Exogenous supplementation of Sulfur (S) and Reduced Glutathione (GSH) Alleviates Arsenic Toxicity in Shoots of Isatis cappadocica Desv and Erysimum allionii L.

The current study was conducted to investigate the role of sulfur (S) and reduced glutathione (GSH) in mitigating arsenic (As) toxicity in Isatis cappadocica and Erysimum allionii. These plants were exposed for 3 weeks to different concentrations (0, 400 and 800 µM) of As to measure fresh weight, total chlorophyll, proline and hydrogen peroxide (H2O2) content, As and S accumulation, and guaiacol peroxidase (POD) and glutathione S-transferase (GST) along with the supplementation of 20 mg L-1 of S and 500 µM of GSH. Results revealed the significant reduction of fresh weight (especially in E. allionii), activities of POD and GST enzymes and proline content as compare to control. However, the application of S and GSH enhanced the fresh weight. Inhibition in H2O2 accumulation and improvement in antioxidant responses were measured with the application of S and GSH. Hence, the supplementation of S and GSH enhanced fresh weight and total chlorophyll in both I. cappadocica and E. allionii by alleviating the adverse effects of As stress via decreased H2O2 content and restricted As uptake.

Elucidating the physiological mechanisms underlying enhanced arsenic hyperaccumulation by glutathione modified superparamagnetic iron oxide nanoparticles in Isatis cappadocica.

Widespread arsenic (As) contamination is a severe environmental and public health concern. Isatis cappadocica, an arsenic hyperaccumulator, holds great potential to clean up As-contaminated soil and groundwater. Iron oxide is one of the most common metal oxides in the natural environment and its nanoparticulate form has been previously utilized for the removal of heavy metals/metalloids from wastewater. However, there is a paucity of information on the impact of iron oxide nanoparticles on the growth and physiological properties of I. cappadocica and its effectiveness on As removal. Current study reports for the first time the impact of superparamagnetic iron oxide nanoparticles and glutathione (GSH) modified superparamagnetic iron oxide nanoparticles (nFe3O4 and nFe3O4@GSH) on the physiological characteristic of I. cappadocica and its accumulation of As under hydroponic condition. nFe3O4@GSH alleviated the harmful impact of As and significantly increased the shoot biomass of I. cappadocica by enhancing the plant defense mechanisms. The application of GSH, nFe3O4 and nFe3O4@GSH all lowered the As concentration in plant shoots as a protective mechanism. However, the substantial shoot biomass increase due to nFe3O4@GSH resulted in a 56% higher As accumulation in plant shoots than in plants exposed to As alone, indicating the strong effectiveness of nFe3O4@GSH as a novel enhancer of the As phytoremediation by I. cappadocica. Our data further showed that the beneficial effect of nFe3O4@GSH on As phytoremediation is due to the enhancement of activities of several enzymatic and nonenzymatic antioxidants.

The role of selenium on mitigating arsenic accumulation, enhancing growth and antioxidant responses in metallicolous and non-metallicolous population of Isatis cappadocica Desv. and Brassica oleracea L.

A hydroponic experiment was conducted to explore the interactive effects of selenium (Se) supplementation (0, 5, and 10 µM) and arsenic (As) toxicity (0, 200, and 400 µM) on the growth, accumulation, and oxidative damage along with defense mechanisms of metallicolous (MP) and non-metallicolous population (NMP) of Isatis cappadocica, an As-hyperaccumulator, and Brassica oleracea as reference brassica. The results revealed that As stress significantly hampered plant growth particularly in B. oleracea. It reduced plant growth due to enhanced oxidative load of As-stressed plants. Between the two Isatis populations, metallicolous plants accumulated significantly higher As, however with considerably low growth defects. Furthermore, Se supplementation counteracted the adverse effects of stress on growth and physiological performance of all studied plants. Addition of Se, particularly at higher dose (10 µM), significantly suppressed root As uptake and slightly its accumulation in shoots of B. oleracea plants treated with 400 µM As, and thus improved growth characteristics of stressed plants. Under As stress, Se supplementation increased the activities of enzymatic (peroxidase (POD) and glutathione reductase (GR)) and non-enzymatic (anthocyanins and total flavonoids) antioxidants, thereby suggesting relieved As stress by reduced oxidative damage. Taken together, these results support the beneficial role of Se in the regulation of As stress by improving growth, physiology, and antioxidant capacity, and highlight its significance for plants grown on such metal-contaminated soils.

