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Most of the reported P-type pentatricopeptide repeat (PPR) proteins play roles in organelle RNA stabilization and splicing. However, P-type PPRs involved in both RNA splicing and editing have rarely been reported, and their underlying mechanism remains largely unknown. Here, we report a rice floury endosperm22 (flo22) mutant with delayed amyloplast development in endosperm cells. Map-based cloning and complementation tests demonstrated that FLO22 encodes a mitochondrion-localized P-type PPR protein. Mutation of FLO22 resulting in defective trans-splicing of mitochondrial nad1 intron 1 and perhaps causing instability of mature transcripts affected assembly and activity of complex Ⅰ, and mitochondrial morphology and function. RNA-seq analysis showed that expression levels of many genes involved in starch and sucrose metabolism were significantly down-regulated in the flo22 mutant compared with the wild type, whereas genes related to oxidative phosphorylation and the tricarboxylic acid cycle were significantly up-regulated. In addition to involvement in splicing as a P-type PPR protein, we found that FLO22 interacted with DYW3, a DYW-type PPR protein, and they may function synergistically in mitochondrial RNA editing. The present work indicated that FLO22 plays an important role in endosperm development and plant growth by participating in nad1 maturation and multi-site editing of mitochondrial messager RNA.

In our daily communication, we communicate both verbally and non-verbally. Nonverbal communication is an indispensable and inseparable part of human interaction and communication. Nonverbal communication system is the result of a certain social culture. It has its own cultural specific features. Nonverbal behavior may convey different meanings in different cultures. Nonverbal differences may influence the effects of cross-cultural communication. This paper intends to give an introduction of the functions and characteristics of nonverbal communication and propose some strategies to improve the efficiency of cross-cultural communication.

Plants have evolved sophisticated mechanisms to coordinate their growth and stress responses via integrating various phytohormone signaling pathways. However, the precise molecular mechanisms orchestrating integration of the phytohormone signaling pathways remain largely obscure. In this study, we found that the rice (Oryza sativa) short internodes1 (shi1) mutant exhibits typical auxin-deficient root development and gravitropic response, brassinosteroid (BR)-deficient plant architecture and grain size as well as enhanced abscisic acid (ABA)-mediated drought tolerance. Additionally, we found that the shi1 mutant is also hyposensitive to auxin and BR treatment but hypersensitive to ABA. Further, we showed that OsSHI1 promotes the biosynthesis of auxin and BR by activating the expression of OsYUCCAs and D11, meanwhile dampens ABA signaling by inducing the expression of OsNAC2, which encodes a repressor of ABA signaling. Furthermore, we demonstrated that 3 classes of transcription factors, AUXIN RESPONSE FACTOR 19 (OsARF19), LEAF AND TILLER ANGLE INCREASED CONTROLLER (LIC), and OsZIP26 and OsZIP86, directly bind to the promoter of OsSHI1 and regulate its expression in response to auxin, BR, and ABA, respectively. Collectively, our results unravel an OsSHI1-centered transcriptional regulatory hub that orchestrates the integration and self-feedback regulation of multiple phytohormone signaling pathways to coordinate plant growth and stress adaptation.

RNA interference is an important technology for gene functional research in many organisms. The pond wolf spider (Pardosa pseudoannulata) is an important natural enemy of rice field pests. To facilitate large-scale gene functional research in this spider species and others, we developed an RNAi method via ingestion of bacteria expressing dsRNA. The dsRNA targeting a cytochrome P450 monooxygenase (cyp41g2) was expressed in E. coli HT115 (DE3). And then the bacterial suspension was fed to 14-20 days old spiderlings. The mRNA abundance of the target gene was significantly reduced after 3-day's ingestion of bacteria expressing dsRNA, and between day 5 and 7, RNAi efficiency remained stable. Thus we selected 5 days as the optimum interference time. Furthermore, the bacteria resuspension containing 20 ng/μL dsRNA was selected as the optimum concentration. To evaluate the applicability of this method, three other genes with different tissue expression pattern were also selected as targets. And the mRNA abundance of all the four target genes were significantly reduced with RNAi efficiency between 66.0% and up to 86.9%. The results demonstrated that the oral delivery of bacteria expressing dsRNA would be an effective RNAi method for the gene functional study in P. pseudoannulata. This article is protected by copyright. All rights reserved.

