Root architecture is one of the most important agronomic traits that determines rice crop yield. The primary root (PR) absorbs mineral nutrients and provides mechanical support; however, the molecular mechanisms of PR elongation remain unclear in rice. Here, the two loss-of-function T-DNA insertion mutants of root length regulator 4 (OsRLR4), osrlr4-1 and osrlr4-2 with longer PR, and three OsRLR4 overexpression lines, OE-OsRLR4-1/-2/-3 with shorter PR compared to the wild type/Hwayoung (WT/HY), were identified. OsRLR4 is one of five members of the PRAF subfamily of the regulator chromosome condensation 1 (RCC1) family. Phylogenetic analysis of OsRLR4 from wild and cultivated rice indicated that it is under selective sweeps, suggesting its potential role in domestication. OsRLR4 controls PR development by regulating auxin accumulation in the PR tip and thus the root apical meristem activity. A series of biochemical and genetic analyses demonstrated that OsRLR4 functions directly upstream of the auxin transporter OsAUX1. Moreover, OsRLR4 interacts with the TRITHORAX-like protein OsTrx1 to promote H3K4me3 deposition at the OsAUX1 promoter, thus altering its transcription level. This work provides insight into the cooperation of auxin and epigenetic modifications in regulating root architecture and provides a genetic resource for plant architecture breeding.

Additional file 3: Table S1. All DEGs in osftsh2 mutants were listed.

Background Filamentation temperature-sensitive H (FtsH) is an ATP-dependent zinc metalloprotease with ATPase activity, proteolysis activity and molecular chaperone-like activity. For now, a total of nine FtsH proteins have been encoded in rice, but their functions have not revealed in detail. In order to investigate the molecular mechanism of OsFtsH2 here, several osftsh2 knockout mutants were successfully generated by the CRISPR/Cas9 gene editing technology. Results All the mutants exhibited a phenotype of striking albino leaf and could not survive through the stage of three leaves. OsFtsH2 was located in the chloroplast and preferentially expressed in green tissues. In addition, osftsh2 mutants could not form normal chloroplasts and had lost photosynthetic autotrophic capacity. RNA sequencing analysis indicated that many biological processes such as photosynthesis-related pathways and plant hormone signal transduction were significantly affected in osftsh2 mutants. Conclusions Overall, the results suggested OsFtsH2 to be essential for chloroplast development in rice.

The chloroplast NAD(P)H dehydrogenase (NDH) complex, as one of the most important photosynthesis protein complexes in thylakoid membrane, is involved in photosystem I (PSI) cyclic electron transport (CEF).Under abiotic environmental stress, the photosynthetic apparatus is susceptible to the damage caused by the strong light illumination.However, the enhancement of NDHdependent CEF could facilitate the alleviation of the damage to the photosynthetic apparatus.The NdhB subunit encoded by chloroplast genome is one of most important subunits of NDH complex and consists of 510 amino acids.Here, according to cloning ndhB from Melrose (cultivated soybean), ACC547 (wild salt-tolerant soybean), S113-6 and S111-9 (hybrid descendant), based on the comparison and analysis of the sequences of NdhB subunits, we found that there is a novel thylakoid transit peptide of NdhB subunit in S111-9.In addition, crosslink immunoprecipitation, immunogold labeling and co-expression of GFP fusion protein indicated that the novel thylakoid transit peptide is favorable to the expression and localization of NdhB subunit in chloroplast.Therefore, we suggest that this novel thylakoid transit peptide plays the same role as chaperonin and contributes to facilitating the expression and localization of NdhB subunit.

By designing and conducting a series of temperature-controllable experiments in greenhouse, this work emphatically investigated how the positive or negative allelochemical effects of a key metabolic compound 3,4dihydroxyacetophenone (DHAP) from the needles of Picea schrenkiana would vary with environmental temperature. It was found that the alleochemical effect of DHAP on the germination, seedling growth, root development, endogenous plant hormone content and antioxidant enzyme activity of P. schrenkiana demonstrated an orientation duality identified by the existence of a compensation concentration at a certain temperature, which set up a boundary between promotional and inhibitory effect. As temperature increased from 10/2 degrees C up to 12/4, 14/6, 16/8, and 18/10 degrees C in turn, the compensation point of DHAP concentration would correspondingly shift from 0.32mM down to 0.28, 0.24, 0.21 and 0.19mM, respectively. Overall, this work will deepen and enrich the understandings to the uncertainty of allelochemical effects on P. schrenkiana regeneration due to climate warming, and provide some helpful suggestions to develop a scientific management strategy for boreal forest.

