Overexpression of IlHMA2, from Iris lactea, Improves the Accumulation of and Tolerance to Cadmium in Tobacco.

Long-distance transport cadmium (Cd) from roots to shoots is a key factor for Cd phytoremediation. Our previous study indicated that heavy metal P1B2-ATPases, IlHMA2, was involved in improving the accumulation of Cd via mediated long-distance transport Cd, contributing to the phytoremediation in Cd accumulator Iris lactea. However, whether the overexpression of IlHMA2 could enhance the accumulation and tolerance to Cd remains unclear in plants. Here, we generated transgenic tobacco overexpressing IlHMA2 and tested its effect on the translocation and accumulation of Cd and zinc (Zn), as well as the physio-biochemical characteristics under 50 mg/L Cd exposure. The overexpression of IlHMA2 significantly increased Cd concentrations in xylem saps, resulting in enhanced root-to-shoot Cd translocation compared with wild-type. Meanwhile, overexpressing IlHMA2 promoted Zn accumulations, accompanied by elevating proline contents and antioxidant enzyme activity (SOD, POD, and CAT) to diminish the overproduction of ROS in transgenic tobacco. These pieces of evidence suggested that higher Zn concentrations and lower ROS levels could tremendously alleviate Cd toxicity for transgenic tobacco, thereby improving the growth and tolerance. Overall, the overexpression of IlHMA2 could facilitate Cd accumulation and enhance its tolerance in tobacco exposed to Cd contaminations. This would provide a valuable reference for improving Cd phytoremediation efficiency.

Chromosome-level reference genome assembly provides insights into the evolution of Pennisetum alopecuroides.

Pennisetum alopecuroides is an important forage grass resource, which plays a vital role in ecological environment improvement. Therefore, the acquisition of P. alopecuroides genome resources is conducive to the study of the adaptability of Pennisetum species in ecological remediation and forage breeding development. Here we assembled a P. alopecuroides cv. 'Liqiu' genome at the chromosome level with a size of approximately 845.71 Mb, contig N50 of 84.83Mb, and genome integrity of 99.13% as assessed by CEGMA. A total of 833.41-Mb sequences were mounted on nine chromosomes by Hi-C technology. In total, 60.66% of the repetitive sequences and 34,312 genes were predicted. The genomic evolution analysis showed that P. alopecuroides cv. 'Liqiu' was isolated from Setaria 7.53-13.80 million years ago and from Cenchrus 5.33-8.99 million years ago, respectively. The whole-genome event analysis showed that P. alopecuroides cv. 'Liqiu' underwent two whole-genome duplication (WGD) events in the evolution process, and the duplication events occurred at a similar time to that of Oryza sativa and Setaria viridis. The completion of the genome sequencing of P. alopecuroides cv. 'Liqiu' provides data support for mining high-quality genetic resources of P. alopecuroides and provides a theoretical basis for the origin and evolutionary characteristics of Pennisetum.

IlNRAMP5 is required for cadmium accumulation and the growth in Iris lactea under cadmium exposures.

Iris lactea is potentially applied for remediating Cd-contaminated soils due to the strong ability of Cd uptake and accumulation. However, its molecular mechanism underlying Cd uptake pathway remains unknown. Here, we report a member of NRAMP (Natural Resistance-Associated Macrophage Protein) family, IlNRAMP5, is involved in Cd/Mn uptake and the growth in I. lactea response to Cd. IlNRAMP5 was localized onto the plasma membrane, and was induced by Cd. It was expressed in the root cortex rather than the central vasculature, and in leaf vascular bundle and mesophyll cells. Heterologous expression in yeast showed that IlNRAMP5 could transport Cd and Mn, but not Fe. Knockdown of IlNRAMP5 triggered a significant reduction in Cd uptake, further diminishing the accumulation of Cd. In addition, silencing IlNRAMP5 disrupted Mn homeostasis by lowering Mn uptake and Mn allocation, accompanied by remarkably inhibiting photosynthesis under Cd conditions. Overall, the findings suggest that IlNRAMP5 plays versatile roles in Cd accumulation by mediating Cd uptake, and contributes to maintain the growth via modulating Mn homeostasis in I. lactea under Cd exposures. This would provide a mechanistic understanding Cd phytoremediation efficiency in planta.

Effects of Graphene Oxide on the Growth and Photosynthesis of the Emergent Plant Iris pseudacorus.

