Allelopathic effects of parthenin against two weedy species,Avena fatua and Bidens pilosa

Parthenin is a natural constituent of Parthenium hysterophorus with phytotoxic and allelopathic properties. Its effect on two weedy species viz. Avena fatua and Bidens pilosa was studied with a view to explore its herbicidal potential. Germination of both the weeds was reduced with increasing concentration of parthenin and a dose-response relationship was observed. This provided information on LC₅₀ and Inhibition threshold concentrations of parthenin that could be useful for future studies. Further, parthenin also inhibited the growth of both the weeds in terms of root and shoot length and seedling dry weight. Inhibition of root growth was greater than that of shoot growth. Similar observations were made when the test weeds were grown in soil amended with different concentrations of parthenin. In addition to growth, there was a reduction of chlorophyll content in the growing seedlings. It also caused water loss in the weedy species. The study, therefore, reveals that parthenin exerts an inhibitory effect on the growth and development of both weeds and can be further explored as a herbicide for future weed management strategies.     

Growth regulatory response of parthenin and its derivatives

Parthenin and its pyrolysed and photo-derivatives exhibit a range of growth regulatory effects depending on their structure. At 5 mg L-1 (the lowest concentration used) the activity of all parthenin derivatives was greater than that of indole-3-acetic acid (IAA) in biotests based on growth and morphogenetic response of hypocotyl cuttings of Phaseolus aureus. Seedling growth was greatly enhanced by pyrolysed parthenin whereas parthenin and photo-parthenin promoted rooting of the hypocotyl cuttings. Structural changes in parthenin resulting from chemical and photochemical reactions, therefore, alter its growth regulatory action.     

Mutagenic activity of structurally related oxiranes and siloranes in Salmonella typhimurium

The in vitro and in vivo genotoxicity of parthenin, a sesquiterpene lactone from Parthenium hysterophorus L. with allergenic and irritant action, was assessed in three short-term tests: bacterial reversion in Salmonella typhimurium and Escherichia coli, in vitro chromosomal aberrations in peripheral blood lymphocytes and micronuclei in mouse peripheral blood. Parthenin was not mutagenic in S. typhimurium strains TA 1535, TA 1537, TA 98 and TA 100 but a weak response was observed in TA 102 (+S9) from 0.19 to 1.22 micromole per plate. Concentrations of 7.62 micromole per plate or higher were toxic, but the effect was reduced when S9 was present. Screening of oxidative mutagenesis with E. coli strains IC 188 and IC 203 gave negative results. Parthenin induced chromosomal aberrations, mainly chromatid breaks, in blood lymphocytes exposed to 10-60 microM during 20 h. An association was found with cytotoxicity, since concomitant nuclear alterations such as pycnosis, micronuclei and karyorrhexis were observed. Sister chromatid exchanges (SCEs) in lymphocytes were not influenced by exposure to parthenin; rather a decrease was observed at 60 microM. On the other hand, a minor increment in polyploid metaphases was found at 40 microM. When a single intraperitoneal (i.p.) dose of 4-31 mg/kg of parthenin was administered to mice, a positive increase in the micronucleated reticulocyte (RET) frequency was observed at 48 h for both sexes at the highest dose.     

Ecological significance of root tip rotation for seedling establishment of Oryza sativa L

