RGA1 alleviates low-light-repressed pollen tube elongation by improving the metabolism and allocation of sugars and energy.

Low-light stress compromises photosynthetic and energy efficiency and leads to spikelet sterility; however, the effect of low-light stress on pollen tube elongation in the pistil remains poorly understood. The gene RGA1, which encodes a Gα-subunit of the heterotrimeric G-protein, enhanced low-light tolerance at anthesis by preventing the cessation of pollen tube elongation in the pistil of rice plants. In this process, marked increases in the activities of acid invertase (INV), sucrose synthase (SUS) and mitochondrial respiratory electron transport chain complexes, as well as the relative expression levels of SUTs (sucrose transporter), SWEETs (sugars will eventually be exported transporters), SUSs, INVs, CINs (cell-wall INV 1), SnRK1A (sucrose-nonfermenting 1-related kinase 1) and SnRK1B, were observed in OE-1 plants. Accordingly, notable increases in contents of ATP and ATPase were presented in OE-1 plants under low-light conditions, while they were decreased in d1 plants. Importantly, INV and ATPase activators (sucrose and Na2 SO3 , respectively) increased spikelet fertility by improving the energy status in the pistil under low-light conditions, and the ATPase inhibitor Na2 VO4 induced spikelet sterility and decreased ATPase activity. These results suggest that RGA1 could alleviate the low-light stress-induced impairment of pollen tube elongation to increase spikelet fertility by promoting sucrose unloading in the pistil and improving the metabolism and allocation of energy.

How Rice Plants Response to Abiotic Stresses.

With the combustion of fossil fuels, unequal and unsustainable energy and land use, and irrational human activities, greenhouse gas emissions remain high, which leads to global warming [...].

The Evolution and Functional Roles of miR408 and Its Targets in Plants.

MicroRNA408 (miR408) is an ancient and highly conserved miRNA, which is involved in the regulation of plant growth, development and stress response. However, previous research results on the evolution and functional roles of miR408 and its targets are relatively scattered, and there is a lack of a systematic comparison and comprehensive summary of the detailed evolutionary pathways and regulatory mechanisms of miR408 and its targets in plants. Here, we analyzed the evolutionary pathway of miR408 in plants, and summarized the functions of miR408 and its targets in regulating plant growth and development and plant responses to various abiotic and biotic stresses. The evolutionary analysis shows that miR408 is an ancient and highly conserved microRNA, which is widely distributed in different plants. miR408 regulates the growth and development of different plants by down-regulating its targets, encoding blue copper (Cu) proteins, and by transporting Cu to plastocyanin (PC), which affects photosynthesis and ultimately promotes grain yield. In addition, miR408 improves tolerance to stress by down-regulating target genes and enhancing cellular antioxidants, thereby increasing the antioxidant capacity of plants. This review expands and promotes an in-depth understanding of the evolutionary and regulatory roles of miR408 and its targets in plants.

Abscisic Acid Improves Rice Thermo-Tolerance by Affecting Trehalose Metabolism.

Heat stress that occurs during the flowering stage severely decreases the rice (Oryza sativa L.) seed-setting rate. This damage can be reversed by abscisic acid (ABA), through effects on reactive oxygen species, carbohydrate metabolism, and heat shock proteins, but the exact role of trehalose and ATP in this process remains unclear. Two rice genotypes, namely, Zhefu802 (heat-resistant plant, a recurrent parent) and its near-isogenic line (faded green leaf, Fgl, heat-sensitive plant), were subjected to 38 °C heat stress after being sprayed with ABA or its biosynthetic inhibitor, fluridone (Flu), at the flowering stage. The results showed that exogenous ABA significantly increased the seed-setting rate of rice under heat stress, by 14.31 and 22.40% in Zhefu802 and Fgl, respectively, when compared with the H2O treatment. Similarly, exogenous ABA increased trehalose content, key enzyme activities of trehalose metabolism, ATP content, and F1Fo-ATPase activity. Importantly, the opposite results were observed in plants treated with Flu. Therefore, ABA may improve rice thermo-tolerance by affecting trehalose metabolism and ATP consumption.

Acid invertase confers heat tolerance in rice plants by maintaining energy homoeostasis of spikelets.

