Upregulation of ATG9b by propranolol promotes autophagic cell death of hepatic stellate cells to improve liver fibrosis.

Proranolol has long been recommended to prevent variceal bleeding in patients with cirrhosis. However, the mechanisms of propranolol in liver fibrosis have not yet been thoroughly elucidated. Autophagic cell death (ACD) of activated hepatic stellate cells (HSCs) is important in the alleviation of liver fibrosis. Our study aims to assess the mechanisms of propranolol regulating HSC ACD and liver fibrosis. ACD of HSCs was investigated using lentivirus transfection. The molecular mechanism was determined using a PCR profiler array. The role of autophagy-related protein 9b (ATG9b) in HSC ACD was detected using co-immunoprecipitation and co-localization of immunofluorescence. Changes in the signalling pathway were detected by the Phospho Explorer antibody microarray. Propranolol induces ACD and apoptosis in HSCs. ATG9b upregulation was detected in propranolol-treated HSCs. ATG9b upregulation promoted ACD of HSCs and alleviated liver fibrosis in vivo. ATG9b enhanced the P62 recruitment to ATG5-ATG12-LC3 compartments and increased the co-localization of P62 with ubiquitinated proteins. The PI3K/AKT/mTOR pathway is responsible for ATG9b-induced ACD in activated HSCs, whereas the p38/JNK pathway is involved in apoptosis. This study provides evidence for ATG9b as a new target gene and propranolol as an agent to alleviate liver fibrosis by regulating ACD of activated HSCs.

Development of dual-visible reporter assays to determine the DNA-protein interaction.

The application of DNA-protein interaction reporter assays for relational or ratiometric measurements within an experimental system is popular in biological research. However, the existing reporter-based interaction assays always require special equipment, expensive chemicals, and a complicated operation. Here, we developed a DNA-protein interaction technology integrating two visible reporters, RUBY and UV-visible GFP (eYGFPuv), which allows the expression of the cassette reporter contained cis-acting DNA element (DE) fused upstream of TATA box and RUBY, and a constitutive promoter regulating eYGFPuv in the same construct. The interaction of transcription factor (TF) and the DE can be detected by co-expressed the cassette reporter and TF in tobacco leaves where the cassette reporter alone serves as a control. We also revealed that eight function-unknown bamboo AP2/ERFs interacted with the DE of ANT-AP2R1R2 (ABE), DRE (DBE), GCC-box (EBE), and RAV1 binding element (RBE), respectively, which are consistent with the results by dual-luciferase reporter assays. Thus, the dual-visible reporters offer a convenient, visible, and cost-saving alternative to other existing techniques for DNA-protein interaction in plants.

A Study on Predicting the Deviation of Jet Trajectory Falling Point under the Influence of Random Wind.

As one of the main external factors affecting the fire extinguishing accuracy of sprinkler systems, it is necessary to analyze and study random wind. However, in practical applications, there is little research on the impact of random wind on sprinkler fire extinguishing points. To address this issue, a new random wind acquisition system was constructed in this paper, and a method for predicting jet trajectory falling points in Random Forest (RF) under the influence of random wind was proposed, and compared with the commonly used prediction model Support Vector Machine (SVM). The method in this article reduces the error in the x direction of the 50 m prediction result from 2.11 m to 1.53 m, the error in the y direction from 0.64 m to 0.6 m, and the total mean absolute error (MAE) from 31.3 to 23.5. Simultaneously, predict the falling points of jet trajectory at different distances under the influence of random wind, to demonstrate the feasibility of the proposed method in practical applications. The experimental results show that the system and method proposed in this article can effectively improve the influence of random wind on the falling points of a jet trajectory. In summary, the image acquisition system and error prediction method proposed in this article have many potential applications in fire extinguishing.

Firefighting Water Jet Trajectory Detection from Unmanned Aerial Vehicle Imagery Using Learnable Prompt Vectors.

