Effect of foliar application of nanoparticles on growth, physiology, and antioxidant enzyme activities of lettuce (Lactuca sativa L.) plants under cadmium toxicity.

Nanotechnology has attracted the interest of scientists due to its wide range of application specifically in agriculture. Nanoparticles (NPs) may act as a promising materials to alleviate cadmium (Cd) stress in plants. This study aims to assess the impact of multiple nanoparticles including nSiO2 (50 mg L-1:100 mg L-1), nTiO2 (20 mg L-1:60 mg L-1), nZnO (50 mg L-1:100 mg L-1), nFe3O4 (100 mg L-1:200 mg L-1), nCuO (50 mg L-1:100 mg L-1), and nCeO2 (50 mg L-1:100 mg L-1) in combination with CdCl2 (5 µM) to mitigate Cd toxicity in lettuce through foliar application in hydroponic solution. Current findings indicate that foliar application of nSiL + Cd (50 mg L-1), nZnL + Cd (50 mg L-1), and nTiL + Cd (20 mg L-1) is more effective in improving growth, biomass, root architecture, and elevated photosynthetic efficiency, which might be attributed to the increasing uptake of essential micronutrient (K, Mg, Ca, Fe, Zn) under Cd stress. Similarly, treatment with nanoparticles leads to reduced accumulation of ROS and MDA in lettuce, while enhancing the SOD, POD, CAT, and APX activities. The results showed that nanoparticles have high tolerance against Cd as depicted by the inhibition in Cd accumulation by 3.2-58% and 10-72% in roots as well as edible parts of lettuce, respectively. In addition, Cd alone reduces the morphological traits, antioxidant enzyme activity, and photosynthetic activity, while increasing the ROS, MDA, and Cd accumulation in lettuce. This comprehensive study suggests the role of nanoparticles in reducing Cd toxicity in lettuce, signifying their importance as stress mitigation agents. However, long-term pot, priming, and field trials are needed to identify the optimal nanoparticle for the lettuce under variable environmental conditions.

The involvement of nitric oxide and ethylene on the formation of endodermal barriers in response to Cd in hyperaccumulator Sedum alfredii.

Nitric oxide (NO) and ethylene are both important signaling molecules which participate in numerous plant development processes and environmental stress resistance. Here, we investigate whether and how NO interacts with ethylene during the development of endodermal barriers that have major consequences for the apoplastic uptake of cadmium (Cd) in the hyperaccumulator Sedum alfredii. In response to Cd, an increased NO accumulation, while a decrease in ethylene production was observed in the roots of S. alfredii. Exogenous supplementation of NO donor SNP (sodium nitroprusside) decreased the ethylene production in roots, while NO scavenger cPTIO (2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide) had the opposite effect. The exogenous addition of NO affected the ethylene production through regulating the expression of genes related to ethylene synthesis. However, upon exogenous ethylene addition, roots retained their NO accumulation. The abovementioned results suggest that ethylene is downstream of the NO signaling pathway in S. alfredii. Regardless of Cd, addition of SNP promoted the deposition of endodermal barriers via regulating the genes related to Casparian strips deposition and suberization. Correlation analyses indicate that NO positively modifies the formation of endodermal barriers via the NO-ethylene signaling pathway, Cd-induced NO accumulation interferes with the synthesis of ethylene, leading to a deposition of endodermal barriers in S. alfredii.

Integrated glycolysis and pyrolysis process for multiple utilization and cadmium collection of hyperaccumulator Sedum alfredii.

Phytoremediation is a cost-effective and environmentally-friendly method to treat cadmium (Cd) contaminated soils, however, there is still a lack of safe disposal methods of harvested hyperaccumulators. In this study, by integrating glycolysis and pyrolysis, we investigated the possibility of bioproduct production and Cd collection from the hyperaccumulator Sedum alfredii. By means of acid-alkali pretreatment, the degree of cellulose polymerization was reduced by 36.24% while the surface accessibility was increased by 115.80%, resulting in a bioethanol yield of 9.29%. Meanwhile, 99.22% of total Cd of biomass could be reclaimed by collecting H2SO4-pretreatment waste. The saccharification residue was subsequently modified by NaOH-pretreatment-filtrate and converted into biochar at 500 °C which possessed a maximum Cd2+ sorption capacity of 60.52 mg g-1 based on the Langmuir model. Furthermore, sustainability analysis indicated that the economic input of this process is acceptable when considering its good environmental benefits. Taken together, our study provides a strategy for simultaneous bioethanol and biochar production during Cd collection from the hyperaccumulator S. alfredii, which could be a promising alternative for the suitable treatment of metal-enriched plants.