Heat stress impairs both pollen germination and pollen tube elongation, resulting in pollination failure caused by energy imbalance. Invertase plays a critical role in the maintenance of energy homoeostasis; however, few studies investigated this during heat stress. Two rice cultivars with different heat tolerance, namely, TLY83 (heat tolerant) and LLY722 (heat susceptible), were subjected to heat stress. At anthesis, heat stress significantly decreased spikelet fertility, accompanied by notable reductions in pollen germination on stigma and pollen tube elongation in ovule, especially in LLY722. Acid invertase (INV), rather than sucrose synthase, contributed to sucrose metabolism, which explains the different tolerances of both cultivars. Under heat stress, larger enhancements in NAD(H), ATP, and antioxidant capacity were found in TLY83 compared with LLY722, whereas a sharp reduction in poly(ADP-ribose) polymerase (PARP) activity was found in the former compared with the latter. Importantly, exogenous INV, 3-aminobenzamide (a PARP inhibitor), sucrose, glucose, and fructose significantly increased spikelet fertility under heat stress, where INV activity was enhanced and PARP activity was inhibited. Therefore, INV can balance the energy production and consumption to provide sufficient energy for pollen germination and pollen tube growth under heat stress.

Epidemiological investigation of feline infectious peritonitis in cats living in Harbin, Northeast China from 2017 to 2019 using a combination of an EvaGreen-based real-time RT-PCR and serum chemistry assays.

Feline infectious peritonitis (FIP) is caused by the FIP virus (FIPV), a highly virulent mutant form of feline coronavirus (FCoV). This disease is one of the most important infectious diseases in cats, and it is associated with high mortality, particularly among younger cats. In this study, we isolated a wild-type FIPV HRB-17 epidemic strain from the blood sample of household pet cat exhibiting the characteristic wet-form FIP symptoms, which has been confirmed further by animal infection. Further, we developed an EvaGreen-based real-time RT-PCR assay for the accurate detection of FCoV based on the amplification of the highly conserved FIPV N gene. Then, using a combination of the real-time RT-PCR approach and a serum chemistry assay, we performed an epidemiological survey of FIPV infection in cats living in Harbin City, Northeast China. The results indicated that the EvaGreen-based real-time RT-PCR assay can be used for screening FCoV infection in the affected cats at an analytical detection limit of 8.2 × 101 viral genome copies/µL, but could not effectively distinguish FIPVs from FECVs. Additionally, the results of the epidemiological survey investigating feline blood samples (n = 1523) collected between July 2017 to July 2019 revealed an FIPV prevalence of approximately 12% (189/1523). Maybe, the prevalence would be less than 12% due to the real-time RT-PCR assay could not accurately differentiate FIPV and FECV. Nevertheless, it still highlighted the severity of the FIP epidemic in cats and reiterated the urgent need to develop effective anti-FIP therapeutic agents and anti-FIPV vaccines. As pet cats are household animals, risk communication and continuous region-extended surveillance cat programs are recommended.

Non-Photochemical Quenching Involved in the Regulation of Photosynthesis of Rice Leaves under High Nitrogen Conditions.

Excess and deficient nitrogen (N) inhibit photosynthesis in the leaves of rice plants, but the underlying mechanism is still unclear. N can improve the chlorophyll content and thus affect photon absorption, but the photosynthetic rate does not increase accordingly. To investigate this mechanism, three concentrations of N treatments were applied to two rice varieties, Zhefu802 and Fgl. The results indicated increased chlorophyll content of leaves with an increased N supply. Little discrepancy was detected in Rubisco enzyme activity and Non-photochemical quenching (NPQ) in the high nitrogen (HN) and moderate nitrogen (MN) treatments. The model that photoinhibition occurs in Zhefu802 due to a lack of balance of light absorption and utilization is supported by the higher malondialdehyde (MDA) content, higher H2O2 content, and photoinhibitory quenching (qI) in HN treatment compared with MN treatment. A lower proportion of N in leaf was used to synthesize chlorophyll for Fgl compared with Zhefu802, reducing the likelihood of photoinhibition under HN treatment. In conclusion, HN supply does not allow ideal photosynthetic rate and increases the likelihood of photoinhibition because it does not sustain the balance of light absorption and utilization. Apart from Rubisco enzyme activity, NPQ mainly contributes to the unbalance. These results of this study will provide reference for the effective N management of rice.

