Improvement of morpho-physiological, ultrastructural and nutritional profiles in wheat seedlings through astaxanthin nanoparticles alleviating the cadmium toxicity.

Heavy metal accumulation in arable lands and water bodies has become one of the serious global issues among multitude of food security challenges. In particular, cadmium (Cd) concentration has been increasing substantially in the environment that negatively affects the growth and yield of important agricultural crops, especially wheat (Triticum aestivum L.). No doubt, nanotechnology is a revolutionary science but the comprehension of nanoparticle-plants interaction and its potential alleviatory role against metal stress is still elusive. Here, we investigated the mechanistic role of astaxanthin nanoparticles (AstNPs) in Cd stress amelioration and their interaction with wheat under Cd-spiked conditions. The AstNPs fabrication was confirmed through ultraviolet visible spectroscopy, where the particles showed characteristic peak at 423 nm. However, Fourier transform infrared, X-ray diffraction, scanning electron microscopy and transmission electron microscopy analyses confirmed the presence of stabilized spherical-shaped nanocrystals of AstNPs within the size range of 12.03-30.37 nm. The hydroponic application of AstNPs (100 mg L-1) to Cd-affected wheat plants increased shoot height (59%), shoot dry weight (31%), nitrogen concentration (42%), and phosphorus concentration (26%) as compared to non-treated Cd affected seedlings. Moreover, AstNPs-treated plants showed reduction in acropetal Cd translocation (29%) in contrast to plants treated with Cd only. Under Cd-spiked conditions, AstNPs-treated plants displayed an improved nutrient profile (P, N, K+ and Ca2+) with a relative decrease in Na+ content in comparison with non-treated plants. Interestingly, it was found that AstNPs restricted the translocation of Cd to aerial plant parts by negatively regulating Cd transporter genes (TaHMA2 and TaHMA3), and relieved plants from oxidative burst by activating antioxidant machinery via triggering expressions of TaSOD and TaPOD genes. Consequently, it was observed that the application of AstNPs helped in maintaining the nutrient acquisition and ionic homeostasis in Cd-affected wheat plants, which subsequently improved the physiochemical profiles of plants under Cd-stress. This study suggests that AstNPs plausibly serve as stress stabilizers for plants under heavy metal-polluted environment.

Physio-ultrastructural footprints and iTRAQ-based proteomic approach unravel the role of Piriformospora indica-colonization in counteracting cadmium toxicity in rice.

Due to its immense capability to concentrate in rice grain and ultimately in food chain, cadmium (Cd) has become the cause of an elevated concern among agriculturists, scientists and the environmental activists. Symbiotic association of Piriformospora indica (P. indica) has been characterized as a potential aid in combating heavy metal stress in plants for sustainable crop production but our scant knowledge regarding ameliorative tendency of P. indica against Cd, specifically in rice, necessitates an in-depth investigation. This study aimed at elaborating the underlying mechanisms involved in P. indica-mediated tolerance against Cd stress in two rice genotypes, IR8 and ZX1H, varying in Cd accumulation pattern. Either colonized or un-inoculated with P. indica, seedlings of both genotypes were subjected to Cd stress. The results showed that P. indica colonization significantly supported plant biomass, photosynthetic attributes and chlorophyll contents in Cd stressed plants. P. indica colonization sustained chloroplast integrity and reduced Cd translocation (46% and 64%), significantly lowering malondialdehyde (MDA) content (11.3% and 50.4%) compared to uninoculated roots under Cd stress in IR8 and ZX1H, respectively. A genotypic difference was evident when a 2-fold enhancement in root peroxidase (POD) activity was recorded in P. indica colonized IR8 plants as compared to ZX1H. The root proteomic analysis was performed using isobaric tags for relative and absolute quantification (iTRAQ) and the results showed that P. indica alleviates Cd stress in rice via down-regulation of key glycolysis cycle enzymes in a bid to reduce energy consumption by the plants and possibly re-directing it to Cd defense response pathways; and up-regulation of glutamine synthetase, a key enzyme in the L-Arg-dependent pathway for nitric oxide (NO) production, which acts as a stress signaling molecule, thus conferring tolerance by reduction of NO-mediated modification of essential proteins in response to Cd stress. Conclusively, both the tested genotypes benefited from P. indica symbiosis at varying levels by an enhanced detoxification capacity and signaling efficiency in response to stress. Hence, a step forward towards the employment of an environmentally sound and self-renewing approach holding the hope for a healthy future.

Modulation of Key Physio-Biochemical and Ultrastructural Attributes after Synergistic Application of Zinc and Silicon on Rice under Cadmium Stress.