The inhibition effect of tea polyphenols on soil nitrification is greater than denitrification in tea garden soil.

Tea polyphenols are the most widely distributed class of secondary metabolites (Camellia sinensis) and account for a considerable proportion of the pruning residues of tea. A large amount of tea polyphenols have fallen down over soil with prunning residues every year. However, the effect of tea polyphenols on soil nitrogen cycle, especially the denitrification process and its related microbial communities, remains unclear. Epigallocatechin gallate (EGCG), the most abundant component of tea polyphenols, was selected to simulate the effects of tea polyphenols on soil nitrification, denitrification, related functional genes and microbial community. The results indicated that addition of EGCG can significantly (p < 0.05) inhibit soil nitrification. Copy numbers of bacterial and archaeal ammonia monooxygenase genes (amoA) decreased as EGCG concentration increased. Further, the ammonia oxidisers exhibited a significantly (p < 0.05) greater niche differentiation under the effect of EGCG compared with the control treatment (no EGCG addition). However, the inhibition effect of EGCG over soil denitrification was most significant at 34 and 36 day of incubation period, and such inhibitory effect was more apparent on nitrification compared with denitrification. EGCG addition increased the diversity of bacterial community. The composition of bacterial community was significantly altered and community variation was primary explained by EGCG, NH4+-N, NO3--N, soil organic carbon contents and potential denitrification rates. EGCG addition significantly increased relative abundance of Proteobacteria and Bacteroidetes phyla whereas decreased Actinobacteria. Overall, tea polyphenols can inhibit soil nitrification to a larger extent than denitrification by reducing the abundance of microorganisms carrying the related functional genes. Our results can serve as important basis of reducing the nitrogen pollution risk in tea orchards and could be considered as a powerful natural nitrification inhibitor to reduce the environmental risks caused by unreasonable nitrogen fertiliser adaptation.

Coordination between root cell wall thickening and pectin modification is involved in cadmium accumulation in Sedum alfredii.

Root cell wall (RCW) modification is a widespread important defense strategy of plant to cope with trace metals. However, mechanisms underlying its remolding in cadmium (Cd) accumulation are still lacking in hyperaccumulators. In this study, changes of RCW structures and components between nonhyperaccumulating ecotype (NHE) and hyperaccumulating ecotype (HE) of Sedum alfredii were investigated simultaneously. Under 25 µM Cd treatment, RCW thickness of NHE is nearly 2 folds than that of HE and the thickened cell wall of NHE was enriched in low-methylated pectin, leading to more Cd trapped in roots tightly. In the opposite, large amounts of high-methylated pectin were assembled around RCW of HE with Cd supply, in this way, HE S. alfredii decreased its root fixation of Cd and enhanced Cd migration into xylem. TEM and AFM results further confirmed that thickened cell wall was caused by the increased amounts of cellulose and lignin while root tip lignification was resulted from variations of sinapyl (S) and guaiacyl (G) monomers. Overall, thickened cell wall and methylated pectin have synchronicity in spatial location of roots, and their coordination contributed to Cd accumulation in S. alfredii.

Ethylene-mediated apoplastic barriers development involved in cadmium accumulation in root of hyperaccumulator Sedum alfredii.