How plant seeds secure root penetration into soil to obtain good seedling establishment is one of the basic ecological problems. In this study, seminal root growth was investigated to clarify the cause of varietal difference of seedling establishment in direct seeding of rice in flooded paddy fields, with special reference to root tip rotation. In a field experiment, seedling establishment percentage had a weak correlation with seminal root elongation rate but was not correlated with apparent seedling weight in water, which has been reported to be the cause of floating seedlings resulting in poor seedling establishment. Root tip rotation was analyzed for indoor-grown seedlings using spectrum analysis: the maximum entropy method (MEM) was used. Maximum entropy method power spectrum analysis clarified that maximum MEM power density (practically corresponds to spiral angle) detected in the frequency range above 0.1 cycles mm^-1 was highly and positively correlated to seedling establishment percentage in the field experiment. Maximum MEM power density in high correlation with seedling establishment was mostly found around frequencies of 0.2 cycles mm^–1, which corresponded to 2.0–3.4 cycles of root tip rotation per day. From these results, root tip rotation (circumnutation) with a larger spiral angle was suggested to play an important role in the establishment of rice seedlings on flooded and very soft soil. A possible explanation for why a larger spiral angle was advantageous for seedling establishment is that if buoyancy and seedling weight are constant, a larger pushing force of the seminal root is available without causing floating of a seedling, due to the upward force being a reaction of the seminal root pushing force.     

Impact of the auxin signaling inhibitor p-chlorophenoxyisobutyric acid on short-term Cd-induced hydrogen peroxide production and growth response in barley root tip

Short-term treatment (30min) of barley roots with a low 10μM Cd concentration induced significant H(2)O(2) production in the elongation and differentiation zone of the root tip 3h after treatment. This elevated H(2)O(2) production was accompanied by root growth inhibition and probably invoked root swelling in the elongation zone of the root tip. By contrast, a high 60μM Cd concentration induced robust H(2)O(2) production in the elongation zone of the root tip already 1h after short-term treatment. This robust H(2)O(2) generation caused extensive cell death 6h after short-term treatment. Similarly to low Cd concentration, exogenously applied H(2)O(2) caused marked root growth inhibition, which at lower H(2)O(2) concentration was accompanied by root swelling. The auxin signaling inhibitor p-chlorophenoxyisobutyric acid effectively inhibited 10μM Cd-induced root growth inhibition, H(2)O(2) production and root swelling, but was ineffective in the alleviation of 60μM Cd-induced root growth inhibition and H(2)O(2) production. Our results demonstrated that Cd-induced mild oxidative stress caused root growth inhibition, likely trough the rapid reorientation of cell growth in which a crucial role was played by IAA signaling in the root tip. Strong oxidative stress induced by high Cd concentration caused extensive cell death in the elongation zone of the root tip, resulting in the cessation of root growth or even in root death.     

Effect of lithium ions on the growth of wheat roots and the role of phosphoinositide cycle in growth regulation]

The influence of lithium ions (LiCl in concentrations of 0.5, 1.0, and 5.0 mM) on the growth processes of roots of 2-5-day old wheat seedlings was studied. It was shown that the inhibition of the root growth increased with the increase of LiCl concentration and seedling age. The membrane potential of root cells was lower and the loss of K+ by cells was greater when roots were treated with 5 mM LiCl, compared with the control. The growth inhibition by lithium was decreased by univalent ions, partially by potassium at the beginning of growth and completely by sodium throughout the experimental period. The divalent ions calcium and barium decreased the Li(+)-induced inhibition of root growth by reducing the rate of lithium uptake by cells. Myoinositol, controlled by Li-sensitive inositolmonophosphatase, reversed the Li-induced root growth inhibition in 2-day old seedlings, but did not prevent the inhibition during subsequent elongation. It can be concluded that lithium effects on wheat root growth are mediated by a partial blockage of signal transduction for proliferation (via the phosphoinositide cycle), because of calcium deficiency and caused by modification of ion transporting systems of the plasmalemma, and by disturbance of ion gradients, primarily H+ and K+.     

The impact of heavy metals on the activity of some enzymes along the barley root

Barley seedlings 48 h after the onset of germination on filter paper treated for 24 h by 1 mM cadmium (Cd), 3 mM nickel (Ni) or 0.5 mM mercury (Hg) showed similar approximately 45% root growth inhibition. Although root growth inhibition was similar, loss of cell viability evaluated, as Evans blue uptake was distinct among Cd, Ni and Hg treated roots. While Cd and Hg caused cell death along the whole barley root (0–8 mm), Ni induced significant loss of cell viability only in root cells 6–8 mm distance from the root tip. Our results suggest that different metabolic processes are activated in different parts of barley root in relation to distance from the root tip during heavy metal (HM) treatment. Some of them are characteristic for several HMs such as inhibition of ascorbic acid oxidase or glutathione-S-transferase stimulation, while others are specific for individual HMs, e.g. activation of acid phosphatase and lipoxygenase by Cd and Hg, or inhibition of ascorbate peroxidase by Ni and Hg treatment.     