Starch accounts for about 80-85 % of the dry weight of grains and determines yield by impact on grain weight. And, the content and composition of starch also determine appearance, eating, cooking and nutritional quality of rice. By coordinating crucial reactions of the primary carbohydrate metabolism in all eukaryotes, fructose-2,6-bisphosphate (Fru-2,6-P

Summary Thioredoxins (TRXs) occur in plant chloroplasts as complex disulphide oxidoreductases. Although many biological processes are regulated by thioredoxins, the regulatory mechanism of chloroplast TRXs are largely unknown.Here we report a rice white panicle2 mutant caused by a mutation in the thioredoxin z gene, an orthologue of AtTRX z in Arabidopsis. white panicle2 (wp2) seedlings exhibited a high‐temperature‐sensitive albinic phenotype.We found that plastid multiple organellar RNA editing factors (MORFs) were the regulatory targets of thioredoxin z. We showed that OsTRX z protein physically interacts with OsMORFs in a redox‐dependent manner and that the redox state of a conserved cysteine in the MORF box is essential for MORF–MORF interactions. wp2 and OsTRX z knockout lines show reduced editing efficiencies in many plastidial‐encoded genes especially under high‐temperature conditions. An Arabidopsis trx z mutant also exhibited significantly reduced chloroplast RNA editing.Our combined results suggest that thioredoxin z regulates chloroplast RNA editing in plants by controlling the redox state of MORFs.

Summary Salt stress dramatically impedes plant growth and development as well as crop yield. The apple production regions are reduced every year, because of the secondary salt damage by improper fertilization and irrigation. To expand the cultivation area of apple (Malus domestica) and select salt‐resistant varieties, the mechanism of salt tolerance in apple is necessary to be clarified. The miR156/SPL regulatory module plays key roles in embryogenesis, morphogenesis, life cycle stage transformation, flower formation and other processes. However, its roles in the mechanisms of salt tolerance are unknown. In order to elucidate the mechanism of 156/SPL regulating salt stress in apple, we performed RLM‐5’ RACE and stable genetic transformation technology to verify that both mdm‐MIR156a and MdSPL13 responded to salt stress in apple and that the latter was the target of the former. MIR156a overexpression weakened salt resistance in apple whereas MdSPL13 overexpression strengthened it. A total of 6094 differentially expressed genes relative to nontransgenic apple plants were found by RNA‐Seq analysis of MdSPL13OE. Further verification indicated that MdSPL13 targeted the MdWRKY100 gene promoter. Moreover, MdWRKY100 overexpression enhanced salt tolerance in apple. Our results revealed that the miR156/SPL module regulates salt tolerance by up‐regulating MdWRKY100 in apple. This study is the first to elucidate the mechanism underlying the miRNA network response to salt stress in apple and provides theoretical and empirical bases and genetic resources for the molecular breeding of salt tolerance in apple.

Salvianolic acid B (Sal-B) is widely used in China for the treatment of numerous diseases. Currently, Salvia miltiorrhiza Bunge is the main source of this compound, but Salvia bowleyana Dunn, a surrogate of S. miltiorrhiza Bge, may provide a novel source for obtaining more Sal-B. In the present study, a simple method for separation and purification of phenolic compounds from S. bowleyana Dunn roots was employed. Sal-B was subsequently purified and its inhibitory effect on the gastric cancer HGC-27 and AGS cell lines was investigated. Sal-B extracted from S. bowleyana Dunn displayed significant antitumor activity in proliferation and apoptosis assays. Overall, it was found that S. bowleyana Dunn has a higher Sal-B content than S. miltiorrhiza Bge and may be used as a novel source of this potential anti-gastric cancer compound.

Over-application of nitrogen fertilizer in fields has had a negative impact on both environment and human health. Domesticated rice varieties with high nitrogen use efficiency (NUE) reduce fertilizer for sustainable agriculture. Here, we perform genome-wide association analysis of a diverse rice population displaying extreme nitrogen-related phenotypes over three successive years in the field, and identify an elite haplotype of nitrate transporter OsNPF6.1HapB that enhances nitrate uptake and confers high NUE by increasing yield under low nitrogen supply. OsNPF6.1HapB differs in both the protein and promoter element with natural variations, which are differentially trans-activated by OsNAC42, a NUE-related transcription factor. The rare natural allele OsNPF6.1HapB, derived from variation in wild rice and selected for enhancing both NUE and yield, has been lost in 90.3% of rice varieties due to the increased application of fertilizer. Our discovery highlights this NAC42-NPF6.1 signaling cascade as a strategy for high NUE and yield breeding in rice., Improving crop nitrogen use efficiency can facilitate sustainable production, however, the genetic mechanisms have not been fully revealed. Here, the authors discover the NAC42-NPF6.1 signaling cascade mainly derives from indica and wild rice and demonstrate the potential of using the allele for cultivar improvement.

