Operating point control method for the Mach-Zehnder modulator in a phase-shift laser range finder.

An operating point control method is proposed for the Mach-Zehnder modulator (MZM) based on a dual-cascaded MZM structure. Unlike traditional methods with dither signals, the proposed method is advantageous because the components monitored in the control process are not masked by the spectrum noise floor and the drift direction is clearly determined at the quadrature point, thus imparting greater control stability. Additionally, the proposed control method is suitable for phase-shift laser range finders (PSLRFs). Compared with traditional methods, experimental results reveal that the proposed method increases the operating point stability of MZM from ±0.59° to ±0.36° within 2 h, resulting in better ranging stability than 17 µm in 1 min and 39 µm in 1 h in a PSLRF with a 200 MHz modulation frequency.

High-speed and high-precision measurement of biaxial in-plane displacements: tens of nanometers principle error suppression in microprobe sensors.

When the microprobe sensor is faced with the demand of high-speed biaxial displacement measurement, due to the characteristics of phase generated carrier (PGC) technology, accompanying optical intensity modulation (AOIM) and unfavorable phase modulation depth (PMD) will bring about the tens of nanometer cyclic nonlinear errors, further hindering high-speed and high-precision measurement. Herein, a light source intensity stabilization system based on semiconductor optical amplifier (SOA) feedback control is achieved to eliminate the error caused by AOIM in the presence of high-frequency and large-amplitude laser modulation. Based on this, the reasons for large nonlinear errors in biaxial measurements and the inability to ensure the stability of the accuracy of multiple measurement axes are methodically examined, and an effective nonlinear error elimination methodology based on the normalized amplitude correction of active temperature scanning is proposed. The continuity and linearity of the temperature scanning are also discussed. The performed experiments show that the above approach is capable of reducing the displacement demodulation error from the nanometer scale to the sub-nanometer scale. Further, the nonlinear error is reduced to within 0.1 nm for both measurement axes and the performance becomes consistent. The dual-axis measurement resolution of the microprobe sensor reaches 0.4 nm and the measurement speed is better than 1.2 m/s with the standard deviation of lower than 0.5 nm.

The Role of mRNA Alternative Splicing in Macrophages Infected with Mycobacterium tuberculosis: A Field Needing to Be Discovered.

Mycobacterium tuberculosis (Mtb) is one of the major causes of human death. In its battle with humans, Mtb has fully adapted to its host and developed ways to evade the immune system. At the same time, the human immune system has developed ways to respond to Mtb. The immune system responds to viral and bacterial infections through a variety of mechanisms, one of which is alternative splicing. In this study, we summarized the overall changes in alternative splicing of the transcriptome after macrophages were infected with Mtb. We found that after infection with Mtb, cells undergo changes, including (1) directly reducing the expression of splicing factors, which affects the regulation of gene expression, (2) altering the original function of proteins through splicing, which can involve gene truncation or changes in protein domains, and (3) expressing unique isoforms that may contribute to the identification and development of tuberculosis biomarkers. Moreover, alternative splicing regulation of immune-related genes, such as IL-4, IL-7, IL-7R, and IL-12R, may be an important factor affecting the activation or dormancy state of Mtb. These will help to fully understand the immune response to Mtb infection, which is crucial for the development of tuberculosis biomarkers and new drug targets.

Embedded micro-probe fiber optic interferometer with low nonlinearity against light intensity disturbance.

A novel low-nonlinearity Michelson microprobe fiber interferometer against light intensity disturbance for high-precision embedded displacement measurements is introduced. To analyze the influence of light intensity disturbance on the microprobe and measurement accuracy of the integrated fiber interferometer, an equivalent model of micro-probe sensing with the tilted target is established. The proposed PGC demodulation and nonlinearity correction method with simple principle helps avoid DC component varying with light intensity. The experiments show that residual displacement errors of the micro-probe fiber interferometer are reduced from 4.36 nm to 0.46 nm, thus allowing embedded displacement detection with sub-nanometer accuracy under low frequency light intensity disturbance.

Tiliroside suppresses triple-negative breast cancer as a multifunctional CAXII inhibitor.