Cd胁迫下黄菖蒲幼苗根系生长与Cd积累的研究

采用水培法研究了Cd胁迫对黄菖蒲（Iris pseudacorus L．）幼苗根系生长和Cd含量的影响,用透射电镜观察了Cd胁迫对根尖细胞亚显微结构的影响并确定了Cd沉积部位。结果表明,10mg·L^-1 Cd处理对黄菖蒲幼苗根系的生长有一定促进作用,根毛数量及不定根上侧根的数量、密度和长度均高于对照;Cd浓度高于25mg·L^-1时,根系的生长发育受到抑制,上述指标均显著低于对照。用10～40mg·L^-1 Cd处理后,黄菖蒲幼苗不定根上侧根中的Cd含量均显著高于未长侧根的不定根和去掉侧根的不定根（P〈0．05）,说明不定根上的侧根对Cd沉积有重要作用。100mg·L^-1 Cd处理能导致根尖细胞质浓缩、质壁分离和液泡变大,且大量Cd颗粒沉积在细胞壁上和细胞间隙中,表明细胞壁和细胞间隙对Cd的沉积作用可能是黄菖蒲幼苗耐Cd胁迫的重要机理之一。     

Effect of water stress on seed germination and seedling growth inOryza sativa L.

The effect of water stress induced by application of polyethylene glycol 6000 during seed germination and seedling growth ofOryza sativa L. cv. IAC 165 was analysed. The seed germination was inhibited by the decrease in the water potential of the medium, the inhibition being greater under white light than under continuous darkness. When the seedling was submitted to water stress (-0.51 MPa) white light inhibited growth of root, coleoptile-and leaf, while under no stress conditions white light caused increase in growth of root and leaf and only inhibition of coleoptile growth.     

Primary stress response induced by different elements is mediated through auxin signalling in barley root tip

Short-term exposure (15 min) of barley roots to different chemical elements revealed that Cd, Cu, Hg and Pb were the most toxic ones causing a marked root growth inhibition even at µM concentrations. Gd, La, Al, Cr, As, Zn, Ni and Se inhibited root growth to a similar extent only at mM concentrations. Despite the high 20 mM concentration, Co caused only a slight, while Mn, Mg or Ca did not evoke any root growth inhibition. Elements at concentrations inhibiting root growth caused a considerable accumulation of indole-3-acetic acid in the root apex. While Cr, As and Zn inhibited, Cd, Cu, Hg, Pb, Gd, La and Al markedly stimulated the generation of reactive oxygen species in the beginning of differentiation zone. Auxin signalling inhibitor alleviated or prevented root growth inhibition, reactive oxygen species generation and the stimulation of lipoxygenase and glutathione peroxidase activity by various elements, indicating a key role of auxin signalling in the stress response of barley root tip. On the other hand, it did not affect or even had an additive effect on dehydroascorbate reductase and ascorbic acid oxidase activity in combination with different elements. Our results indicate that the primary response of barley roots to the presence of various chemical elements during the short-term treatment is not a specific but rather a general adaptive stress response enabling the plant to survive adverse conditions.     

