NAD pattern and NADH oxidase activity in pea (Pisum sativum L.) under cadmium toxicity

Seeds of pea (Pisum sativum L.) were germinated for 5 days by soaking in distilled water or 5 mM cadmium chloride. Compared to the control, cadmium (Cd) caused a reduction in percent germination and embryo growth. Pyridine nucleotide coenzyme concentrations were determined in cotyledons and embryonic axis. Nicotinamide adenine dinucleotide (NADH) oxidase activity was examined. Cd treatment caused a restriction in levels of reduced coenzyme form in the mitochondria and the post-mitochondrial fraction of cotyledons, and embryonic axis. The oxidized coenzyme form has been accumulated by Cd-treated mitochondria of both tissues. It was also found that NADH oxidase activity was stimulated. The relationship between coenzyme levels, seed germination, pea growth, and Cd stress has been reported.     

A first report of water hyacinth (Eichhornia crassipes) soil seed banks in South Africa

We investigated water hyacinth seed banks in several aquatic systems in South Africa. Fifteen sites, mainly in the Eastern and Western Cape provinces, were surveyed from August to October 2009. Soil seed density varied between 0 and 2534 seeds/m² but did not differ significantly between the type of waterbody (impoundment vs. river) or the history of control carried out at a site. Average germination was 54.17% with very fast velocities (Vigour Index = 36.66) and maximum germination around three days. Although we demonstrated the existence of an important reservoir of seeds, results from this study indicated that a combination of factors such as water fluctuation, eutrophication and seed decomposition might have had a great influence on dispersal and persistence of seeds.     

粗茎秦艽种子粗提物与破眠方法研究

为了探究影响粗茎秦艽（Gentiana crassicaulis Duthie ex Burk.）种子休眠的因素,破除休眠,寻找其种子快速萌发的方法,以干燥的粗茎秦艽种子为材料,测定种子吸水率及粗提物的活性,使用不同浓度的赤霉素（GA₃）、高锰酸钾（KMnO₄）、聚乙二醇（PEG6000）和过氧化氢（H₂O₂）溶液进行浸种处理,比较不同处理条件对粗茎秦艽种子萌发的影响。结果显示,粗茎秦艽种皮对种子吸水没有明显阻碍作用;不同浓度的种子粗提物对白菜、小麦的萌发和生长均表现出抑制作用;不同浓度的粗提物对粗茎秦艽种子自身的萌发也表现出一定的抑制作用,当粗提物浓度达到0.1 g/mL时,抑制作用最显著（P < 0.05）;高锰酸钾处理可提高粗茎秦艽种子的萌发率,浓度为1.5%时效果较显著（P < 0.05）,而过氧化氢处理对粗茎秦艽种子的萌发效果不如前者,此外,用500 mg/L的赤霉素浸种和300 mg/L的聚乙二醇预处理也可显著打破粗茎秦艽种子休眠（P < 0.01）。研究结果表明粗茎秦艽种子的内源抑制物是影响其休眠的因素之一;种皮的机械阻碍也在一定程度上影响了种子萌发;粗茎秦艽种子具有综合性休眠特性。高锰酸钾预处理、赤霉素浸种和聚乙二醇引发均可打破种子休眠、缩短种子出芽时间,提高种子的发芽能力。     

温水浸种对蒙古黄芪种子萌发特性的影响

研究温水浸种破除蒙古黄芪（Astragalus membranaceus Bge.var.mongholicus（Bge.） Hsiao）种子休眠的适宜条件,采用恒温和变温2种浸种方式,测定蒙古黄芪种子在不同时间、不同温度梯度条件下的发芽率、发芽势和发芽指数。结果表明,温水浸种破除蒙古黄芪种子休眠的效果显著,变温浸种效果略优于恒温浸种。不同温度破除休眠效果排序依次为60℃> 70℃> 80℃,其中60℃变温浸种2 min和5 min的发芽率分别为对照的2.87倍和2.31倍。浸种时间、浸种温度均可显著影响蒙古黄芪种子萌发,与对照相比,随着浸种时间增加,发芽率总体呈现出先急速升高后逐渐降低的趋势;发芽指数与浸种温度极显著正相关;浸种温度与发芽势和发芽指数均显著正相关。本研究结果表明温水浸种可以提高种子发芽率,保持种子活力。     