BACKGROUND: To investigate the therapeutic effect of total Astragalus saponins (AST) against viral myocarditis in animal and cell models. METHODS: Primary myocardiocytes (PMCs) were stimulated by the coxsackie B (CVB) 3 virus to prepare the cell model of viral myocarditis. Cell viability, apoptosis and the mRNA expression of C-Myc, tumor necrosis factor (TNF)-α and Fas were detected to evaluate the protective effects of AST on CVB3-induced PMC damage. RESULTS: AST could significantly increase survival and decrease the ratio of heart weight: body weight (P

Rice WSL6 is involved in chloroplast ribosome biogenesis and is essential for early chloroplast development. Construction of the genetic translation system is a prerequisite for chloroplast development in plants. However, the molecular mechanism underlying this process is largely unknown. Here, we isolated a white stripe leaf6 (wsl6) mutant in rice. The mutant seedlings displayed white-striped leaves that were more severe under low-temperature conditions. Transmission electron microscopy analysis showed that the wsl6 mutant was defective in early chloroplast development. Map-based cloning revealed that WSL6 encodes an Era-type guanosine-5'-triphosphate (GTP)-binding protein located in chloroplasts. Immunoblotting and quantitative real-time polymerase chain reaction (qRT-PCR) analyses demonstrated an absence of 70S ribosomes in wsl6 chloroplasts. Further research showed that WSL6 binds to the 16S ribosomal RNA (rRNA) subunit of chloroplast ribosome 30S. In summary, these results show that WSL6 is essential for chloroplast ribosome biogenesis during early chloroplast development in rice.

Colouration in spider mites is due to the presence of carotenoids with diverse colours, including yellows, oranges and reds. Tetranychus urticae has two main colour forms, red and green. Although a ketolase has been implicated in determining the colour, the underlying genetic basis of body colour divergence between the two forms has remained unclear. Based on a combination of comparative transcriptomes and RNA interference, we found that a gene encoding a cytochrome P450 enzyme of the CYP4 clan (CYP389B1) had remarkably high expression in adult females of the red T. urticae, as well as in hybrids obtained by crossing the red and green forms. Down-regulation of this gene by RNA interference resulted in decreased accumulation of red pigment. Up-regulation of the expressions of a scavenger receptor gene (SCARB1) and a mitochondrial glycine transporter (SLC25A38) also strongly contributed to red colour development in adult females. Suppressing the mRNA levels of these genes also resulted in reduced accumulation of red pigment in the three other spider mites with red body colour. Our results provide evidence that the body colour divergence between the two forms is caused by different expressions of pigmentation-related genes, and point to a possible role of a novel cytochrome P450 gene (CYP389B1) in regulating red-orange body colour. These findings expand the number of candidate cytochrome P450 genes involved in endogenous pigmentation and will help to understand their roles in determining colour patterns in mites and other species.

Dense vesicles (DVs) are Golgi-derived plant-specific carriers that mediate post-Golgi transport of seed storage proteins in angiosperms. How this process is regulated remains elusive. Here, we report a rice (Oryza sativa) mutant, named glutelin precursor accumulation8 (gpa8) that abnormally accumulates 57-kDa proglutelins in the mature endosperm. Cytological analyses of the gpa8 mutant revealed that proglutelin-containing DVs were mistargeted to the apoplast forming electron-dense aggregates and paramural bodies in developing endosperm cells. Differing from previously reported gpa mutants with post-Golgi trafficking defects, the gpa8 mutant showed bent Golgi bodies, defective trans-Golgi network (TGN), and enlarged DVs, suggesting a specific role of GPA8 in DV biogenesis. We demonstrated that GPA8 encodes a subunit E isoform 1 of vacuolar H+-ATPase (OsVHA-E1) that mainly localizes to TGN and the tonoplast. Further analysis revealed that the luminal pH of the TGN and vacuole is dramatically increased in the gpa8 mutant. Moreover, the colocalization of GPA1 and GPA3 with TGN marker protein in gpa8 protoplasts was obviously decreased. Our data indicated that OsVHA-E1 is involved in endomembrane luminal pH homeostasis, as well as maintenance of Golgi morphology and TGN required for DV biogenesis and subsequent protein trafficking in rice endosperm cells.

