Effects of cadmium on growth of Helianthus annuus seedlings: physiological aspects

Sunflower seedlings ( Helianthus annuus hybrid Select) were grown in a complete nutrient solution in the absence or presence of Cd²⁺ (10 and 20 μM). Analyses were performed to establish whether there was a differential effect of Cd²⁺ on mature and young leaves. After 7 d the growth parameters as well as the leaf area had decreased in both mature and young leaves. Accumulation of Cd²⁺ in the roots exceeded that in the shoots. Seedlings treated with Cd²⁺ exhibited reduced contents of chlorophyll and CO₂ assimilation rate, with a greater decrease in young leaves. The photochemical efficiency of photosystem II (PSII) was not altered by Cd²⁺ treatment in either mature or young leaves, although during steady-state photosynthesis in young leaves there was a significant alteration in the following parameters: quantum yield of electron transport by PSII (Φ_PSII), photochemical quenching ( q _P), non-photochemical quenching ( q _NP), and excitation capture efficiency of PSII (Φ_exc).     

Localisation of nickel and mineral nutrients Ca,K, Fe,Mg by Scanning Electron Microscopy microanalysis in tissues of the nickel-hyperaccumulator Alyssum bertolonii Desv. and the non-accumulator Alyssum montanum L.

Plants of the nickel-hyperaccumulator Alyssum bertolonii Desv. and of the non-accumulator A. montanum L. growing on a serpentine site in Tuscany, Italy, and plants of A. montanum from a nearby non-serpentine site were analysed for metal concentration and localisation. The leaves of A. bertolonii contained 160 times more nickel than those of A. montanum from the same site, thus demonstrating its hyperaccumulation capacity towards this metal. On the other hand, both species showed an inversion of the Ca/Mg ratio in their organs relative to the soil. Nickel localisation in plant tissues was examined by Scanning Electron Microanalysis (SEM/EDX). In A. bertolonii, a specific pattern of nickel distribution was detected, with the highest concentrations present in parenchyma and sclerenchyma cells for the roots; in the shoots, the highest amounts of nickel were found in the stem epidermis, the leaf epidermal surface, and the leaf trichome base. This particular nickel tissue distribution pattern was not found in the non-accumulator A. montanum growing on serpentine soil. Other mineral nutrients, namely Mg, Ca, K, Fe, instead, had a similar distribution in the two species. The A. montanum plants from the non-serpentine site had very low nickel levels in their tissues, and these were of the same magnitude as those found in A. bertolonii plants grown in a greenhouse on commercial horticultural soil with low nickel concentration. In A. bertolonii plants, the tissue-specific allocation patterns appeared to depend on the degree of nickel hyperaccumulation, which is, in turn, directly linked to the soil characteristics.     

美人蕉(Canna indica Linn)镉胁迫的抗氧化机理

采用水培的方式,探讨了不同Cd2+水平(0、1、2.5、7.5、15 mg·L-1)对美人蕉生物量、超氧化物歧化酶(SOD)、过氧化物酶(POD)和过氧化氢酶(CAT)活性以及丙二醛(MDA)、谷胱甘肽(GSH)、植物螯合肽(PCs)、酸溶性SH、Cd含量的影响.结果表明,1 mg·L-1的Cd显著提高了美人蕉的生物量(p<0.05),促进了美人蕉的生长.随着Cd2+浓度的提高,SOD、 POD、CAT活性以及MDA含量显著增加(p<0.05),表明美人蕉受到了活性氧物质的胁迫.美人蕉中GSH、PCs、SH含量也随Cd2+含量的增加而增加,表明Cd胁迫诱导了PCs 的产生,有利于降低Cd对植物体本身的毒害,且根系中的含量均高于叶片.美人蕉中Cd含量随着Cd浓度的增加而显著增加,在15 mg·L-1处理时,地上部Cd含量达到555.4 mg·kg-1,表明美人蕉对Cd有较强的富集能力.     