In the plant, plasmodesmata (PDs) permit direct cytoplasmic connections and facilitate the local molecular exchange of neighboring cells. Cell-to-cell communications through PD play critical roles in the small RNAs mobile, transport of metabolites, cell fate specification and development. The trafficking of PD is under tight control and it is also constantly fine-tuned to fit in the requirements of cell-to-cell communication. However, how PD involved in the transition from the dormant to the germination of buds is not yet clearly known. Here, we provide experimental evidence that PD formation and permeability are correlated to the germination of buds in North-American holly. Moreover, the increased PD number and permeability in the germinating buds are associated with the down-regulated SA signaling, which also facilitates starch degradation. Altogether, our study stresses the fundamental role of PD in governing the transition from dormant to germination of floral buds, unveiling the regulation of PD during the flowering of North-American holly. Understanding the mechanism of SA-mediated PD regulation during the flowering will provide an effective way to enhance the improvement of agronomical tree traits by the application of PD engineering.

The chaperonin 60 (Cpn60) protein is of great importance to plants due to its involvement in modulating the folding of numerous chloroplast protein polypeptides. In chloroplasts, Cpn60 is differentiated into two subunit types—Cpn60α and Cpn60β and the rice genome encodes three α and three β plastid chaperonin subunits. However, the functions of Cpn60 family members in rice were poorly understood. In order to investigate the molecular mechanism of OsCpn60β1, we attempted to disrupt the OsCpn60β1 gene by CRISPR/Cas9-mediated targeted mutagenesis in this study. We succeeded in the production of homozygous OsCpn60β1 knockout rice plants. The OsCpn60β1 mutant displayed a striking albino leaf phenotype and was seedling lethal. Electron microscopy observation demonstrated that chloroplasts were severely disrupted in the OsCpn60β1 mutant. In addition, OsCpn60β1 was located in the chloroplast and OsCpn60β1 is constitutively expressed in various tissues particularly in the green tissues. The label-free qualitative proteomics showed that photosynthesis-related pathways and ribosomal pathways were significantly inhibited in OsCpn60β1 mutants. These results indicate that OsCpn60β1 is essential for chloroplast development in rice.

SummaryThe rapid selection of salinity‐tolerant crops to increase food production in salinized lands is important for sustainable agriculture. Recently, high‐throughput plant phenotyping technologies have been adopted that use plant morphological and physiological measurements in a non‐destructive manner to accelerate plant breeding processes. Here, a hyperspectral imaging (HSI) technique was implemented to monitor the plant phenotypes of 13 okra (Abelmoschus esculentus L.) genotypes after 2 and 7 days of salt treatment. Physiological and biochemical traits, such as fresh weight, SPAD, elemental contents and photosynthesis‐related parameters, which require laborious, time‐consuming measurements, were also investigated. Traditional laboratory‐based methods indicated the diverse performance levels of different okra genotypes in response to salinity stress. We introduced improved plant and leaf segmentation approaches to RGB images extracted from HSI imaging based on deep learning. The state‐of‐the‐art performance of the deep‐learning approach for segmentation resulted in an intersection over union score of 0.94 for plant segmentation and a symmetric best dice score of 85.4 for leaf segmentation. Moreover, deleterious effects of salinity affected the physiological and biochemical processes of okra, which resulted in substantial changes in the spectral information. Four sample predictions were constructed based on the spectral data, with correlation coefficients of 0.835, 0.704, 0.609 and 0.588 for SPAD, sodium concentration, photosynthetic rate and transpiration rate, respectively. The results confirmed the usefulness of high‐throughput phenotyping for studying plant salinity stress using a combination of HSI and deep‐learning approaches.

