.) as affected by gastric pH conditions

Phenolic compounds from soft and hard wheat and their milling fractions were extracted into distilled deionized water, and their in vitro antioxidant activities were evaluated. Wheat samples were used as such (nontreated) or subjected to pH adjustment (treated) in order to simulate gastrointestinal pH conditions. The total phenolic content (TPC) was determined using Folin-Ciocalteu's procedure. The total antioxidant activity (TAA) was determined using Trolox equivalent antioxidant capacity assay and expressed as Trolox equivalents. The antioxidant activity of wheat extracts was also evaluated using the beta-carotene bleaching assay, scavenging of 2,2-diphenyl-1-picrylhydrazyl radical, and inhibition of oxidation of human low density lipoprotein cholesterol. The TPC, TAA, and antioxidant potential, evaluated using different methods of wheat samples, were significantly increased following gastrointestinal tract-simulated pH changes. Thus, digestion taking place in the gastrointestinal tract in vivo may also enhance the antioxidant properties of the extracts.     

Glucosinolate concentration in turnip (Brassica rapa ssp. rapifera L.) roots as affected by nitrogen and sulfur supply

Three greenhouse pot experiments were conducted with four different nitrogen (N) treatments (80, 160, 240, and 320 kg ha (-1)) in combination with three sulfur (S) treatments (10, 20, and 60 kg ha (-1)) to study the effects of combined N and S supply on glucosinolate concentration and composition in turnip roots. Total glucosinolate concentration varied widely from 9.7 (N 320S 10) to 91.6 (N 160S 60) mg (100 g) (-1) root fresh weight (FW) and individual glucosinolate concentrations were increased with increasing S supply regardless of the N treatment, whereas enhanced N supply (160 - 320 N ha (-1)) at the high S level (60 kg ha (-1)) did not affect total glucosinolate concentration. In contrast, assumingly attributed to the individual glucosinolate biosynthesis concentration of N-containing tryptophan-derived indole glucosinolate was highest with increased N supply, whereas S-containing methionine-derived aromatic and aliphatic glucosinolates decreased with increasing N supply combined at low S level (10-20 kg ha (-1)).     

Long-term in situ Toxicity Bioassays of the Reconquista River (Argentina) Water with Cyprinus carpio as Sentinel Organism

Since the early 1970s, more than 25% of Japanese red pine forests have died or declined in Hiroshima Prefecture, Japan. Mass mortality phenomenon of pine trees has appeared throughout Japan, expanding northward from the Prefectures in southern Japan such as Nagasaki, Kogashima. The mortality was above 80% in Nagasaki, Kumamoto, Miyazaki and Tokyo. This study shows that the massive mortality of pines forest mainly occurred in south-facing slopes, where mortality was 90% in Gokurakujisann Mount, Hiroshima. The mortality was very low, < 3.7%, on north-facing slopes and mountain peaks. On west-facing slopes, the mortality was 20–40%. The foliar phenological development on south-facing slopes was 14 days earlier than that on north-facing slopes, 18 days earlier than that on mountain peaks. The mortality of pine trees in Gokurakujisann Mount increased with earlier phenological development and was significantly correlated (R = 0.98^**). The mortality of pine trees in each Prefecture increased from south to north with earlier phenological progress and was significantly correlated (R = 0.56^**). The outbreak of pine tree mortality was closely related to appearance of abnormally extreme lowest air temperature in February and March. The annual ring width in Gokurakujisann Mount was decreased from 1972 to 1984, concomitantly the abnormally extreme lowest air temperature in February and March. Based on these, it may be concluded that the combined effects of accelerated phenological progress and reduced frost hardiness caused by acid deposition, mainly nitrogen deposition, and extreme abnormally lowest air temperature in late winter and early spring in February and March are the original main factors of mortality of pine trees on a large scale.     

Sweet potato [Ipomoea batatas (L.) Lam.] cultivated as tuber or leafy vegetable supplier as affected by elevated tropospheric ozone

Sweet potato cultivars respond differently to elevated tropospheric ozone concentrations of ca. 130 mug m (-3), 8 h a day for 4 weeks, which affects their selection for cultivation. In the first cultivar presented here, an adequate leafy vegetable supplier, the ozone load resulted in a shift of biomass to maintain the canopy at the expense of tuber development. Starch content of leaves was reduced, indicating an impairment of quality, but carotenoid content remained stable. The second cultivar may be grown for tuber production. Although the ratio tuber/plant remained stable under ozone, tuber yield and its starch content were significantly reduced. The lower starch content indicated a worse quality for certain industrial processing, but it is desirable for chip production. Elevated tropospheric ozone concentrations also influenced free amino acids and macronutrient contents of tubers, but these modifications were of minor significance for tuber quality in the second cultivar.     