The extensive applications of graphene oxide (GO) inevitably lead to entry into the natural aquatic environment. However, information on its toxicity to emergent plants is still lacking. In this study, an emergent plant, Iris pseudacorus, was exposed to GO (1, 20, 80, and 140 mg·L-1) under hydroponic conditions for 15 weeks. Changes in plant growth were assessed by analyzing plant biomass and photosynthetic pigment contents; the photosynthesis response was verified by measuring chlorophyll a fluorescence; and the nutrient levels of the plant were evaluated. Results showed that GO at 20-140 mg·L-1 significantly increased plant dry weight by 37-84% and photosynthetic pigment contents by 26-178%, and 80 mg·L-1 was the optimal concentration. PSII activity, adjustment capacities of electron transport in PSII, the grouping or energetic connectivity between PSII units, light energy conversion efficiency, photosynthesis performance indexes (by 11-51%), and contents of several nutrient elements (N, Fe, and Cu) were increased by 49-69%, 34-84%, and 11-38%, respectively. These findings indicate that GO can enhance plant growth by promoting plant photosynthesis performance and improving plant nutrient levels, and has great application potential in promoting the growth and development of this emergent plant as a phytoremediation agent.

An All-in-One, Bioderived, Air-Permeable, and Sweat-Stable MXene Epidermal Electrode for Muscle Theranostics.

Muscle fatigue is a common symptom experienced by many people and associated with less maximal force production of fatigued muscle. It is highly desirable to simultaneously and imperceivably diagnose muscle fatigue and restore muscle function using one skin electrode, yet no such electrode has been developed so far. Herein, we report an all-in-one, bioderived, air-permeable, and sweat-stable MXene electrode that can concurrently and comfortably record electromyographic (EMG) signals and achieve electrostimulation and electrothermal therapy for muscle theranostics. Leveraging the structural arrangement of perennial herbs and ion cross-linking of MXene in sweat, MXene-based electrodes (MBE) exhibit high breathability, are ultralightweight (∼0.25 mg/cm3), and have low and stable electrode-skin interfacial impedance at a variety of environments, facilitating the long-term reliable monitoring of electrophysiology. Taken together with electrostimulation and electrothermal therapy at the skin surface, MBE can diagnose muscle fatigue and restore muscle function by stimulating blood circulation. In addition, it can also be used for muscle rehabilitation training and prosthesis control via human-computer interaction. Our all-in-one, bioderived, air-permeable, and sweat-stable MXene electrode has a great potential for daily wearable healthcare of muscle fatigue.

ROS scavenging and ion homeostasis is required for the adaptation of halophyte Karelinia caspia to high salinity.

The halophyte Karelinia caspia has not only fodder and medical value but also can remediate saline-alkali soils. Our previous study showed that salt-secreting by salt glands is one of main adaptive strategies of K. caspia under high salinity. However, ROS scavenging, ion homeostasis, and photosynthetic characteristics responses to high salinity remain unclear in K. caspia. Here, physio-biochemical responses and gene expression associated with ROS scavenging and ions transport were tested in K. caspia subjected to 100-400 mM NaCl for 7 days. Results showed that both antioxidant enzymes (SOD, APX) activities and non-enzymatic antioxidants (chlorogenic acid, α-tocopherol, flavonoids, polyamines) contents were significantly enhanced, accompanied by up-regulating the related enzyme and non-enzymatic antioxidant synthesis gene (KcCu/Zn-SOD, KcAPX6, KcHCT, KcHPT1, Kcγ-TMT, KcF3H, KcSAMS and KcSMS) expression with increasing concentrations of NaCl. These responses are beneficial for removing excess ROS to maintain a stable level of H2O2 and O2 - without lipid peroxidation in the K. caspia response to high salt. Meanwhile, up-regulating expression of KcSOS1/2/3, KcNHX1, and KcAVP was linked to Na+ compartmentalization into vacuoles or excretion through salt glands in K. caspia. Notably, salt can improve the function of PSII that facilitate net photosynthetic rates, which is helpful to growing normally in high saline. Overall, the findings suggested that ROS scavenging systems and Na+/K+ transport synergistically contributed to redox equilibrium, ion homeostasis, and the enhancement of PSII function, thereby conferring high salt tolerance.

Defining key metabolic roles in osmotic adjustment and ROS homeostasis in the recretohalophyte Karelinia caspia under salt stress.