This research presents an innovative methodology aimed at monitoring jet trajectory during the jetting process using imagery captured by unmanned aerial vehicles (UAVs). This approach seamlessly integrates UAV imagery with an offline learnable prompt vector module (OPVM) to enhance trajectory monitoring accuracy and stability. By leveraging a high-resolution camera mounted on a UAV, image enhancement is proposed to solve the problem of geometric and photometric distortion in jet trajectory images, and the Faster R-CNN network is deployed to detect objects within the images and precisely identify the jet trajectory within the video stream. Subsequently, the offline learnable prompt vector module is incorporated to further refine trajectory predictions, thereby improving monitoring accuracy and stability. In particular, the offline learnable prompt vector module not only learns the visual characteristics of jet trajectory but also incorporates their textual features, thus adopting a bimodal approach to trajectory analysis. Additionally, OPVM is trained offline, thereby minimizing additional memory and computational resource requirements. Experimental findings underscore the method's remarkable precision of 95.4% and efficiency in monitoring jet trajectory, thereby laying a solid foundation for advancements in trajectory detection and tracking. This methodology holds significant potential for application in firefighting systems and industrial processes, offering a robust framework to address dynamic trajectory monitoring challenges and augment computer vision capabilities in practical scenarios.

Efficacy and Safety of Triple Therapy with SGLT-2 Inhibitor, DPP-4 Inhibitor, and Metformin in Type 2 Diabetes: A Meta-Analysis.

Objective: Type 2 diabetes poses significant pain, economic burden, and health risks. This meta-analysis evaluates the efficacy and safety of triple therapy with SGLT-2 inhibitor add-on to DPP-4 inhibitor plus metformin for type 2 diabetes treatment. Methods: A comprehensive search was conducted in PubMed, Embase, and the Cochrane Library to identify randomized controlled trials evaluating the efficacy and safety of triple therapy (SGLT-2 inhibitor + DPP-4 inhibitor + metformin) compared to dual therapy (DPP-4 inhibitor + metformin) in type 2 diabetes. The search covered the period from inception to December 2018. Two reviewers independently screened the literature, extracted data, and assessed study quality. Meta-analysis was performed using RevMan 5.3 software. Results: A total of eight randomized controlled trials were included in this meta-analysis. The results showed that compared to dual therapy with DPP-4 inhibitor add-on to metformin, triple therapy with SGLT-2 inhibitor add-on to DPP-4 inhibitor plus metformin was associated with greater reductions in HbA1c, fasting blood glucose, postprandial blood glucose, body weight, and blood pressure (P < .05). However, the risk of genital tract infection was higher in the triple therapy group (OR = 4.43, 95% CI (2.26, 8.70), P < .0001), while there were no statistically significant differences in the incidence of adverse events, hypoglycemia episodes, urinary tract infection, and fractures between the two groups (P > .05). Conclusions: Based on current evidence, triple therapy was found to significantly improve blood glucose, body weight, and blood pressure when compared to dual therapy. Safety indicators did not show significant differences, except for an increased risk of genital tract infection.

De novo assembly of a fruit transcriptome set identifies AmMYB10 as a key regulator of anthocyanin biosynthesis in Aronia melanocarpa.

Aronia is a group of deciduous fruiting shrubs, of the Rosaceae family, native to eastern North America. Interest in Aronia has increased because of the high levels of dietary antioxidants in Aronia fruits. Using Illumina RNA-seq transcriptome analysis, this study investigates the molecular mechanisms of polyphenol biosynthesis during Aronia fruit development. Six A. melanocarpa (diploid) accessions were collected at four fruit developmental stages. De novo assembly was performed with 341 million clean reads from 24 samples and assembled into 90,008 transcripts with an average length of 801 bp. The transcriptome had 96.1% complete according to Benchmarking Universal Single-Copy Orthologs (BUSCOs). The differentially expressed genes (DEGs) were identified in flavonoid biosynthetic and metabolic processes, pigment biosynthesis, carbohydrate metabolic processes, and polysaccharide metabolic processes based on significant Gene Ontology (GO) biological terms. The expression of ten anthocyanin biosynthetic genes showed significant up-regulation during fruit development according to the transcriptomic data, which was further confirmed using qRT-PCR expression analyses. Additionally, transcription factor genes were identified among the DEGs. Using a transient expression assay, we confirmed that AmMYB10 induces anthocyanin biosynthesis. The de novo transcriptome data provides a valuable resource for the understanding the molecular mechanisms of fruit anthocyanin biosynthesis in Aronia and species of the Rosaceae family.