Ethylene is an important phytohormone for plant adaptation to heavy metal stress. However, the effects of ethylene on radial apoplastic transport of Cd remain elusive. This study investigated the role of ethylene on apoplastic barriers development and consequences for Cd uptake in Sedum alfredii. In response to Cd, endogenous ethylene production in hyperaccumulating ecotype (HE) roots was decreased due to the down-regulated expressions of ethylene biosynthesis genes, while the opposite result was observed in non-hyperaccumulating ecotype (NHE). Interestingly, the ethylene emission in HE was always higher than that in NHE, regardless of Cd concentrations. Results of exogenous application of ethylene biosynthesis precursor/inhibitor indicate that ethylene with high level would delay the formation of apoplastic barriers in HE through restraining phenylalanine ammonia lyase activity and gene expressions related to lignin/suberin biosynthesis. Simultaneously, correlation analyses suggest that Cd-induced apoplastic barriers formation may be also regulated by ethylene signaling. By using an apoplastic bypass tracer and scanning ion-selected electrode, we observed that the delayed deposition of apoplastic barriers significantly promoted Cd influx in roots. Taken together, high endogenous ethylene in HE postponed the formation of apoplastic barriers and thus promoted the Cd accumulation in the apoplast of roots.

A comparative study of root cadmium radial transport in seedlings of two wheat (Triticum aestivum L.) genotypes differing in grain cadmium accumulation.

The radial transport of cadmium (Cd) is essential for Cd influx in roots. The role of radial transport pathway on the Cd translocation from root to shoot among wheat genotypes are still poorly understood. This study explored the role of apoplastic and symplastic pathway on root Cd uptake and root-to-shoot translocation in Zhenmai 10 (ZM10, high Cd in grains) and Aikang 58 (AK58, low Cd in grains). Under Cd treatment, the deposition of Casparian strips (CSs) and suberin lamellae (SL) initiated closer to the root apex in ZM10 than that in AK58, which resulted in the lower Cd concentration in apoplastic fluid of ZM10. Simultaneously, Cd-induced expression levels of genes related to Cd uptake in roots were significantly higher in AK58 by contrast with ZM10, contributing to the symplastic Cd accumulation in AK58 root. Moreover, the addition of metabolic inhibitor CCCP noticeably decreased the Cd accumulation in root of both genotypes. Intriguingly, compared to ZM10, greater amounts of Cd were sequestrated in the cell walls and vacuoles in roots of AK58, limiting the translocation of Cd from root to shoot. Furthermore, the elevated TaHMA2 expression in ZM10 indicates that ZM10 had a higher capacity of xylem loading Cd than AK58. All of these results herein suggest that the radial transport is significant for Cd accumulation in roots, but it cannot explain the difference in root-to-shoot translocation of Cd in wheat genotypes with contrast Cd accumulation in grains.

Ochrobactrum intermedium and saponin assisted phytoremediation of Cd and B[a]P co-contaminated soil by Cd-hyperaccumulator Sedum alfredii.

Pot-culture experiments were conducted to investigate the potential of microorganism-saponin assisted phytoremediation of cadmium (Cd) and benzo(a)pyrene (B[a]P) co-contaminated soil using Cd-hyperaccumulator Sedum alfredii. Results showed that B[a]P-degrading bacterium (Ochrobactrum intermedium B[a]P-16) inoculation significantly increased root (by 22.1-24.1%) and shoot (by 20.5-23.4%) biomass of S. alfredii, whereas the application of saponin had no effect on the growth of S. alfredii. The saponin solution at 2 g L-1 extracted more Cd and B[a]P than water, saponin enhanced Cd and B[a]P bioavailability in soil and thus promoted their uptake and accumulation in S. alfredii. The activity of B[a]P-16, dehydrogenase and polyphenol oxidase in co-contaminated soil was promoted by growing S. alfredii, and the application of B[a]P-16 and saponins caused a significant (P < 0.05) increase in both enzyme activities. The maximum B[a]P removal rate (82.0%) and Cd phytoextraction rate (19.5%) were obtained by co-application of S. alfredii with B[a]P-16 and saponin. The B[a]P-16 and plant promoted biodegradation were the predominant contributors towards removal of B[a]P from soil. A significant (P < 0.05) synergistic effect of B[a]P-16 and saponin on B[a]P and Cd removal efficiency was observed in this study. It is suggested that planting S. alfredii with application of B[a]P-16 and saponin would be an effective method for phytoremediation of soil co-contaminated with Cd and PAHs.

Unique root exudate tartaric acid enhanced cadmium mobilization and uptake in Cd-hyperaccumulator Sedum alfredii.