Global warming in conjunction with various biotic or abiotic interferences has been jeopardizing the ecosystem of boreal forests. By integrating field inspection with experimental simulation, this work comprehensively investigated the allelopathic effects of a key allelochemical 3,4-dihydroxyacetophenone (DHAP) in the exudates of P. schrenkiana needles on its seed and seedling growth, endogenous hormone metabolism and antioxidant enzyme activity, identified the existence of DHAP allelopathy duality at a certain temperature with an inflection concentration point (e.g. about 0.25 mM at dark/light temperature of 4/12 °C) as the boundary between promotional and inhibitory effect, and verified that the inflection point of DHAP concentration would inevitably shift to a lower level as temperature increased. Consequently, this paper gives a scientific explanation into the intrinsic mechanism of P. schrenkiana degradation due to allelopathy, but also presents a new approach to explore the relationship between forest evolution and global warming. Highlight A quantitative description on the duality of 3, 4-dihydroxyacetophenone (DHAP) as a promoter or an inhibitor to affect the seed germination, seedling growth and root development of P. schrenkiana , as well as the antioxidant enzyme activities and hormone contents. The new findings of DHAP inflection concentration as boundary to divide the promotional and inhibitory effect of allelopathy which would decrease as environment temperatures rise. An explanation into the intrinsic mechanism of P. schrenkiana degradation due to allelopathy, and a new approach to explore the relationship between forest evolution and global warming. Abbreviations : DHAP : 3, 4-dihydroxyacetophenone FDA : fluorescein diacetate PI : propidium iodide ZT : Zeatin GA3 : Gibberellin IAA : Indoleacetic acid ABA : Abscisic acid SOD : Speroxide dismutase POD : Peroxidase CAT : Catalase GR : Glutathione reductase

Stomata control gas exchange and water transpiration and are one of the most important physiological apparatuses in higher plants. The regulation of stomatal aperture is closely coordinated with photosynthesis, nutrient uptake, plant growth, development, and so on. With advances in scanning electron microscopy (SEM), high-resolution images of plant stomata and cell surfaces can be obtained from detached plant tissues. However, this method does not allow for rapid analysis of the dynamic variation of plant stomata and cell surfaces in situ under nondestructive conditions. In this study, we demonstrated a novel plant surface impression technique (PSIT, Silagum-Light as correction impression material based on A-silicones for all two-phase impression techniques) that allows for precise analysis of plant stomata aperture and cell surfaces. Using this method, we successfully monitored the dynamic variation of stomata and observed the nanoscale microstructure of soybean leaf trichomes and dragonfly wings. Additionally, compared with the analytical precision and the time used for preparing the observation samples between PSIT and traditional SEM, the results suggested that the analytical precision of PSIT was the same to traditional SEM, but the PSIT was more easy to operate. Thus, our results indicated that PSIT can be widely applied to the plant science field.

The major problem for industrial application of bioflocculants is its high production cost. Here, a novel bacterium Pseudomonas boreopolis G22, which can secret a cellulase-free xylanase and simultaneously produce bioflocculants (MBF-G22) through directly converting untreated biomass, was isolated. The bioflocculants' production of G22 was closely related to its xylanase activity, hydrolysis ability of biomass and the hemicellulose loss caused by G22. The optimal fermentation conditions with the highest bioflocculants' yield (3.75 mg g(-1) dry biomass) were obtained at the fermentation time of 96 h, incubation temperature of 30 degrees C, inoculum concentration of 1.0% and biomass concentration of 1.0% in an initial pH value of 7.0. MBF-G22 mainly consisted of polysaccharides (63.3%) with a molecular weight of 3.982x10(6) Da and showed the highest flocculating efficiency of 97.1% at a dosage of 3.5 mg L-1. In addition, MBF-G22 showed high flocculating efficiency of microalgae (95.7%) at a dosage of 80 mg L-1.

The yellow stripe-like (YSL) family of transporters mediates the uptake, translocation, and distribution of various mineral elements in vivo by transferring metal ions chelated with phytosiderophore or nicotianamine (NA). However, little is known about the roles of the YSL genes against cadmium in planta. In this study, we first cloned and characterized a vital member of the YSL gene family, MsYSL1, from the bioenergy plant Miscanthus sacchariflorus. MsYSL1 localized in the plasma membrane and was widely expressed throughout the whole seedling with the highest expression level in the stem. In addition, its expression in the root was stimulated by excess manganese (Mn), cadmium (Cd), and lead, and a shortage of iron (Fe), zinc (Zn), and copper. Functional complementation in yeast indicated that MsYSL1 showed transport activity for Fe(II)–NA and Zn–NA, but not for Cd–NA. Although they exhibited no significant differences versus the wild type under normal cultivation conditions, MsYSL1-overexpressing Arabidopsis lines displayed a higher resistance to Cd accompanied by longer root lengths, lower Cd, Zn, and Mn levels in roots, and higher Cd, Fe, and Mn translocation ratios under Cd stress. Moreover, genes related to NA synthesis, metal translocation, long-distance transport, and Cd exclusion were highly induced in transgenic lines under Cd stress. Thus, MsYSL1 may be an essential transporter for diverse metal–NAs to participate in the Cd detoxification by mediating the reallocation of other metal ions.