丛枝菌根真菌对万寿菊生长、吸镉及生理特性的影响

A pot experiment was carried out to study the effects of three arbuscular mycorrhizal fungi (AMF), including Glomus intraradices, Glomus constrictum and Glomus mosseae, on the growth, root colonization and Cd accumulation of marigold (Tagetes erecta L.) at Cd addition levels of 0, 5 and 50 mg kg-1 in soil. The physiological characteristics, such as chlorophyll content, soluble sugar content, soluble protein content and antioxidant enzyme activity, of Tagetes erecta L. were also investigated. The symbiotic relationship between the marigold plant and arbuscular mycorrhizal fungi was well established under Cd stress. The symbiotic relationship was reffected by the better physiobiochemical parameters of the marigold plants inoculated with the three AMF isolates where the colonization rates in the roots were between 34.3% and 88.8%. Compared with the non-inoculated marigold plants, the shoot and root biomass of the inoculated marigold plants increased by 15.2%- 47.5% and 47.8%-130.1%, respectively, and the Cd concentration and accumulation decreased. The chlorophyll and soluble sugar contents in the mycorrhizal marigold plants increased with Cd addition, indicating that AMF inoculation helped the marigold plants to grow by resisting Cd stress. The antioxidant enzymes reacted differently with the three AMF under Cd stress. For plants inoculated with G. constrictum and G. mosseae, the activities of superoxide dismutase (SOD) and catalase (CAT) increased with increasing Cd addition, but peroxidase (POD) activity decreased with increasing Cd addition. For plants inoculated with G. intraradices, three of the antioxidant enzyme activities were significantly decreased at high levels of Cd addition. Overall, the activities of the three antioxidant enzymes in the plants inoculated with AMF were higher than those of the plants without AMF inoculation under Cd stress. Our results support the view that antioxidant enzymes have a great influence on the biomass of plants, and AMF can improve the capability of reactive oxygen species (ROS) scavenging and reduce Cd concentration in plants to alleviate Tagetes erecta L. from Cd stress.     

Soil infiltrability and water content as affected by Baobab (Adansonia digitata L.) and Néré (Parkia biglobosa (Jacq.) Benth.) trees in farmed parklands of West Africa

The effect of baobab ( Adansonia digitata L.) and néré ( Parkia biglobosa (Jacq.) Benth.) trees on soil water content and water infiltration was investigated in farmed parkland at Nobéré, Burkina Faso, West Africa. Soil infiltrability was measured using tension head infiltrometers under trees and in an open field. Soil water content was determined gravimetrically. Soil infiltrability was higher below the edge of tree crowns compared with the open. There were no differences in infiltrability between soil under both tree species and in the open field. However, soil infiltrability increased from tree trunks to crown edges. This is probably due to tillage and/or higher compaction under trees than in the open field. In contrast to soil infiltrability, soil moisture was higher under tree crowns compared with the open field, perhaps due to higher organic matter content and reduced evaporation under trees.     

Lead-Induced Histological and Ultrastructural Changes in the Leaves of Soybean (Glycine max (L.) Merr.)

The seedlings of the soybean ( Glycine max. (L.) Merr.) cv. Polan were investigated by subjecting them to water culture for a period of 14 d. To the Knop nutrient solution, lead was added as PbCl₂ at four concentrations: 0, 10, 20 and 40 mg dm⁻³. Observations of soybean leaf tissues were carried out by light microscopy, transmission electron microscopy and scanning electron microscopy. The Pb levels used in the present study reduced the area of cotyledons and leaf blades of the soybean plants. Pb-induced changes in the leaf epidermis structure involved a reduction in the cell size, more abundant wax coating, and an increase in the number of stomata and trichomes per unit area with simultaneous reduction in the size of the guard cells. The lead treatment resulted in the reduction in the thickness of the leaf blades, reduction in the area of xylem and phloem in the vascular bundles and in the diameter of the xylem vessels. Under Pb stress, the leaf mesophyll cells were characterized by the presence of altered chloroplasts with a reduced lamellar system and multidirectional pattern of the thylakoid system. Burst stroma of the thylakoid system and cracked chloroplast envelopes were also observed. The importance of the increase in the number of stomata and trichomes for plants under the metal stress was examined.