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The mechanisms that balance plant growth and stress responses are poorly understood, but they appear to involve abscisic acid (ABA) signaling mediated by protein kinases. Here, to explore these mechanisms, we examined the responses of Arabidopsis thaliana protein kinase mutants to ABA treatment. We found that mutants of BRASSINOSTEROID INSENSITIVE 1-ASSOCIATED RECEPTOR KINASE 1 (BAK1) were hypersensitive to the effects of ABA on both seed germination and primary root growth. The kinase OPEN STOMATA 1 (OST1) was more highly activated by ABA in bak1 mutant than the wild type. BAK1 was not activated by ABA treatment in the dominant negative mutant abi1-1 or the pyr1 pyl4 pyl5 pyl8 quadruple mutant, but it was more highly activated by this treatment in the abi1-2 abi2-2 hab1-1 loss-of-function triple mutant than the wild type. BAK1 phosphorylates OST1 T146 and inhibits its activity. Genetic analyses suggested that BAK1 acts at or upstream of core components in the ABA signaling pathway, including PYLs, PP2Cs, and SnRK2s, during seed germination and primary root growth. Although the upstream brassinosteroid (BR) signaling components BAK1 and BR INSENSITIVE 1 (BRI1) positively regulate ABA-induced stomatal closure, mutations affecting downstream components of BR signaling, including BRASSINOSTEROID-SIGNALING KINASEs (BSKs) and BRASSINOSTEROID-INSENSITIVE 2 (BIN2), did not affect ABA-mediated stomatal movement. Thus, our study uncovered an important role of BAK1 in negatively regulating ABA signaling during seed germination and primary root growth, but positively modulating ABA-induced stomatal closure, thus optimizing the plant growth under drought stress.

A two-step sequential deposition has been a reliable method to synthesize large-area and high-quality perovskite films due to its better reproducibility. However, the long-term performance and stability of thin films are adversely affected by the slow and incomplete transformation of PbI2 to perovskites. Here, we propose a nanosecond laser shock annealing process to induce ultrafast organic salt diffusion into the PbI2 layer to modulate the crystalline structure, residual tensile strain, and electron transport kinetics in two-step fabricated halide perovskite films. We found that pulse-laser induced ultrafast diffusion reduces the thickness of the residual PbI2 layer at the bottom of the perovskite film fabricated by a two-step method, resulting in the reduction of residual tensile strain by over seven times. The shocking moment of the nanosecond laser promotes the diffusion of the organic salt to the PbI2 layer. Compared to traditional thermal annealing, ultrafast laser shock annealing enhances the molecular interaction, which significantly affects the orbital overlap resulting in band structure changes. Laser shock annealing induced modulation of band structures in perovskites leads to remarkable improvement in their carrier lifetime, producing a responsivity (R) and detectivity (D*) of 2.45 A W−1 and 1.48 × 1012 Jones, respectively. Besides, the stability testing under various harsh thermal and humid thermal conditions shows that laser shock annealing improves the stability of perovskite thin films as a result of the reduced PbI2 layer and residual tensile strain. The presented technology that utilizes laser shock annealing to modulate the ultrafast diffusion in the PbI2 layer provides a guideline for future improvement in the device performance and stability of hybrid organic–inorganic halide perovskites.

Legumes, unlike other plants, have the ability to establish symbiosis with nitrogen-fixing rhizobia. It has been theorized that a unique property of legume root cortical cells enabled the initial establishment of rhizobial symbiosis1–3. Here we show that a SHORTROOT–SCARECROW (SHR–SCR) stem cell program in cortical cells of the legume Medicago truncatula specifies their distinct fate. Regulatory elements drive the cortical expression of SCR, and stele-expressed SHR protein accumulates in cortical cells of M. truncatula but not Arabidopsis thaliana. The cortical SHR–SCR network is conserved across legume species, responds to rhizobial signals, and initiates legume-specific cortical cell division for de novo nodule organogenesis and accommodation of rhizobia. Ectopic activation of SHR and SCR in legumes is sufficient to induce root cortical cell division. Our work suggests that acquisition of the cortical SHR–SCR module enabled cell division coupled to rhizobial infection in legumes. We propose that this event was central to the evolution of rhizobial endosymbiosis. Repurposing of an SHR–SCR stem cell program in the legume root cortex enables rhizobial symbiosis.