Abscisic acid synergizes with sucrose to enhance grain yield and quality of rice by improving the source-sink relationship.

BACKGROUND: Abscisic acid (ABA) and sucrose act as molecular signals in response to abiotic stress. However, how their synergy regulates the source-sink relationship has rarely been studied. This study aimed to reveal the mechanism underlying the synergy between ABA and sucrose on assimilates allocation to improve grain yield and quality of rice. The early indica rice cultivar Zhefu802 was selected and planted in an artificial climate chamber at 32/24 °C (day/night) under natural sunlight conditions. Sucrose and ABA were exogenously sprayed (either alone or in combination) onto rice plants at flowering and 10 days after flowering. RESULTS: ABA plus sucrose significantly improved both the grain yield and quality of rice, which was mainly a result of the higher proportion of dry matter accumulation and non-structural carbohydrates in panicles. These results were mainly ascribed to the large improvement in sucrose transport in the sheath-stems in response to the ABA plus sucrose treatment. In this process, ABA plus sucrose significantly enhanced the contents of starch, gibberellic acids, and zeatin ribosides as well as the activities and gene expression of enzymes involved in starch synthesis in grains. Additionally, remarkable increases in trehalose content and expression levels of trehalose-6-phosphate synthase1, trehalose-6-phosphate phosphatase7, and sucrose non-fermenting related protein kinase 1A were also found in grains treated with ABA plus sucrose. CONCLUSION: The synergy between ABA and sucrose increased grain yield and quality by improving the source-sink relationship through sucrose and trehalose metabolism in grains.

Dual priming oligonucleotide (DPO)-based real-time RT-PCR assay for accurate differentiation of four major viruses causing porcine viral diarrhea.

Currently in China, porcine epidemic diarrhea virus (PEDV), transmissible gastroenteritis virus (TGEV), porcine rotavirus (PoRV), and porcine deltacoronavirus (PDCoV) are the major causes of porcine viral diarrhea, and mixed infections in clinics are common, resulting in significant economic losses in pig industry. Here, a dual priming oligonucleotide (DPO)-based multiplex real-time SYBR Green RT-PCR assay were developed for accurately differentiating PEDV, TGEV, PoRV, and PDCoV in clinical specimens targeting the N gene of TGEV, PEDV, and PDCoV, and the VP7 gene of PoRV. Results showed that the DPO primer allowed a wider annealing temperature range (40-65 °C) and had a higher priming specificity compared to conventional primer, in which more than 3 nucleotides in the 3'- or 5'-segment of DPO primer mismatched with DNA template, PCR amplification efficiency would decrease substantially or extension would not proceed. DPO-based multiplex real-time RT-PCR method had analytical detection limit of 8.63 × 102 copies/µL, 1.92 × 102 copies/µL, 1.74 × 102 copies/µL, and 1.76 × 102 copies/µL for PEDV, TGEV, PoRV, and PDCoV in clinical specimens, respectively. A total of 672 clinical specimens of piglets with diarrheal symptoms were collected in Northeastern China from 2017 to 2018 followed by analysis using the assay, and epidemiological investigation results showed that PEDV, TGEV, PoRV, and PDCoV prevalence was 19.05%, 5.21%, 4.32%, and 3.87%, respectively. The assay developed in this study showed higher detection accuracy than conventional RT-PCR method, suggesting a useful tool for the accurate differentiation of the four major viruses causing porcine viral diarrhea in practice.

N-linked glycosylation sites in G protein of infectious hematopoietic necrosis virus (IHNV) affect its virulence and immunogenicity in rainbow trout.

