Auxinic herbicide conjugates with an α-amino acid function: Structural requirements for biological activity on motor cells.

Two Fabaceae exhibiting rapid osmocontractile pulvinar movements were used in this study because this activity is modified by natural auxin and dramatically by 2,4D. A short chain with a carboxylic group being required for auxinic properties, a critical point to analyze is whether the recently synthesized proherbicide ε-(2,4-dichlorophenoxyacetyl)-L-Lys (2-4D-L-Lys) maintains some biological activity despite the increase in length of the chain and the substitution of the carboxyl group by an α-amino acid function. No trace of 2,4D could be detected in the pulvinar tissues treated for 1 h with 2,4D-L-Lys. Complementary approaches (electrophysiology, pH measurements, use of plasma membrane vesicles) suggest that it was less efficient than 2,4D to activate the plasma membrane H+-ATPase (PM-H+-ATPase). However, it modified the various ion-driven reactions of Mimosa pudica and Cassia fasciculata pulvini in a similar way as 2,4D. Additionally, it was much more effective than fusicoccin to inhibit seismonastic movements of M. pudica leaves and, at low concentrations, to promote leaflet opening in dark, indicating that its mode of action is more complex than the only activation of the PM-H+-ATPase. Various substitutions on 2,4D-L-Lys affected its activity in correlation with the molecular descriptor "halogen ratio" of these derivatives. Conjugation with D-Lys also led to a decrease of pulvinar reaction, suggesting that 2,4D-Lys maintains the main signaling properties of 2,4D involved in pulvinar movements providing that the terminal zwitterion is in a suitable orientation. Our data guide future investigations on the effect of 2,4D and 2,4D-L-Lys on the vacuolar pump activity of motor cells.

Characteristic early membrane effects induced by tryptophan in pulvinar motor cells.

Tryptophan at concentrations higher than 0.1 mM, triggered characteristic early physiological effects such as rapid (within 5 min) dose-dependent membrane hyperpolarization in Mimosa pudica motor cells and modification of the time course of the spontaneous proton efflux monitored in the incubation medium of pulvinar tissues. The rapid modifications of the leaf turgor-mediated movements seen on the primary pulvini of M. pudica following a shock and on Cassia fasciculata leaflets during a transition from light to darkness indicate that tryptophan disturbed the ionic migrations involved in the electrophysiological events and in the osmocontractile reaction of the motor cells. These reactions were specific to tryptophan compared to those induced by serine and 5-hydroxytryptophan. The tryptophan mode of action cannot be linked to a direct modification of the plasma membrane H+-ATPase activity as monitored on purified pulvinar plasma membrane vesicles. The tryptophan metabolism-linked products tryptamine and indole also inhibited the motile reactions, activated in a continuous manner the H+ secretion of pulvinar tissues and showed properties of a protonophore and an ATPase activity inhibitor on plasma membrane vesicles, respectively. The specific behavior of tryptophan in the reaction studies here is discussed in light of the previously reported action of phytohormones.

Cytotoxic allelochemicals induce ultrastructural modifications in Cassia tora L. and mitotic changes in Allium cepa L.: a weed versus weed allelopathy approach.

The stress induced by allelochemicals present in stem aqueous extract (SAE) of Nicotiana plumbaginifolia on alterations in growth, ultrastructure on Cassia tora L., and mitotic changes on Allium cepa L. were inspected. Application of SAE at different concentrations (0.5, 1, 2, and 4%) expressively reduced the growth of C. tora in terms of seedling length and dry biomass. Moreover, the ultrastructural variations induced in the epidermis of Cassia leaf (adaxial and abaxial surface) of 15-day-old saplings were analyzed through scanning electron microscopy (SEM). The variations noticed are rupturing and shrinking of cells along epidermis; damaged margins, extensively curled leaf apex along with the appearance of puff-like structures, grooves, and thread-like structures on the leaf surface. The epidermal cells of samples exposed to treatment no longer appear smooth relative to control, besides showing necrosis as well. Upon exposure to different concentrations of extract, A. cepa root tip cells showed aberrations in chromosome arrangement and disparity in the shape of the interphase and prophase nuclei along various phases of mitotic cycle as compared to control. The mitotic index (MI) showed a concentration-dependent decline in onion root tips exposed to SAE. The aberrations appearing frequently were formation of multinucleated cells, sticky metaphase and anaphase with bridges, sticky telophase, disturbed polarity, etc. The results also show the induction of elongated cells, giant cells, and cells with membrane damage by extract treatment. To our knowledge, this is the first gas chromatography-mass spectrometry (GC-MS) analysis of the methanolic extract of N. plumbaginifolia stem. Overall, 62 compounds were reported, covering 99.61% of the entire constituents, which can be considered responsible for the allelopathic suppression of C. tora. The chief component was 4-tert-butylcalix[4]arene with the highest composition of 19.89%, followed by palmitic acid (12.25%), palmitoleic acid (8.23%), precocene 2 (7.53%), isophytyl acetate (4.01%), and betastigmasterol (3.95%).