Salinity induced nuclear and DNA degradation in meristematic cells of soybean (Glycine max (L.)) roots

Changes in the nuclei of meristematic root cells of soybean (Glycine max (L.) Merr. cv. Acme) in response to severe salinity were studied. Root growth was inhibited by 200 mM NaCl, when 1 mM CaCl_2 was present in the culture media. Increasing CaCl_2 up to 5 mM partially prevented this inhibition. However, inhibition also occurred with 100~mM NaCl without CaCl_2. We examined the meristematic cells under a series of NaCl treatments. Nuclear deformation of the cells occurred with 24 h of 150 mM or higher NaCl, and was followed by degradation of nuclei in the apical region of the root. TEM observation and agarose gel electrophoretic analysis confirmed that root tip nuclear DNA deformed or degraded with 150 mM or higher NaCl concentrations.     

Effects of 2-benzoxazolinone on the germination, early growth and morphogenetic response of mung bean (Phaseolus aureus)

2‐Benzoxazolinone (BOA), a type of hydroxamic acid present in cereals and implicated in allelopathy, is now being viewed as a potential candidate for the development of natural herbicides. A study was conducted to determine the effect of BOA on mung bean (Phaseolus aureus) through a multitude of bioassays to understand its physiological and biochemical action. It was observed that BOA significantly decreased the germination of mung bean and its early growth (measured in terms of seedling length and dry weight). A typical dose–response relationship was observed with BOA treatment, and I₅₀ values (concentrations at which 50% inhibition occurs) for germination, seedling length and seedling dry weight were calculated to be 4.3, 0.71 and 0.77 mM , respectively. There was therefore a greater inhibitory effect on seedling growth than on germination. Treated seedlings were characterised by a loss of chlorophyll and decreased respiratory activity, indicating a possible adverse effect of BOA on photosynthetic and respiratory metabolism. Mitotic activity in root‐tip cells of onion (Allium cepa) was completely arrested in response to BOA treatment, and the cells exhibited abnormality in shape and size. BOA also adversely affected rhizogenesis in hypocotyl cuttings of mung bean, indicating an impact on morphogenetic potential. It was associated with significant changes in the protein content and activities of proteases and polyphenol oxidases during the root development phase. This study concludes that BOA interfered with essential biochemical processes in mung bean. Such studies provide useful information on the biochemical and physiological modes of actions of BOA, with a view to its use as a herbicidal compound.     

Alleviation of salt stress by plant growth regulators and IAA producing bacteria in wheat

The action of phytohormone producing bacteria and plant growth regulators on germination and seedling growth of wheat under saline conditions were studied. Seed dormancy enforced by salinity (100 mM NaCl) was substantially alleviated and the germination was promoted by gibberellin, auxin, zeatin, and ethephon from 54 to 97%. The IAA producing bacterial strains Pseudomonas aureantiaca TSAU22, Pseudomonas extremorientalis TSAU6 and Pseudomonas extremorientalis TSAU20 significantly increased seedling root growth up to 25% in non-salinated conditions and up to 52% at 100 mM NaCl, compared to control plants. It is concluded that growth regulators considerably alleviated salinity-induced dormancy of wheat seeds. The facts mentioned above make it possible to recommend root colonizing bacteria that produce phytohormone to alleviate salt stress of wheat grown under conditions of soil salinity.     

A mathematical model to predict the tissue response to parthenin — An allelochemical

Parthenin — a sesquiterpene lactone fromParthenium hysterophorus L. is an allelochemical that prevents the germination ofPhaseolus aureus Roxb. cv. ML-5 seeds. The response of the seed has been attributed to the inhibition of the respiratory electron transport ability of its embryo. It has been shown to depend directly not only upon concentration of parthenin, but simultaneously on the duration of exposure of the seeds to the chemical as well. A strong correlation exists between the quantum of the response and the product of the period of exposure and the concentration of parthenin. In order to predict the maximum possible germination ability of the seed exposed for a given period to a given concentration of parthenin, an expression X = 10000 Y³ / [e^Y/0.31-1] was formulated from the equation X = AY³/[e^Y/Y0-1], where X represents values of maximum respiratory activity, Y represents the product of concentration and time in units mg cn^-3 h, A represents a dimensional constant. The trend and nature of response that is calculated on the basis of formulation coincides with that of measured response through spectrophotometry.     