Infectious hematopoietic necrosis virus (IHNV) causes infectious hematopoietic necrosis in salmonid fish, resulting in substantial economic losses to the aquaculture industry worldwide. The G protein, which harbors the major antigenic determinants of IHNV, is an envelope glycoprotein that plays an important role in both pathogenicity and immunogenicity of IHNV. Previous studies have demonstrated that changes to viral glycosylation sites may affect replication and immunogenicity, but little is known about the specific contributions of G protein glycosylation to IHNV replication and pathogenicity. In this study, we predicted four N-linked glycosylation sites at position 56, 379, 401, and 438 Asp (N) in G protein, and using a reverse genetics system developed in our laboratory, constructed nine recombinant viruses with single, triple, or quadruple glycosylation site disruptions using alanine substitutions in the following combinations: rIHNV-N56A, rIHNV-N379A, rIHNV-N401A, rIHNV-N438A, rIHNV-N56A-N379A-N401A, rIHNV-N56A-N379A-N438A, rIHNV-N56A-N401A-N438A, rIHNV-N379A-N401A-N438A, and rIHNV-N56A-N379A-N401A-N438A. Our results confirmed that all four asparagines are sites of N-linked glycosylation, and Western blot confirmed that mutation of each predicted N-glycosylation sited impaired glycosylation. Among the nine recombinant IHNVs, replication levels decreased significantly in vitro and in vivo in the triple and quadruple mutants that combined mutation of asparagines 401 and 438, indicating the importance of glycosylation at these sites for efficient replication. Moreover, juvenile rainbow trout mortality after challenge by each of the nine mutants showed that, while eight mutants suffered almost 100% cumulative mortality over 30 days, the mutant with a single alanine substitution at position 438 resulted in cumulative mortality of less than 50% over 30 days. This mutant also elicited specific anti-IHNV IgM production earlier than other mutants, suggesting that glycosylation of asparagine 438 may be important for viral immune escape. In conclusion, our study reveals the effect of G protein glycosylation on the pathogenicity and immunogenicity of IHNV and provides a foundation for developing a live-attenuated vaccine.

Salicylic acid reverses pollen abortion of rice caused by heat stress.

BACKGROUND: Extremely high temperatures are becoming an increasingly severe threat to crop yields. It is well documented that salicylic acid (SA) can enhance the stress tolerance of plants; however, its effect on the reproductive organs of rice plants has not been described before. To investigate the mechanism underlying the SA-mediated alleviation of the heat stress damage to rice pollen viability, a susceptible cultivar (Changyou1) was treated with SA at the pollen mother cell (PMC) meiosis stage and then subjected to heat stress of 40 °C for 10 d until 1d before flowering. RESULTS: Under control conditions, no significant difference was found in pollen viability and seed-setting rate in SA treatments. However, under heat stress conditions, SA decreased the accumulation of reactive oxygen species (ROS) in anthers to prevent tapetum programmed cell death (PCD) and degradation. The genes related to tapetum development, such as EAT1 (Eternal Tapetum 1), MIL2 (Microsporeless 2), and DTM1 (Defective Tapetum and Meiocytese 1), were found to be involved in this process. When rice plants were exogenously sprayed with SA or paclobutrazol (PAC, a SA inhibitor) + H2O2 under heat stress, a significantly higher pollen viability was found compared to plants sprayed with H2O, PAC, or SA + dimethylthiourea (DMTU, an H2O2 and OH· scavenger). Additionally, a sharp increase in H2O2 was observed in the SA or PAC+ H2O2 treatment groups compared to other treatments. CONCLUSION: We suggest that H2O2 may play an important role in mediating SA to prevent pollen abortion caused by heat stress through inhibiting the tapetum PCD.

A substitution mutation in OsPELOTA confers bacterial blight resistance by activating the salicylic acid pathway.

We previously reported a spotted-leaf mutant pelota (originally termed HM47) in rice displaying arrested growth and enhanced resistance to multiple races of Xanthomonas oryzae pv. oryzae. Here, we report the map-based cloning of the causal gene OsPELOTA (originally termed splHM47 ). We identified a single base substitution from T to A at position 556 in the coding sequence of OsPELOTA, effectively mutating phenylalanine to isoleucine at position 186 in the translated protein sequence. Both functional complementation and over-expression could rescue the spotted-leaf phenotype. OsPELOTA, a paralogue to eukaryotic release factor 1 (eRF1), shows high sequence similarity to Drosophila Pelota and also localizes to the endoplasmic reticulum and plasma membrane. OsPELOTA is constitutively expressed in roots, leaves, sheaths, stems, and panicles. Elevated levels of salicylic acid and decreased level of jasmonate were detected in the pelota mutant. RNA-seq analysis confirmed that genes responding to salicylic acid were upregulated in the mutant. Our results indicate that the rice PELOTA protein is involved in bacterial leaf blight resistance by activating the salicylic acid metabolic pathway.