Somatic Hybrids of Sunflower (Helianthus annuus L.) Identified at the Callus Stage by Isoenzyme Analysis

Dark grown hypocotyl protoplasts from Helianthus annuus L. cvs. Cerflor and Euroflor were electrically fused to produce somatic hybrids. Following fusion, the protoplasts were cultivated in agarose droplets for four weeks. Macroscopic visible calli (0.1-0.3 mm) were transferred onto solid medium and calli reaching a size of 3 mm were collected. Their isoenzyme patterns were analysed based on two different isoenzymes that allow discrimination between the two cultivars used for fusion. From the examined calli, about 26 % showed an isoenzyme pattern of putative binary heterokaryocytes. The isoenzyme pattern of the non-fused control revealed less than 5% of possibly chimeric colonies.     

Behaviour of antioxidant defense system in the adaptive response to salt stress in Helianthus annuus L. cells

A relationship between the antioxidant defense system and salt tolerance in two types of sunflower calli differing in salt sensitivity was studied. No reduction in growth occurred in the NaCl-salt-adapted cell line (T) when grown on 175 mM NaCl but growth of the salt-stressed cell line (S) was reduced by 83%. Lipid peroxidation and protein oxidation increased during acute stress of salt stressed cells at 14 and 28 d of the experiment, while salt-adapted calli (T) remained similar to non-shocked (C) values. The antioxidant defense system of callus adapted to growth under NaCl responded differently to 175 mM of salt compared with the corresponding controls under shock treatment. Salt-adapted and salt-stressed calli showed a similar pattern in GSH content at day 14 but at day 28 in S calli, GSH content was increased 100% over the non-shocked calli, while T calli returned to the initial values. In the salt-stressed calli, a general decrease in all the antioxidant enzymes studied (except for glutathione reductase and dehydroascorbate reductase activities) was observed at day 28. Except for catalase, the antioxidant enzymes were elevated constitutively in adapted calli as compared to stressed cells, when both were grown in the absence of NaCl (time 0), and remained unaltered until 28 d after the beginning of the experiment. These results suggest the involvement of an enzymatic antioxidant defense system in the adaptive response to salt stress in Helianthus annuus L. cells.     

Microscopical Observations on the Embryoid Formation in Cultured Unfertilized Ovules of Helianthus annuus L.

A recent advance in plant experimental embryology is the induction of haploid plants via in vitro culture of unfertilized ovules. Using float culture method on liquid media, we have raised haploid as well as diploid embryoids in sunflower cultivars by ovule culture. The present investigation was aimed to know the origin and developmental processes of these embryoids. Young flowers 1–4 days before anthesis were dissected and ovules were inoculated on N6 medium supplemented with 0.5–2 ppm MCPA and 6% sucrose. During culture period, samples were collected at intervals, fixed, stained and sectioned by paraffin method. Fifty one gynogenie embryoids of various sizes were observed among Ca. 2000 ovules. They were located at the micropylar end of the embryo sacs and proved to be originated from the unfertilized egg cells. At the early stages, they bore a strong resemblance to the zygotic proembryos in vivo, but after a considerable enlargement, they grew into globular, ovoid or elongated big bodies without polarized organ differentiation. Chromosome counts on some mitotic figures in these embyoids revealed their haploid nature. Embryoids were also produced from the endothelial tissue, which proliferated markedly after inoculation, especially at the chalazal parts, resulting in massive multilayered irregular folds and then degenerated. In some eases, cell divisions at one or several places led to embryoid or callus formation. The problems of how to regulate the growth of in vitro ovules in order to promote the gynogenic embryoids and inhibit the somatic embryoids or calli are left for future research.     