Excessive industrialization and the usage of pesticides plague the farming soils with heavy metals, reducing the quality of arable land. Assessing phytoavailability of cadmium (Cd) from growth medium to plant system is crucial and necessitates precise and timely monitoring of Cd to ensure food safety. Zinc (Zn) and silicon (Si) have singularly demonstrated the potential to ameliorate Cd toxicity and are important for agricultural production, human health, and environment in general. However, Zn-Si interaction on Cd toxicity alleviation, their effects and underlying mechanisms are still fragmentarily understood. Seven treatments were devised besides control to evaluate the single and combined effects of Zn and Si on the physio-biochemical attributes and ultrastructural fingerprints of Cd-treated rice genotypes, i.e., Cd tolerant "Xiushui-110" and Cd sensitive "HIPJ-1". Supplementation of both Zn and Si promoted plant biomass, photosynthetic parameters, ionic balance, and improved chloroplast ultrastructure with minimized Cd uptake and malondialdehyde (MDA) content due to the activation of antioxidant enzymes in Cd stressed plants. The combined effects of 10 µM Zn and 15 µM Si on 15 µM Cd displayed a greater reduction in Cd uptake and root-leaf MDA content, while enhancing photosynthetic activity, superoxide dismutase (SOD) activity and root-leaf ultrastructure particularly in HIPJ-1, whilst Xiushui-110 had an overall higher leaf catalase (CAT) activity and a higher root length and shoot height was observed in both genotypes compared to the Cd 15 µM treatment. Alone and combined Zn and Si alleviation treatments reduced Cd translocation from the root to the stem for HIPJ-1 but not for Xiushui-110. Our results confer that Zn and Si singularly and in combination are highly effective in reducing tissue Cd content in both genotypes, the mechanism behind which could be the dilution effect of Cd due to improved biomass and competitive nature of Zn and Si, culminating in Cd toxicity alleviation. This study could open new avenues for characterizing interactive effects of simultaneously augmented nutrients in crops and provide a bench mark for crop scientists and farmers to improve Cd tolerance in rice.

Highly Conserved Evolution of Aquaporin PIPs and TIPs Confers Their Crucial Contribution to Flowering Process in Plants.

Flowering is the key process for the sexual reproduction in seed plants. In gramineous crops, the process of flowering, which includes the actions of both glume opening and glume closing, is directly driven by the swelling and withering of lodicules due to the water flow into and out of lodicule cells. All these processes are considered to be controlled by aquaporins, which are the essential transmembrane proteins that facilitate the transport of water and other small molecules across the biological membranes. In the present study, the evolution of aquaporins and their contribution to flowering process in plants were investigated via an integration of genome-wide analysis and gene expression profiling. Across the barley genome, we found that HvTIP1;1, HvTIP1;2, HvTIP2;3, and HvPIP2;1 were the predominant aquaporin genes in lodicules and significantly upregulated in responding to glume opening and closing, suggesting the importance of them in the flowering process of barley. Likewise, the putative homologs of the above four aquaporin genes were also abundantly expressed in lodicules of the other monocots like rice and maize and in petals of eudicots like cotton, tobacco, and tomato. Furthermore, all of them were mostly upregulated in responding to the process of floret opening, indicating a conserved function of these aquaporin proteins in plant flowering. The phylogenetic analysis based on the OneKP database revealed that the homologs of TIP1;1, TIP1;2, TIP2;3, and PIP2;1 were highly conserved during the evolution, especially in the angiosperm species, in line with their conserved function in controlling the flowering process. Taken together, it could be concluded that the highly evolutionary conservation of TIP1;1, TIP1;2, TIP2;3 and PIP2;1 plays important roles in the flowering process for both monocots and eudicots.

Calcium Plays a Double-Edged Role in Modulating Cadmium Uptake and Translocation in Rice.

Cadmium (Cd) contamination in soils poses great risks to both agricultural production and human health. Calcium (Ca) is an essential element playing a significant role in protecting plants against Cd toxicity. However, how Ca affects Cd uptake and translocation in rice is still not fully elucidated. In this study, the regulatory role of Ca in Cd uptake and upward translocation was investigated in rice at different growth stages. Our results showed that the supplement of 5 mM Ca significantly reduced Cd uptake by rice roots, because of their competition for Ca-permeable channels as an absorption site and Ca-induced downregulation of OsNRAMP1 and OsNRAMP5. However, Ca application facilitated the upward translocation of Cd by both upregulating OsHMA2 to induce xylem loading of Cd and downregulating OsHMA3 to reduce vacuolar sequestration of Cd. Such contrary results suggested a double-edged role of Ca in regulating root Cd uptake and root-to-shoot Cd translocation in rice. Although it increased Cd content in the aboveground vegetative tissues during the whole growth period, the addition of 5 mM Ca eventually decreased Cd content in rice grains at the ripening stage. All these results suggest that Ca-based amendments possess great potential for the production of low-Cd rice grains.