Fusarium wilt caused byFusarium oxysporumf. sp.momordicae(FoM) is an important fungal disease that affects the production of bitter gourd. Hypovirulence-associated mycoviruses have great potential and application prospects for controlling the fungal disease. In this study, a novel ourmia-like virus, namedFusarium oxysporumourmia-like virus 1 (FoOuLV1), was isolated fromFoMstrain HuN8. The viral genomic RNA is 2,712 nucleotides (nt) in length and contains an open reading frame (ORF) encoding a putative RNA-dependent RNA polymerase (RdRp) using either standard or mitochondrial codes. In strain HuN8, there was also a FoOuLV1-associated RNA segment with 1,173 nt in length with no sequence homology. Phylogenetic analysis showed that FoOuLV1 is a member of the genusMagoulivirusof the familyBotourmiaviridae. FoOuLV1 was found to be associated with hypovirulence inFoM. Moreover, FoOuLV1 and its hypovirulence trait can be transmitted horizontally to otherFoMstrains and also to other formae speciale strains ofF. oxysporum. In addition, FoOuLV1 showed significant biological control effect against the bitter gourd Fusarium wilt. To our knowledge, this study reveals the first description of a hypovirulence-associated ourmia-like mycovirus, which has the potential to the biological control of Fusarium wilt.

A series of nucleotide sugar interconversion enzymes (NSEs) generate the activated sugar donors required for biosynthesis of cell wall matrix polysaccharides and glycoproteins. UDP-glucose 4-epimerases (UGEs) are NSEs that function in the interconversion of UDP-glucose (UDP-Glc) and UDP-galactose (UDP-Gal). The roles of UDP-glucose 4-epimerases in monocots remain unclear due to redundancy in the pathways. Here, we report a brittle plant (bp1) rice mutant that exhibits brittle leaves and culms at all growth stages. The mutant culms had reduced levels of rhamnogalacturonan I, homogalacturonan, and arabinogalactan proteins. Moreover, the mutant had altered contents of uronic acids, neutral noncellulosic monosaccharides, and cellulose. Map-based cloning demonstrated that OsBP1 encodes a UDP-glucose 4-epimerase (OsUGE2), a cytosolic protein. We also show that BP1 can form homo- and hetero-protein complexes with other UGE family members and with UDP-galactose transporters 2 (OsUGT2) and 3 (OsUGT3), which may facilitate the channeling of Gal to polysaccharides and proteoglycans. Our results demonstrate that BP1 participates in regulating the sugar composition and structure of rice cell walls.

Mutations of stromal Hsp70 cause chloroplast developmental abnormalities and knockout mutants of stromal Hsp70 usually exhibit protein import deficiencies. However, their effects have not been studied in amyloplast development. Here, we identified an amyloplast abnormal development mutant, floury endosperm11 (flo11) that exhibited an opaque phenotype in the inner core and the periphery of grains. Semi-thin section revealed defective amyloplast development in the flo11 endosperm. Map-based cloning and subsequent complementation test demonstrated that FLO11 encoded a plastid-localized heat shock protein 70 (OsHsp70cp-2). OsHsp70cp-2 was abundantly expressed in developing endosperm, whereas its paralogous gene OsHsp70cp-1 was mainly expressed in photosynthetic tissues. Ectopic expression of OsHsp70cp-1 under the control of OsHsp70cp-2 promoter rescued the mutant phenotype of flo11. Moreover, simultaneous knockdown of both OsHsp70cp genes resulted in white stripe leaves and opaque endosperm. BiFC and Co-IP assays revealed that OsHsp70cp-2 was associated with Tic complex. Taken together, OsHsp70cp-2 may regulate protein import into amyloplasts, which is essential for amyloplast development in rice.

Maslinic acid (2α,3β-dihydroxyolean-12-en-28-oic acid) is a naturally occurring pentacyclic triterpenic compound. Maslinic acid is gradually gaining attention as an excellent pharmacologically active product because of its premium biological properties. In this review we will focus on chemopreventive properties of Maslinic acid against different cancers. Seemingly, available data is limited and we have yet to unravel how Maslinic acid therapeutically targeted oncogenic cell signal transduction cascades in different cancers. Moreover, there are visible knowledge gaps about the ability of Maslinic acid to modulate oncogenic and tumor suppressor microRNAs in various cancers.