The recretohalophyte Karelinia caspia is of forage and medical value and can remediate saline soils. We here assess the contribution of primary/secondary metabolism to osmotic adjustment and ROS homeostasis in Karelinia caspia under salt stress using multi-omic approaches. Computerized phenomic assessments, tests for cellular osmotic changes and lipid peroxidation indicated that salt treatment had no detectable physical effect on K. caspia. Metabolomic analysis indicated that amino acids, saccharides, organic acids, polyamine, phenolic acids, and vitamins accumulated significantly with salt treatment. Transcriptomic assessment identified differentially expressed genes closely linked to the changes in above primary/secondary metabolites under salt stress. In particular, shifts in carbohydrate metabolism (TCA cycle, starch and sucrose metabolism, glycolysis) as well as arginine and proline metabolism were observed to maintain a low osmotic potential. Chlorogenic acid/vitamin E biosynthesis was also enhanced, which would aid in ROS scavenging in the response of K. caspia to salt. Overall, our findings define key changes in primary/secondary metabolism that are coordinated to modulate the osmotic balance and ROS homeostasis to contribute to the salt tolerance of K. caspia.

Cadmium uptake by a hyperaccumulator and three Pennisetum grasses with associated rhizosphere effects.

Pennisetum grasses (P. purpureum Schumach. 'Purple', P. alopecuroides (L.) Spreng. 'Liren' and P. alopecuroides (L.) Spreng. 'Changsui'), and a cadmium (Cd) hyperaccumulator (Thlaspi caerulescens J.Presl & C.Presl), were grown in soil with four Cd addition levels of 0, 2, 20 and 200 mg/kg. Toxicity symptoms were not observed although growth of all plants decreased as Cd addition increased. Shoot bioconcentration factor (BCFS), the translocation factor (TF) and shoot accumulation of Cd for most plants first increased and then declined as Cd concentrations increased. In contrast, the root bioconcentration factor (BCFR) for T. caerulescens declined and root Cd accumulation for T. caerulescens and two P. alopecuroides cultivars increased consistently as Cd levels increased. P. purpureum had the largest biomass with shoot Cd accumulation similar to that of T. caerulescens, despite lower foliar Cd concentration. Although shoot Cd concentrations of two P. alopecuroides cultivars were lower than for P. purpureum, root Cd concentrations were greater. P. purpureum had Cd BCFS and TF (> 1) at 2- and 20-mg/kg Cd addition treatments, similar to T. caerulescens. P. alopecuroides cultivars had Cd BCFR (> 1) and TF (< 1) at all Cd levels. Roots did not affect rhizosphere pH. However, concentrations of acid extractable Cd in rhizosphere soil were lower than those of corresponding non-rhizosphere soil at all Cd levels for T. caerulescens and P. purpureum; T. caerulescens and P. purpureum did not affect less bioavailable Cd fractions. Concentrations of acid extractable Cd in the rhizosphere of the P. alopecuroides cultivars were not reduced at any Cd level. Differences in Cd accumulation among the three Pennisetum grasses were mainly attributable to root biomass and Cd TFs rather than rhizosphere Cd mobility.

[Uptake and Accumulation of Cadmium and Zinc by Two Energy Grasses: A Field Experiment].

The remediation potential of large biomass energy grasses in cadmium-contaminated soil remains ambiguous. A field experiment was carried out in a cadmium-contaminated farmland using two energy grasses and two control plants. The two energy grasses were hybrid pennisetum (Pennisetum americanum×P. purpureum, PAP) and purple elephant grass (P. purpureum 'Purple', PPP), and the two control plants were Iris lactea var. chinensis (ILC) and a cadmium hyperaccumulator, Noccaea caerulescens (NC). The results showed that the aboveground biomass of PAP was the largest among the four plants, and 126 and 36 times that of NC and ILC, respectively, but no significant difference with that of PPP. The concentrations of cadmium and zinc in the shoots and roots of NC were significantly higher than in the other plants. Zinc concentrations in the shoots and roots of ILC were lower than in the other plants, while cadmium concentrations were significantly higher than in PAP and PPP (P<0.05). The amounts of cadmium and zinc accumulated in the shoots of PPP were the highest among the four plants, while cadmium concentrations in the shoots and roots of PPP were significantly lower than in ILC and NC (P<0.05). Cadmium amounts accumulated in PPP shoots were 7.0 and 4.1 times that of ILC and NC, respectively. Zinc amounts accumulated in PPP shoots were 41 and 11 times that of ILC and NC, respectively (P<0.05). Cadmium accumulation in the shoots of PAP was 19.4% lower than in PPP, and zinc accumulation had no significant difference with that of PPP. NC, having a bioconcentration factor of shoot (BCFS) and a translocation factor (TF) for cadmium and zinc both larger than 1, is usable for phytoextraction of soils contaminated by cadmium and zinc. ILC, having a bioconcentration factor of root (BCFR) larger than 1 and a TF lower than 1 for cadmium, is usable for the phytostabilization of soils contaminated by cadmium. PPP, having a BCFR larger than 1 and a TF lower than 1 for zinc, can be used in the phytostabilization of soils contaminated by zinc. Under field conditions, PPP and PAP showed great potential for the extraction and removal of cadmium and zinc from soil due to their large biomass and ability to produce economic benefits, have good application prospects.