Geographical variation in arsenic, cadmium, and lead of soils and rice in the major rice producing regions of China.

Rapid industrialization and urbanization have accelerated the contamination of paddy soils with potentially toxic elements (PTEs). However, the status and the key factors responsible for the geographical variation in PTE concentrations in rice remain poorly understood. Here, a total of 113 pairs of soil and rice plant samples were collected from 19 provinces in four major rice producing areas of China to assess the geographical variation in total arsenic (As), cadmium (Cd) and lead (Pb) concentrations in the soil-rice system. Average total concentrations of As, Cd and Pb were 11.8, 0.45 and 25.7 mg kg-1, respectively, in the soils and 0.089, 0.087 and 0.036 mg kg-1 in the polished rice. The national maximum allowable concentrations of total soil As and Cd were exceeded in 6.19 and 33.6% of soils and that of Cd was exceeded in 7.96% of polished rice and no polished rice exceed the Pb limit. The As, Cd and Pb concentrations of rice were significantly and positively correlated (p < 0.05) with their corresponding soil available concentrations rather than with their soil total concentrations. Due to the combined effects of local rice varieties, cultivation of varieties with high Cd translocation factors and high Cd availability in acid soils, the highest rice Cd risk occurred in south China. The Cd concentrations in polished rice exceeded the maximum allowable by 4.0 and 15.8% in uncontaminated and contaminated soils, respectively. Results from 113 fixed samples may represent the actual current As, Cd and Pb status of rice in the main rice production areas nationally as they were very consistent with 574 random samples. In view of the high Cd contamination risk in acid soils of south China, countermeasures are needed to minimize Cd accumulation in rice crops in this region.

The pathway of transmembrane cadmium influx via calcium-permeable channels and its spatial characteristics along rice root.

To develop elite crops with low cadmium (Cd), a fundamental understanding of the mechanism of Cd uptake by crop roots is necessary. Here, a new mechanism for Cd2+ entry into rice root cells was investigated. The results showed that Cd2+ influx in rice roots exhibited spatially and temporally dynamic patterns. There was a clear longitudinal variation in Cd uptake along rice roots, with the root tip showing much higher Cd2+ influx and concentration than the root mature zone, which might be due to the much higher expression of the well-known Cd transporter genes OsIRT1, OsNRAMP1, OsNRAMP5, and OsZIP1 in the root tip. Both the net Cd2+ influx and the uptake of Cd in rice roots were highly inhibited by ion channel blockers Gd3+ and TEA+, supplementation of Ca2+ and K+, and the plasma membrane H+-ATPase inhibitor vanadate, with Gd3+ and Ca2+ showing the most inhibitory effects. Furthermore, Ca2+- or Gd3+-induced reduction in Cd2+ influx and Cd uptake did not coincide with the expression of Cd transporter genes, but with that of two Ca channel genes, OsAAN4 and OsGLR3.4. These results indicate that Cd transporters are in part responsible for Cd2+ entry into rice root, and provide a new perspective that the Ca channels OsAAN4 and OsGLR3.4 might play an important role in rice root Cd uptake.

Cadmium remobilization from shoot to grain is related to pH of vascular bundle in rice.

The remobilization of cadmium (Cd) from shoots to grain is the key process to determine the Cd accumulation in grain. The apoplastic pH of plants is an important factor and signal in influencing on plant responding to environmental variation and inorganic elements uptake. It is proposed that pH of rice plants responds and influences on Cd remobilization from shoots to grain when rice is exposed to Cd stress. The results of hydroponic experiment showed that: pH of the rice leaf vascular bundles among 3 cultivars was almost increased, pH value of 1 cultivar was slightly increasing when rice plants were treated with Cd. The decrease degree of H+ concentration in leaf vascular bundles was different among cultivars. The cultivar with higher decreasing in H+ concentration, showed higher Cd transfer efficiency from shoots to grain. The H+ concentration of leaf vascular bundles under normal condition was negatively correlated to cadmium accumulation in leaf. Moreover, pH change was related to Cd accumulation in shots and remobilization from shoots to grain. Uncovering the role of pH response is a key component for the understanding Cd uptake and remobilization mechanism for rice production.