Efficient synthesis of isoindolones by intramolecular cyclisation of pyridinylbenzoic acids.

A straightforward one-pot method for the synthesis of unreported pyrido-[2,1-a]isoindolones in excellent yield is described. Two novel isoindolones were synthesized and fully characterized. The alkyl substituents on the pyridine play an important role in the outcome of the reaction. The mechanism, investigated through DFT calculations, features an unprecendented intramolecular cyclization reaction involving a carboxylic acid activated by tosyl chloride and an electron-poor pyridinic nitrogen. This protocol completes the known strategies to obtain functionalized isoindolones.

A bamboo leaf-specific aquaporin gene PePIP2;7 is involved in abiotic stress response.

KEY MESSAGE: PePIP2;7, a leaf-specific aquaporin gene in bamboo, is upregulated under abiotic stresses. Overexpressing PePIP2;7 confers abiotic stresses tolerance in transgenic Arabidopsis plant and yeast. Aquaporins (AQPs) participate in the regulation of water balance in plants. However, the function of AQPs in bamboo remains unclear. Here, PePIP2;7 was identified as a leaf-specific aquaporin gene in moso bamboo based on the expression analysis of transcriptome data and PCR. In situ hybridization further indicated that PePIP2;7 was mainly expressed in mesophyll cells of mature leaves, while in immature leaves it was dominant in blade edge cells followed by mesophyll cells. Interestingly, PePIP2;7 was strongly expressed in the mesophyll cells near bulliform cells of immature leaves, suggesting that PePIP2;7 might function in water transport and contribute to leaf unfolding. The transient expression assay showed that PePIP2;7 was a plasma membrane intrinsic protein. Furthermore, PePIP2;7 was upregulated under abiotic stresses such as high light, drought, and NaCl. Compared with Col-0, transgenic Arabidopsis plants overexpressing PePIP2;7 had better seed germination rate, longer taproot length, higher SOD activity, and lower MDA content under abiotic stresses. Besides, yeasts expressing PePIP2;7 also had higher tolerance to stress compared to the control. Taken together, our results show that PePIP2;7 is leaf-specific and involved in stress response, which provides new insights into aquaporin function in bamboo.

Activation of ACS7 in Arabidopsis affects vascular development and demonstrates a link between ethylene synthesis and cambial activity.

Ethylene is a gaseous hormone that affects many processes of plant growth and development. During vascular development, ethylene positively regulates cambial cell division in parallel with tracheary element differentiation inhibitory factor (TDIF) peptide signaling. In this study, we identified an ethylene overproducing mutant, acs7-d, exhibiting enhanced cambial activity and reduced wall development in fiber cells. Using genetic analysis, we found that ethylene signaling is necessary for the phenotypes of enhanced cambial cell division as well as defects in stem elongation and fiber cell wall development. Further, the cambial cell proliferation phenotype of acs7-d depends on WOX4, indicating that the two parallel pathways, ethylene and TDIF signaling, converge at WOX4 in regulating cambium activity. Gene expression analysis showed that ethylene impedes fiber cell wall biosynthesis through a conserved hierarchical transcriptional regulation. These results advance our understanding of the molecular mechanisms of ethylene in regulating vascular meristem activity.

Genome-wide analysis of laccase genes in moso bamboo highlights PeLAC10 involved in lignin biosynthesis and in response to abiotic stresses.