Comparative studies of nickel,cobalt, and copper uptake by some nickel hyperaccumulators of the genus Alyssum

The uptake of Ni, Co, and Cu by the nickel hyperaccumulator Alyssum troodii Boiss and the non-accumulator Aurinia saxatilis (L.) Desv. were studied in pot trials using artificial rooting media with varying concentrations of the metals added as soluble salts, singly and in combination. The ability of five other Ni hyperaccumulating species of Alyssum to hyperaccumulate Co was also investigated.Leaves and stems of A. troodii accumulated Ni to almost the same extent (8000–10 000 g g^-1). In roots, the highest Ni concentration was 2000 g g^-1. In leaves of Au. saxatilis, the maximum Ni concentration was only 380 g g^-1 and the level in roots was even lower.In media containing Co, the maximum concentration of this element in A. troodii (2325 g g^-1) was ten times higher than in the non-accumulator species. Slightly less Co was found in stems and roots of both species. Among the other Ni hyperaccumulators, the maximum concentration of Co in leaves ranged from about 1000–8000 g g^-1.Copper concentrations were the same in all organs of both species when they were grown in copper-rich media and were in the range 40–80 g g^-1, showing that neither plant was capable of taking up Cu at levels comparable to those of Ni and Co.When both plants were grown in media containing equal amounts of both Co and Ni, the Co concentrations in plant organs were the same as for specimens grown in media containing Co only. However, the Ni levels were lower in both species. Uptake of Co therefore appeared to suppress Ni uptake.Pot trials showed that the order of tolerance was Ni>Cu>Co for A. troodii and Ni>CoCu for Au. saxatilis, whereas the seedling tests showed the order to be Co>Ni>Cu. At metal concentrations 10 000 g g^-1, the overall tolerance of A. troodii was greater than that of Au. saxatilis which exhibited equally low tolerance to Ni and Cu.We conclude that in A. troodii, A. corsicum Duby, A. heldreichii Hausskn., A. murale Waldstein & Kitaibel, A. pintodasilvae T.R. Dudley, and A. tenium Hálácsy, Ni tolerance and hyperaccumulation conveys the same character towards Co. This behaviour should be investigated in other hyperaccumulators of Ni and/or Co.     

Relations between heterosis and enzymatic polymorphism in populations of cultivated sunflowers (Helianthus annuus L.)

Nine polymorphic isoenzymatic systems were studied in 39 cultivated sunflower populations originating from ten countries. Analysis of combining abilities with four tester lines was also performed on these populations for seed yield, seed moisture and seed oil content. The MDH, PGI, PGD and GOT systems appeared to provide the best discrimination of specific combining ability effects with the four testers. The MDH and GOT systems provided a between-population structure that was consistent with the country of origin.Abbreviations MDH Malate dehydrogenase - PGD phosphogluconate dehydrogenase - PGI phosphoglucoisomerase - PGM phosphoglucomutase - ACO aconitase hydratase - ADH2 alcohol dehydrogenase - GOT glutamate oxaloacetate transaminase - LAP leucine amino peptidase - EST esterases     

向日葵品种鉴定和纯度分析的AFLP研究

从杂交油葵A15及其亲本的1/2粒干种子中提取基因组DNA,选用17对引物组合进行AFLP分析,构建了它们的指纹图谱,17对引物在A系与R系当中共扩增出1125条扩增产物,其中144条带表现出多态性,平均每对引物扩增66条带。不同引物组合产生的DNA片段数目在50-70之间,大小分布于100bp-500bp,多态性比率为12.8%。从中筛选出的2对引物E-AAC/M-CTC和E-ACG/M-CTG可将亲本和子代区分开;引物对E-AAC/M-CTC在A系中扩增出440bp,190bp,160bp3条特征谱带,在R系中扩增出380bp,350bp,225bp,180bp4条特征谱带,E-ACG/M-CTG在A系中扩增出了2条特征带480bp和265bp,在R系中扩增出490bp,220bp,205bp,125bp4条特征谱带,且上述谱带均在子代中出现,用引物组合E-ACG/M-CTG对A15,双亲以及与A15外型十分相似的10个常用油葵杂交种进行AFLP分析,不仅表现出良好的多态性,并能够清楚地将它们加以区分,以其对50粒A15杂交种子进行纯度鉴定,得到与大田纯度检测一致的结果。说明使用AFLP标记检测油用向日葵的品种和纯度是可行的,对行种子纯度和品种鉴定的方法进行了讨论。     