北柴胡和狭叶柴胡的萌发试验研究

目的：提高北柴胡和狭叶柴胡种子的发芽率,加大黑龙江省道地药材的开发力度。方法：采用40℃温水浸种8h后,流水冲洗4h;150w微波辐射25s（北柴胡）、20s（狭叶柴胡）;柳枝浸出液为萌发剂;浓H2SO4浸种2s,同时培养未经处理的种子作为对照组。结果流水组两种柴胡的发芽率分别为82%、86%;微波组发芽率分别为77%、75%;柳浸组发芽率分别为67%、72%;浓硫酸组发芽率分别为45%和35%;对照组发芽率41%、47%。结论：大规模播种可采用40℃温水浸种8h,流水冲洗4h,150w微波辐射25s（北柴胡）、20s（狭叶柴胡）,采用柳枝浸出液为萌发剂均可提高发芽率。     

香圆种子萌发特征及催芽技术

以香圆（Citrus wilsonii Tanaka）种子为实验材料,通过测定种皮透水性、种皮机械障碍、发芽温度等对种子萌发的影响从而探讨种子萌发特征;通过切割种子、热水浸种、赤霉素浸种、低温层积等处理方式探索种子的催芽方法。结果显示：香圆种皮不存在透水性障碍,但其机械障碍对种子萌发具有抑制作用,且内种皮对萌发影响较大;种子在15℃、25℃、35℃恒温及15℃/30℃变温条件下均可萌发,其最适萌发温度为25℃;采用低温层积、热水浸种、切除1/3种子等方法均可一定程度上促进种子萌发。生产中宜采用始温60℃热水浸种24h的方法对香圆种子进行催芽。     

Water limitation effect on seed development and germination in Trigonella coerulea (Fabaceae)

The annual garden spice legume Trigonella coerulea was subjected to controlled drought conditions to investigate the influence of resource limitation on flowering, seed production and germination. Limitations in water availability significantly reduced plant height and the number of fruits produced. However, treatments had no significant effect on seed set within the fruit. Fruit number per plant, rather than seed number per fruit was affected by resource limitation. Plants growing under water deficiency had higher flower abortion rates. Simultaneously, the size and germination rate of the seeds were decreased. In terms of reproductive success T. coerulea was not able to adapt reproduction to water shortage. Increase in moisture had no significant effect on seed quantity and quality when compared to the control.     

Water Relations of Tomato Seeds During Imbibition and Osmotic Priming

Water uptake of tomato (Lycopersicon esculentum Mill. cv. Moneymaker) seeds during germination was obviously triphasic. The completion of the first phase of water uptake by whole seed could not be realized until 10~12 h later after sowing though varies in different parts of seed. The mechanical resistance of endosperm and seed coat restricted water uptake of the embryo envoleped by the endosperm. Water potential of the intact embryo was still 0. 6~0. 9 Mpa lower than the whole seed when the equilibrium between seed and imbibing solution was established. GA and ABA had no direct effects on the water uptake of tomato seeds. The water potential of embryo was positively correlated with its moisture content. The osmotic potential of tomato embryos decreased slowly during imbibition in water and osmotic solution as well.     

A comparative study of water distribution, free radical production and activation of antioxidative metabolism in germinating pea seeds