KEY MESSAGE: Twenty-three PeLACs have been identified in moso bamboo, overexpression of PeLAC10 increases the lignin content and confers drought and phenolic acid tolerance in transgenic Arabidopsis. Laccases (LACs) have multifunction involved in the processes of cell elongation, lignification and stress response in plants. However, the function of laccases in bamboo remain unclear. Here, a total of 23 laccase genes (PeLAC1-PeLAC23) were identified in moso bamboo (Phyllostachys edulis). The diverse gene structure and expression pattern of PeLACs suggested that their function should be spatiotemporal and complicated, which was supported by the expression profiles in different tissues of moso bamboo. Eighteen PeLACs were identified as the targets of ped-miR397. The putative ped-miR397-binding site in the coding region of PeLAC10 was further confirmed by RLM-5' RACE, indicating that PeLAC10 was regulated by ped-miR397 after transcription. With the increasing shoot height, the expression abundance of PeLAC10 was up-regulated and reached the maximum in 15 cm shoots, while that of ped-miR397 was relative lower and showed the minimum in 15 cm shoots. PeLAC10 was up-regulated obviously under both ABA (100 µmol L-1) and NaCl (400 mmol L-1) treatments, and it was down-regulated under the GA3 (100 µmol L-1) treatment. The transgenic Arabidopsis plants over-expressing PeLAC10 became slightly smaller and their petioles were shorter than those of Col-0. However, they had a stronger capacity in resistance to phenolic acids and drought besides higher lignin content in stems. These results indicated that overexpression of PeLAC10 was helpful to increase the content of lignin in transgenic Arabidopsis and improve the adaptability to phenolic acid and drought stresses.

NaCl-responsive ROS scavenging and energy supply in alkaligrass callus revealed from proteomic analysis.

BACKGROUND: Salinity has obvious effects on plant growth and crop productivity. The salinity-responsive mechanisms have been well-studied in differentiated organs (e.g., leaves, roots and stems), but not in unorganized cells such as callus. High-throughput quantitative proteomics approaches have been used to investigate callus development, somatic embryogenesis, organogenesis, and stress response in numbers of plant species. However, they have not been applied to callus from monocotyledonous halophyte alkaligrass (Puccinellia tenuifora). RESULTS: The alkaligrass callus growth, viability and membrane integrity were perturbed by 50 mM and 150 mM NaCl treatments. Callus cells accumulated the proline, soluble sugar and glycine betaine for the maintenance of osmotic homeostasis. Importantly, the activities of ROS scavenging enzymes (e.g., SOD, APX, POD, GPX, MDHAR and GR) and antioxidants (e.g., ASA, DHA and GSH) were induced by salinity. The abundance patterns of 55 salt-responsive proteins indicate that salt signal transduction, cytoskeleton, ROS scavenging, energy supply, gene expression, protein synthesis and processing, as well as other basic metabolic processes were altered in callus to cope with the stress. CONCLUSIONS: The undifferentiated callus exhibited unique salinity-responsive mechanisms for ROS scavenging and energy supply. Activation of the POD pathway and AsA-GSH cycle was universal in callus and differentiated organs, but salinity-induced SOD pathway and salinity-reduced CAT pathway in callus were different from those in leaves and roots. To cope with salinity, callus mainly relied on glycolysis, but not the TCA cycle, for energy supply.

Genome-wide identification and expression analysis of brassinosteroid action-related genes during the shoot growth of moso bamboo.

Brassinosteroids (BRs) are a group of plant steroid hormones that play crucial roles in a range of plant growth and development processes. BR action includes active BR formation by a complex biosynthesis process and driving BR biological function through signal transduction. Although the characterization of several BR action-related genes has been conducted in a few model plants, systematic information about these genes in bamboo is still lacking. We identified 64 genes related to BR action from the genome of moso bamboo (Phyllostachys edulis), including twenty that participated in BR biosynthesis and forty-four involved in BR signal transduction. The characteristics of all these candidate genes were identified by bioinformatics methods, including the gene structures, basic physical and chemical properties of proteins, conserved domains and evolutionary relationships. Based on the transcriptome data, the candidate genes demonstrated different expression patterns, which were further validated by qRT-PCR using templates from bamboo shoots with different heights. Thirty-four positive and three negative co-expression modules were identified by 44 candidate genes in the newly emerging bamboo shoot. The gene expression patterns and co-expression modules of BR action-related genes in bamboo shoots indicated that they might function to promote bamboo growth through BR biosynthesis and signal transduction processes. This study provides the first step towards the cloning and functional dissection of the role of BR action-related genes in moso bamboo, which also presents an excellent opportunity for genetic engineering using the candidate genes to improve bamboo quantity and quality.