Circadian Modulation of Circumnutation Length, Period, and Shape in Helianthus annuus

Modulation of a turgor-growth movement called circumnutation in sunflower (Helianthus annuus L.) was investigated using a picture analysis system. Two photoperiod conditions were applied: light–darkness conditions (LD) 8:8 and LD 20:10. After about 3 weeks of these regimes, the plants were placed under constant light to determine whether circadian regulation of circumnutation existed or not. The rhythms of movement activity with regard to the trajectory length, period, and shape of individual circumnutations were examined. Data were processed by Fourier spectral analysis. All the parameters, trajectory length, period, and shape, revealed the ability to entrain to the administered daily cycles (16 h or 30 h). We observed diurnal fluctuations of the circumnutation parameters with the phase of the highest trajectory length, the shortest period, and the highest shape coefficient (the most circular form) during the dark period. After the LD–LL transition, the parameters revealed periodicity, which was close to 24 h. After several days of a clear circadian free running rhythm, a gradual decrease of the amplitude of the rhythm was observed. However, the rhythm did not disappear completely. The trajectory length manifested the strongest entrainment; the circumnutation period and the circumnutation shape were less modulated by photoperiod. These findings indicate for the first time that different parameters of circumnutation in sunflower are circadian-regulated rhythms, not solely ultradian as had been thought previously.     

Isolation of phenolic constituents and characterization of antioxidant markers from sunflower (Helianthus annuus) seed extract

A new compound, benzyl alcohol β-d-apiofuranosyl-(1→6)-β-d-(4-O-caffeoyl) glucopyranoside (1), was isolated from the seed of sunflower (Helianthus annuus), together with eight known phenolic compounds: caffeic acid (2), methyl caffeoate (3), chlorogenic acid (4), 4-O-caffeoylquinic acid (5), 3-O-caffeoylquinic acid (6), methyl chlorogenate (7), 3,5-di-O-caffeoylquinic acid (8), and eriodictyol 5-O-β-d-glucoside (9). Their structures were elucidated on the basis of spectroscopic methods and chemical evidence. The antioxidative effect of the phenolic constituents from the sunflower seeds was also evaluated based on the oxygen-radical absorbance capacity (ORAC), and the fraction containing caffeic acid derivatives showed a high antioxidant potency.     

Assessment of EDDS and vermicompost for the phytoextraction of Cd and Pb by sunflower (Helianthus annuus L.)

The effects of Ethylenediamine disuccinic acid (EDDS) (0 and 5?mmol·kg^?1) as a synthetic chemical amendment, vermicompost (0 and 5%w/w) as an organic amendment and their combined application were evaluated for the phytoextraction by sunflower (Helianthus annuus L.) of cadmium (Cd) and lead (Pb) at three artificial contamination levels in soils (0, 50, and 100?mg·kg^?1 for Cd and 0, 100, and 200?mg·kg^?1 for Pb). The results showed that the application of EDDS was the most effective method to increase Pb and Cd concentrations in both parts of the plant. The results also showed that the application of EDDS increased 9.27% shoot Pb content at 200?mg·kg^?1 but decreased 15.95% shoot Cd content at 100?mg·kg^?1 contamination level with respect to the respective controls. The bioavailable concentrations of Cd at 100?mg·kg^?1 and Pb at 200?mg·kg^?1 contamination level in the soil at the end of experiment increased 25% and 26%, respectively after the application of EDDS but vermicompost decreased 43.28% the bioavailable Pb concentration relative to their controls. Vermicompost increased the remediation factor index of Cd, thus making it the best treatment for the phytoextraction of Cd. The combined application of EDDS and vermicompost was the best amendment for Pb phytoextraction.     