Dual-wavelength Nd:YAG laser operation at 1319 and 1338 nm by direct pumping at 885 nm.

This paper presents a continuous wave and a Q-switched Nd:YAG laser pumped by diode laser at 885 nm. The maximum output power of the CW laser is 8.28 W with an absorbed slope efficiency of 35.01%. The Q-switching is achieved using a V³âº:YAG crystal as the saturable absorber. The maximum output power of the passively Q-switched laser is 3.55 W with an absorbed pumping power of 28.65 W operated with a dual wavelength at 1319 and 1338 nm. The shortest pulse widths of the Q-switched laser are 20.20 and 20.86 ns, with a maximum repetition rate of 64.10 kHz.

Single base substitution in OsCDC48 is responsible for premature senescence and death phenotype in rice.

A premature senescence and death 128 (psd128) mutant was isolated from an ethyl methane sulfonate-induced rice IR64 mutant bank. The premature senescence phenotype appeared at the six-leaf stage and the plant died at the early heading stage. psd128 exhibited impaired chloroplast development with significantly reduced photosynthetic ability, chlorophyll and carotenoid contents, root vigor, soluble protein content and increased malonaldehyde content. Furthermore, the expression of senescence-related genes was significantly altered in psd128. The mutant trait was controlled by a single recessive nuclear gene. Using map-based strategy, the mutation Oryza sativa cell division cycle 48 (OsCDC48) was isolated and predicted to encode a putative AAA-type ATPase with 809 amino-acid residuals. A single base substitution at position C2347T in psd128 resulted in a premature stop codon. Functional complementation could rescue the mutant phenotype. In addition, RNA interference resulted in the premature senescence and death phenotype. OsCDC48 was expressed constitutively in the root, stem, leaf and panicle. Subcellular analysis indicated that OsCDC48:YFP fusion proteins were located both in the cytoplasm and nucleus. OsCDC48 was highly conserved with more than 90% identity in the protein levels among plant species. Our results indicated that the impaired function of OsCDC48 was responsible for the premature senescence and death phenotype.

Q-switched 1329 nm Nd:CNGG laser.

We demonstrate a laser-diode-pumped Q-switched 1329 nm neodymium-doped calcium-niobium-gallium-garnet (Nd:CNGG) laser using a V:YAG crystal as a saturable absorber. An average output power of 353 mW and a repetition rate of 13.43 kHz for Q-switched pulses were obtained. The pulse width was from 124 to 151.4 ns under different pump powers. Output power of 685 mW was obtained without the V:YAG crystal inserted.

Characterization and genetic analysis of a novel rice spotted-leaf mutant HM47 with broad-spectrum resistance to Xanthomonas oryzae pv. oryzae.

A stable inherited rice spotted-leaf mutant HM47 derived from an EMS-induced IR64 mutant bank was identified. The mutant expressed hypersensitive response (HR)-like symptoms throughout its whole life from the first leaf to the flag leaf, without pathogen invasion. Initiation of the lesions was induced by light under natural summer field conditions. Expression of pathogenesis-related genes including PAL, PO-C1, POX22.3 and PBZ1 was enhanced significantly in association with cell death and accumulation of H2 O2 at and around the site of lesions in the mutant in contrast to that in the wild-type (WT). Disease reaction to Xanthomonas oryzae pv. oryzae from the Philippines and China showed that HM47 is a broad-spectrum disease-resistant mutant with enhanced resistance to multiple races of bacterial blight pathogens tested. An F2 progeny test showed that bacterial blight resistance to race HB-17 was co-segregated with the expression of lesions. Genetic analysis indicated that the spotted-leaf trait was controlled by a single recessive gene, tentatively named spl(HM47) , flanked by two insertion/deletion markers in a region of approximately 74 kb on the long arm of chromosome 4. Ten open reading frames are predicted, and all of them are expressed proteins. Isolation and validation of the putative genes are currently underway.

RGA1 Negatively Regulates Thermo-tolerance by Affecting Carbohydrate Metabolism and the Energy Supply in Rice.