Xylan removal by bacterial pretreatments has been confirmed to increase the digestibility of biomass. Here, an effective xylan removal technique has been developed to enhance the digestibility of wheat straw and simultaneously produce bioflocculants by a cellulase-free xylanase-secreting strain, Pseudomonas boreopolis G22.

Extensive research revealed tremendous details about how plants sense pathogen effectors during effector-triggered immunity (ETI). However, less is known about downstream signaling events. In this report, we demonstrate that prolonged activation of MPK3 and MPK6, two Arabidopsis pathogen-responsive mitogen-activated protein kinases (MPKs), is essential to ETI mediated by both coiled coil-nucleotide binding site-leucine rich repeats (CNLs) and toll/interleukin-1 receptor nucleotide binding site-leucine rich repeats (TNLs) types of R proteins. MPK3/MPK6 activation rapidly alters the expression of photosynthesis-related genes and inhibits photosynthesis, which promotes the accumulation of superoxide ([Formula: see text]) and hydrogen peroxide (H2O2), two major reactive oxygen species (ROS), in chloroplasts under light. In the chemical-genetically rescued mpk3 mpk6 double mutants, ETI-induced photosynthetic inhibition and chloroplastic ROS accumulation are compromised, which correlates with delayed hypersensitive response (HR) cell death and compromised resistance. Furthermore, protection of chloroplasts by expressing a plastid-targeted cyanobacterial flavodoxin (pFLD) delays photosynthetic inhibition and compromises ETI. Collectively, this study highlights a critical role of MPK3/MPK6 in manipulating plant photosynthetic activities to promote ROS accumulation in chloroplasts and HR cell death, which contributes to the robustness of ETI. Furthermore, the dual functionality of MPK3/MPK6 cascade in promoting defense and inhibiting photosynthesis potentially allow it to orchestrate the trade-off between plant growth and defense in plant immunity.

In rice, there are five members of the auxin carrier AUXIN1/LIKE AUX1 family; however, the biological functions of the other four members besides OsAUX1 remain unknown. Here, by using CRISPR/Cas9, we constructed two independent OsAUX3 knock-down lines, osaux3-1 and osaux3-2, in wild-type rice, Hwayoung (WT/HY) and Dongjin (WT/DJ). osaux3-1 and osaux3-2 have shorter primary roots (PRs), decreased lateral root (LR) density, and longer root hairs (RHs) compared with their WT. OsAUX3 expression in PRs, LRs, and RHs further supports that OsAUX3 plays a critical role in the regulation of root development. OsAUX3 locates at the plasma membrane and functions as an auxin influx carrier affecting acropetal auxin transport. OsAUX3 is up-regulated in the root apex under aluminium (Al) stress, and osaux3-2 is insensitive to Al treatments. Furthermore, 1-naphthylacetic acid accented the sensitivity of WT/DJ and osaux3-2 to respond to Al stress. Auxin concentrations, Al contents, and Al-induced reactive oxygen species-mediated damage in osaux3-2 under Al stress are lower than in WT, indicating that OsAUX3 is involved in Al-induced inhibition of root growth. This study uncovers a novel pathway alleviating Al-induced oxidative damage by inhibition of acropetal auxin transport and provides a new option for engineering Al-tolerant rice species.

Laccases from bacteria have been widely studied in the past 2 decades due to the higher growth rate of bacteria and their excellent thermal and alkaline pH stability. In this study, a novel laccase gene was cloned from Bacillus sp., analyzed, and functionally expressed in Escherichia coli. The laccase was highly induced in the E. coli expression system with a maximum intracellular activity of 16 U mg-1 protein. The optimal temperature and pH of the purified laccase were 40°C and 4.6, respectively, when ABTS (2,2'-azino-bis[3-ethylbenzothiazoline-6-sulfonate]) was used as the substrate. The purified laccase showed high stability in the pH range of 3.0-9.0, and retained more than 70% of its activity after 24 h of incubation at 40°C with a pH value of 9.0. Furthermore, the enzyme exhibited extremely high temperature and ion metal tolerance. The half-life of the purified laccase at 70°C was 15.9 h. The purified laccase could efficiently decolorize 3 chemical dyes, especially in the presence of ABTS as a mediator. The high production of this laccase in E. coli and exceptional characteristics of the recombinant enzyme protein make it a promising candidate for industrial applications.