The aim of this study was to investigate whether there is a relationship between hydration of the embryo axes and cotyledons and the resumption of the oxidative metabolism in both organs of germinating seeds of pea (Pisum sativum L. cv. Piast). Nuclear magnetic resonance (¹H-NMR) spectroscopy and imaging were used to study temporal and spatial water uptake and distribution in pea seeds. The observations revealed that water penetrates into the seed through the hilum, micropyle and embryo axes, and cotyledons hydrate to different extents. Thus, inhomogeneous water distribution may influence the resumption of oxidative metabolism. Electron paramagnetic resonance (EPR) measurements showed that seed germination was accompanied by the generation of free radicals with g₁ and g₂ values of 2.0032 and 2.0052, respectively. The values of spectroscopic splitting coefficients suggest that they are quinone radicals. The highest content of free radicals was observed in embryo axes immediately after emergence of the radicle. Glutathione content decreased during the entire germination period in both embryo axes and cotyledons. A different profile was observed for ascorbate, with significant increases in embryo axes, coinciding with radicle protrusion. Electrophoretic analysis showed that superoxide dismutase (SOD, EC 1.15.1.1), catalase (CAT, EC 1.11.1.6), ascorbate peroxidase (APX, EC 1.11.1.11), dehydroascorbate reductase (DHAR, EC 1.8.5.1) and glutathione reductase (GR, EC 1.6.4.2) were present in dry seeds and were activated later during germination, especially in embryo axes. The presence of all antioxidative enzymes as well as low molecular antioxidants in dry seeds allowed the antioxidative machinery to be active as soon as the enzymes were reactivated by seed imbibition. The observed changes in free radical levels, antioxidant contents and enzymatic activities in embryo axes and cotyledons appear to be more closely related to metabolic and developmental processes associated with preparation for germination, and do not correspond directly to the hydration of the tissues.     

The involvement of polyphenols and peroxidase activities in heavy-metal accumulation by epidermal glands of the waterlily (Nymphaeaceae)

Co-localization of polyphenols and peroxidase activity was demonstrated in epidermal glands of the waterlily (Nymphaea) by histochemistry. Total phenols, tannins and peroxidase activity were determined quantitatively in plant extracts. Polyphenols were partially identified and were found to consist mainly of hydrolyzable tannins, gallic and tannic acid derivatives.Nymphaea polyphenols were shown to chelate Cr, Hg, and Pb in vitro, and Cd-binding by polymerized polyphenols was demonstrated in leaves exposed to Cd in vivo. Both polyphenols and peroxidases were found at very high constitutive levels, which were not induced or altered by external conditions, such as light and heavy-metal stress. It is suggested that the polymerization of polyphenols by peroxidases, enhanced after heavy-metal uptake and detoxification, is responsible for the binding of heavy metals in Nymphaea epidermal glands. Received: 29 April 2000 / Accepted: 8 June 2000     

Oxidative damage in liver after perinatal intoxication with lead and/or cadmium

Lead acetate (300 mg Pb/L) and/or cadmium acetate (10mg Cd/L) in blood and liver were administrated as drinking water to pregnant Wistar rats from day 1 of pregnancy to parturition (day 0) or until weaning (day 21), to investigate the toxic effects in blood and in the liver. Both metals produced mycrocitic anaemia in the pups as well as oxidative damage in the liver, as suggested by the significant increase in TBARS production and the high catalase activity. Moreover, intense alkaline and acid phosphatase activity, used as biomarkers of liver adaptation to damaging factors, was observed. In addition, the toxikinetics are different for Pb and Cd: while Cd is a hepatotoxic from day 0, Pb is not until day 21. Finally, simultaneous perinatal administration of both metals seems to protect, at least, in the liver TBARS production against the toxicity produced by Cd or Pb separately.     

干藏和淹水对三峡库区21种草本植物种子萌发的影响

研究了三峡库区河岸带21种常见草本植物种子在干藏以及不同淹水深度（0.5、1.0和2.0m）处理下的萌发特性。结果表明：以新采集种子的萌发率作为对照,干藏后8个物种的萌发率增加（占总数的38.1%）,1种萌发率不变（4.8%）,12种萌发率降低（57.1%）。与对照相比,淹水后3个物种的萌发率增加（占总数的14.3%）,2个物种的萌发率不变（9.5%）,16个物种的萌发率下降（76.2%）。水淹处理后,占总数76.2%的物种的萌发率下降,其中11种显著下降;但淹水深度对种子的萌发没有显著影响。干藏后的繁穗苋（Amaranthus paniculatus）、皱果苋（Amaranthus viridis）和红刺果苋（Amaranthus sp.）以及水淹后的鳢肠（Eclipta prostrata）萌发率〉50%,而且能够利用库区退水期完成生活史形成种子,表明这些植物可作为三峡库区消落带植被恢复时优先考虑的物种源。     