Phenolic acid allelochemicals induced morphological, ultrastructural, and cytological modification on Cassia sophera L. and Allium cepa L.

The allelopathic potential of leaf aqueous extract (LAE) of Calotropis procera on growth behavior, ultrastructural changes on Cassia sophera L., and cytological changes on Allium cepa L. was investigated. LAE at different concentrations (0.5, 1, 2, and 4 %) significantly reduced the root length, shoot length, and dry biomass of C. sophera. Besides, the ultrastructural changes (through scanning electron microscopy, SEM) induced in epidermal cells of 15-day-old seedlings of Cassia leaf were also noticed. The changes induced were shrinking and contraction of epidermal cells along with the formation of major grooves, canals, and cyst-like structures. The treated samples of epidermal cells no longer seem to be smooth as compared to control. LAE at different concentrations induces chromosomal aberrations and variation in shape of the interphase and prophase nucleus in A. cepa root tip cells when compared with control groups. The mitotic index in treated onion root tips decreased with increasing concentrations of the extracts. The most frequent aberrations were despiralization at prophase with the formation of micronuclei, sticky anaphase with bridges, sticky telophase, C-metaphase, etc. The results also show the induction of ghost cells, cells with membrane damage, and cells with heterochromatic nuclei by extract treatment. Upon HPLC analysis, nine phenolic acids (caffeic acid, gentisic acid, catechol, gallic acid, syringic acid, ellagic acid, resorcinol, p-coumaric acid, and p-hydroxy benzoic acid) were identified. Thus, the phenolic acids are mainly responsible for the allelopathic behavior of C. procera.

In vitro formation of the anthranoid scaffold by cell-free extracts from yeast-extract-treated Cassia bicapsularis cell cultures.

The anthranoid skeleton is believed to be formed by octaketide synthase (OKS), a member of the type III polyketide synthase (PKS) superfamily. Recombinant OKSs catalyze stepwise condensation of eight acetyl units to form a linear octaketide intermediate which, however, is incorrectly folded and cyclized to give the shunt products SEK4 and SEK4b. Here we report in vitro formation of the anthranoid scaffold by cell-free extracts from yeast-extract-treated Cassia bicapsularis cell cultures. Unlike field- and in vitro-grown shoots which accumulate anthraquinones, cell cultures mainly contained tetrahydroanthracenes, formation of which was increased 2.5-fold by the addition of yeast extract. The elicitor-stimulated accumulation of tetrahydroanthracenes was preceded by an approx. 35-fold increase in OKS activity. Incubation of cell-free extracts from yeast-extract-treated cell cultures with acetyl-CoA and [2-(14)C]malonyl-CoA led to formation of torosachrysone (tetrahydroanthracene) and emodin anthrone, beside two yet unidentified products. No product formation occurred in the absence of acetyl-CoA as starter substrate. To confirm the identities of the enzymatic products, cell-free extracts were incubated with acetyl-CoA and [U-(13)C(3)]malonyl-CoA and (13)C incorporation was analyzed by ESI-MS/MS. Detection of anthranoid biosynthesis in cell-free extracts indicates in vitro cooperation of OKS with a yet unidentified factor or enzyme for octaketide cyclization.

Foliar nitrogen, phosphorus and potassium content in trees in environmentally toxic plastic industry area.

In plants, nitrogen deficiency causes stunted growth and chlorosis or yellowing of the leaves due to decreased levels of chlorophyll, while excess nitrogen uptake may cause dark green overly vigorous foliage which may have increased susceptibility to disease and insect attacks. Phosphorus is an important nutrient in crop production, since many soils in their native state do not have sufficient available phosphorus to maximize crop yield. Potassium deficiency may cause necrosis or interveinal chlorosis. Plastics are synthetic or semi-synthetic moldable organic solids that are organic polymers of high molecular mass, most commonly derived from petrochemicals; these polymers are based on chains of carbon atoms alone or with oxygen, sulfur, or nitrogen. Plastic is a non- biodegradable major toxic pollutant. It pollutes earth and leads to air pollution and water pollution. Merely there is any safe way to dispose the hazardous plastic wastes. The study was targeted to estimate foliar level of NPK content of three plant species, viz. Cassia tora (Herb), Ailanthus excelsa (Tree) and Dalbergia sissoo (Tree) from polluted areas associated to polythene-industries as well as control areas having least pollution, where all the parameters were found to be higher than the control experiments.