Combining-groups in cultivated sunflower populations (Helianthus annuus L.) and their relationships with the country of origin

Using four tester lines an analysis of combining abilities for seed yield, seed moisture content and seed oil content was performed on 39 cultivated sunflower populations originating from ten countries. A between-populations structure based on specific combining abilities (SCA) was designed, defining separate combining-groups for each of the four testers. This structure corresponds to the country in which the populations originated.     

Modifying of calcareous soil with some acidifying materials and its effect on Helianthus annuus (L.) growth

Calcareous soils contain a high concentration of calcium carbonate (CaCO₃), which influences soil properties related to plant growth. Humic acid (HA) and ammonium molybdate (AM) were added as treatments for calcareous soils at concentrations of 0.1, 0.5 and 1 g/l respectively. The pots were divided into three groups. The first set of groups were irrigated with AM, while the second set of groups were irrigated with HA. As a control, the third group was irrigated using only tap water. Many soil properties and plant characteristics were measured during the experiment. The results showed that most of the studied treatments aided to increase organic carbon of calcareous soil and improved sunflower height, leaf area and shoot and root biomass. All investigated treatments significantly enhanced carbohydrates content in the sunflower shoots, except the treatment with 0.1 g/l AM, while only the with AM (under all studied concentrations) significantly enhanced carbohydrates content in roots higher than untreated. Proteins content in the shoots and roots of sunflower significantly increased when treated only with 1 g/l HA higher than control. The amino acid content of sunflower roots enhanced when treated with 0.1 and 1 m/l HA and 0.5 g/l AM Evidently, acidifying materials enhanced the calcareous soil and increased productivity.     

Changes in mass and dimensions of sunflower (Helianthus annuus L.) Achenes and seeds due to carbonization

When analyzing sunflower (Helianthus annuus L.) remains, which are often carbonized, archaeobotanists commonly differentiate between wild and domesticated achenes and seeds based on the measured length (L) and width (W) or the calculated index L*W. Carbonization reduces the dimensions. To compensate for these reductions, archaeobotanists use a single correction factor proposed by Richard Yarnell (1978) for all cases. The use of a single correction factor can bias the reconstructed dimensions as carbonization is a highly variable process. The current study determines the relationship between carbonization and the dimensions of length and width. Measurements established that a decrease of 2.5-22.5% in achene length and 10-29% in achene width can occur, depending on temperature, heating rate, and variety. For seeds, temperature is of most importance, and shrinkage ranges from 0-27% for the length and from 0-20% for the width. These ranges make the use of a single correction factor problematic. A method is developed in which reflectance (an optical property applied in coal technology to determine coal rank) is used to measure the carbonization temperature, and in turn the shrinkage can be calculated. Subsequently, correction factors are calculated to reconstruct the original length and width. When applied to an assemblage of carbonized sunflower achenes, the newly developed method shows that the Yarnell single correction factor may bias the dimensions towards classifications of “wild” or “ruderal” forms of sunflower     

Combining-ability analysis of somatic embryogenesis from epidermal layers in the sunflower (Helianthus annuus L.)