Plants adapt to their ever-changing environment via positive and negative signals induced by environmental stimuli. Drought stress, for instance, induces accumulation of the plant hormone abscisic acid (ABA), triggering ABA signal transduction. However, the molecular mechanisms for switching between plant growth promotion and stress response remain poorly understood. Here we report that RAF (rapidly accelerated fibrosarcoma)-LIKE MITOGEN-ACTIVATED PROTEIN KINASE KINASE KINASE 22 (RAF22) in Arabidopsis thaliana physically interacts with ABA INSENSITIVE 1 (ABI1) and phosphorylates ABI1 at Ser416 residue to enhance its phosphatase activity. Interestingly, ABI1 can also enhance the activity of RAF22 through dephosphorylation, reciprocally inhibiting ABA signaling and promoting the maintenance of plant growth under normal conditions. Under drought stress, however, the ABA-activated OPEN STOMATA1 (OST1) phosphorylates the Ser81 residue of RAF22 and inhibits its kinase activity, restraining its enhancement of ABI1 activity. Taken together, our study reveals that RAF22, ABI1, and OST1 form a dynamic regulatory network that plays crucial roles in optimizing plant growth and environmental adaptation under drought stress.

Stomatal opening is largely promoted by light-activated plasma membrane-localized proton ATPases (PM H+-ATPases), while their closure is mainly modulated by abscisic acid (ABA) signaling during drought stress. It is unknown whether PM H+-ATPases participate in ABA-induced stomatal closure. We established that BRI1-ASSOCIATED RECEPTOR KINASE 1 (BAK1) interacts with, phosphorylates and activates the major PM Arabidopsis H+-ATPase isoform 2 (AHA2). Detached leaves from aha2-6 single mutant Arabidopsis thaliana plants lost as much water as bak1-4 single and aha2-6 bak1-4 double mutants, with all three mutants losing more water than the wild-type (Columbia-0 [Col-0]). In agreement with these observations, aha2-6, bak1-4, and aha2-6 bak1-4 mutants were less sensitive to ABA-induced stomatal closure than Col-0, whereas the aha2-6 mutation did not affect ABA-inhibited stomatal opening under light conditions. ABA-activated BAK1 phosphorylated AHA2 at Ser-944 in its C-terminus and activated AHA2, leading to rapid H+ efflux, cytoplasmic alkalinization, and reactive oxygen species (ROS) accumulation, to initiate ABA signal transduction and stomatal closure. The phosphorylation-mimicking mutation AHA2S944D driven by its own promoter could largely compensate for the defective phenotypes of water loss, cytoplasmic alkalinization, and ROS accumulation in both aha2-6 and bak1-4 mutants. Our results uncover a crucial role of AHA2 in cytoplasmic alkalinization and ABA-induced stomatal closure during the plant’s response to drought stress.

In plants, the spatiotemporal expression of circadian oscillators provides adaptive advantages in diverse species. However, the molecular basis of circadian clock in soybean is not known. In this study, we used soybean hairy roots expression system to monitor endogenous circadian rhythms and the sensitivity of circadian clock to environmental stimuli. We discovered in experiments with constant light and temperature conditions that the promoters of clock genes GmLCLb2 and GmPRR9b1 drive a self-sustained, robust oscillation of about 24-h in soybean hairy roots. Moreover, we demonstrate that circadian clock is entrainable by ambient light/dark or temperature cycles. Specifically, we show that light and cold temperature pulses can induce phase shifts of circadian rhythm, and we found that the magnitude and direction of phase responses depends on the specific time of these two zeitgeber stimuli. We obtained a quadruple mutant lacking the soybean gene GmLCLa1, LCLa2, LCLb1, and LCLb2 using CRISPR, and found that loss-of-function of these four GmLCL orthologs leads to an extreme short-period circadian rhythm and late-flowering phenotype in transgenic soybean. Our study establishes that the morning-phased GmLCLs genes act constitutively to maintain circadian rhythmicity and demonstrates that their absence delays the transition from vegetative growth to reproductive development.

Plant stomata phenotypic traits can provide a basis for enhancing crop tolerance in adversity. Manually counting the number of stomata and measuring the height and width of stomata obviously cannot satisfy the high-throughput data. How to detect and recognize plant stomata quickly and accurately is the prerequisite and key for studying the physiological characteristics of stomata. In this research, we consider stomata recognition as a multi-object detection problem, and propose an end-to-end framework for intelligent detection and recognition of plant stomata based on feature weights transfer learning and YOLOv4 network. It is easy to operate and greatly facilitates the analysis of stomata phenotypic traits in high-throughput plant epidermal cell images. For different cultivars, multi-scales, rich background features, high density, and small stomata object images, the proposed method can precisely locate multiple stomata in microscope images and automatically give phenotypic traits of stomata. Users can also adjust the corresponding parameters to maximize the accuracy and scalability of automatic stomata detection and recognition. Experimental results on actual data provided by the National Maize Improvement Center show that the proposed method is superior to the existing methods in high stomata automatic detection and recognition accuracy, low training cost, strong generalization ability.