[Effect of exogenous carbon monoxide donor hematin on seed germination and physiological characteristics of Cassia obtusifolia seedlings under NaCl stress].

OBJECTIVE: In order to get the method to improve the salt resistance of seeds and seedlings for Cassia obtusbifolia under NaCl stress, seed germination and physiological characteristics of C. obtusifolia seedlings were studied. METHOD: Several physiological indexes of C. obtusifolia seeds treated with exogenous carbon monoxide donor hematin under NaCl stress like the germination vigor, germination rate, germination index and vigor index were measured. And other indexes like the relative water content, the contents of photosynthetic pigment, chlorophyll fluorescence parameters, the contents of soluble sugar, protein and proline, malondialdehyde (MDA), the activities of superoxide (SOD), peroxidase (POD) and catalase (CAT) were also measured. RESULT: The germination indexes of C. obtusifolia seeds under NaCl stress had been inhibited obviously. But after the treatment of hematin, every germination indexes were all increased. The result showed that the treatment of exogenous CO donor hematin obviously improved the germination vigor, germination rate, germination index and vigor index, increased the content of chlorophyll a, chlorophyll b, total chlorophyll, improved the photochemical efficiency of photosystem II (Fv/Fm), photochemical efficiency (Fv'/Fm'), PS II actual photochemical efficiency (phiPS II), photochemical quench coefficient (qP), decreased non-photochemical quenching coefficient (NPQ) and the content of malondialdehyde (MDA) , increased the relative water content of leaves and the content of soluble surge, protein and proline. Meanwhile, the results also indicated that CO improved the activities of superoxide (SOD), peroxidase (POD) and catalase (CAT). The effects of CO could be reversed when CO scavenger Hb is added. CONCLUSION: Exogenous CO donor hematin with appropriate concentration could significantly alleviate the damages to the seeds and seedlings of C. obtusifolia under NaCl stress and promote the salt resistance of the seeds and seedlings through improving the germination indexes, the photochemical efficiency and the antioxidase activities of the seedlings.

Biochemical changes in plant leaves as a biomarker of pollution due to anthropogenic activity.

The air pollution due to anthropogenic activities seriously affected human life, vegetation, and heritage as well. The vegetation cover in and around the city mitigates the air pollution by acting as a sink for pollution. An attempt was made to evaluate biochemical changes occurred in four selected plant species, namely Azadirachta indica, Mangifera indica, Delonix regia, and Cassia fistula of residential, commercial, and industrial areas of Nagpur city in India. It was observed that the correlated values of air pollutants and plant leaves characteristics alter foliar biochemical features (i.e., chlorophyll and ascorbic acid content, pH and relative water content) of plants due to air pollution. The changes in air pollution tolerance index of plants was also estimated which revealed that these plants can be used as a biomarker of air pollution.

Protecting cell walls from binding aluminum by organic acids contributes to aluminum resistance.

Aluminum-induced secretion of organic acids from the root apex has been demonstrated to be one major Al resistance mechanism in plants. However, whether the organic acid concentration is high enough to detoxify Al in the growth medium is frequently questioned. The genotypes of Al-resistant wheat, Cassia tora L. and buckwheat secrete malate, citrate and oxalate, respectively. In the present study we found that at a 35% inhibition of root elongation, the Al activities in the solution were 10, 20, and 50 muM with the corresponding malate, citrate, and oxalate exudation at the rates of 15, 20 and 21 nmol/cm(2) per 12 h, respectively, for the above three plant species. When exogenous organic acids were added to ameliorate Al toxicity, twofold and eightfold higher oxalate and malate concentrations were required to produce the equal effect by citrate. After the root apical cell walls were isolated and preincubated in 1 mM malate, oxalate or citrate solution overnight, the total amount of Al adsorbed to the cell walls all decreased significantly to a similar level, implying that these organic acids own an equal ability to protect the cell walls from binding Al. These findings suggest that protection of cell walls from binding Al by organic acids may contribute significantly to Al resistance.

Phytotoxicity of Phytolacca americana leaf extracts on the growth, and physiological response of Cassia mimosoides.