Expression and functional analysis of two PsbS genes in bamboo (Phyllostachys edulis).

Higher plants have an array of photoprotection mechanisms alleviating the harmful effects of light. Non-photochemical quenching (NPQ) is one of the photoprotective mechanisms, which dissipates the excess of light energy absorbed in the light-harvesting complexes (LHCs) into thermal energy. The photosystem II subunit S (PsbS), a member of the LHC family thought to be present exclusively in higher plants, is supposed to activate NPQ through interactions with antenna proteins. However, the roles of PsbS in bamboo remain unclear. Here, two genes of bamboo (Phyllostachys edulis), PePsbS1 and PePsbS2, are investigated and functionally analyzed. PePsbS1 and PePsbS2 have a similar gene structure with three introns separated by two exons, which encode 269 and 268 amino acid residues, respectively. Tissue-specific analysis showed that PePsbS1 and PePsbS2 are highly expressed in leaf blade. Besides, they are both upregulated in the leaf blade when plantlets are submitted to an increased and prolonged light intensity, suggesting that they are light-induced. Western blot analysis indicated that the accumulation level of total PePsbSs is consistent with what obtained by quantitative real-time polymerase chain reaction for PePsbS1 and PePsbS2. Transgenic Arabidopsis plants overexpressing PePsbS1 and PePsbS2 both displayed an enhanced photoprotection. Moreover, the expression of PePsbS1 and PePsbS2 could both rescue the NPQ of Arabidopsis npq4 mutant, indicating that the PsbSs are functionally conserved between monocots and dicots. These results indicated that both PePsbS1 and PePsbS2 could circumvent photoinhibition and enhance photoprotection, which are key factors for bamboo's adaptation to different light environment.

Carvedilol Ameliorates Intrahepatic Angiogenesis, Sinusoidal Remodeling and Portal Pressure in Cirrhotic Rats.

BACKGROUND Carvedilol is the first-line drug for the primary prophylaxis of variceal bleeding due to portal hypertension (PHT) in liver cirrhosis. This study aimed to investigate the effects of carvedilol on intrahepatic angiogenesis and sinusoidal remodeling in cirrhotic rats and explore the underlying mechanisms of carvedilol in PHT. MATERIAL AND METHODS For in vivo experiments, carbon tetrachloride was used to induce liver cirrhosis in rats, and carvedilol was simultaneously administered by gavage. The portal pressure was measured in rats, and liver tissues were examined by immunohistochemistry. Sinusoidal remodeling was observed by transmission electron microscopy. For in vitro experiments, the effects of carvedilol on fibronectin (FN) synthesis in human umbilical vein endothelial cells (HUVECs) were explored by quantitative real-time polymerase chain reaction and western blot analysis. RESULTS Portal vein pressure measurements showed that carvedilol reduced portal pressure in cirrhotic rats. Immunohistochemistry assays indicated that carvedilol ameliorated intrahepatic angiogenesis. Transmission electron microscopy examination demonstrated that carvedilol improved sinusoidal remodeling. In the in vitro experiments, carvedilol suppressed transforming growth factor ß1 (TGFß1)-induced FN synthesis in HUVECs by inhibition of the TGFß1/Smads pathway. CONCLUSIONS Carvedilol ameliorated intrahepatic angiogenesis, sinusoidal remodeling and portal pressure in cirrhotic rats. Carvedilol improved sinusoidal remodeling by suppressing FN synthesis in endothelial cells. Carvedilol has potential utility for treating early-stage liver cirrhosis.

Early thawing after snow removal and no straw mulching accelerates organic carbon cycling in a paddy soil in Northeast China.