Seed germination is a physiological process regulated by multiple factors. Abscisic acid (ABA) can inhibit seed germination to improve seedling survival under conditions of abiotic stress, and this process is often regulated by light signals. Constitutive Photomorphogenic 1 (COP1) is an upstream core repressor of light signals, and is involved in several ABA responses. Here, we demonstrate that COP1 is a negative regulator of the ABA-mediated inhibition of seed germination. Disruption of COP1 enhanced Arabidopsis seed sensitivity to ABA and increased ROS levels. In seeds, ABA induced the translocation of COP1 to the cytoplasm, resulting in enhanced ABA-induced ROS levels. Genetic evidence indicated that HY5 and ABI5 act downstream of COP1 in the ABA-mediated inhibition of seed germination. ABA-induced COP1 cytoplasmic localization increased HY5 and ABI5 protein levels in the nucleus, leading to increased expression of ABI5 target genes and ROS levels in seeds. Together, our results reveal that ABA-induced cytoplasmic translocation of COP1 activates the HY5-ABI5 pathway to promote the expression of ABA-responsive genes and the accumulation of ROS during ABA-mediated inhibition of seed germination. These findings enhance the role of COP1 in the ABA signal transduction pathway.

Background When developing CRISPR/Cas9 systems for crops, it is crucial to invest time characterizing the genome editing efficiency of the CRISPR/Cas9 cassettes, especially if the transformation system is difficult or time-consuming. Cotton is an important crop for the production of fiber, oil, and biofuel. However, the cotton stable transformation is usually performed using Agrobacterium tumefaciens taking between 8 and 12 months to generate T0 plants. Furthermore, cotton is a heterotetraploid and targeted mutagenesis is considered to be difficult as many genes are duplicated in this complex genome. The application of CRISPR/Cas9 in cotton is severely hampered by the long and technically challenging genetic transformation process, making it imperative to maximize its efficiency. Results In this study, we provide a new system to evaluate and validate the efficiency of CRISPR/Cas9 cassettes in cotton using a transient expression system. By using this system, we could select the most effective CRISPR/Cas9 cassettes before the stable transformation. We have also optimized the existing cotton CRISPR/Cas9 system to achieve vastly improved mutagenesis efficiency by incorporating an endogenous GhU6 promoter that increases sgRNA expression levels over the Arabidopsis AtU6-29 promoter. The 300 bp GhU6.3 promoter was cloned and validated using the transient expression system. When sgRNAs were expressed under the control of the GhU6.3 promoter in CRISPR/Cas9 cassettes, expression levels were 6–7 times higher than those provided by the AtU6-29 promoter and CRISPR/Cas9-mediated mutation efficiency was improved 4–6 times. Conclusions This study provides essential improvements to maximize CRISPR/Cas9-mediated mutation efficiency by reducing risk and workload for the application of CRISPR/Cas9 approaches in the targeted mutagenesis of cotton. Electronic supplementary material The online version of this article (10.1186/s13007-018-0353-0) contains supplementary material, which is available to authorized users.

Background Brassica napus (B. napus) encompasses diverse transcription factors (TFs), but thorough identification and characterization of TF families, as well as their transcriptional responsiveness to multifarious stresses are still not clear. Results Totally 2167 TFs belonging to five families were genome-widely identified in B. napus, including 518 BnAP2/EREBPs, 252 BnbZIPs, 721 BnMYBs, 398 BnNACs and 278 BnWRKYs, which contained some novel members in comparison with existing results. Sub-genome distributions of BnAP2/EREBPs and BnMYBs indicated that the two families might have suffered from duplication and divergence during evolution. Synteny analysis revealed strong co-linearity between B. napus and its two ancestors, although chromosomal rearrangements have occurred and 85 TFs were lost. About 7.6% and 9.4% TFs of the five families in B. napus were novel genes and conserved genes, which both showed preference on the C sub-genome. RNA-Seq revealed that more than 80% TFs were abiotic stress inducible and 315 crucial differentially expressed genes (DEGs) were screened out. Network analysis revealed that the 315 DEGs are highly co-expressed. The homologous gene network in A. thaliana revealed that a considerable amount of TFs could trigger the differential expression of targeted genes, resulting in a complex clustered network with clusters of genes responsible for targeted stress responsiveness. Conclusions We identified and characterized five TF families in B. napus. Some crucial members and regulatory networks involved in different abiotic stresses have been explored. The investigations deepen our understanding of TFs for stress tolerance in B. napus.

