PROTEIN PHOSPHATASE95 Regulates Phosphate Homeostasis by Affecting Phosphate Transporter Trafficking in Rice.

Phosphate (Pi) uptake in plants depends on plasma membrane (PM)-localized phosphate transporters (PTs). OsCK2 phosphorylates PTs and inhibits their trafficking from the endoplasmic reticulum (ER) to the PM in rice (Oryza sativa), but how PTs are dephosphorylated is unknown. We demonstrate that the protein phosphatase type 2C (PP2C) protein phosphatase OsPP95 interacts with OsPT2 and OsPT8 and dephosphorylates OsPT8 at Ser-517. Rice plants overexpressing OsPP95 reduced OsPT8 phosphorylation and promoted OsPT2 and OsPT8 trafficking from the ER to the PM, resulting in Pi accumulation. Under Pi-sufficient conditions, Pi levels were lower in young leaves and higher in old leaves in ospp95 mutants than in those of the wild type, even though the overall shoot Pi levels were the same in the mutant and the wild type. In the wild type, OsPP95 accumulated under Pi starvation but was rapidly degraded under Pi-sufficient conditions. We show that OsPHO2 interacts with and induces the degradation of OsPP95. We conclude that OsPP95, a protein phosphatase negatively regulated by OsPHO2, positively regulates Pi homeostasis and remobilization by dephosphorylating PTs and affecting their trafficking to the PM, a reversible process required for adaptation to variable Pi conditions.

OsbHLH6 interacts with OsSPX4 and regulates the phosphate starvation response in rice.

Low soil phosphorus (P) availability is a major limitation for crop production. The molecular mechanisms underlying plant responses and adaptation to phosphate (Pi) deficiency are unclear. OsbHLH6 (hereafter bHLH6), an uncharacterized rice (Oryza sativa) Pi starvation response gene encoding a basic helix-loop-helix protein, was identified by yeast two-hybrid screening using the phosphate response repressor OsSPX4 (hereafter SPX4) as bait. bHLH6 is expressed in shoots and roots, and its expression is significantly induced in shoots by Pi deficiency. bHLH6 overexpression lines showed Pi accumulation and enhanced Pi starvation responses, including upregulation of Pi starvation-induced genes and longer root hairs. A bhlh6 mutant showed no significant phenotype variation at the seedling stage. A pull-down assay indicated that bHLH6 had higher binding affinity with SPX4 compared to OsPHR2; therefore, bHLH6 competitively inhibited the interaction of SPX4 and OsPHR2. SPX4 overexpression rescued the Pi accumulation caused by bHLH6 overexpression under high- and low-P conditions. Moreover, overexpression of bHLH6 in an spx4 background did not affect the Pi content of spx4 under high- and low-P conditions. The bhlh6 spx4 double mutant showed lower shoot Pi concentrations and transcript levels of OsPT3 and OsPT10 compared with the spx4 mutant under high-P conditions. RNA sequencing results indicated that bHLH6 overexpression and spx4 mutant lines share many differentially expressed Pi-responsive genes. Therefore, bHLH6 is an important regulator for Pi signaling and homeostasis which antagonizes SPX4. This knowledge helps elucidate the molecular regulation of plant adaptation to Pi deficiency and will promote efforts toward the creation of low Pi-tolerant crops.

OsSPL3, an SBP-Domain Protein, Regulates Crown Root Development in Rice.

The major root system of cereals consists of crown roots (or adventitious roots), which are important for anchoring plants in the soil and for water and nutrient uptake. However, the molecular basis of crown root formation is largely unknown. Here, we isolated a rice (Oryza sativa) mutant with fewer crown roots, named lower crown root number1 (lcrn1). Map-based cloning revealed that lcrn1 is caused by a mutation of a putative transcription factor-coding gene, O. sativa SQUAMOSA PROMOTER BINDING PROTEIN-LIKE3 (OsSPL3). We demonstrate that the point mutation in lcrn1 perturbs theO. sativa microRNA156 (OsmiR156)-directed cleavage of OsSPL3 transcripts, resulting in the mutant phenotype. Chromatin immunoprecipitation sequencing assays of OsSPL3 binding sites and RNA sequencing of differentially expressed transcripts in lcrn1 further identified potential direct targets of OsSPL3 in basal nodes, including a MADS-box transcription factor, OsMADS50. OsMADS50-overexpressing plants produced fewer crown roots, phenocopying lcrn1, while knocking out OsMADS50 in the lcrn1 background reversed this phenotype. We also show that OsSPL12, another OsmiR156 target gene, regulates OsMADS50 and crown root development. Taken together, our findings suggest a novel regulatory pathway in which the OsmiR156-OsSPL3/OsSPL12 module directly activates OsMADS50 in the node to regulate crown root development in rice.