Low molecular weight organic acids (LMWOA) involved in heavy metal tolerance, translocation, and accumulation in plants. However, underlying mechanism of LMWOA secretion in metal mobilization and uptake in hyperaccumulator still need to be identified. In this study, a 13C labeling rhizobox was designed to investigate the composition and distribution of LMWOA in the rhizosphere of S. alfredii. The result showed that about 2.30%, 2.25% and 2.35% of the assimilated 13C was incorporated into oxalic acid, malic acid, and tartaric acid in rhizosphere of S. alfredii after 13CO2 assimilation, respectively. Oxalic acid, malic acid, and tartaric acid were the predominant LMWOA in rhizosphere soil solution of hyperaccumulating ecotype (HE) S. alfredii, however, almost no tartaric acid was detected for non-hyperaccumulating ecotype (NHE). Tartaric acid was identified as the unique root exudate from HE S. alfredii which was mainly distributed within the range of rhizosphere 0-6 mm. Tartaric acid significantly increased the solubility of four Cd minerals. HE S. alfredii treated with tartrate + CdCO3 had higher Cd contents and larger biomass than CdCO3 treatment. Cadmium accumulation in HE S. alfredii was promoted by the exudation of tartaric acid, which was highly efficient in Cd solubilization due to the formation of soluble Cd-tartrate complexes.

Bioavailability and cytotoxicity of Cerium- (IV), Copper- (II), and Zinc oxide nanoparticles to human intestinal and liver cells through food.

Anthropogenic nanoparticles (NPs) are emitted to the environment and may be present in vegetables for human consumption. However, the toxicity of NPs exposure through food lack systematical investigations. In order to propose a systematical study, lettuce grown in a Cerium- (IV), Copper- (II) and Zinc oxide NP contaminated environment were digested. This digestate was used to culture human intestine cells (i.e. epithelial colorectal adenocarcinoma cells, Caco-2). The basolateral juice produced by the intestinal cells was then used to culture normal human liver (HL-7702) cells. Bioavailability and biotoxicity of the NPs in the vitro models were assessed. NPs were found to be taken up from the environment by vegetables, and may thus be transferred to humans through oral exposure. Bioavailability and the effect of their concentration in the digestate medium differed in regards to NP materials. The levels of NPs found in the digestate were detrimental to intestine cells, while the liver cells exposed to lower concentrations of NP in the bodily fluid showed no statically significant change in cell necrosis. A closer assessment of the detrimental effect of the studied NPs to Caco-2 cells revealed that the damage was mainly related to the solubility of the NPs. This may partly be due to that the more soluble NP material (ZnO > CuO > CeO2) render higher metal ion release and thus higher bioavailability. This appeared to cause more cell death, and even lead to local intestinal inflammation. Although no liver cells died, there was an increase of ROS level, causing ROS-related DNA damage prior to cell necrosis. The findings in this study enhances our understanding of the relative detrimental effect of different types of NPs, and the mechanisms causing their biotoxicity in human cells through food.

Abscisic acid-mediated modifications of radial apoplastic transport pathway play a key role in cadmium uptake in hyperaccumulator Sedum alfredii.

Abscisic acid (ABA) is a key phytohormone underlying plant resistance to toxic metals. However, regulatory effects of ABA on apoplastic transport in roots and consequences for uptake of metal ions are poorly understood. Here, we demonstrate how ABA regulates development of apoplastic barriers in roots of two ecotypes of Sedum alfredii and assess effects on cadmium (Cd) uptake. Under Cd treatment, increased endogenous ABA level was detected in roots of nonhyperaccumulating ecotype (NHE) due to up-regulated expressions of ABA biosynthesis genes (SaABA2, SaNCED), but no change was observed in hyperaccumulating ecotype (HE). Simultaneously, endodermal Casparian strips (CSs) and suberin lamellae (SL) were deposited closer to root tips of NHE compared with HE. Interestingly, the vessel-to-CSs overlap was identified as an ABA-driven anatomical trait. Results of correlation analyses and exogenous applications of ABA/Abamine indicate that ABA regulates development of both types of apoplastic barriers through promoting activities of phenylalanine ammonialyase, peroxidase, and expressions of suberin-related genes (SaCYP86A1, SaGPAT5, and SaKCS20). Using scanning ion-selected electrode technique and PTS tracer confirmed that ABA-promoted deposition of CSs and SL significantly reduced Cd entrance into root stele. Therefore, maintenance of low ABA levels in HE minimized deposition of apoplastic barriers and allowed maximization of Cd uptake via apoplastic pathway.