Cadmium stress affects seed germination and seedling growth in <Emphasis Type="Italic">Sorghum bicolor</Emphasis> (L.) Moench by changing the activities of hydrolyzing enzymes

Seed germination, one of the most important phases in the life cycle of a plant, is highly responsive to existing environment. Hydrolyzing enzymes play a major role in the mobilization of food reserves by hydrolyzing carbohydrates, proteins and fats. This paper reports on the effect of Cd toxicity on seed germination and the activities of hydrolyzing enzymes, like acid phosphatases (ACPs), proteases and α-amylases in Sorghum bicolor (L.) Moench. The metal uptake by embryonic axes and seeds was quantified. We found that sorghum could tolerate up to 0.5 mM Cd. At concentrations above 3.0 mM, seed germination was adversely affected with a complete cessation of seedling growth. All investigated hydrolyzing enzymes exhibited a significant decrease in activity with increasing Cd concentrations. The isozyme profiles indicated the loss of one or two isozymes of ACP, induction of a new isozyme for total protease (at 3.0 mM Cd) and a decline in the intensity of α-amylase isozymes. SEM studies revealed that Cd affected a change in root hair density. SEM investigations also confirmed the assay results of the inhibition of starch mobilization from endosperm. This suggested an inhibition of the hydrolysis of reserve carbohydrates and translocation of hydrolyzed sugars, ultimately resulting in decreased germination and disruption of seedling growth. Because sorghum is an important dryland crop, its response to the presence of Cd in agro-ecosystems and Cd-induced phytotoxicity during seed germination and seedling growth needs critical investigation.     

Testing the potential seed availability in dung samples: comparison of two seedling emergence methods

To optimize the estimation of species composition and viable seed content of herbivore faeces and to make different approaches comparable, two seedling emergence methods are evaluated. The Ter Heerdt method (TH) employs concentrated samples, potentially increasing and accelerating seedling emergence, as shown for soil samples (95% of all seedlings emerged within 6 weeks). Samples are kept under controlled conditions (glasshouse or climate room). Secondly, a common garden method (CG) using unconcentrated samples so that seasonal changes could fulfill the germination requirements of a broad species spectrum (experiment duration approximately 15 months) was applied. The methods were tested by the use of sheep faeces samples, collected during a six-day grazing period in a threatened dry grassland (Allio-Stipetum capillatae).Both methods proved largely similar in species composition (QS=0.81) and viable seed content (QS=0.69). More species (e.g. monocotyls) and a higher seedling emergence of hard-seeded species (Fabaceae and Cistaceae) were found in the CG method. Besides a higher emergence of some small-seeded winter annuals, few other species emerged exclusively by use of the TH method. Nevertheless, all species detected by only one method were found in low individual numbers (4).Depending on research interest and availability of space and time, the most appropriate method can be chosen. If the main focus is on the species composition, unconcentrated faeces samples can be studied by CG. In case the overall viable seed content is more important and/or a shorter time period is available, TH serves as a suitable alternative.     

Soil seed banks in a heavy-metal polluted grassland at Prayon (Belgium)