Effects of water management on arsenic and cadmium speciation and accumulation in an upland rice cultivar.

Pot and field experiments were conducted to investigate the effects of water regimes on the speciation and accumulation of arsenic (As) and cadmium (Cd) in Brazilian upland rice growing in soils polluted with both As and Cd. In the pot experiment constant and intermittent flooding treatments gave 3-16 times higher As concentrations in soil solution than did aerobic conditions but Cd showed the opposite trend. Compared to arsenate, there were more marked changes in the arsenite concentrations in the soil solution as water management shifted, and therefore arsenite concentrations dominated the As speciation and bioavailability in the soil. In the field experiment As concentrations in the rice grains increased from 0.14 to 0.21 mg/kg while Cd concentrations decreased from 0.21 to 0.02 mg/kg with increasing irrigation ranging from aerobic to constantly flooding conditions. Among the various water regimes the conventional irrigation treatment produced the highest rice grain yield of 6.29 tons/ha. The As speciation analysis reveals that the accumulation of dimethylarsinic acid (from 11.3% to 61.7%) made a greater contribution to the increase in total As in brown rice in the intermittent and constant flooding treatments compared to the intermittent-aerobic treatment. Thus, water management exerted opposite effects on Cd and As speciation and bioavailability in the soil and consequently on their accumulation in the upland rice. Special care is required when irrigation regime methods are employed to mitigate the accumulation of metal(loid)s in the grain of rice grown in soils polluted with both As and Cd.

Water management affects arsenic and cadmium accumulation in different rice cultivars.

Paddy rice (Oryza sativa L.) is a staple food and one of the major sources of dietary arsenic (As) and cadmium (Cd) in Asia. A field experiment was conducted to investigate the effects of four water management regimes (aerobic, intermittent irrigation, conventional irrigation and flooding) on As and Cd accumulation in seven major rice cultivars grown in Zhejiang province, east China. With increasing irrigation from aerobic to flooded conditions, the soil HCl-extractable As concentrations increased significantly and the HCl-extractable Cd concentrations decreased significantly. These trends were consistent with the As and Cd concentrations in the straw, husk and brown rice. Water management both before and after the full tillering stage affected As and Cd accumulation in the grains. The intermittent and conventional treatments produced higher grain yields than the aerobic and flooded treatments. Cd concentrations in brown rice varied 13.1-40.8 times and As varied 1.75-8.80 times among the four water management regimes. Cd and As accumulation in brown rice varied among the rice cultivars, with Guodao 6 (GD6) was a low Cd but high-As-accumulating cultivar while Indonesia (IR) and Yongyou 9 (YY9) were low As but high-Cd-accumulating cultivars. Brown rice Cd and As concentrations in the 7 cultivars were significantly negatively correlated. The results indicate that As and Cd accumulated in rice grains with opposite trends that were influenced by both water management and rice cultivar. Production of 'safe' rice with respect to As and Cd might be possible by balancing water management and rice cultivar according to the severity of soil pollution.

[Determination of 2-acetyl-1-pyrroline in aroma rice using gas chromatography-mass spectrometry].

A gas chromatography-mass spectrometry method was developed for the determination of 2-acetyl-1-pyrroline in aroma rice. 2,4,6-Trimethylpyridine (TMP) was added into the sample of aroma rice as internal standard, and the compound was extracted by the mixture of anhydrous ethyl alcohol and methylene chloride (1: 1, v/v) at 80 degrees C for 3 h under sealed condition in water bath. The effects of the temperature and time on the extraction were investigated. The analyte was separated and determined using gas chromatography-mass spectrometry on an HP-5MS capillary column (30 m x 0.25 mm x 0.25 microm) in scan monitoring mode. Calculated as 2,4, 6-trimethylpyridine, the average recovery of the method was 82.57% with the relative standard deviation of 5.09%, and the detection limit of method was 0.01 mg/kg. The method was employed for the determination of 2-acetyl-1-pyrroline in 11 aroma rice breeding varieties. The results showed that 2-acetyl-1-pyrroline was detected in 5 aroma rice varieties, including Qingxiangmi, Taixiang R207, Texmati, Guixiangsinuo and Zhongjian 2, with the contents of 0.097, 0.098, 0.699, 0.045 and 0.047 mg/kg, respectively. The method is simple, rapid and sensitive with low sample and reagent consumption. It is suitable for screening a lot of aroma rice varieties in breeding through the determination of 2-acetyl-1-pyrroline content.