Effects of elevated CO2 and endophytic bacterium on photosynthetic characteristics and cadmium accumulation in Sedum alfredii.

Elevated CO2 and use of endophytic microorganisms have been considered as efficient and novel ways to improve phytoextraction efficiency. However, the interactive effects of elevated CO2 and endophytes on hyperaccumulator is poorly understood. In this study, a hydroponics experiment was conducted to investigate the combined effect of elevated CO2 (eCO2) and inoculation with endophyte SaMR12 (ES) on the photosynthetic characteristics and cadmium (Cd) accumulation in hyperaccumulator Sedum alfredii. The results showed that eCO2 × ES interaction promoted the growth of S. alfredii, shoot and root biomass net increment were increased by 264.7 and 392.3%, respectively, as compared with plants grown in ambient CO2 (aCO2). The interaction of eCO2 and ES significantly (P < 0.05) increased chlorophyll content (53.2%), Pn (111.6%), Pnmax (59.8%), AQY (65.1%), and Lsp (28.8%), but reduced Gs, Tr, Rd, and Lcp. Increased photosynthetic efficiency was associated with higher activities of rubisco, Ca2+-ATPase, and Mg2+-ATPase, and linked with over-expression of two photosystem related genes (SaPsbS and SaLhcb2). PS II activities were significantly (P < 0.05) enhanced with Fv/Fm and Φ(II) increased by 12.3 and 13.0%, respectively, compared with plants grown in aCO2. In addition, the net uptake of Cd in the shoot and root tissue of S. alfredii grown in eCO2 × ES treatment was increased by 260.7 and 434.9%, respectively, due to increased expression of SaHMA2 and SaCAX2 Cd transporter genes. Our results suggest that eCO2 × ES can promote the growth of S. alfredii due to increased photosynthetic efficiency, and improve Cd accumulation and showed considerable potential of improving the phytoextraction ability of Cd by S. alfredii.

Endophytic bacterium Buttiauxella sp. SaSR13 improves plant growth and cadmium accumulation of hyperaccumulator Sedum alfredii.

Inoculation with endophytic bacterium has been considered as a prospective application to improve the efficiency of phytoextraction. In this study, the effect of Buttiauxella sp. SaSR13 (SaSR13), a novel endophytic bacterium isolated from the root of hyperaccumulator Sedum alfredii, on plant growth and cadmium (Cd) accumulation in S. alfredii was investigated. Laser scanning confocal microscopic (LSCM) images showed that SaSR13 was mainly colonized in the root elongation and mature zones. The inoculation with SaSR13 to Cd-treated plants significantly enhanced plant growth (by 39 and 42% for shoot and root biomass, respectively), chlorophyll contents (by 38%), and Cd concentration in the shoot and root (by 32 and 22%, respectively). SaSR13 stimulated the development of roots (increased root length, surface area, and root tips number) due to an increase in the indole-3-acid (IAA) concentrations and a decrease in the concentrations of superoxide anion (O2.-) in plants grown under Cd stress. Furthermore, inoculation with SaSR13 enhanced the release of root exudates, especially malic acid and oxalic acid, which might have facilitated the uptake of Cd by S. alfredii. It is suggested that inoculation with endophytic bacterium SaSR13 is a promising bioaugmentation method to enhance the Cd phytoextraction efficiency by S. alfredii.

[Re-study on characteristic fingerprint of volatile oil from Herba Pogostemonis by GC].

OBJECTIVE: To establish the characteristic fingerprint and parameter of volatile oil from Herba Pogostemonis. METHODS: The dffferent collecting time and county samples of Herba Pogostemonis were determined by GC. RESULTS: Accuracy, stability and repeatability of the method were good. The same region samples had good stability and main constituents were same in different collection time, but content were different. The samples collected from different region were more different in constituents and content. CONCLUSION: The 11 components in common buildup the characterisitic fingerprint of volatile oil from Herba Pogostemonis. The ratiio of pogostone to patchoulic alcohol were used index parameter for quality evaluation and characteristics of producing region.