Rapeseed (Brassica napus L.) is an important oil crop worldwide. For current B. napus production, it is urgent to develop new varieties with higher seed productivity and increased stress tolerance for better adaptation to the abiotic stresses as a result of global climate change. Genetic engineering, to some extent, can overcome the limitations of genetic exchange in conventional breeding. Consequently, it considered as an effective method for improving modern crop breeding for B. napus. Since crop stress resistance is a polygenic complex trait, only by multi-gene synergistic effects can effectively achieve the comprehensive stress resistance of crops. Hence, in the present study, five stress resistance genes, NCED3, ABAR, CBF3, LOS5, and ICE1 were transferred into B. napus. Compared with wildtype (WT) plants, the multi-gene transformants K15 exhibited pronounced growth advantage under both normal growth and stress conditions. Additionally, K15 plants also showed significantly higher resistance response to multiple stresses at seed germination and seedling stages than WT plants. Furthermore, K15 plants had significantly higher leaf temperature and significantly lower stomatal aperture and water loss rate than WT plants, which indicated that the water-holding capacity of K15 plants was significantly superior to that of WT plants after stress treatment. In addition, K15 plants had significantly higher abscisic acid (ABA) content and significantly lower malondialdehyde (MDA) content than WT plants. In conclusion, the above results suggested that multi-gene co-expression could rapidly trigger plant stress resistance, reduce the stress injury on plants and synergistically improve the comprehensive resistance of B. napus.

Anthocyanins are secondary metabolites that play important roles in plant adaption to adverse environments. The anthocyanin biosynthetic pathway is conserved in high plants. Previous studies revealed the significant role of anthocyanins in natural-colorized cotton. However, little is known about the involvement of anthocyanins in the interaction of cotton and pathogen. In this study, a pathogen-induced gene was isolated from Gossypium barbadense that encodes an anthocyanidin synthase protein (GbANS) with dioxygenase structures. GbANS was preferentially expressed in colored tissue. Silencing of GbANS significantly reduced the production of anthocyanins, as well as the cotton’s resistance to Verticillium dahliae. Biochemical studies revealed that GbANS-silenced cotton accumulated more hydrogen peroxide compared to control plants during the V. dahliae invasion process. This accumulation of hydrogen peroxide corresponded with increased cell death around the invasion sites, which in turn accelerated the V. dahliae infection. Taken together, we found that GbANS contributes to the biosynthesis of anthocyanins in cotton and anthocyanins positively regulate cotton’s resistance to V. dahliae.

Background Genetically modified cotton accounts for 64% of the world’s cotton growing area (22.3 million hectares). The genome sequencing of the diploid cotton progenitors Gossypium raimondii and Gossypium arboreum as well as the cultivated Gossypium hirsutum has provided a wealth of genetic information that could be exploited for crop improvement. Unfortunately, gene functional characterization in cotton is lagging behind other economically important crops due to the low efficiency, lengthiness and technical complexity of the available stable transformation methods. We present here a simple, fast and efficient method for the transient transformation of G. hirsutum that can be used for gene characterization studies. Results We developed a transient transformation system for gene characterization in upland cotton. Using β-glucuronidase as a reporter for Agrobacterium-mediated transformation assays, we evaluated multiple transformation parameters such as Agrobacterium strain, bacterial density, length of co-cultivation, chemicals and surfactants, which can affect transformation efficiency. After the initial characterization, the Agrobacterium EHA105 strain was selected and a number of binary constructs used to perform gene characterization studies. 7-days-old cotton seedlings were co-cultivated with Agrobacterium and transient gene expression was observed 5 days after infection of the plants. Transcript levels of two different transgenes under the control of the cauliflower mosaic virus (CaMV) 35S promoter were quantified by real-time reverse transcription PCR (qRT-PCR) showing a 3–10 times increase over the levels observed in non-infected controls. The expression patterns driven by the promoters of two G. hirsutum genes as well as the subcellular localization of their corresponding proteins were studied using the new transient expression system and our observations were consistent with previously published results using Arabidopsis as a heterologous system. Conclusions The Agrobacterium-mediated transient transformation method is a fast and easy transient expression system enabling high transient expression and transformation efficiency in upland cotton seedlings. Our method can be used for gene functional studies such as promoter characterization and protein subcellular localization in cotton, obviating the need to perform such studies in a heterologous system such as Arabidopsis. Electronic supplementary material The online version of this article (10.1186/s13007-018-0319-2) contains supplementary material, which is available to authorized users.

