Generation of low-cadmium rice germplasms via knockout of OsLCD using CRISPR/Cas9

The OsLCD gene,which has been implicated in cadmium (Cd) accumulation in rice,might be a useful target for CRISPR/Cas9 editing.However,the effects of Os LCD gene editing on Cd accumulation,plant growth,and yield traits remain unknown.Here,we used CRISPR/Cas9to generate oslcd single mutants from indica and japonica rice cultivars.We also generated osnramp5 single mutants and oslcd osnramp5 double mutants in the indica background.When grown in Cd-contaminated paddy soils,all oslcd single mutants accumulated less Cd than the wild types (WTs).Consistent with this,oslcd single mutants grown in Cd-contaminated hydroponic culture accumulated significantly less Cd in the shoots as compared to WTs.This decrease in accumulation probably resulted from the reduction of Cd translocation under Cd stress.Oxidative damage also decreased,and plant growth increased in all oslcd single mutant seedlings as compared to WTs in the presence of Cd.Plant growth and most yield traits,as well essential element concentrations in rice seedling shoots,brown rice,and rice straw,were similar between oslcd single mutants and WTs.In the presence of Cd,Cd concentrations in the brown rice and shoots of oslcd osnramp5 double mutants were significantly decreased compared with WTs as well as osnramp single mutants.Our results suggested that OsL CD knockout may reduce Cd accumulation alone or in combination with other knockout mutations in a variety of rice genotypes;unlike Os Nramp5 mutations,Os LCD knockout did not reduce essential element contents.Therefore,Os LCD knockout might be used to generate low-Cd rice germplasms.

Ru/Pt@BSA nanoparticles for efficient photo-catalytic oxidation of NAD(P)H and targeted cancer treatment under hypoxic conditions

Photo-catalytic oxidation of intracellular nicotinamide adenine dinucleotide(2'-phosphate)(NAD(P)H)has attracted much attention for cancer therapy.However,the general oxygen-dependent mechanism heavily depresses the efficacy in hypoxic tumors.To solve this problem,herein platinum nanoparticles(Pt NPs)with catalase-like(CAT-like)and catalytic H_(2)evolution activities were introduced as a powerful assistant to enhance the photo-catalytic NAD(P)H oxidation of Ru1([Ru(phen)2(PIP-OCH_(3))]^(2+),phen=1,10-phenanthroline,PIP-OCH_(3)=2-(4-methoxy phenyl)-1H-imidazo[4,5-f][1,10]phenanthroline)under hypoxic and even oxygen-free conditions.Firstly,Pt NPs can transform the original and in situ formed H_(2)O_(2)once again into O_(2)by the CAT-like activity,thus relieving tumor hypoxia and realizing cyclic utilization(at least in part)of the precious oxygen in hypoxia.Secondly,Pt NPs can also be served as H_(2)evolution catalysts while using Ru1 as the photosensitizer and NAD(P)H as the electron and proton donor.In this process,NAD(P)H is oxidized without the participation of oxygen,which can provide an effective way even under oxygen-free conditions.Via co-encapsulation of Rul and Pt NPs in bovine serum albumin(BSA)with tumor targeting ability,the resultant Ru/Pt@BSA could photo-catalyze intracellular NAD(P)H oxidation under hypoxic conditions(3%O_(2)),and exhibited an efficient and selective anticancer activity both in vitro and in vivo.Our results may provide new sights for efficient and targeted cancer treatment underhypoxic conditions.

Comparative physiological and transcriptomic analyses illuminate common mechanisms by which silicon alleviates cadmium and arsenic toxicity in rice seedlings

The inessential heavy metal/loids cadmium(Cd)and arsenic(As),which often co-occur in polluted paddy soils,are toxic to rice.Silicon(Si)treatment is known to reduce Cd and As toxicity in rice plants.To better understand the shared mechanisms by which Si alleviates Cd and As stress,rice seedlings were hydroponically exposed to Cd or As,then treated with Si.The addition of Si significantly ameliorated the inhibitory effects of Cd and As on rice seedling growth.Si supplementation decreased Cd and As translocation from roots to shoots,and significantly reduced Cd-and As-induced reactive oxygen species generation in rice seedlings.Transcriptomics analyses were conducted to elucidate molecular mechanisms underlying the Si-mediated response to Cd or As stress in rice.The expression patterns of the differentially expressed genes in Cd-or As-stressed rice roots with and without Si application were compared.The transcriptomes of the Cd-and As-stressed rice roots were similarly and profoundly reshaped by Si application,suggesting that Si may play a fundamental,active role in plant defense against heavy metal/loid stresses by modulating whole genome expression.We also identified two novel genes,0s01g0524500 and 0s06g0514800,encoding a myeloblastosis(MYB)transcription factor and a thionin,respectively,which may be candidate targets for Si to alleviate Cd and As stress in rice,as well as for the generation of Cd-and/or As-resistant plants.This study provides valuable resources for further clarification of the shared molecular mechanisms underlying the Si-mediated alleviation of Cd and As toxicity in rice.

Comparative transcriptomics provide new insights into the mechanisms by which foliar silicon alleviates the effects of cadmium exposure in rice

Silicon(Si)has been shown to alleviate Cd stress in rice.Here,we investigated the beneficial effects of foliar Si in an indica rice Huanghuazhan(HHZ).Our results showed that foliar Si in-creases the dry weight and decreases Cd translocation in Cd-exposed rice at the grain-filling stage only,implying that the filling stage is critical for foliar Si to reduce Cd accumulation.We also investigated the transcriptomics in flag leaves(FLs),spikelets(SPs),and node Is(NIs)of Cd-exposed HHZ after foliar Si application at the filling stage.Importantly,the gene expression profiles associated with the Si-mediated alleviation of Cd stress were tissue spe-cific,while shared pathways were mediated by Si in Cd-exposed rice tissues.Furthermore,after the Si treatment of Cd-exposed rice,the ATP-binding cassette(ABC)-transporters were mostly upregulated in FL and SP,while the bivalent cation transporters were mostly down-regulated in FL and NI,possibly helping to reduce Cd accumulation.The genes associated with essential nutrient transporters,carbohydrate and secondary metabolite biosynthesis,and cytochrome oxidase activity were mostly upregulated in Cd-exposed FL and SP,which may help to alleviate oxidative stress and improve plant growth under Cd exposure.Inter-estingly,genes responsible for signal transduction were negatively regulated in FL,but pos-itively regulated in SP,by foliar Si.Our results provide transcriptomic evidence that foliar Si plays an active role in alleviating the effects of Cd exposure in rice.In particular,foliar Si may alter the expression pattern of genes associated with transport,biosynthesis and metabolism,and oxidation reduction.