[Chemical composition analysis of Eucalyptus essential oil and allelopathic effects of α-terpineol]. / æ¡‰æ ‘æ²¹åŒ–å­¦æˆåˆ†åˆ†æžåŠα-松油醇的化感作用.

We extracted Eucalyptus essential oil by steam distillation and analyzed its chemical components by GC-MS. The inhibitory effects of α-terpineol, a component in Eucalyptus essential oil, were tested in Petri dishes on the germination and growth of Amaranthus retroflexus. The allelopathic effects of α-terpineol on A. retroflexus were examined with a pot experiment by measuring germination rate and biomass. The yield of essential oil was 0.04%. Ninety-two chemical components were detected from the essential oil. In Petri dish experiment, the germination inhibitory effect under 5 µL and 7.5 µL of α-terpineol per dish reached 100%. In pot experiment, germination, aboveground fresh and dry weights of A. retroflexus were significantly affected by α-terpineol. At the concentration of 1.6 µL·cm-3, the inhibition effect was strongest, with the allelopathic response index being -0.51 for germination, -0.62 for aboveground fresh weight, and -0.44 for aboveground dry weight, and the inhibition ratio being 51%, 62% and 44%, respectively.

Extended light exposure increases stem digestibility and biomass production of switchgrass.

Switchgrass is a photoperiod-sensitive energy grass suitable for growing in the marginal lands of China. We explored the effects of extended photoperiods of low-irradiance light (7 µmol·m-2·s-1, no effective photosynthesis) on the growth, the biomass dry weight, the biomass allocation, and, especially, the stem digestibility and cell wall characteristics of switchgrass. Two extended photoperiods (i.e., 18 and 24 h) were applied over Alamo. Extended light exposure (18 and 24 h) resulted in delayed heading and higher dry weights of vegetative organs (by 32.87 and 35.94%, respectively) at the expense of reducing the amount of sexual organs (by 40.05 and 50.87%, respectively). Compared to the control group (i.e., natural photoperiod), the yield of hexoses (% dry matter) in the stems after a direct enzymatic hydrolysis (DEH) treatment significantly increased (by 44.02 and 46.10%) for those groups irradiated during 18 and 24 h, respectively. Moreover, the yield of hexoses obtained via enzymatic hydrolysis increased after both basic (1% NaOH) and acid (1% H2SO4) pretreatments for the groups irradiated during 18 and 24 h. Additionally, low-irradiance light extension (LILE) significantly increased the content of non-structural carbohydrates (NSCs) while notably reducing the lignin content and the syringyl to guaiacyl (S/G) ratio. These structural changes were in part responsible for the observed improved stem digestibility. Remarkably, LILE significantly decreased the cellulose crystallinity index (CrI) of switchgrass by significantly increasing both the arabinose substitution degree in xylan and the content of ammonium oxalate-extractable uronic acids, both favoring cellulose digestibility. Despite this LILE technology is not applied to the cultivation of switchgrass on a large scale yet, we believe that the present work is important in that it reveals important relationships between extended day length irradiations and biomass production and quality. Additionally, this study paves the way for improving biomass production and digestibility via genetic modification of day length sensitive transcription factors or key structural genes in switchgrass leaves.

Exploring surface characterization and electrostatic property of Hybrid Pennisetum during alkaline sulfite pretreatment for enhanced enzymatic hydrolysability.