Background Soil salinization is one of the major environmental constraints to plant growth and agricultural production worldwide. Signaling components involving calcium (Ca2+) and the downstream calcium-dependent protein kinases (CPKs) play key roles in the perception and transduction of stress signals. However, the study of CPKs in cotton and their functions in response to salt stress remain unexplored. Results A total of 98 predicted CPKs were identified from upland cotton (Gossypium hirsutum L. ‘TM-1’), and phylogenetic analyses classified them into four groups. Gene family distribution studies have revealed the substantial impacts of the genome duplication events to the total number of GhCPKs. Transcriptome analyses showed a wide distribution of CPKs’ expression among different organs. A total of 19 CPKs were selected for their rapid responses to salt stress at the transcriptional level, most of which were also incduced by the thylene-releasing chemical ethephon, suggesting a partal overlap of the salinity and ethylene responses. Silencing of 4 of the 19 CPKs (GhCPK8, GhCPK38, GhCPK54, and GhCPK55) severely compromised the basal cotton resistance to salt stress. Conclusions Our genome-wide expression analysis of CPK genes from up-land cotton suggests that CPKs are involved in multiple developmental responses as well as the response to different abiotic stresses. A cluster of the cotton CPKs was shown to participate in the early signaling events in cotton responses to salt stress. Our results provide significant insights on functional analysis of CPKs in cotton, especially in the context of cotton adaptions to salt stress. Electronic supplementary material The online version of this article (doi: 10.1186/s12870-018-1230-8) contains supplementary material, which is available to authorized users.

The ordered aggregation of non-fullerene small molecular acceptors (SMAs) plays a key role in determining the charge transport and bimolecular recombination in polymer/SMA solar cells. However, due to the competition between crystallization and phase separation, the polymers are prone to form a network first, which inhibits the molecular diffusion of SMAs, resulting in weak crystallinity of SMAs. Here, we demonstrated a sequential crystallization method for high performance PBDB-T/ITIC solar cells with much improved crystallinity of ITIC. By tuning the sequence of thermal annealing (TA) and solvent vapor annealing (SVA), sequential crystallization of ITIC and PBDB-T can be fine controlled to grow a highly crystalline ITIC and PBDB-T network. The crystallization kinetics results indicate that when the crystallization of ITIC occurred prior to the formation of the PBDB-T crystallized network, the crystallinity of ITIC is significantly improved due to high molecular diffusion. However, if the crystallization of PBDB-T occurred first, the diffusion of ITIC was restricted by the crystalline network of PBDB-T, resulting in a low crystallinity of ITIC. The enhanced crystallinity of ITIC is beneficial to the electron transport and suppressed the bimolecular recombination, which helps boost the device performance from 8.14% to 10.95%. This work demonstrates that manipulation of the crystallization sequence of the donor and acceptor may be key to further boost the efficiency of polymer/SMA solar cells.

Heavy metal contamination has been threatening the health of human beings. To decrease the bio-toxicity of heavy metals, a thiol-functionalized nano-silica (SiO2-SH) was adopted to remediate the soil contaminated by lead (Pb), cadmium (Cd) and copper (Cu). The remediation effect of SiO2-SH on contaminated soils was investigated by the uptake of the heavy metals into lettuce and pakchoi in pot experiment. The bio-toxicity of the SiO2-SH was evaluated, and its immobilization mechanisms were proposed by the fraction distribution of Cd, Pb and Cu. It was found that the SiO2-SH can significantly reduce the uptake of Cd, Pb, Cu into pakchoi by 92.02%, 68.03%, 76.34% and into lettuce by 89.81%, 43.41%, 5.76%, respectively. The chemical species analyses of Cd, Pb, Cu indicate SiO2-SH can transform the heavy metal in acid soluble states into reducible fraction and oxidizable fraction, thereby inhibiting the extraction of heavy metals into soil solution. The concentrations of microbial biomass carbon, organic matter, and cation exchange capacity of the soil increased while the soil bulk density decreased after remediation. Those changes demonstrate that SiO2-SH not only has no bio-toxic impact on the soil environment but also improves the soil environment, which proves the prepared SiO2-SH is environmental-friendly. The SiO2-SH could be a promising amendment for heavy metal contaminated soils.

MYB proteins form one of the largest transcription factor families in plants. Growing evidence has suggested that MYB has important roles in the regulation of plant growth and development, as well as responses to abiotic and biotic stresses. In the present study, the GbMYB60 gene isolated from Gossypium barbadense was heterogeneously expressed in Arabidopsis. GbMYB60 encodes a R2R3-MYB protein with transcriptional activation activity. Histochemical staining revealed that GbMYB60 was preferential expressed in the leaf vascular and meristem tissues of young seedlings and in the axillary bud tissue of mature plants. Expression pattern analysis indicated that GbMYB60 is induced by ABA, mannitol, and salt, but not H2O2. Transgenic Arabidopsis with constitutively higher expression of GbMYB60 exhibited reduced germination rates under salt and mannitol treatments as well as reduced tolerance to salt at the adult-plant stage. Transgenic Arabidopsis accumulates more reactive oxygen species compared to wild-type plants under stress conditions, corresponding with more severe cell death in transgenic plants. Additionally, ABA-related genes, including ABI5 and ABF3, were down-regulated in GbMYB60-overexpressing plants. This research has revealed that cotton GbMYB60 negatively regulates plant tolerance to salt stress, possibly through the inhibition of ABA signaling pathways and enhancing reactive oxygen species over-production.