Variations in soil organic carbon (SOC) have implications for atmospheric CO2 concentrations and the greenhouse effect. However, the effects of snow cover and straw mulching on the variations in SOC fractions across winter remain largely unknown. In this study, soil samples were collected during different stages of winter from an in situ experiment comprising three treatments: 1) snow removal with no straw mulching (Sn-SM-); 2) snow cover with no straw mulching (SC), and; 3) snow cover with straw mulching (SC + SM+). Results showed that labile organic carbon, semi-labile organic carbon, recalcitrant organic carbon (ROC), the light fraction of organic carbon (LFOC), and easily oxidized organic carbon (EOC) contents did not vary significantly (P > .05) during the unfrozen to hard frost stages. Compared to the unfrozen stage, microbial biomass carbon (MBC) contents decreased by 519.03 mg kg-1, 325.21 mg kg-1, and 244.09 mg kg-1 and dissolved organic carbon (DOC) contents increased by 473.36 mg kg-1, 348.10 mg kg-1, and 258.89 mg kg-1 at the hard frost stage in Sn-SM-, SC, and SC + SM + treatments, respectively. Throughout all thawing stages, > 61% and 59% of SOC and ROC accumulation, respectively in the three treatments were observed in thawing stage II, indicating that higher temperatures and microbial activities in thawing stage II accelerated the inputs of SOC and ROC. ROC accumulation accounted for >65% of the SOC accumulation and the proportions of ROC in SOC increased in the three treatments during the thawing stages. SC + SM + treatment maintained lower EOC contents during thawing stages than other treatments. The observation of lowest SOC and LFOC accumulation and contents in the SC + SM + treatment during thawing stages showed that SC + SM + experienced the least inputs of SOC in the soil.

The bamboo aquaporin gene PeTIP4;1-1 confers drought and salinity tolerance in transgenic Arabidopsis.

KEY MESSAGE: PeTIP4;1-1, an aquaporin gene involved in bamboo shoot growth, is regulated by abiotic stresses. Overexpression of PeTIP4;1-1 confers drought and salinity tolerance in transgenic Arabidopsis. Aquaporins play a central role in numerous physiological processes throughout plant growth and development. PeTIP4;1-1, an aquaporin gene isolated from moso bamboo (Phyllostachys edulis), comprises an open reading frame (ORF) of 756 bp encoding a peptide of 251 amino acids. The genomic sequence corresponding to the ORF of PeTIP4;1-1 was 1777 bp and contained three exons separated by two introns. PeTIP4;1-1 was constitutively expressed at the highest level in culms, and the expression level was elevated with increasing height of the bamboo shoot. PeTIP4;1-1 was significantly up-regulated in response to drought and salinity stresses in bamboo roots and leaves. To investigate the role of PeTIP4;1-1 in response to drought and salinity stresses, transgenic Arabidopsis plants overexpressing PeTIP4;1-1 under the control of CaMV 35S promoter were generated and subjected to morphological and physiological assays. Compared with Col-0, the transgenic plants showed enhanced tolerance to drought and salinity stresses and produced longer taproots, which had more green leaves, higher F v/F m and NPQ values, higher activities of SOD, POD and CAT, lower MDA concentration and higher water content. Transcript levels of three stress-related genes (AtP5CS, AtNHX1 and AtLEA) were enhanced. These results indicated that PeTIP4;1-1 might play an important function in response to drought and salinity stresses, and is a candidate gene for breeding of stress tolerance in other crops through genetic engineering.

Efficacy of dupilumab plus topical corticosteroids in children with atopic dermatitis: A meta-analysis of randomized controlled trials.

INTRODUCTION: Children with atopic dermatitis (AD) bear a significant burden of illness that adversely affects their quality of life. OBJECTIVE: To determine the efficacy of dupilumab and topical corticosteroids for the treatment of pediatric AD. METHODS: A comprehensive literature search was conducted using three prominent databases: Web of Science, PubMed, and Embase. Using a fixed-effects or random-effects model, the standard mean difference or risk ratios with 95% confidence intervals were calculated, and the trial protocol was listed as CRD42023408546. RESULTS: A total of 3 studies were included, and 896 participants met the inclusion criteria. The combined estimate showed that dupilumab plus topical corticosteroids had numerically greater efficacy in terms of Eczema Area and Severity Index (EASI)-50, EASI-75, EASI-90, and Investigator Global Assessment (IGA) score of 0 or 1. Children who received topical corticosteroids and dupilumab achieved significantly higher Children's Dermatological Life Quality Index scores compared to those who received placebo. The number of individuals who achieved IGA 0/1 increased with the use of dupilumab and topical corticosteroids. CONCLUSIONS: Dupilumab and topical corticosteroids can be used to treat symptoms in children with AD. However, given the substantial variation in treatment outcomes among studies, the findings should be interpreted with caution.