CASEIN KINASE2-Dependent Phosphorylation of PHOSPHATE2 Fine-tunes Phosphate Homeostasis in Rice.

Plants have evolved complex physiological and biochemical mechanisms to adapt to a heterogeneous soil phosphorus environment. PHOSPHATE2 (PHO2) is a phosphate (Pi) starvation-signaling regulator involved in maintaining Pi homeostasis in plants. Arabidopsis (Arabidopsis thaliana) PHO2 targets PHOSPHATE TRANSPORTER1 (PHT1) and PHO1 for degradation, whereas rice (Oryza sativa) PHO2 is thought to mediate PHOSPHATE TRANSPORTER TRAFFIC FACILITATOR1 degradation. However, it is unclear whether and how PHO2 is post-translationally regulated. Here, we show that in rice, the CASEIN KINASE2 (OsCK2) catalytic subunit OsCK2α3 interacts with OsPHO2 in vitro and in vivo in vascular tissues cells, and phosphorylates OsPHO2 at Ser-841. Phosphorylated OsPHO2 is degraded more rapidly than native OsPHO2 in cell-free degradation assays. OsPHO2 interacts with OsPHO1 and targets it for degradation through a multivesicular body-mediated pathway. PHO1 mutation partially rescued the pho2 mutant phenotype. Further genetic analysis showed that a nonphosphorylatable version of OsPHO2 rescued the Ospho2 phenotype of high Pi accumulation in leaves better than native OsPHO2. In addition to the previously established role of OsCK2 in negatively regulating endoplasmic reticulum exit of PHT1 phosphate transporters, this work uncovers a role for OsCK2α3 in modulating Pi homeostasis through regulating the phosphorylation status and abundance of OsPHO2 in rice.

Rice SPX6 negatively regulates the phosphate starvation response through suppression of the transcription factor PHR2.

Phosphorus (P) is an essential macronutrient for plant growth and development, but the molecular mechanism determining how plants sense external inorganic phosphate (Pi) levels and reprogram transcriptional and adaptive responses is incompletely understood. In this study, we investigated the function of OsSPX6 (hereafter SPX6), an uncharacterized member of SPX domain (SYG1, Pho81 and XPR1)-containing proteins in rice, using reverse genetics and biochemical approaches. Transgenic plants overexpressing SPX6 exhibited decreased Pi concentrations and suppression of phosphate starvation-induced (PSI) genes. By contrast, transgenic lines with decreased SPX6 transcript levels or spx6 mutant showed significant Pi accumulation in the leaf and upregulation of PSI genes. Overexpression of SPX6 genetically suppressed the overexpression of PHOSPHATE STARVATION RESPONSE REGULATOR 2 (PHR2) in terms of the accumulation of high Pi content. Moreover, direct interaction of SPX6 with PHR2 impeded PHR2 translocation into the nucleus, and inhibited PHR2 binding to the P1BS (PHR1 binding sequence) element. SPX6 protein was degraded in leaves under Pi-deficient conditions, whereas it accumulated in roots. We conclude that rice SPX6 is another important negative regulator in Pi starvation signaling through the interaction with PHR2. SPX6 shows different responses to Pi starvation in shoot and root, which differ from those of other SPX proteins.

[Research Progress of Raman Spectroscopy on Dyestuff Identification of Ancient Relics and Artifacts].