Artificial induction of polyploidy is an efficient technique for improving the biological properties and developing new varieties of many plants. Autotetraploid plants were generated through in vitro chromosome doubling of the ‘Hanfu’ apple cultivar (Malus × domestica) using colchicine treatment. In the present study, we analysed and compared the differences in the morphological and biological characteristics of diploid and autotetraploid ‘Hanfu’ apple cultivars from 2009 to 2014. We found that the autotetraploid plants flowered at 5 years of age, which was 2 years later than in the diploid plants. Compared with the diploid plants, the autotetraploid plants showed retarded growth and significant dwarfing characteristics before flowering, but their growth was restored after flowering. The stomata were larger and the leaves were not only larger and heavier but also darker green in the autotetraploid than in the diploid plants. The chlorophyll content of the leaves was significantly higher and the photosynthetic efficiency and fluorescence performance were increased in the autotetraploid plants. Additionally, the volume and size of the flowers, fruits and seeds were greater and the soluble solid contents were higher in the autotetraploid than its diploid counterparts. In this study, we provide the first detailed analysis of the morphological and biological characteristics associated with growth and development in diploid and autotetraploid ‘Hanfu’ apple, and the results will make a meaningful contribution to the breeding of polyploid apples.

PurposeThe design of a macro-prudential framework and its interaction with monetary policy has been at the forefront of the policy agenda since the global financial crisis. However, most advanced economies (AEs) have little experience using macroprudential policies. As a result, relatively little is known empirically about macroprudential instruments’ effectiveness in mitigating systemic risks in these countries, about their channels of transmission, and about how these instruments would interact with monetary policy. This paper aims to fill in the gap.Design/methodology/approachThe authors develop a new approach using the euro area bank lending survey to assess the effectiveness of macro-prudential policies in containing credit growth and house price appreciation in mortgage markets. Estimation is performed under the panel regressions (OLS, GLS) and panel VAR setup. Endogeneity issues arising from measures of macro-prudential policies are addressed by introducing GMM estimation and various instruments.FindingsThe authors find instruments targeting the cost of bank capital most effective in slowing down mortgage credit growth, and that the impact is transmitted mainly through price margins, the same banking channel as monetary policy. Limits on loan-to-value ratios are also effective, especially when monetary policy is excessively loose.Originality/valueWith limited data on macroprudential policy measures in the AEs, this paper proposed a new methodology of using answers from bank lending survey as proxies to assess the effectiveness of specific macroprudential measures and their transmission channels.

Insecticide resistance frequently results from target-site insensitivity, such as point mutations in acetylcholinesterases (AChEs) for resistance to organophosphates and carbamates. From a field-originated population of Nilaparvata lugens, a major rice pest, a resistant population (R9) was obtained by nine-generation continuous selection with chlorpyrifos. From the same field population, a relatively susceptible population (S9) was also constructed through rearing without any insecticides. Compared to the susceptible strain, Sus [medium lethal dose (LC50) = 0.012 mg/l], R9 had a resistance ratio (RR) of 253.08-fold, whereas the RR of S9 was only 2.25-fold. Piperonyl butoxide and triphenyl phosphate synergized chlorpyrifos in R9 less than three-fold, indicating other important mechanisms for high resistance. The target-site insensitivity was supported by the key property differences of crude AChEs between R9 and S9. Compared to S9, three mutations (G119S, F331C and I332L) were detected in NlAChE1 from individuals of the R9 and field populations, but no mutation was detected in NlAChE2. G119S and F331C could decreased insecticide sensitivities in recombinant NlAChE1, whereas I332L took effect through increasing the influence of F331C on target insensitivity. F331C might be deleterious because of its influence on the catalytic efficiency of NlAChE1, whereas I332L would decrease these adverse effects and maintain the normal functions of AChEs.