Hydrogels consisting of reactive monomers can respond to external stimuli, which is promising for many applications such as sensors, soft robotics, and actuators, etc . However, most of responsive hydrogels are hard to fabricate into smart devices due to their poor mechanical properties. In this paper, vinyl-modified graphene oxide (GO-VTES) nanosheets were facilely prepared through the coupling reaction between hydroxyl or carboxylic acid groups from graphene oxide (GO) and hydroxyl groups from hydrolyzed vinyltriethoxysilane (VTES). By using the multifunctional GO-VTES nanosheets as crosslinkers, N -isopropylacrylamide (NIPAM) and acrylamide (AAm) were copolymerized to generate a series of tough and thermo-responsive composite hydrogels. The structures and morphologies of GO-VTES were investigated by FTIR, Raman, XPS, XRD, AFM and TEM. The obtained thermo-responsive composite hydrogels exhibit enhanced mechanical properties, which can be tuned by varying the content of GO-VTES and AAm. Moreover, these hydrogels were thermo-responsive with the LCST tunable by manipulating the contents of GO-VTES and/or AAm in the hydrogels. This strategy to prepare responsive composite hydrogels with enhanced mechanical properties has attractive applications for fabricating smart soft actuators.

An interpenetrating bulk-heterojunction structure with a domain size of 10–20 nm is the ideal morphology for carrier generation, separation and transportation in organic solar cells. However, depending on the blend composition, the phase-separation behavior of the crystalline small molecule blend system of donor 7,7′-(4,4-bis(2-ethylhexyl)-4H-silolo[3,2-b:4,5-b′]dithiophene-2,6-diyl)bis(6-fluoro-4-(5′-hexyl-[2,2′-bithiophen]-5-yl)benzo[c][1,2,5]thiadiazole) (p-DTS(FBTTh2)2) and acceptor N,N′-bis(1-ethylpropyl)-perylene-3,4,9,10-tetracarboxylic diimide (EP-PDI) is quite different. When the weight ratio of p-DTS(FBTTh2)2/EP-PDI is greater than 8 : 2 or smaller than 4 : 6, a large phase separation structure is observed induced by p-DTS(FBTTh2)2 or EP-PDI crystallization. When the ratio of p-DTS(FBTTh2)2 : EP-PDI changes from 7 : 3 to 5 : 5, no obvious phase separation can be detected due to the interaction between p-DTS(FBTTh2)2 and EP-PDI. In order to obtain the interpenetrating bulk-heterojunction structure with a domain size of 10–20 nm, we proposed to tune the molecule diffusion of p-DTS(FBTTh2)2 and EP-PDI by different thermal annealing temperature to control the phase separation domain size and phase purity. When the annealing temperature is T Tc p-DTS(FBTTh2)2 − Tb (T is the thermal annealing temperature, Ta and Tb are constants), the molecule diffusion rate of both p-DTS(FBTTh2)2 and EP-PDI is too slow or too fast, resulting in no phase separation or large phase separation morphology. When T is between Tm EP-PDI + Ta and Tc p-DTS(FBTTh2)2 − Tb, EP-PDI is in a melting state, but p-DTS(FBTTh2)2 self-assembles into crystals. As a result, p-DTS(FBTTh2)2 crystallized and formed a framework, which inhibited the massive crystallization of EP-PDI due to the spatial confinement of the p-DTS(FBTTh2)2 crystallization framework, leading to the formation of a bi-continuous phase separation structure with suitable domain size and phase purity. Based on the above phase separation structure we got, a power conversion efficiency of 4.25% was obtained, which is relatively high in this system without any additives.

Legumes, unlike other plants, have the ability to establish symbiosis with nitrogen-fixing rhizobia. It has been theorized that a unique property of legume root cortical cells enabled the initial establishment of rhizobial symbiosis

Stress-associated proteins (SAPs) containing the A20/AN1 zinc-finger domain play important roles in response to both biotic and abiotic stresses in plants. Nevertheless, few studies have focused on the SAP gene family in cotton. To explore the distributions and expression patterns of these genes, we performed genome-wide identification and characterization of SAPs in tetraploid Gossypium hirsutum L. TM-1 (AD1). A total of 37 genes encoding SAPs were identified, 36 of which were duplicated in the A and D sub-genomes. The analysis of gene architectures and conserved protein motifs revealed that nearly all A20-AN1-type SAPs were intron-free, whereas AN1-AN1-type SAPs contained one intron. The cis-elements of the SAP promoters were studied, as were the expression levels of cotton SAP genes under different stresses based on RNA-seq data and validated by qRT-PCR. Most cotton SAP genes were induced by multiple stresses and phytohormones, particularly salt stress, indicating that SAP genes may play important roles in cotton's response to unfavorable environmental changes. Among these identified SAPs, the expression of GhSAP17A/D is suppressed in cotton response to Vertillium dahliae, and the GhSAP17A/D-silenced cotton exhibits more resistance to V. dahliae. This study provides insight into the evolution of SAP genes in upland cotton and may aid in efforts at further functional identification of A20/AN1-type proteins and cotton's response to different stresses.