Phytotoxicity of the volatile monoterpene citronellal against some weeds

A study was undertaken to assess the phytotoxicity of citronellal, an oxygenated monoterpenoid with an aldehyde group, towards some weedy species [Ageratum conyzoides L., Chenopodium album L., Parthenium hysterophorus L., Malvastrum coromandelianum (L.), Garcke, Cassia occidentalis L. and Phalaris minor Retz.]. A significant effect on weed emergence and early seedling growth was observed in a dose-response based laboratory bioassay in a sand culture. Emergence of all test weeds was completely inhibited at 100 micro/g sand content of citronellal. Seeds of A. conyzoides and P. hysterophorus failed to emerge even at 50 microg/g content. Root length was inhibited more compared to shoot length. The failure of root growth was attributed to the effect of citronellal on the mitotic activity of growing root tips cells as ascertained by the onion root tip bioassay. At 2.5 mM treatment of citronellal, mitosis was completely suppressed and at higher concentrations cells showed various degrees of distortion and were even enucleated. The post-emergent application of citronellal also caused visible injury in the form of chlorosis and necrosis, leading to wilting and even death of test weeds. Among the test weeds, the effect was severe on C. album and P. hysterophorus. There was loss of chlorophyll pigment and reduction in cellular respiration upon citronellal treatment indicating the impairment of photosynthetic and respiratory metabolism. Scanning electron microscopic studies in C. occidentalis leaves upon treatment of citronellal revealed disruption of cuticular wax, clogging of stomata and shrinkage of epidermal cells at many places. There was a rapid electrolyte leakage in the leaf tissue upon exposure to citronellal during the initial few hours. In P. minor electrolyte leakage in response to 2 mM citronellal was closer to the maximum leakage that was obtained upon boiling the tissue. The rapid ion leakage is indicative of the severe effect of citronellal on the membrane structure and loss of membrane integrity. In all, the study concludes that citronellal causes a severe phytotoxicity on the weeds.     

The Inhibition of Growth by Gravitropic Stimulation in Darkness in Agravitropic Primary Roots of Zea and the Role of Calcium

The primary roots of the "Golden Cross Bantam 70" cultivar ofZea mays are agravitropic in darkness and their orthogravitropismis light-dependent. Analysis of the agravitropic roots providesimportant information about the mechanism of orthogravitropism.However, the underlying mechanism of the agravitropic responsein darkness is unknown. We found that the growth of intact primaryroots was inhibited by gravitropic stimulation (i.e., changingthe orientation of the roots from vertical to horizontal) indarkness, but that of detipped roots was not. The role of calciumin this gravistimulation-dependent inhibition of growth wasinvestigated using apical 5-mm segments of the primary roots.The gravistimulation-dependent inhibition of growth was preventedby applying 10 mM MES-KOH buffer at pH 6.0 to the root cap.By contrast, the application of 0.1–1 mM buffer at pH6.0 and 10 mM buffer at pH 4.5–5.0 allowed the gravistimulation-dependentinhibition of growth. Furthermore, when the buffer of 10 mM(pH 6.0) contained 1–5 mM CaCl₂, the gravistimulation-dependentinhibition of growth was apparent. By contrast, when weak (1mM) buffer at pH 6.0 or 10 mM buffer at pH 4.5 contained 5 mMEGTA, no gravistimulation-dependent inhibition of growth wasobserved. Thus, the gravistimulation-dependent inhibition ofgrowth in darkness seemed to be mediated by an increase in thelevel of free Ca²⁺ in the root tip. These results suggest thatfree Ca²⁺ in the apoplast of the root tip plays an importantrole in the agravitropic response in darkness as well as inorthogravitropism under light of the roots of this cultivarof Zea mays. (Received March 21, 1994; Accepted July 25, 1994)     

Improving vigour assessment of pine (Pinus nigra Arnold) seedlings before their use in reforestation