Low impact development (LID) facilities have ability to increase evapotranspiration (ET), and thus moderate the hydrological cycle and mitigate urban heat island (UHI) effects. However, it is difficult to measure ET of LID facilities by traditional methods (e.g. weighing lysimeter, Bowen ratio system and eddy covariance system) in urban environment. A recently developed method, called three-temperature model (3T model), has been used to measure actual ET of natural underlying surfaces in situ. However, its applicability in ET measurement for LID facilities has not been investigated. In order to investigate the accuracy of this method to estimate ET of LID facilities and its influencing factors, the experimental devices for vegetated LID (green roof with Zoysia tenuifoli (GRZ) and green roof with Callisia repens (GRC)) and non-vegetated LID (permeable concrete pavement (PCP) and permeable brick pavement (PBP)) were installed. The method was validated by comparing the ET measured by 3T model with that measured by weighing platform (hourly data, n = 223–302 for each facility). After validation, the method was used to measure ET variations of typical LID facilities under moist sub-tropical weather condition in Shenzhen, China. The results indicate the coefficient of determination (R2) between ET measured by 3T model and weighing platform is 0.63, 0.60, 0.70 and 0.65 for GRZ, GRC, PCP and PBP, respectively, and the slopes of regression equations are close to 1 and their intercepts are close to 0. Furthermore, the 3T model has higher absolute relative error (ARE) of ET estimation in the early morning and the late afternoon than that in the rest of daytime. This is mainly due to the effect of energy related factors, such as solar radiation, air temperature and the difference between reference surface temperature and air temperature. In addition, ET rates of LID facilities can maintain at a relatively high level in the first few days after rainfall. However, ET of PCP decreases faster than that of other LID facilities during the drying period. These results demonstrate the ability and accuracy of 3T model to measure variations of ET of LID facilities in situ.

Figure S6. Complementation of gpa6 mutant phenotypes by p35S:OsNHX5-GFP. (a) Immunoblot analysis with monoclonal GFP antibodies. (b) p35S:OsNHX5-GFP transgene rescued the grain phenotype of the gpa6 mutant. Barsâ =â 1â mm. (c) p35S:OsNHX5-GFP transgene in the gpa6 mutant reduce the amount of 57-KD proglutelins to a level comparable to the wild type. GL1 to GL3 denote the grains from three independent T1 transgenic lines. Red arrows indicate the 57-kD proglutelins. (DOCX 186 kb)

Table S4. Primers used for vector construction. (DOCX 14 kb)

Table S2. Segregation of mutant phenotypes in reciprocal crosses between the wild type and gpa6 mutant. (DOCX 13 kb)

Figure S7. Subcellular Localization of OsNHX6 in N. benthamiana protoplasts. (a) to (c) Confocal microscopy images showing that OsNHX6-GFP is localized as punctate signals in the cytosol and its distribution partially overlaps with the markers for Golgi (ST-mRFP [a]), TGN (mRFP-SYP61 [b]) and PVC (mRFP-VSR2 [c]). Bars = 10 μm (a-c). (DOCX 112 kb)

Figure S2. RT-qPCR assay of the expression of representative genes coding for storage proteins in 12-DAF endosperm. Glutelin genes: GluA1, GluB2, GluC1, GluD1; prolamin genes: pro10.1, pro16.2, pro13a.2, pro13b.2. Values are means Âą SD. n =â 3. (DOCX 81 kb)

Figure S3. Immunofluorescence microscopy of protein bodies in the subaleurone cells of the wild type and gpa6 mutant. (a) to (f) Immunofluorescence microscopy images of storage proteins in wild-type (a-c) and gpa6 (d-f) 12 DAF seeds. (a, d) Secondary antibodies conjugated with Alexa fluor 555 (red) were used to trace the antigens recognized by the anti-α-globulin antibodies. (b, e) Secondary antibodies conjugated with Alexa fluor 488 (green) were used to trace the antigens recognized by the anti-glutelin antibodies. (c, f) Merged images. White arrowheads in (f) indicate the mis-sorted α-globulins in the PMB. Bars = 10 μm (a-f). (DOCX 135 kb)

Table S1. Properties of wild-type and gpa6 seeds. (DOCX 13 kb)

Figure S4. Distribution of cell wall materials in 12 DAF endosperm cells. (a) to (f) Sections of 12 DAF endosperms from wild type (a-c) and gpa6 (d-f) plant were incubated with pectin (JIM7) or glutelin antibodies, followed by secondary antibodies conjugated to Alexa-555 or Alexa-488. Bars = 10 μm (a-f). (DOCX 152 kb)

Figure S5. Amino acid sequences alignment of OsNHX5 and OsNHX6. (DOCX 421 kb)