As the birthplace of Silk Road, China has a long dyeing history. The valuable information about the production time, the source of dyeing material, dyeing process and preservation status were existed in organic dyestuff deriving from cultural relics and artifacts. However, because of the low contents, complex compositions and easily degraded of dyestuff, it is always a challenging task to identify the dyestuff in relics analyzing field. As a finger-print spectrum, Raman spectroscopy owns unique superiorities in dyestuff identification. Thus, the principle, characteristic, limitation, progress and development direction of micro-Raman spectroscopy (MRS/µ-Raman), near infrared reflection and Fourier transform Raman spectroscopy (NIR-FT-Raman), surface-enhanced Raman spectroscopy (SERS) and resonance raman spectroscopy (RRS) have been introduced in this paper. Furthermore, the features of Raman spectra of gardenia, curcumin and other natural dyestuffs were classified by MRS technology, and then the fluorescence phenomena of purpurin excitated with different wavelength laser was compared and analyzed. At last, gray green silver colloidal particles were made as the base, then the colorant of madder was identified combining with thin layer chromatography (TLC) separation technology and SERS, the result showed that the surface enhancement effect of silver colloidal particles could significantly reduce fluorescence background of the Raman spectra. It is pointed out that Raman spectroscopy is a rapid and convenient molecular structure qualitative methodology, which has broad application prospect in dyestuff analysis of cultural relics and artifacts. We propose that the combination of multi-Raman spectroscopy, separation technology and long distance transmission technology are the development trends of Raman spectroscopy.

[Study of Paints and Drawing Techniques of Fine Brushwork Yunlong Ripples Painting in Qing Dynasty].

In order to study the paints and techniques of decorative patterns of dragon among clouds and water waves, the materials based on a Qing Dynasty meticulous painting were measured by three-dimensional video microscopy, Raman microscopy and energy dispersive X-ray fluorescence spectroscopy. The results showed that the green clothes was firstly colored by Paris green, the decorative patterns of dragon among clouds and water waves were then painted by hematite, the edge was delineated by brass powder at last. The dark yellow area within the decorative patterns was presented due to the interaction of green and red paints. In addition, ultramarine blue was checked in the painting. According to the first synthesized time of ultramarine blue and Paris green, we can make sure the time limit of the painting finished.

[Raman microspectroscopic analysis of polychrome wooden artifact from the Astana tomb of the tang dynasty, Xinjiang, China].

The Turpan District is characterized by a typical dry climate, so that many organic relics have been well preserved. A piece of wooden artifact with 9 colors was unearthed from Astana graveyard which is rich in cultural connotations. In the present paper, Raman microscopy was employed for in-situ, nondestructive analysis of pigments that remained on this artifact, and many mineral pigments (gypsum, red lead, carbon black, haematite, atacamite) and vegetable dyes (gamboges and indigo) were identified. It is noteworthy that this is the earliest example that gamboges were used as yellow dye in China at present. The results show that the Gaochang people had mastered skills proficiently, including the preparation, deployment and usage of pigment. The investigation of pigments could provide a basis for the restoration and conservation of relies, and more evidence for pigments trade business and cultural exchanges.

Copper assisted sequence-specific chemical protein conjugation at a single backbone amide.

Direct, site-specific methods of protein functionalization are highly desirable for biotechnology. However, such methods are challenging due to the difficulty of chemically differentiating a single site within a large protein. Herein, we propose "metal binding targeting" strategy and develop a Copper Assisted Sequence-specific conjugation Tag (CAST) method to achieve rapid (second order rate 8.1 M-1 s-1), site-specific protein backbone chemical modification with pinpoint accuracy. We demonstrate the versatility of CAST conjugation by preparing various on-demand modified recombinant proteins, including a homogeneous antibody-drug conjugate with high plasma stability and potent efficacy in vitro and in vivo. Thus, CAST provides an efficient and quantitative method to site-specifically attach payloads on large, native proteins.

The first record of the monospecific genus Rhinopalpa (Lepidoptera: Nymphalidae) from China.

BACKGROUND: The family Nymphalidae is the largest group of butterflies with high species richeness. Rhinopalpapolynice (Cramer, [1779]), a forest species, was discovered in the mid-stream of the Yuanjiang-Red River Valley of Yunnan Province for the first time, which represents the first record of the genus Rhinopalpa in China. NEW INFORMATION: The species R. polynice (Cramer, [1779]) is the first record of the genus Rhinopalpa from China. The specimen was collected in the mid-stream of the Yuanjiang-Red River Valley of Yunnan Province. The female genitalia are described for the first time.