We examined the allelochemical effects of control soil, native soil (treated soil), and leaf extracts of Phytolacca americana (pokeweed) on the germination rate and seedling growth of Cassia mimosoides var. nomame. We also studied the resulting changes in root-tip ultrastructure and peroxidase isozyme biochemistry. P. americana leaf extract inhibited seed germination, seedling growth, and biomass when compared to control and treated soil. Root and shoot growth in treated soil was stimulated relative to control soil, but root growth was inhibited by 50% in the leaf extract treatment. Biomass of C. mimosoides seedlings grown on leaf extract was reduced sevenfold when compared to the control seedlings. The amounts of total phenolic compounds in the leaf extract, treated soil, and control soil were 0.77, 0.14, and 0.03 mg l(-1), respectively. The root tips of C. mimsoides treated with leaf extracts of P. americana showed amyloplasts and large central vacuoles with electron-dense deposits inside them when compared to control root tips. The activity of guaiacol peroxidase (GuPOX) in whole plant, roots, and shoots of C. mimosoides increased as leaf extract increased; maximum activity was observed in extract concentrations of 75% and higher. Root GuPOX activity was three times higher than in shoots. Therefore, we conclude that inhibition of C. mimosoides growth is related to the phenolic compounds in the P. americana leaf extract and the ultrastructure changes in root-tip cells and increased GuPOX activity is a response to these allelochemicals.

Nitric oxide reduces aluminum toxicity by preventing oxidative stress in the roots of Cassia tora L.

Nitric oxide (NO) as a key signaling molecule has been involved in mediation of various biotic and abiotic stress-induced physiological responses in plants. In the present study, we investigated the effect of NO on Cassia tora L. plants exposed to aluminum (Al). Plants pre-treated for 12 h with 0.4 mM sodium nitroprusside (SNP), an NO donor, and subsequently exposed to 10 microM Al treatment for 24 h exhibited significantly greater root elongation as compared with the plants without SNP treatment. The NO-promoted root elongation was correlated with a decrease in Al accumulation in root apexes. Furthermore, oxidative stress associated with Al treatment increased lipid peroxidation and reactive oxygen species, and the activation of lipoxygenase and antioxidant enzymes was reduced by NO. Such effects were confirmed by the histochemical staining for the detection of peroxidation of lipids and loss of membrane integrity in roots. The ameliorating effect of NO was specific, because the NO scavenger cPTIO [2-(4-carboxy-2-phenyl)-4,4,5,5-tetramethylinidazoline-1-oxyl-3-oxide] completely reversed the effect of NO on root growth in the presence of Al. These results indicate that NO plays an important role in protecting the plant against Al-induced oxidative stress.

Efficacy of various amendments for amelioration of fly-ash toxicity: growth performance and metal composition of Cassia siamea Lamk.

Plants of Cassia siamea Lamk were grown in garden soil (control), fly-ash (100%) and fly-ash amended by various ameliorants (cowdung manure, press-mud, garden soil; 1:1, w/w). The plants survived in fly-ash (100%) though their growth was less in comparison to the treatments. Fly-ash+press-mud (1:1, w/w) proved to be the best combination as growth (total biomass, leaf number, photosynthetic area, total chlorophyll and protein) was significantly high in this treatment followed by cowdung manure and garden soil. Leaves and roots accumulated significant amount of Cu, Zn, Ni and and Fe. However, the concentration of all the metals was more in roots than leaves except Ni. Although, fly-ash contains high amount of metals but the metal uptake was more in the plants grown in fly-ash+press-mud mixture. Inspite of high metal availability in fly-ash and press-mud mixture, plant growth was good. This might be attributed to the some metal detoxification mechanism active in this treatment. The present study concluded that C. siamea seems to be a suitable plant for developing a vegetation cover on fly-ash dumps.

Salicylic acid-induced aluminum tolerance by modulation of citrate efflux from roots of Cassia tora L.

Aluminum-induced exudation of organic acids from roots has been proposed as a mechanism for Al tolerance in plants. To better understand the regulatory process leading to efflux of organic acids, the possible involvement of salicylic acid (SA) in regulating Al-induced citrate release in Cassia tora L. was identified. The response of citrate efflux to exogenous SA was concentration-dependent. Application of SA at 5 microM in solution containing 20 microM Al increased citrate efflux to levels 1.76-fold higher than in controls (20 microM Al alone). However, inhibition of citrate release was observed when SA concentrations increased to more than 20 microM. Increased citrate efflux due to the SA treatment was associated with decreased inhibition of root growth and Al content in root tips, suggesting that exogenous SA could confer Al tolerance by increasing citrate efflux. We also examined citrate synthase activities (EC 4.1.3.7) and citrate concentrations in root tips exposed to Al and/or SA. However, both citrate synthase activities and citrate accumulation remained unaffected. These results indicate that SA-promotion of Al-induced citrate efflux is not correlated with increase in citrate production. Total endogenous SA concentrations were measured in root tips and the SA concentrations were significantly enhanced by Al at levels of 10-50 microM.