Starch synthesized and stored in amyloplasts serves as the major energy storage molecule in cereal endosperm. To elucidate the molecular mechanisms underlying amyloplast development and starch synthesis, we isolated a series of floury endosperm mutants in rice (Oryza sativa). We identified the rice mutant floury shrunken endosperm1 (fse1), which exhibited obvious defects in the development of compound starch grains, decreased starch content, and altered starch physicochemical features. Map-based cloning showed that FSE1 encodes a phospholipase-like protein homologous to phosphatidic acid-preferring phospholipase A(1). FSE1 was expressed ubiquitously with abundant levels observed in developing seeds and roots. FSE1 was localized to both the cytosol and intracellular membranes. Lipid profiling indicated that total extra-plastidic lipids and phosphatidic acid were increased in fse1 plants, suggesting that FSE1 may exhibit in vivo phospholipase A(1) activity on phosphatidylcholine, phosphatidylinositol, phosphatidyl-Ser, phosphatidylethanolamine, and, in particular, phosphatidic acid. Additionally, the total galactolipid content in developing fse1 endosperm was significantly reduced, which may cause abnormal amyloplast development. Our results identify FSE1 as a phospholipase-like protein that controls the synthesis of galactolipids in rice endosperm and provide a novel connection between lipid metabolism and starch synthesis in rice grains during endosperm development.

Maslinic acid (2α,3β-dihydroxyolean-12-en-28-oic acid) is a naturally occurring pentacyclic triterpenic compound. Maslinic acid is gradually gaining attention as an excellent pharmacologically active product because of its premium biological properties. In this review we will focus on chemopreventive properties of Maslinic acid against different cancers. Seemingly, available data is limited and we have yet to unravel how Maslinic acid therapeutically targeted oncogenic cell signal transduction cascades in different cancers. Moreover, there are visible knowledge gaps about the ability of Maslinic acid to modulate oncogenic and tumor suppressor microRNAs in various cancers.

An albinic rice is caused by mutation of threonyl-tRNA synthetase, which is essential for plant development by stabilizing of NEP and PEP gene expressions and chloroplast protein synthesis. Chloroplast biogenesis and development depend on complex genetic mechanisms. Apart from their function in translation, aminoacyl-tRNA synthetases (aaRSs) play additional role in gene expression regulation, RNA splicing, and cytokine activity. However, their detailed functions in plant development are still poorly understood. We isolated a lethal albinic seedling (las) mutant in rice. Physiological and ultrastructural analysis of las mutant plants revealed weak chlorophyll fluorescence, negligible chlorophyll accumulation, and defective thylakoid membrane development. By map based cloning we determined that the LAS allele gene encodes threonyl-tRNA synthetase (ThrRS). LAS was constitutively expressed with relatively high level in leaves. NEP-dependent gene transcripts accumulated in the developing chloroplasts, while PEP-dependent transcripts were reduced in the las mutant. This result indicated that PEP activity was impaired. Chloroplast-encoded protein levels were sharply reduced in the las mutant. Biogenesis of chloroplast rRNAs (16S and 23S rRNA) was arrested, leading to impaired translation and protein synthesis. Together, our findings indicated that LAS is essential not only for chloroplast development by stabilizing the NEP and PEP gene expression, but also for protein synthesis and construction of the ribosome system in rice chloroplasts.

A number of East Asian and oil-exporting countries have generated a large inflow of foreign currencies as a result of their continued trade surplus and surging foreign investments in recent decades. In China, the booming foreign inflow was accompanied by a modest appreciation of the real exchange rate. This paper argues that the failure of the real exchange rate to appreciate in China is more the result of a higher demand for real monetary balances than of exchange rate manipulation. Such "sterilization by the people" is more evident in earlier episodes than in more recent ones. This can be due to the emergence of competitive financial instruments, a deeper financial market, and a more developed social security system.

Urban sprawl is a major driving force that alters local and regional hydrology and increases non-point source pollution. Using the Bao’an District in Shenzhen, China, a typical rapid urbanization area, as the study area and land-use change maps from 1988 to 2014 that were obtained by remote sensing, the contributions of different land-use types to NPS pollutant production were assessed with a localized long-term hydrologic impact assessment (L-THIA) model. The results show that the non-point source pollution load changed significantly both in terms of magnitude and spatial distribution. The loads of chemical oxygen demand, total suspended substances, total nitrogen and total phosphorus were affected by the interactions between event mean concentration and the magnitude of changes in land-use acreages and the spatial distribution. From 1988 to 2014, the loads of chemical oxygen demand, suspended substances and total phosphorus showed clearly increasing trends with rates of 132.48 %, 32.52 % and 38.76 %, respectively, while the load of total nitrogen decreased by 71.52 %. The immigrant population ratio was selected as an indicator to represent the level of rapid urbanization and industrialization in the study area, and a comparison analysis of the indicator with the four non-point source loads demonstrated that the chemical oxygen demand, total phosphorus and total nitrogen loads are linearly related to the immigrant population ratio. The results provide useful information for environmental improvement and city management in the study area.