Seed Germination Ecology of the Cold Desert Annual Isatis violascens (Brassicaceae): Two Levels of Physiological Dormancy and Role of the Pericarp.

The occurrence of various species of Brassicaceae with indehiscent fruits in the cold deserts of NW China suggests that there are adaptive advantages of this trait. We hypothesized that the pericarp of the single-seeded silicles of Isatis violascens restricts embryo expansion and thus prevents germination for 1 or more years. Thus, our aim was to investigate the role of the pericarp in seed dormancy and germination of this species. The effects of afterripening, treatment with gibberellic acid (GA3) and cold stratification on seed dormancy-break were tested using intact silicles and isolated seeds, and germination phenology was monitored in an experimental garden. The pericarp has a role in mechanically inhibiting germination of fresh seeds and promotes germination of nondormant seeds, but it does not facilitate formation of a persistent seed bank. Seeds in silicles in watered soil began to germinate earlier in autumn and germinated to higher percentages than isolated seeds. Sixty-two percent of seeds in the buried silicles germinated by the end of the first spring, and only 3% remained nongerminated and viable. Twenty to twenty-five percent of the seeds have nondeep physiological dormancy (PD) and 75-80% intermediate PD. Seeds with nondeep PD afterripen in summer and germinate inside the silicles in autumn if the soil is moist. Afterripening during summer significantly decreased the amount of cold stratification required to break intermediate PD. The presence of both nondeep and intermediate PD in the seed cohort may be a bet-hedging strategy.

[Effects of drought stress on growth and water use efficiency of two medicinal plants].

Growth characteristics, stable carbon isotope discrimination (Delta13C), water use efficiency (WUE), and their correlation of Cassia obtusifolia and Isatis indigotica were measured at three soil water levels, i. e., 30%, 50% and 75% of field water holding capacity (FWHC), and at three growth stages. The growth indices of the two medicinal plants at 75% of FWHC were higher than those at 30% and 50% of FWHC, suggesting that the two medicinal plants could obtain high production under sufficient moisture condition. The Delta13C(A) (aboveground biomass-based Delta13C) and Delta13C(T) (total biomass-based Delta13C) decreased, and the WUE(A) (aboveground biomass-based WUE) and WUE(T) (total biomass-based WUE) of C. obtusifolia and I. indigotica increased with the increasing degree of drought stress. The growth indices of the two medicinal plants had little difference in the different water treatments, which indicated that the two medicinal plants were insensitive to drought stress. Water use efficiency of I. indigotica had significant negative relationships with aboveground biomass and total biomass, while that of C. obtusifolia had a significant positive correlation with the root/shoot ratio.

Photosynthetic physiological response of Radix Isatidis (Isatis indigotica Fort.) seedlings to nicosulfuron.