[Two chemotypes of Pogostemon cablin and influence of region of cultivation and harvesting time on volatile oil composition].

AIM: To analyze and compare the constituents of the volatile oil of Pogostemon cablin collected from different regions of cultivation and harvesting times in order to evaluate the quality of Shipai Huoxiang and to expound the chemical intension of Pogostemon cablin. METHODS: The combination of GC and MS. RESULTS: The volatile oil compositions of Herba Pogostemonis collected from various of cultivation regions and harvesting times are obviously different. Based on the chemical differences of the volatile oil compositions, Pogostemon cablin is divided into two chemotypes, Pogostone-type and Patchouliol-type. The former was cultivated in Guangzhou and Gaoyao regions, locally named as "Shipai Huoxiang"; the latter was locally named as "Hainan Huoxiang", cultivated in Wuchuan, Suixi and Leizhou regions of Guangdong Province and Wanning region of Hainan Province. The Pogostone-type contains rich oxygenated components, especially pogostone in the volatile oil compositions and poor non-oxygenated composition with patchouliol. The above chemical data may be used as evaluation standard for the authentic Shipai Huoxiang. The Patchouliol-type contains similar quantities of oxygenated and non-oxygenated composition, especially rich patchouliol with poor pogostone in oxygenated compositions, rich delta-guaiene and alpha-guaiene in non-oxygenation compositions. The contents of volatile oil and their constituents were varied due to different harvesting time. The yields of pogostone and volatile oil of Shipai Huoxiang was higher in July. The quality of the samples collected in this month was better. CONCLUSION: According to the volatile oil compositions, there are two chemotypes (Pogostone-type and Patchouliol-type) in Pogostemon cablin plant. These two chemotypes correspond to the genotypes of plastid matK gene and nuclear 18s rRNA gene by cluster analysis.

[DNA profiling of Pogostemon cablin chemotypes differing in essential oil composition].

AIM: To provide molecular evidence for quality evaluation and GAP production of Pogostemon cablin (Blanco) Benth. cultivated in different regions in Guangdong and Hainan provinces, China, by comparing two sequences (1.2 kb of plastid matK gene and 1.8 kb of nuclear 18S rRNA gene) and two chemotypes (Pogostone-type and Patchouliol-type in essential oil composition). METHODS: PCR direct sequencing was applied to detemine the matK and 18S rRNA sequences for six samples of Pogostemon cablin from different localities. RESULTS: The matK sequences of six samples of Pogostemon cablin from different regions of cultivation are 1,245 bp in length, which coding 415 amino acids of protein (maturase), and 18S rRNA sequences are 1,803-1,805 bp in size. Based on multiple sequence alignment, there are 47 variable sites in the matK sequence of these six samples, 17 in the 18S rRNA sequence. The cluster tree reconstructed by UPGMA method shows that the sequence divergence both in matK and 18S rRNA genes among six samples of Pogostemon cablin was well correlative with their regions of cultivation and intraspecific chemotypes of essential oil composition. CONCLUSION: Combining with chemical and biogeographical data, DNA sequencing can become a powerful tool in the key technique-species identification of quality evaluation and GAP production of Pogostemon cablin.

[Constituents analysis on volatile oil of Pogostemon cablin from different collection time cultivated in Hainan].

The constituents of the volatile oil of Pogostemon cablin collected from Wanning city, Hainan province and the influence owing to different collection time were undertaken by GC/MS combination technology. The results showed that main compounds was patchouli alcohol in stem oil and leaf oil, its contents were respectively 36.06% and 37.74%; pogostone was poor, its contents in stem oil and leaf oil were respectively 17.08% and 0.85%; the contents of ten compounds were over 1%, which was respectively beta-patchoulene, beta-elemene, transcaryophyllene, delta-guaiene, seychellene, alpha-patchoulene, aciphyllene, alpha-guaiene, patchouli alcohol and pogostone; the contents of volatile oil from June to August were respectively 0.8%, 0.7% and 0.6%; patchouli alcohol, higher in July and June(42.62% and 40.84%), lower in August(31.40%).