The surface characterization and electrostatic property of Hybrid Pennisetum (HP) after alkaline sulfite pretreatment were explored for enhanced enzymatic hydrolysability. The O/C ratio in HP increased from 0.34 to 0.60, and C1 concentration decreased from 62.5% to 31.6%, indicating that alkaline sulfite pretreatment caused poorer lignin but richer carbohydrate on HP surface. Zeta potential and sulfur element analysis indicated that more enzymes would preferably adsorb on the carbohydrate surface of alkaline sulfite pretreated HP because the lignin was sulfonated, which facilitated the decrease of non-productive adsorption. Glucose yield of alkaline sulfite pretreated HP reached to 100% by synergistic action of cellulase and xylanase in the hydrolysis, which was significantly higher than that of NaOH pretreated, and the concentration of glucose released was 1.52times higher. The results suggested that alkaline sulfite pretreatment had potential for improving the HP hydrolysability, and the surface characterization and electrostatic property facilitated the enzymatic digestibility.

Structural properties and hydrolysabilities of Chinese Pennisetum and Hybrid Pennisetum: Effect of aqueous ammonia pretreatment.

The effects of aqueous ammonia pretreatment on structural properties and hydrolysabilities of Chinese Pennisetum and Hybrid Pennisetum were investigated. Aqueous ammonia pretreatment increased cellulose crystallinities and hydrolysabilities of Chinese Pennisetum and Hybrid Pennisetum. Compared with Chinese Pennisetum, Hybrid Pennisetum showed better enzymatic digestibility. Xylanase supplementation was more effective than the increase of cellulase loadings in the hydrolysis of aqueous ammonia pretreated Chinese Pennisetum and Hybrid Pennisetum. After supplementation of 2mg of xylanase/g dry matter to 5 FPU of cellulases/g dry matter, the hydrolysis yields of cellulose of aqueous ammonia pretreated Chinese Pennisetum and Hybrid Pennisetum were 92.3-95.4%, and the hydrolysis yields of xylan were 86.9-94.2%. High hydrolysability and low dosage of enzyme loadings together with the advantages of high yield and widely distribution demonstrated the potential of Chinese Pennisetum and Hybrid Pennisetum for the production of platform sugars.

Tassel removal positively affects biomass production coupled with significantly increasing stem digestibility in switchgrass.

In this study, tassels of Cave-in-Rock (upland) and Alamo (lowland) were removed at or near tassel emergence to explore its effects on biomass production and quality. Tassel-removed (TR) Cave-in-Rock and Alamo both exhibited a significant (P<0.05) increase in plant heights (not including tassel length), tiller number, and aboveground biomass dry weight (10% and 12%, 30% and 13%, 13% and 18%, respectively by variety) compared to a control (CK) treatment. Notably, total sugar yields of TR Cave-in-Rock and Alamo stems increased significantly (P<0.05 or 0.01) by 19% and 19%, 21% and 14%, 52% and 18%, respectively by variety, compared to those of control switchgrass under 3 treatments by direct enzymatic hydrolysis (DEH), enzymatic hydrolysis after 1% NaOH pretreatment (EHAL) and enzymatic hydrolysis after 1% H2SO4 pretreatment (EHAC). These differences were mainly due to significantly (P<0.05 or 0.01) higher cellulose content, lower cellulose crystallinity indexes (CrI) caused by higher arabinose (Ara) substitution in xylans, and lower S/G ratio in lignin. However, the increases of nitrogen (N) and sulphur (S) concentration negatively affects the combustion quality of switchgrass aboveground biomass. This work provides information for increasing biomass production and quality in switchgrass and also facilitates the inhibition of gene dispersal of switchgrass in China.

[Impacts of salt stress on the growth and physiological characteristics of Panicum virgatum seedlings].

An experiment was conducted in an artificial climate chamber to study the growth and physiological characteristics of switchgrass (Panicum virgatum) seedlings exposed to 0, 50, 100, 150, and 200 mmol x L(-1) of NaCl solutions. With the increasing concentration of the NaCl, the seedling growth was obviously inhibited. The plant height decreased, leaves became smaller, photosynthetic leaf area and net photosynthetic rate reduced, and dry matter accumulation decreased significantly, presenting the general traits of glycophyte. Meanwhile, the seedlings also showed relatively high salt tolerance. After exposed to 200 mmol x L(-1) of NaCl for 30 days, the seedlings still survived, with the green leaf area per plant and net photosynthetic rate being 491.9 cm2 and 0.93 micromol CO2 x m(-2) x s(-1), respectively. In this experiment, the salt tolerance threshold for P. virgatum was 178.6 mmol L(-1) when taking 50% drop in biomass as the standard.