Seed banks were investigated in a grassland dominated byAgrostis capillaris and Thlaspicaerulescens, established on soils polluted by atmospheric depositsof SO₂ and heavy-metal rich dust (Zn, Pb, Cd) from an ore-smelter inPrayon (E Belgium). A total of 155 soil cores (0–7 depth)distributed among 10 plots were sampled in November and March. Each samplingplot was characterised for the botanical composition and standing biomass of theestablished vegetation as well as for four environmental parameters (soil pH andextractable zinc, volume percentage of stones, relative altitude within thesite). Very high densities of germinable seeds were found (grand mean ±SE, pooled over dates and plots: 28,950 ± 6830 ). Agrostis capillaris represented 95% of theseeds, followed by Thlaspi caerulescens (3%); the remaining2% were shared among 24 other species. Two thirds of the seeds were concentratedin the upper 3 of the soil. The high seed density might be dueto a low rate of microbial decay of seeds due to heavy metal toxicity. The seedbank is almost entirely derived from those few heavy-metal tolerant species thathave re-colonised the polluted area in the last three decades. Only two species(Anagallis arvensis and Urtica dioica)not represented in the established vegetation and with seeds concentrated in thelower soil layer are probably derived from seed buried prior to the onset ofheavy-metal pollution. This study provides the first data suggesting that theheavy-metal hyperaccumulator Thlaspi caerulescens possessespersistent seed banks. Seed densities of Agrostiscapillaris, Thlaspi caerulescens andRumex acetosa were positively correlated with the standingbiomass of grasses and negatively correlated with the volume percentage occupiedby stones in the soil. It is argued that small-scale variation in microclimaticfactors that cue germination (fluctuations in soil temperature and moisture) canaccount for the observed pattern of spatial variation in seed density. From thestandpoint of restoration management, the results indicate that little change inthe floristic composition of the heavy-metal tolerant grasslands of Prayon canbe expected from the recruitment of buried seeds.

     

Soil seed bank dynamics in post-fire heathland succession in south-eastern Australia

Soil seed banks can exert a strong influence on the path of vegetation succession following fire, with species varying in their capacity to persist in the seed bank over time, leading to changes in seed bank composition and propagules available for post-fire colonisation. This study examined the effect of time since fire on soil seed bank dynamics in a chronosequence of seven sites spanning 26 years in a south-eastern Australian sand heathland. No significant change was evident in the species richness and density of the germinable soil seed bank, but species composition differed significantly among young (0–6 years since fire), intermediate (10–17 years since fire) and old-aged (24–26 years since fire) sites (using presence/absence data). No significant trend was observed in the similarity between the extant vegetation and the soil seed bank with time since fire. A total of 32% of the species recorded in the soil seed bank were not present in the above-ground vegetation at the same site, which suggests that species requiring fire for germination may be present in the seed bank. Most species present in the extant vegetation were not recorded (63%) or were in very low abundances in the soil seed bank (29%). The mode of regeneration appears to be the major determinant of species absence in the soil seed bank, as 66% of species occurring in the extant vegetation but not in the seed bank have the capacity to regenerate by resprouting. This study shows that a major shift in the successional pathway after fire due to altered seed bank composition is unlikely in this vegetation; most species not recorded in the seed bank are either resprouters (obligate or facultative) or serotinous, suggesting that they will readily regenerate following fire. Unless fire frequencies are high and kill fire-sensitive obligate seeders before they reach maturity, the chance that the soil seed bank could substantially alter vegetation composition within the study area after fire is low. However, it is unclear how successional pathways may alter in response to severe fires with the potential to kill both seeders and resprouters.     

Influence of environmental factors on Vallisneria americana seed germination

Over the course of a growing season (April–October) water quality (water temperature, light, salinity, dissolved oxygen) and reproductive phenology (biomass, production of flowering shoots and seed pods, seed bank densities) were quantified in three Vallisneria americana beds in Nanjemoy Creek, MD, a tributary to the Chesapeake Bay. Clonal production of V. americana biomass increased at all sites when water temperatures rose above 25 °C. Flowering occurred during peak biomass (August–September) and resulted in the production of up to 16,000 seeds m⁻² at the end of the growing season. However, observed seed bank densities represented <1% of seed production. Laboratory experiments quantified the effects of dissolved oxygen (0.29–8.00 mg l⁻¹), light (0–160 μmol m² s⁻¹), temperature (13–29 °C), salinity (0.1–17.4 psu), sediment composition (3–86% sand; 0.9–8.3% sediment organic content), and burial depth (0.2–10 cm) on V. americana seed germination. Germination of V. americana seeds was enhanced (greater overall germination and shorter time to germination) under oxygenated conditions (8.00 mg l⁻¹), temperatures >22 °C, salinities of <1 psu, and in sediments composed of ≤3% organic content and >40% sand. Light (<160 μmol m⁻² s⁻¹) and burial depth (0.2–10 cm) had no significant effects on germination. Temperatures most favorable for seed germination (>22 °C) occurred in June, 2 months in the growing season just prior to development of peak vegetative standing stock. Seedlings were therefore at a distinct disadvantage to plants developed from over wintering buds. A lack of viable seed retention and inadequate environmental conditions at critical times in the growing season may be limiting seed germination success and subsequent seedling establishment within V. americana beds in the Chesapeake Bay. However, ungerminated seeds were found to maintain high viability, especially at salinities of 10 psu that can have significant negative effects of shoot growth survival. This suggests that seeds may serve as a source of reproductive material for bed recovery after periods of drought or other stressful conditions in estuarine systems.     