The complete mitochondrial genome of Pycanum ochraceum Distant 1893 (Hemiptera: Tessaratomidae).

In the present study, the complete mitochondrial genome of Pycanum ochraceum was identified for the first time. The entire genome is 17,198 bp in length with 73.6% A + T content. It contains 22 transfer RNA genes (tRNAs), 2 ribosomal RNA genes (rRNAs), 13 protein-coding genes (PCGs) and 1 noncoding control region (D-loop). Phylogenetic analysis showed that Tessaratomidae bugs are monophyletic. This study can provide essential DNA molecular data for further phylogenetic and evolutionary analysis for Heteroptera.

Constructing an unprecedented {MnII38} matryoshka doll with a [Mn18(CO3)9] inorganic core and magnetocaloric effect.

An unprecedented inner [Mn18(CO3)9] inorganic core and [Mn20] metal-organic periphery compose a high-nuclearity homometallic single-valent {MnII38} molecular aggregate with a [Mn6] ⊂ [Mn12] ⊂ [Mn8] ⊂ [Mn12] matryoshka doll-like skeleton that displays a significant magnetocaloric effect (MCE).

Complete mitochondrial genome of the Endangered long-armed scarab Cheirotonus gestroi (Coleoptera: Euchiridae).

The long-armed scarab Cheirotonus gestroi is an endangered large insect in southwestern China and neighboring countries. Herein, we present the first complete mitochondrial genome of C. gestroi. The 16,899 bp long circular genome consists of 2 ribosomal RNA genes, 22 transfer RNA genes, 13 protein-coding genes, and a non-coding control region. Phylogenetic analysis showed that C. gestroi shared the closest evolutionary relationship with C. jansoni and that Scarabaeidae and Lucanidae were correctly identified within superfamily Scarabaeoidea. The complete mitogenome sequence will provide a basis for further phylogenetic studies and conservation genetics of the genus Cheirotonus.

Strong genetic differentiation among populations of Cheirotonus gestroi (Coleoptera: Euchiridae) in its native area sheds lights on species conservation.

The long-armed scarab (Cheirotonus gestroi) is an endangered large insect in southwestern China and neighboring countries; however, limited information is available regarding its population genetics, hindering conservation efforts. Therefore, we investigated the population genetic structure and evolutionary history of C. gestroi in southwestern China. Twenty-five haplotypes were obtained from 47 specimens across five populations. The Dawei Mountain (DWS) population differed from other populations by a high genetic distance. Population structure analysis generated three distinct clades, corresponding to Hengduan Mountains (HM), Ailao Mountains (AM), and Dawei Mountains (DM), and high-level genetic diversity was found in two HM populations. Collectively, the strong genetic differentiation among populations might be due to limited gene flow, geographical isolation, and habitat fragmentation. Therefore, while developing a conservation strategy, HM, AM, and DM groups should be defined as separate management units. Additionally, the DWS population should be given priority protection due to its uniqueness and low genetic diversity.

Aqueous compatible boron nitride nanosheets for high-performance hydrogels.

Hexagonal boron nitride nanosheets (BNNSs) possess ultimate thermal and chemical stabilities and mechanical strengths. However, the unmodified BNNSs are hydrophobic and insoluble in water, which hinders their use in many technological areas requiring aqueous compatibility. In this work, h-BN was treated with molten citric acid to produce aqueous dispersible boron nitride sheets (ca-BNNSs). The resultant ca-BNNSs were used to fabricate ca-BNNS/polyacrylamide (i.e., BNNS2.5/PAAm) nanocomposite hydrogels, targeting high water retentivity and flexibility. The BNNS2.5/PAAm hydrogel (initially swollen in water) largely remained swollen (water content ∼94 wt%) even after one-year storage under ambient conditions. Importantly, the swollen BNNS2.5/PAAm hydrogel (water content ∼95 wt%) was highly flexible. Its elongation and compressive strength exceeded 10,000% and 8 MPa at 97% strain, respectively. Moreover, the aforementioned hydrogel recovered upon the removal of compression force, without obvious damage. The substantially improved water retentivity and flexibility revealed that BNNSs can serve as a promising new platform in the development of high-performance hydrogels.