BACKGROUND: Signal transduction mediated by heterotrimeric G proteins, which comprise the α, ß, and γ subunits, is one of the most important signaling pathways in rice plants. RGA1, which encodes the Gα subunit of the G protein, plays an important role in the response to various types of abiotic stress, including salt, drought, and cold stress. However, the role of RGA1 in the response to heat stress remains unclear. RESULTS: The heat-resistant mutant ett1 (enhanced thermo-tolerance 1) with a new allele of the RGA1 gene was derived from an ethane methyl sulfonate-induced Zhonghua11 mutant. After 45 °C heat stress treatment for 36 h and recovery for 7 d, the survival rate of the ett1 mutants was significantly higher than that of wild-type (WT) plants. The malondialdehyde content was lower, and the maximum fluorescence quantum yield of photosystem II, peroxidase activity, and hsp expression were higher in ett1 mutants than in WT plants after 12 h of exposure to 45 °C. The RNA-sequencing results revealed that the expression of genes involved in the metabolism of carbohydrate, nicotinamide adenine dinucleotide, and energy was up-regulated in ett1 under heat stress. The carbohydrate content and the relative expression of genes involved in sucrose metabolism indicated that carbohydrate metabolism was accelerated in ett1 under heat stress. Energy parameters, including the adenosine triphosphate (ATP) content and the energy charge, were significantly higher in the ett1 mutants than in WT plants under heat stress. Importantly, exogenous glucose can alleviate the damages on rice seedling plants caused by heat stress. CONCLUSION: RGA1 negatively regulates the thermo-tolerance in rice seedling plants through affecting carbohydrate and energy metabolism.

Optical trapping of gold nanoparticles by cylindrical vector beam.

Optical trapping of gold nanoparticles is experimentally demonstrated using radially and azimuthally polarized beams. The transverse optical trapping stiffness of gold nanoparticles is measured. The radially polarized beam exhibits a higher trapping efficiency than the azimuthally polarized beam and the Gaussian beam. The transverse stiffness of particles with different diameters is measured experimentally and calculated via the discrete-dipole approximation method, and good agreement between theory and experiment is found.

Ultrasensitive detection of amplified spontaneous emission at 710 nm by means of picosecond collinear optical parametric amplification near degeneracy.

We report the experimental results on improving the detection of an ultraweak optical signal using a 355 nm pumped picosecond collinear optical parametric amplification (OPA). The OPA is seeded by the amplified spontaneous emission (ASE) generated in a solution of pyridine-1 dye in ethanol. The gain factor of this amplifier is determined as ~1.5×10(8), and the detection limit is ~1.25 aJ per pulse, corresponding to five photons at 710 nm within the 15 ps pulse width of the pump beam. This is achieved by reducing superfluorescence background noise by means of signal/idler double seeding near degeneracy, amplification under a slightly phase-mismatched condition, and space filtering with increasing observation distance up to 3.2 m. Compared with previous reports, the detection limit is significantly enhanced. The pulse shape of the ASE is also measured with OPA, and it agrees well with that measured by an ultrafast oscilloscope.

Increased ATPase activity promotes heat-resistance, high-yield, and high-quality traits in rice by improving energy status.

Heat stress during the reproductive stage results in major losses in yield and quality, which might be mainly caused by an energy imbalance. However, how energy status affected heat response, yield and quality remains unclear. No relationships were observed among the heat resistance, yield, and quality of the forty-nine early rice cultivars under normal temperature conditions. However, two cultivars, Zhuliangyou30 (ZLY30) and Luliangyou35 (LLY35), differing in heat resistance, yield, and quality were detected. The yield was higher and the chalkiness degree was lower in ZLY30 than in LLY35. Decreases in yields and increases in the chalkiness degree with temperatures were more pronounced in LLY35 than in ZLY30. The accumulation and allocation (ratio of the panicle to the whole plant) of dry matter weight and non-structural carbohydrates were higher in ZLY30 than in LLY35 across all sowing times and temperatures. The accumulation and allocation of dry matter weight and non-structural carbohydrates in panicles were higher in ZLY30 than in LLY35. Similar patterns were observed in the relative expression levels of sucrose unloading related genes SUT1 and SUT2 in grains. The ATP content was higher in the grains of LLY35 than in ZLY30, whereas the ATPase activity, which determined the energy status, was significantly lower in the former than in the latter. Thus, increased ATPase activity, which improved the energy status of rice, was the factor mediating the balance among heat-resistance, high-yield, and high-quality traits in rice.