Triple negative breast cancer (TNBC) is an aggressive subtype of breast cancer characterized by poor prognosis, early recurrence, and the lack of durable chemotherapy responses and specific targeted treatments. In this preclinical study, we examines Tiliroside (TS, C30H26O13), as one of the major compounds of Tribulus terrestris L. which has been used as an alternative therapy in clinic practice of breast cancer treatment, for its therapeutic use in TNBC. The association between CAXII expression level and survival probability of TNBC patients, and the difference of CAXII expression level between TNBC and normal samples were evaluated by using publicly accessible databases. To determine the anticancer efficacy of TS on TNBC cells, cell proliferation, wound healing, cell invasion, and 3D spheroid formation assays were performed and excellent anticancer activities of TS were displayed. Mouse models further demonstrated that TS significantly reduced the tumor burden and improved survival rate. The properties of TS as a novel CAXII inhibitor have also been evaluated by CAXII activity assay, pHi, pHe and lactate level assay. Further RT-PCR and Caspase-3 activity analyses also revealed the positive regulating effects of TS on E2F1,3/Caspase-3 axis in TNBC cells cultured in 2D or 3D systems. The findings indicate that TS suppresses TNBC progression as a potential novel CAXII inhibitor in preclinical experiments, which warrants further investigation on its therapeutic implications.

Lysobacter lactosilyticus sp. nov., a Novel ß-Galactosidase Producing Bacterial Strain Isolated from Farmland Soil Applied with Amino Acid Fertilizer.

To isolate ß-galactosidase producing bacterial resources, a novel Gram-stain-negative, strictly aerobic bacterial strain designated as A6T was obtained from a farmland soil sample. Cells of the strain were rod-shaped (0.4-0.7 µm × 1.8-2.2 µm) without flagella and motility. Strain A6T grew optimally at 30 °C, pH 7.0 with 0% (w/v) NaCl. Based on phylogenetic analysis, strain A6T clustered within the genus Lysobacter clade and branched with Lysobacter dokdonensis KCTC 12822T (99.5%, 16S rRNA gene sequence similarity) and Lysobacter caseinilyticus KACC 19816T (98.5%). The average nucleotide identity (ANI) and digital DNA-DNA hybridization (dDDH) values between strain A6T and Lysobacter dokdonensis KCTC 12822T were 82.7% and 26.2%, and the values for strain A6T and KACC 19816T were 81.4% and 23.8%, respectively. Iso-C16:0, iso-C15:0, summed feature 9 (C17:1 iso ω9c and/or C16:0 10-methyl) and summed feature 3 (C16:1ω7c and/or C16:1 ω6c) were the major fatty acids, diphosphatidylglycerol, phosphatidylglycerol, and phosphatidylethanolamine were the major polar lipids, and ubiquinone 8 (Q-8) was the major ubiquinone. The genomic DNA G+C content was 67.2 mol%. Furthermore, under the condition of 30 °C, pH 7.0, 4% inoculation with 10.0 g L-1 lactose, the ß-galactosidase activity produced by strain A6T was highest, reaching 95.3 U mL-1, indicating that this strain could be applied as a potential strain for ß-galactosidase production. Strain A6T represents a novel species of the genus Lysobacter, and Lysobacter lactosilyticus sp. nov. is proposed on the basis of phenotypic, genotypic, and chemotaxonomic analysis. The type strain is A6T (=KCTC 82184T=CGMCC 1.18582T).

Comparative Transcriptomic Analysis to Identify Brassinosteroid Response Genes.

Brassinosteroids (BRs) are plant growth-promoting steroid hormones. BRs affect plant growth by regulating panels of downstream genes. Much effort has been made to establish BR-regulated gene expression networks, but there is little overlap among published expression networks. In this study, we built an optimal BR-regulated gene expression network using the model plant Arabidopisis (Arabidopisis thaliana). Seven- and 24-d-old seedlings of the constitutive photomorphogenesis and dwarfism mutant and brassinosteroid-insensitive 1-701 (bri1-701) BRI1-like receptor genes1 (brl1) brl3 triple mutant seedlings were treated with brassinolide and RNA sequencing (RNA-seq) was used to detect differentially expressed genes. Using this approach, we generated a transcriptomic database of 4,498 differentially expressed genes and identified 110 transcription factors that specifically respond to BR at different stages. We also found that, among the identified BR-responsive transcription factors, ABSCISIC ACID-INSENSlTIVE4 (ABI4), an ethylene response factor transcription factor, inhibits BR-regulated growth. Compared to wild-type plants, the abi4-102 mutant was less sensitive to brassinazole and more sensitive to BR. Next, we performed a chromatin immunoprecipitation followed by high-throughput sequencing assay and established that ABI4 binds directly to the BRI1-associated receptor kinase1 promoter and inhibits transcription. These results provide insight into BR-responsive gene functions in regulating plant growth at different stages and may serve as a basis for predicting gene function, selecting candidate genes, and improving the understanding of BR regulatory pathways.

The ancestral duplicated DL/CRC orthologs, PeDL1 and PeDL2, function in orchid reproductive organ innovation.