ABSTRACT

We investigated whether changes in the root system of pine seedlings induced by stress (lifting of bare-root seedlings from the nursery bed irrespective of dormancy; prolonged storage of bare-root seedlings in a cold room) could provide a measure of plant vigour. Physiological parameters, such as growth potential and root electrolyte leakage, and morphological parameters, such as root length and number of root tips, were calculated. Computerised image analysis was used to measure root growth, overall and based on root-diameter class (0–0.5 mm, 0.5–1.0 mm and 1.0–1.5 mm). The efficiency of vigour assessment was evaluated by correlating the data for each parameter with percentage seedling survival. Root growth potential was more efficient than root electrolyte leakage, but both parameters were affected by seedling age. Total root length was a more efficient indicator of plant vigour than root tip number, particularly when referred to roots of the same diameter class. A comparative analysis of physiological and morphological parameters referred to the root systems improves their relative effciency.     

Induction of DNA single-strand breaks in barley by sodium azide applied at pH 3.

Sodium azide (1 to 50 mM), adjusted to pH 3 and applied for 2 h to presoaked barley seeds, induced a dose-dependent frequency of single-strand breaks in DNA of non-germinating embryos. This was demonstrated by sedimentation analyses of isolated DNA samples in alkaline sucrose gradients and in neutral sucrose gradients with 80% formamide. The doses applied also inhibited dose dependently the root length, seed germination and partially the seedling height. Only the sub-lethal doses (10 and 12.5 mM) induced a low frequency of chromatid breaks and translocations in the root tip metaphases. The sedimentation rate (in alkaline sucrose gradients) of calf thymus DNA treated with sodium azide at pH 3, was similar to that of the control DNA treated with buffer (pH 3) alone.     

Effects of periodic flooding and root pruning on Quercus nuttallii seedlings

In the southern United States, much of the emphasis in bottomland restoration is placed on establishing an oak-dominated forest. Artificial regeneration is an alternative for restoration on cleared lands and where a desirable seed source is not present. Currently the standard procedure for seedling preparation is to prune the roots prior to transplanting in the field. It is not fully known what effect(s) root pruning has on transplanted seedlings. In addition, bottomland restoration efforts inherently take place on floodplains. The potential interaction between root pruning and flooding on seedling performance is not known. This study consisted of two separate but related laboratory experiments. The purpose of the first experiment was to quantify the effects of various percentages of root removal and varying soil moisture regimes on transplanted Nuttall oak seedlings (Quercus nuttallii Palmer). Root pruning treatments consisted of removal of roots at 0%, 25% and 75% while soil moisture regime was maintained at non-flooded or periodically flooded conditions. Plant gas exchange, growth, and survival were measured. Root pruning alone had adverse effects on height growth during the first 72 days following transplanting. Periodic flooding also produced adverse effects on stomatal conductance (p = 0.0002), height growth (p = 0.005), and survival (p = 0.02). Photosynthetic data indicated that as pruning intensified in the periodically flooded seedlings, photosynthetic rates decreased. In contrast, as pruning intensified in the non-flooded seedlings, photosynthesis increased. This demonstrated that pruning rate had a varying effect on photosynthesis dependent upon soil moisture condition. Experiment 2 focused on the effects of varying degrees of root pruning on new root formation. The seedlings were grown under laboratory conditions, harvested at 0, 10, 20, and 30 days after treatment initiation, and analyzed for new root formation. Results of Experiment 2 indicated no difference in new root formation, root length, or root biomass due to the pruning treatment. Overall, our results from both experiments indicated that root pruning had no detectable long-term adverse effects on growth and survival of seedlings under drained soil conditions; however, as results from Experiment 1 demonstrated, if seedlings were planted in periodically flooded conditions, root pruning produced adverse effects. Thus, in restoration efforts utilizing Nuttall oak seedlings, the planting strategy and pruning rate should be carefully evaluated based on the knowledge of sites' hydrology. Alternatively, on sites with unpredictable flooding both pruned and unpruned seedlings may be utilized to ensure survival.