Grid-plainification enables medium-temperature PbSe thermoelectrics to cool better than Bi2Te3.

Thermoelectric cooling technology has important applications for processes such as precise temperature control in intelligent electronics. The bismuth telluride (Bi2Te3)-based coolers currently in use are limited by the scarcity of Te and less-than-ideal cooling capability. We demonstrate how removing lattice vacancies through a grid-design strategy switched PbSe from being useful as a medium-temperature power generator to a thermoelectric cooler. At room temperature, the seven-pair device based on n-type PbSe and p-type SnSe produced a maximum cooling temperature difference of ~73 kelvin, with a single-leg power generation efficiency approaching 11.2%. We attribute our results to a power factor of >52 microwatts per centimeter per square kelvin, which was achieved by boosting carrier mobility. Our demonstration suggests a path for commercial applications of thermoelectric cooling based on Earth-abundant Te-free selenide-based compounds.

In Planta Gene Expression and Gene Editing in Moso Bamboo Leaves.

A novel in planta gene transformation method was developed for bamboo, which avoids the need for time-consuming and labor-intensive callus induction and regeneration processes. This method involves Agrobacterium-mediated gene expression via wounding and vacuum for bamboo seedlings. It successfully demonstrated the expression of exogenous genes, such as the RUBY reporter and Cas9 gene, in bamboo leaves. The highest transformation efficiency for the accumulation of betalain in RUBY seedlings was achieved using the GV3101 strain, with a percentage of 85.2% after infection. Although the foreign DNA did not integrate into the bamboo genome, the method was efficient in expressing the exogenous genes. Furthermore, a gene editing system has also been developed with a native reporter using this method, from which an in situ mutant generated by the edited bamboo violaxanthin de-epoxidase gene (PeVDE) in bamboo leaves, with a mutation rate of 17.33%. The mutation of PeVDE resulted in decreased non-photochemical quenching (NPQ) values under high light, which can be accurately detected by a fluorometer. This makes the edited PeVDE a potential native reporter for both exogenous and endogenous genes in bamboo. With the reporter of PeVDE, a cinnamoyl-CoA reductase gene was successfully edited with a mutation rate of 8.3%. This operation avoids the process of tissue culture or callus induction, which is quick and efficient for expressing exogenous genes and endogenous gene editing in bamboo. This method can improve the efficiency of gene function verification and will help reveal the molecular mechanisms of key metabolic pathways in bamboo.

A new biotechnology for in-planta gene editing and its application in promoting flavonoid biosynthesis in bamboo leaves.

BACKGROUND: Bamboo is a perennial and renewable biomass forest resource and its leaf flavonoid is an antioxidant for biological and pharmacological research. The established genetic transformation and gene editing systems in bamboo are significantly limited by the dependence on bamboo regeneration capability. The way to improve the flavonoid content in bamboo leaves through biotechnology is still not feasible. RESULTS: Here, we developed an in-planta, Agrobacterium-mediated gene expression method for exogenous genes via wounding and vacuum in bamboo. We demonstrated that the RUBY served as a reporter efficiently expressed in bamboo leaves and shoots, albeit unable to integrate into the chromosome. We have also developed a gene editing system by creating an in situ mutant of the bamboo violaxanthin de-epoxidase (PeVDE) gene in bamboo leaves, with lower NPQ values under the fluorometer, which can serve as a native reporter for gene editing. Furthermore, the bamboo leaves with increased flavonoid content were achieved by knocking out the cinnamoyl-CoA reductase genes. CONCLUSIONS: Our method can be applied for the functional characterization of novel genes in a short time and is helpful for bamboo leaf flavonoid biotechnology breeding in the future.