Genesis of a soil seed bank on a primary succession in the Central Alps (Ötztal,Austria)

Soil seed bank and standing vegetation were investigated on the Rotmoos Glacier foreland (Ötztal, Tyrol, Austria) along the chronosequence (i.e. on the pioneer, early, and late successional stage) as well as on a subalpine pasture beyond the glacier foreland (old successional stage). We aimed to answer the following questions: (1) How large are soil seed banks along the successional gradient? (2) Do the seed banks reflect the actual standing vegetation or do they remember earlier successional stages or do they represent already the next successional stage?     

Biofortification of chromium in fenugreek seeds

BackgroundFenugreek and chromium are known to have anti-diabetic properties and this has been well demonstrated by multiple studies. Researches have been undertaken to study thebiofortification of chromium (Cr³⁺⁾ in fenugreek. Some of the researchers have studied the biofortification from the soil irrigated with tannery waste water or from soil amended by tannery-sludge, with a view of enhancing the anti-diabetic effect of fenugreek plants. The present research work was also undertaken to increase the chromium content of fenugreek seeds, but through direct treatment of chromium picolinate solution to fenugreek seeds.MethodsFenugreek seeds were procured and divided in four groups having three batches of 10 g seeds- treatment₁, treatment₂, control₁ and control₂. Control₁ group was kept raw and was given no treatment and control₂ group was germinated using double distilled water. Treatment₁ and treatment₂, on the other hand, were given treatment of 0.02 g and 0.04 g chromium picolinate solution, on first day and 0.01 g and 0.02 g chromium picolinate solution on second day, respectively. The germinated samples were then completely dried, powdered, digested with di-acid mixture and assayed using Inductively Coupled Plasma optical emission spectrometry method for chromium content. The treatment₁ sample was selected for further nutritional analysis along with control₁ and control₂ group to compare the nutritional composition of raw, germinated and chromium treated fenugreek seed flour. Fifteen sprouts from treatment₁ group (treatment_1A group) and fifteen sprouts from control₂ group (control₃ group) were sown in earthen pots for the analysis of chromium content in seeds of new plants.ResultsThe fenugreek seeds treated with two different concentrations of chromium picolinate viz. treatment₁ and treatment₂ group attained 55 and 80 times higher chromium content as compared to control₂ group, respectively. All the estimated minerals and bioactive compounds were significantly high (p ≤ 0.01) in germinated fenugreek seed flour and chromium treated fenugreek seed flour compared to raw fenugreek seed flour. Germinated fenugreek seed flour and chromium treated fenugreek seed flour were statistically comparable to each other in respect of all the parameters analysed. Hence, it was evident that enriching fenugreek seeds with chromium, did not affect the nutritional content of fenugreek seed by any mean. Also, there was no significant difference between the chromium content in seeds of control₃ group and T_1A group.ConclusionsTreatment of fenugreek seeds with chromium solution seems to be an efficient and safe method for increasing their chromium concentration as compared to application of chromium to the soil for biofortification with minimal to no chance of chromium accumulation and inheritance in next generation plants. However, there is a need of more research to see how reliable these observations would be when different chromium salts and/or varied chromium concentration are used.