Orchid gynostemium, the fused organ of the androecium and gynoecium, and ovule development are unique developmental processes. Two DROOPING LEAF/CRABS CLAW (DL/CRC) genes, PeDL1 and PeDL2, were identified from the Phalaenopsis orchid genome and functionally characterized. Phylogenetic analysis indicated that the most recent common ancestor of orchids contained the duplicated DL/CRC-like genes. Temporal and spatial expression analysis indicated that PeDL genes are specifically expressed in the gynostemium and at the early stages of ovule development. Both PeDLs could partially complement an Arabidopsis crc-1 mutant. Virus-induced gene silencing (VIGS) of PeDL1 and PeDL2 affected the number of protuberant ovule initials differentiated from the placenta. Transient overexpression of PeDL1 in Phalaenopsis orchids caused abnormal development of ovule and stigmatic cavity of gynostemium. PeDL1, but not PeDL2, could form a heterodimer with Phalaenopsis equestris CINCINNATA 8 (PeCIN8). Paralogous retention and subsequent divergence of the gene sequences of PeDL1 and PeDL2 in P. equestris might result in the differentiation of function and protein behaviors. These results reveal that the ancestral duplicated DL/CRC-like genes play important roles in orchid reproductive organ innovation.

Aquamicrobium zhengzhouense sp. nov., a Bacterium Isolated from Farmland Soil Applied with Amino Acid Fertilizer.

A novel Gram-stain-negative, rod-shaped, strictly aerobic, non-motile bacterium, designated strain cd-1T, was isolated from a farmland soil applied with amino acid fertilizer in Zhengzhou, Henan province, China. The optimum growth of strain cd-1T occurred at 30 °C, pH 7.0 in Luria-Bertani (LB) broth without NaCl supplement. Phylogenetic analysis based on 16S rRNA gene sequences indicated that cd-1T is member of the genus Aquamicrobium, and formed a separate branch with Aquamicrobium aerolatum DSM 21857T (96.5%) and Aquamicrobium soli KCTC 52165T (95.7%). The draft genome sequencing revealed a DNA G + C content of 59.2 mol% and Q-10 was the predominant respiratory quinone. The major cellular fatty acids were identified as C18:1 ω7c (35.8%), C19:0 cyclo ω8c (32.1%), and C18:1 ω7c 11-methyl (5.2%). The polar lipids consisted of phosphatidylethanolamine, diphosphatidylglycerol, phosphatidylglycerol, phosphatidylcholine and phosphatidylmonomethylethanolamine. Average nucleotide identity (ANI) and the digital DNA-DNA hybridizations (dDDH) for draft genomes between strain cd-1T and KCTC 52165T were 71.0% and 19.9%, respectively, the values for strain cd-1T and DSM 21857T were 73.4% and 20.6%. Based on the physiological and biochemical characteristics, phylogenetic and chemotaxonomic analysis, strain cd-1T is considered to represent a novel species of the genus Aquamicrobium, for which the name Aquamicrobium zhengzhouense sp. nov. is proposed. The type strain is cd-1T (= KCTC 82182T = CCTCC M 2018904T).

Bioaugmentation of acetamiprid-contaminated soil with Pigmentiphaga sp. strain D-2 and its effect on the soil microbial community.

Bioaugmentation, a strategy based on microbiome engineering, has been proposed for bioremediation of pollutant-contaminated environments. However, the complex microbiome engineering processes for soil bioaugmentation, involving interactions among the exogenous inoculum, soil environment, and indigenous microbial microbiome, remain largely unknown. Acetamiprid is a widely used neonicotinoid insecticide which has caused environmental contaminations. Here, we used an acetamiprid-degrading strain, Pigmentiphaga sp. D-2, as inoculum to investigate the effects of bioaugmentation on the soil microbial community and the process of microbiome reassembly. The bioaugmentation treatment removed 94.8 and 92.5% of acetamiprid within 40 days from soils contaminated with 50 and 200 mg/kg acetamiprid, respectively. A decrease in bacterial richness and diversity was detected in bioaugmentation treatments, which later recovered with the removal of acetamiprid from soil. Moreover, the bioaugmentation treatment significantly influenced the bacterial community structure, whereas application of acetamiprid alone had little influence on the soil microbial community. Furthermore, the bioaugmentation treatment improved the growth of bacteria associated with acetamiprid degradation, and the inoculated and recruited taxa significantly influenced the keystone taxa of the indigenous microbiome, resulting in reassembly of the bacterial community yielding higher acetamiprid-degrading efficiency than that of the indigenous and acetamiprid-treated communities. Our results provide valuable insights into the mechanisms of microbiome engineering for bioaugmentation of acetamiprid-contaminated soils.