As the diploid progenitor of common wheat, Aegilops tauschii Cosson (DD, 2n = 2x = 14) is considered to be a promising genetic resource for the improvement of common wheat. In this work, we demonstrated that the efficiency of transferring A. tauschii segments to common wheat was clearly improved through the use of synthetic octaploid wheat (AABBDDDD, 2n = 8x = 56) as a “bridge.” The synthetic octaploid was obtained by chromosome doubling of hybrid F1 (A. tauschii T015 × common wheat Zhoumai 18). A set of introgression lines (BC1F8) containing 6016 A. tauschii segments was developed and displayed significant phenotype variance among lines. Twelve agronomic traits, including growth duration, panicle traits, grain traits, and plant height (PH), were evaluated. And transgressive segregation was identified in partial lines. Additionally, better agronomic traits could be observed in some lines, compared to the recurrent parent Zhoumai 18. To verify that the significant variance of those agronomic traits was supposedly controlled by A. tauschii segments, 14 quantitative trait loci (QTLs) for three important agronomic traits (thousand kernel weight, spike length, and PH) were further located in the two environments (Huixian and Zhongmou), indicating the introgression of favorable alleles from A. tauschii into common wheat. This study provides an ameliorated strategy to improve common wheat utilizing a single A. tauschii genome.

Saline-alkali soil is a major environmental constraint impairing plant growth and crop productivity. In this study, we identified a Ca(2+) sensor/kinase/plasma membrane (PM) H(+)-ATPase module as a central component conferring alkali tolerance in Arabidopsis (Arabidopsis thaliana). We report that the SCaBP3 (SOS3-LIKE CALCIUM BINDING PROTEIN3)/CBL7 (CALCINEURIN B-LIKE7) loss-of-function plants exhibit enhanced stress tolerance associated with increased PM H(+)-ATPase activity and provide fundamental mechanistic insights into the regulation of PM H(+)-ATPase activity. Consistent with the genetic evidence, interaction analyses, in vivo reconstitution experiments, and determination of H(+)-ATPase activity indicate that interaction of the Ca(2+) sensor SCaBP3 with the C-terminal Region I domain of the PM H(+)-ATPase AHA2 (Arabidopsis thaliana PLASMA MEMBRANE PROTON ATPASE2) facilitates the intramolecular interaction of the AHA2 C terminus with the Central loop region of the PM H(+)-ATPase to promote autoinhibition of H(+)-ATPase activity. Concurrently, direct interaction of SCaPB3 with the kinase PKS5 (PROTEIN KINASE SOS2-LIKE5) stabilizes the kinase-ATPase interaction and thereby fosters the inhibitory phosphorylation of AHA2 by PKS5. Consistently, yeast reconstitution experiments and genetic analysis indicate that SCaBP3 provides a bifurcated pathway for coordinating intramolecular and intermolecular inhibition of PM H(+)-ATPase. We propose that alkaline stress-triggered Ca(2+) signals induce SCaBP3 dissociation from AHA2 to enhance PM H(+)-ATPase activity. This work illustrates a versatile signaling module that enables the stress-responsive adjustment of plasma membrane proton fluxes.

Detailed information for the 153 common DEGs. (PDF 61 kb)

Final statistics for synteny analsyis. (PDF 85 kb)

Supplementary materials that supporting the main text. (PDF 61,360 kb)

The finally identified 315 crucial DEGs. (PDF 128 kb)

Responsive genes of the five TF families. (PDF 531 kb)

Surface plasmon resonance (SPR) nanosensors based on metallic nanohole arrays have been widely reported to detect binding interactions in biological specimens. A simple and effective method for constructing nanoscale arrays is essential for the development of SPR nanosensors. In this work, we report a one-step method to fabricate nanohole arrays by thermal nanoimprinting in the matrix of IPS (Intermediate Polymer Stamp). No additional etching process or supporting substrate is required. The preparation process is simple, time-saving and compatible for roll-to-roll process, potentially allowing mass production. Moreover, the nanohole arrays were integrated into detection platform as SPR sensors to investigate different types of biological binding interactions. The results demonstrate that our one-step method can be used to efficiently fabricate large-area and uniform nanohole arrays for biochemical sensing.