The changes in content and form of polyamines (PAs) in the leaves of two wheat ( Triticum aestivum L.) cultivar seedlings, differing in drought tolerance, were investigated under osmotic stress. Yumai No. 18 cv. (drought-tolerant) showed marked increases of free-spermidine (free-Spd) and free-spermine (free-Spm) levels in leaves after polyethylene glycol (PEG) 6000 treatment for 7 days, while Yangmai No. 9 cv. (drought-sensitive) showed a significant increase of free-putrescine (free-Put) level. Thus, Yumai No. 18 cv. gave a higher ratio of (free-Spd+free-Spm)/free-Put than Yangmai No. 9 cv. in leaves in response to osmotic stress. Exogenous treatment of seedlings with methylglyoxal-bis-(guanyl-hydrazone) (MGBG), an S -adenosylmethionine decarboxylase (SAMDC) inhibitor, resulted in reduction of free-Spd and -Spm levels in leaves and aggravation of PEG-induced injury to Yumai No. 18 cv. seedlings, while exogenous Spd treatment resulted in an increase of free-Spd+free-Spm in leaves, in concert with an alleviation of PEG-induced injury to Yangmai No. 9 cv. seedlings. There was no statistically significant difference between the contents of perchloric-acid soluble conjugated PAs (PS-conjugated PAs) in leaves of both cultivars under PEG osmotic stress, suggesting that the level of PS-conjugated PAs in leaves has no relationship with the osmotic stress tolerance of wheat seedlings. In contrast, osmotic stress induced a significant increase of perchloric acid insoluble bound putrescine (PIS-bound Put) level in seedling leaves of Yumai No. 18 cv., whereas osmotic stress affected slightly the PIS-bound Put level in Yangmai No. 9 cv. seedlings. Exogenous treatment of seedlings with phenanthrolin ( o -Phen), an inhibitor of transglutaminase (TGase), also aggravated PEG-induced injury to Yumai No. 18 cv. seedlings, coupled with a decrease of PIS-bound Put level. The results suggested that free-Spd, -Spm and PIS-bound Put facilitated the osmotic stress tolerance of wheat seedlings.

Neonicotinoid insecticides, such as imidacloprid, are selective agonists of the insect nicotinic acetylcholine receptors (nAChRs) and extensively used in areas of crop protection and animal health to control a variety of insect pest species. Here we describe that two cis-nitromethylene neonicotinoids (IPPA152002 and IPPA152004), recently synthesized in our laboratory, discriminated between the high and low affinity imidacloprid binding sites in the brown planthopper, Nilaparvata lugens, a major insect pest of rice crops in many parts of Asia. [(3)H]imidacloprid has two binding sites with different affinities (Kd value of 0.0035 +/- 0.0006 nM for the high-affinity site and 1.47 +/- 0.22 nM for the low-affinity site). Although the cis-nitromethylene neonicotinoids showed low displacement ability (Ki values of 0.15 +/- 0.03 microM and 0.42 +/- 0.07 microM for IPPA152002 and IPPA152004, respectively) against [(3)H]imidacloprid binding, low concentrations (0.01 microM) of IPPA152002 completely inhibited [(3)H]imidacloprid binding at its high-affinity site. In Xenopus oocytes co-injected with cRNA encoding Nlalpha1 and rat beta2 subunits, obvious inward currents were detected in response to applications of IPPA152002 and IPPA152004, although the agonist potency is reduced to that of imidacloprid. The previously identified Y151S mutation in Nlalpha1 showed significant effects on the agonist potency of IPPA152002 and IPPA152004, such as a 75.8% and 70.6% reduction in Imax, and a 2.4- and 2.1-fold increase in EC(50). This data clearly shows that the two newly described cis-nitromethylene neonicotinoids act on insect nAChRs and like imidacloprid, discriminated between high and low affinity binding sites in N. lugens native nAChRs. These compounds may be useful tools to further elucidate the pharmacology and nature of neonicotinoid binding sites.