Pigmentiphaga sp. Strain D-2 Uses a Novel Amidase To Initiate the Catabolism of the Neonicotinoid Insecticide Acetamiprid.

Acetamiprid, a chloronicotinyl neonicotinoid insecticide, is among the most commonly used insecticides worldwide, and its environmental fate has caused considerable concern. The compound 1-(6-chloropyridin-3-yl)-N-methylmethanamine (IM 1-4) has been reported to be the main intermediate during acetamiprid catabolism in microorganisms, honeybees, and spinach. However, the molecular mechanism underlying the hydrolysis of acetamiprid to IM 1-4 has not yet been elucidated. In this study, a novel amidase (AceAB) that initially hydrolyzes the C-N bond of acetamiprid to generate IM 1-4 was purified and characterized from the acetamiprid-degrading strain Pigmentiphaga sp. strain D-2. Based on peptide profiling of the purified AceAB and the draft genome sequence of strain D-2, aceA (372 bp) and aceB (2,295 bp), encoding the α and ß subunits of AceAB, respectively, were cloned and found to be necessary for acetamiprid hydrolysis in strain D-2. The characteristics of AceAB were also systematically investigated. Though AceA and AceB showed 35% to 56% identity to the α and ß subunits of the N,N-dimethylformamidase from Paracoccus aminophilus, AceAB was specific for the hydrolysis of acetamiprid and showed no activities to N,N-dimethylformamide or its structural analogs.IMPORTANCE Acetamiprid, among the top neonicotinoid insecticides used worldwide, is one of the most important commercial insecticides. Due to its extensive use, the environmental fate of acetamiprid, especially its microbial degradation, has caused considerable concern. Although the catabolic pathways of acetamiprid in microorganisms have been extensively studied, the molecular mechanisms underlying acetamiprid biodegradation (except for a nitrile hydratase) remain largely unknown, and the enzyme responsible for the biotransformation of acetamiprid into its main intermediate, IM 1-4, have not yet been elucidated. The amidase AceAB and its encoding genes, aceA and aceB, characterized in this study, were found to be necessary and specific for the initial hydrolysis of the C-N bond of acetamiprid to generate IM 1-4 in Pigmentiphaga sp. strain D-2. The finding of the novel amidase AceAB will greatly enhance our understanding of the microbial catabolism of the widely used insecticide acetamiprid at the molecular level.

Correction of nonlinear errors from PGC carrier phase delay and AOIM in fiber-optic interferometers for nanoscale displacement measurement.

In fiber-optic interferometers with laser frequency modulation, carrier phase delay and accompanied optical intensity modulation (AOIM) in phase-generated-carrier (PGC) demodulation inevitably produce nonlinear errors that can seriously hamper displacement measurement accuracy. As for the existing improved PGC scheme, they are only capable to compensate for one of these effects. As the only method that is effective in eliminating the two effects simultaneously, typical ellipse fitting methods require target movements λlaser/4, and fail when the PGC carrier phase delay is proximate to certain values (e.g., nπ +π/4, nπ +π/2). Herein, a modified nonlinear-error correction method for errors due to PGC carrier phase delay and AOIM is proposed. Active laser-wavelength scanning by constant variation of the laser drive temperature is used to replace the target movement. A fiber-optic Michelson interferometer is constructed and experiments are performed to verify the feasibility of the proposed method. The experimental results show that after correction, the nonlinear error is reduced to less than 1nm, and nanoscale displacement measurement is achieved.

Phase modulation depth setting technique of a phase-generated-carrier under AOIM in fiber-optic interferometer with laser frequency modulation.

The phase modulation depth (PMD) in phase-generated-carrier demodulation is determined by the laser frequency modulation amplitude and working distance of a fiber-optic interferometer and must be set at a certain value. Active setting of the amplitude is unsuitable, especially for high-speed modulation, owing to variations in the laser source tuning coefficients. Existing calculation schemes for passive setting cannot work both owing to carrier phase delay (CPD) and the accompanied optical-intensity modulation (AOIM). Herein, a modified phase modulation depth calculation and setting technique is proposed. Double photoelectric detection and signal division are optimized to eliminate AOIM using a fiber delay chain and phase-locked amplifier module. Fast Fourier-transform and look-up table methods are used to calculate phase modulation depth without adding the carrier, which is unaffected by CPD. A fiber-optic Michelson interferometer is constructed to verify the feasibility of the proposed method. The experimental results show that AOIM can be eliminated; moreover, PMD can be calculated and set precisely. The displacement deviation is less than 1.03 nm. The resolution of measurement is considerably lesser than 1 nm and nanoscale accuracy is achieved in displacement measurement.