Crosses were made between three cytoplasmic male-sterile and five restorer sunflower inbred lines. F₁ hybrids, including their parents, were studied for their embryogenetic ability. Sterilized seeds were germinated in culture tubes on agar-solidified basal medium. Seven days after germination, epidermal layers from hypocotyls were transferred into MS and B5 liquid-midia for 5 and 8 days respectively. Then they were transferred to MS-120 solid-medium with a high level of sucrose (120 g/l). The experimental design was complete randomized blocks with three replications. Each replication per genotype consisted of three Erlenmeyer flasks with 20–25 epidermal layers (explants). Analysis of variance indicated the presence of significant variation among genotypes for all traits studied. General combining ability (GCA) and specific combining ability (SCA) showed significant effects for the studied traits. The highest value of GCA for the number of embryogenic explants per 100 explants plated (EE/100EP) belongs to parental female line ’CMS-PET1 B9’. This inberd line also gave a high positive GCA effect for the number of embryos per ten embryogenic explants (E/10EE). Additionally, it had the highest values for EE/100EP and E/10EE (41.70 and 19.28 respectively) and should be a promising parent in crossing programmes for the enhancement of somatic embryogenesis in the sunflower. The highest values of specific combining ability (SCA) for EE/100EP and E/10EE belong to the F₁ hybrid ’CMS-PET1B9 ’x’ RT1B11’ which has produced 53.45 embryogenic explants/100 explants and 9.67 embryos per ten embryogenic explants. Received: 4 December 1998 / 16 June 1999     

植物对镉胁迫响应的研究进展

总结了近年来植物对一重要环境污染物镉的反应研究报道。探讨了镉对植物的各种毒性效应,并论述植物对镉胁迫采取的相应防御机制,如络合（PC、MT）、应激（应激乙烯、应激蛋白）,及额外防御机制,如细胞壁固定化、原生质膜排除,区域化、过氧化物酶等。     

Ruditapes decussatus and Ruditapes philippinarum exposed to cadmium: toxicological effects and bioaccumulation patterns

Since differences in metal accumulation may exist between bivalve species, the aim of this study was to assess the impact of cadmium (Cd) on Ruditapes decussatus and Ruditapes philippinarum. For this, the Cd accumulation, mortality rates and biochemical responses were analysed in the two species after 5days of exposure, under laboratory-controlled conditions. The concentration of Cd that caused 50% of mortality on clams was two-times higher in R. decussatus than in R. philippinarum. For both species, higher percentage (84.5-98.2%) of the Cd was in the insoluble fraction, but the Cd concentration in solution was 3 to 8 times higher in R. decussatus. Nevertheless, R. philippinarum presented higher oxidative stress and higher CAT activity. The paradox observed between the two clams can be explained by the higher capacity of R. decussatus to increase the expression of MTs when exposed to Cd.     

Pollen aroma fingerprint of two sunflower (Helianthus annuus L.) genotypes characterized by different pollen colors

Samples of fresh pollen grains, collected from capitula in full bloom from two genotypes of sunflower (Helianthus annuus L.) and characterized by a different color, i.e., white‐cream (WC) and orange (O), were analyzed by the HS‐SPME (headspace solid phase microextraction)/GC/MS technique. This study defined for the first time the fingerprint of the sunflower pollen, separated from the disc flowers, to define its contribution to the inflorescence aroma. In the GC/MS fingerprints of the WC and O genotypes, 61 and 62 volatile compounds were identified, respectively. Monoterpene hydrocarbons (34% in O vs. 28% in WC) and sesquiterpene hydrocarbons (37% in O vs. 31% in WC) were ubiquitous in all samples analyzed and represented the main chemical classes. α‐Pinene (21% in O vs. 20% in WC) and sabinene (11% in O vs. 6% in WC) were the dominant volatiles, but also a full range of aliphatic hydrocarbons and their oxygenated derivatives gave a decisive contribution to the aroma composition (10% in O vs. 12% in WC). In addition, dendrolasin (3% in O vs. 4% in WC) and some minor constituents such as (E)‐hex‐2‐en‐1‐ol (0.4% in O vs. 0.1% in WC) were pointed out not only for their contribution to the pollen scent, but also for their well‐known role in the plant ecological relationships. Having evaluated two pollen morphs with different carotenoid‐based colors, the study sought to highlight also the presence of some volatile precursors or derivatives of these pigments in the aroma. However, the pollen aroma of the two selected genotypes made a specific chemical contribution to the sunflower inflorescence scent without any influence on carotenoid derivatives.