Genome editing is an important tool for gene functional studies as well as crop improvement. The recent development of the CRISPR/Cas9 system using single guide RNA molecules (sgRNAs) to direct precise double strand breaks in the genome has the potential to revolutionize agriculture. Unfortunately, not all sgRNAs are equally efficient and it is difficult to predict their efficiency by bioinformatics. In crops such as cotton (Gossypium hirsutum L.), with labor-intensive and lengthy transformation procedures, it is essential to minimize the risk of using an ineffective sgRNA that could result in the production of transgenic plants without the desired CRISPR-induced mutations. In this study, we have developed a fast and efficient method to validate the functionality of sgRNAs in cotton using a transient expression system. We have used this method to validate target sites for three different genes GhPDS, GhCLA1, and GhEF1 and analyzed the nature of the CRISPR/Cas9-induced mutations. In our experiments, the most frequent type of mutations observed in cotton cotyledons were deletions (∼64%). We prove that the CRISPR/Cas9 system can effectively produce mutations in homeologous cotton genes, an important requisite in this allotetraploid crop. We also show that multiple gene targeting can be achieved in cotton with the simultaneous expression of several sgRNAs and have generated mutations in GhPDS and GhEF1 at two target sites. Additionally, we have used the CRISPR/Cas9 system to produce targeted gene fragment deletions in the GhPDS locus. Finally, we obtained transgenic cotton plants containing CRISPR/Cas9-induced gene editing mutations in the GhCLA1 gene. The mutation efficiency was very high, with 80.6% of the transgenic lines containing mutations in the GhCLA1 target site resulting in an intense albino phenotype due to interference with chloroplast biogenesis.

The plant glutathione peroxidase (GPX) family consists of multiple isoenzymes with distinct subcellular locations, tissue-specific expression patterns and environmental stress responses. In this study, 13 putative GPXs from the genome of Gossypium hirsutum (GhGPXs) were identified and a conserved pattern among plant GPXs were exhibited, besides this they also responded to multiple environmental stresses and we predicted that they had hormone responsive cis-elements in their promoter regions. Most of the GhGPXs on expression in yeast can scavenge H2O2. Our results showed that different members of the GhGPX gene family were co-ordinately regulated under specific environmental stress conditions, and suggested the importance of GhGPXs in hormone treatments and abiotic stress responses.

Based on transcriptomic data from four experimental settings with drought-resistant and drought-sensitive cultivars under drought and well-watered conditions, statistical analysis revealed three categories encompassing 169 highly differentially expressed genes (DEGs) in response to drought in Brassica napus L., including 37 drought-resistant cultivar-related genes, 35 drought-sensitive cultivar-related genes and 97 cultivar non-specific ones. We provide evidence that the identified DEGs were fairly uniformly distributed on different chromosomes and their expression patterns are variety specific. Except commonly enriched in response to various stimuli or stresses, different categories of DEGs show specific enrichment in certain biological processes or pathways, which indicated the possibility of functional differences among the three categories. Network analysis revealed relationships among the 169 DEGs, annotated biological processes and pathways. The 169 DEGs can be classified into different functional categories via preferred pathways or biological processes. Some pathways might simultaneously involve a large number of shared DEGs, and these pathways are likely to cross-talk and have overlapping biological functions. Several members of the identified DEGs fit to drought stress signal transduction pathway in Arabidopsis thaliana. Finally, quantitative real-time PCR validations confirmed the reproducibility of the RNA-seq data. These investigations are profitable for the improvement of crop varieties through transgenic engineering.

The architecture of the cotton plant, including fruit branch formation and flowering pattern, is the most important characteristic that directly influences light exploitation, yield and cost of planting. Nulliplex branch is a useful phenotype to study cotton architecture. We used RNA sequencing to obtain mRNA and miRNA profiles from nulliplex- and normal-branch cotton at three developmental stages. The differentially expressed genes (DEGs) and miRNAs were identified that preferentially/specifically expressed in the pre-squaring stage, which is a key stage controlling the transition from vegetative to reproductive growth. The DEGs identified were primarily enriched in RNA, protein and signalling categories in Gossypium barbadense and Gossypium hirsutum. Interestingly, during the pre-squaring stage, the DEGs were predominantly enriched in transcription factors in both G. barbadense and G. hirsutum and these transcription factors were mainly involved in branching and flowering. Related miRNAs were also identified. The results showed that fruit branching in cotton is controlled by molecular pathways similar to those in Arabidopsis and that multiple regulated pathways may affect the development of floral buds. Our study showed that the development of fruit branches is closely related to flowering induction and provides insight into the molecular mechanisms of branch and flower development in cotton.