Radix Isatidis (Isatis indigotica Fort.) is one of the most important traditional Chinese medicine plants. However, there is no suitable herbicide used for weed control in Radix Isatidis field during postemergence stage. To explore the safety of sulfonylurea herbicide nicosulfuron on Radix Isatidis (Isatis indigotica Fort.) seedlings and the photosynthetic physiological response of the plant to the herbicide, biological mass, leaf area, photosynthetic pigment content, photosynthetic rate, chlorophyll fluorescence characteristics, and P700 parameters of Radix Isatidis seedlings were analyzed 10 d after nicosulfuron treatment at 5th leaf stage in this greenhouse research. The results showed that biological mass, total chlorophyll, chlorophyll a, and carotenoids content, photosynthetic rate, stomatal conductance, PS II maximum quantum yield, PS II effective quantum yield, PS II electron transport rate, photochemical quenching, maximal P700 change, photochemical quantum yield of PS I, and PS I electron transport rate decreased with increasing herbicide concentrations, whereas initial fluorescence, quantum yield of non-regulated energy dissipation in PS II and quantum yield of non-photochemical energy dissipation due to acceptor side limitation in PS I increased. It suggests that nicosulfuron ≥1 mg L-1 causes the damage of chloroplast, PS II and PS I structure. Electron transport limitations in PS I receptor side, and blocked dark reaction process may be the main cause of the significantly inhibited growth and decreased photosynthetic rate of Radix Isatidis seedlings.

Effects of fully open-air [CO2] elevation on leaf photosynthesis and ultrastructure of Isatis indigotica fort.

Traditional Chinese medicine relies heavily on herbs, yet there is no information on how these herb plants would respond to climate change. In order to gain insight into such response, we studied the effect of elevated [CO2] on Isatis indigotica Fort, one of the most popular Chinese herb plants. The changes in leaf photosynthesis, chlorophyll fluorescence, leaf ultrastructure and biomass yield in response to elevated [CO2] (550±19 µmol mol(-1)) were determined at the Free-Air Carbon dioxide Enrichment (FACE) experimental facility in North China. Photosynthetic ability of I. indigotica was improved under elevated [CO2]. Elevated [CO2] increased net photosynthetic rate (P N), water use efficiency (WUE) and maximum rate of electron transport (J max) of upper most fully-expended leaves, but not stomatal conductance (gs), transpiration ratio (Tr) and maximum velocity of carboxylation (V c,max). Elevated [CO2] significantly increased leaf intrinsic efficiency of PSII (Fv'/Fm') and quantum yield of PSII(ΦPS II ), but decreased leaf non-photochemical quenching (NPQ), and did not affect leaf proportion of open PSII reaction centers (qP) and maximum quantum efficiency of PSII (Fv/Fm). The structural chloroplast membrane, grana layer and stroma thylakoid membranes were intact under elevated [CO2], though more starch grains were accumulated within the chloroplasts than that of under ambient [CO2]. While the yield of I. indigotica was higher due to the improved photosynthesis under elevated [CO2], the content of adenosine, one of the functional ingredients in indigowoad root was not affected.

IiSDD1, a gene responsive to autopolyploidy and environmental factors in Isatis indigotica.

In plants, stomata play a pivotal role in the regulation of gas exchange and are distributed throughout the aerial epidermis. SDD1, a gene isolated from Arabidopsis thaliana has been demonstrated to specialize in stomatal density and distribution. In our present study, a comprehensive survey of global gene expression performed by using an A. thaliana whole genome Affymetrix gene chip revealed SDD1 tends to be significantly lower in tetraploid Isatis indigotica than in diploid ones. To intensively investigate different SDD1 expression in response to polyploidy, a full-length cDNA clone (IiSDD1) encoding SDD1 was isolated from the traditional Chinese medicinal herb I. indigotica cDNA library. IiSDD1 shared a high level of identity with that from A. thaliana, containing some basic features of subtilases: D, H and S regions, as well as a substrate-binding site. Real-time quantitative PCR analysis indicated that IiSDD1 was constitutively expressed in all tested tissues, including roots, stems and leaves, both in tetraploid and diploid I. indigotica, and with the highest expression in leaves. In addition, IiSDD1 was also found to be down-regulated by signalling molecules for plant defence responses, such as abscisic acid (100 microM) and gibberellin (100 mg/L), as well as by environmental stresses including salt, darkness, coldness and drought. Our study, for the first time, indicates SDD1 participates not only in the defense/stress responsive pathways, but also probably involves in plants polyploidy evolution.