riceXIP , a XIP-Type xylanase inhibitor gene identified in rice, was cloned and expressed in Escherichia coli . Recombinant protein riceXIP was active against xylanase from Aspergillus niger , suggesting correct expression. By using transgenic techniques, we achieved the overexpression of the riceXIP gene and gene knock-down plants and elucidated that riceXIP may likely participate in plant defence against herbivores. The defence-related genes were significantly elevated in riceXIP -overexpressing transgenic plants treated with rice brown planthopper infestation. The full-length promoter (2009 bp, RP1) of riceXIP gene and 1534 (RP2), 1179 (RP3), 891 (RP4), 491 (RP5), 400 (RP6), 214 (RP7) and 112 bp (RP8) 5′ deletion constructs were fused with β-glucuronidase ( GUS ) gene and transgenic rice plants were used to clarify its function. The transgenic lines transformed with promoter fragments of riceXIP differentially responded to methyl jasmonate and wounding stress by quantitative GUS analysis. The 5′ deletion analysis also showed that two repressor elements exist between −1502 and −1147 bp and between −182 and −80 bp, respectively, whereas an enhancer element exists between −1147 and −859 bp. The study strengthens the possibility that riceXIP participates in the resistance against herbivory in rice and provides a helpful insight for understanding the cis-regulation of the riceXIP gene.

Malic acid (MA) plays an important role in the regulation of plant growth, stomatal aperture, nutrition elements homeostasis and toxic metals tolerance. However, little is known about the effects of exogenous MA on physiological and biochemical responses to toxic metals in plants. To measure the alleviation roles of exogenous MA against cadmium (Cd), we determined the effects of MA on plant growth, net photosynthetic rate (Pn), reactive oxygen species (ROS) accumulation and the activities of anti-oxidant enzymes in the leaves of Miscanthus sacchariflorus (M. sacchariflorus) under Cd stress. The Cd exposure alone significantly inhibited plant growth and Pn, but increased the accumulation of ROS even though the anti-oxidant enzymes were markedly activated in the leaves of M. sacchariflorus. Treatment with MA significantly enhanced plant growth and decreased Cd accumulation accompanied by increasing Pn under Cd stress as compared to Cd stress alone, especially when treatment with high concentration of MA (200μM) was used. In addition, Cd and MA indicated synergistic effects by further increasing the activities and genes expression of partial anti-oxidant enzymes, thus resulting in higher glutathione accumulation and reduction of ROS production. The results showed that application of MA alleviated Cd-induced phytotoxicity and oxidant damage through the regulation of both enzymatic and non-enzymatic anti-oxidants under Cd stress in M. sacchariflorus.

Bioflocculation has been developed as a cost-effective and environment-friendly method to harvest multiple microalgae. However, the high production cost of bioflocculants makes it difficult to scale up. In the current study, low-cost bioflocculants were produced from untreated corn stover by a biomass-degrading bacterium Pseudomonas sp. GO2.

The toughness of microalgal cell walls makes lipid extraction and large-scale biodiesel production difficult. This study investigated the enzymatic hydrolysis of algal cell walls, in which the enzymes were produced by eight biomass-degrading bacterial strains. The bacteria were first cultured in mineral salt medium containing 5% (w/v) wheat bran and various lignocellulolytic enzymes, including exoglucanases (CMCase), endoglucanases (FPase), xylanase, and laccase were monitored in order to obtain an enzymatic extract. All the strains showed marked CMCase activity, with a range of 3.0-6.9 U ml(-1) after incubation for 2-5 d. Some strains also produced FPase, xylanase, and laccase. The enzymatic extract was directly added to fresh algae culture at a ratio of 1:3 (v/v) for 48 h. All the bacterial enzymatic extracts significantly disrupted algal cell walls, according to the enhancement of reducing sugar content in the culture. The lipid extraction yield was markedly increased by 10.4-43.9%, depending on the bacteria strains used. Due to its high reducing sugar production and lipid extraction efficiency, Bacillus sp. K1 was selected for a time-course experiment. Maximum lipid yield was obtained after 24 h of incubation at the room temperature, with about 40% of the cells were disrupted. These results showed that enzymes produced by biomass-degrading bacteria can weaken and disrupt cell walls and components of algae and facilitate the release of lipids from algae. (C) 2017 Elsevier Ltd. All rights reserved.