Tissue-Specific Transcriptomics Reveals an Important Role of the Unfolded Protein Response in Maintaining Fertility upon Heat Stress in Arabidopsis.

High temperatures have a great impact on plant reproductive development and subsequent fruit and seed set, but the underlying molecular mechanisms are not well understood. We used transcriptome profiling to investigate the effect of heat stress on reproductive development of Arabidopsis thaliana plants and observed distinct response patterns in vegetative versus reproductive tissues. Exposure to heat stress affected reproductive developmental programs, including early phases of anther/ovule development and meiosis. Also, genes participating in the unfolded protein response (UPR) were enriched in the reproductive tissue-specific genes that were upregulated by heat. Moreover, we found that the UPR-deficient bzip28 bzip60 double mutant was sensitive to heat stresses and had reduced silique length and fertility. Comparison of heat-responsive wild type versus bzip28 bzip60 plants identified 521 genes that were regulated by bZIP28 and bZIP60 upon heat stress during reproductive stages, most of which were noncanonical UPR genes. Chromatin immunoprecipitation coupled with high-throughput sequencing analyses revealed 133 likely direct targets of bZIP28 in Arabidopsis seedlings subjected to heat stress, including 27 genes that were also upregulated by heat during reproductive development. Our results provide important insights into heat responsiveness in Arabidopsis reproductive tissues and demonstrate the protective roles of the UPR for maintaining fertility upon heat stress.

Synthesis of Hydroxylatopillar[6]arene-Controlled Gold Nanoparticles-Cellulose Nanocrystals and Their Applications.

In this work, two macrocyclic hosts, named hydroxylatopillar[6]arene and dihydroxylatopillar[6]arene (HP6, 2HP6), are proposed. We found that the reduction of Au3+ to Au0 can success by using HP6 or 2HP6 as a reductant and stabilizing agent. At the end of HP6/2HP6, hydroxyl (-OH) groups were used as a reductant to reduce Au3+ to Au0. At the same time, -OH on HP6/2HP6 was oxidized to -COOH, and then the formed -COOH can be used as the stabilizer to prevent the infinite growth of AuNPs. The cellulose nanocrystals (CNCs) prepared by a clean and nonpolluting method were used as carriers to load AuNPs on them. The CNCs were applied for the adsorption of methylene blue (MB), and then the MB was catalytically degraded by HP6/2HP6-AuNPs-CNC. Besides, the HP6/2HP6-AuNPs-CNC showed remarkable catalytic performance for reducing nitro to the amino group in 4-nitrophenol. The advantages of clean and green synthesis make the HP6/2HP6-AuNPs-CNC a hybrid material and its application sustainable.

Analysis of a highly efficient phase-locking stabilization method for electro-optic comb generation.

We theoretically show that a slightly modified Pound-Drever-Hall (PDH) stabilization scheme can lead to the optimum time-domain characteristics for electro-optic comb generators (EOCG). The ideal locking point is located by analyzing the EOCG output pulse width. By summing up the electrical field reflected by the EOCG front mirror, a model of the phase-locking error signal is derived with the Jacobi-Anger identical transformation. The simulation and experiment show that the zero-locking point of the error signal of the modified scheme coincides well with the ideal locking point in contrast with the direct application of the PDH scheme. Finally, a power efficiency of up to 2.9% is achieved with this EOCG stabilization scheme. A relative instability of better than 2.6×10-8 is demonstrated by a dual comb interferometer with fixed paths. The Allan deviations of the comb mode frequencies are smaller than 2.8×10-9 and 1.1×10-10 for average times of 1 and 100 s, respectively.

Design for A Highly Stable Laser Source Based on the Error Model of High-Speed High-Resolution Heterodyne Interferometers.

Heterodyne interferometers with two opposite Doppler shift interference signals have been proposed for high-resolution measurement with high measurement speed, which can be used in the background of high-speed high-resolution measurement. However, a measurement error model for high-speed high-resolution heterodyne interferometers (HSHR-HIs) has not yet been proposed. We established a HSHR-HI measurement error model, analyzed the influence of beat frequency stability with a simplified optical structure, and then designed an offset-locked dual-frequency laser source with a digital control system to reduce the impact of beat frequency drift. Experiments were used to verify the correction of the measurement error model and the validity of the laser source. The results show that the new laser source has a maximum beat frequency range of 45 MHz, which shows the improvements in the measuring speed and resolution.

Ultrastable Offset-Locking Continuous Wave Laser to a Frequency Comb with a Compound Control Method for Precision Interferometry.

A simple and robust analog feedforward and digital feedback compound control system is presented to lock the frequency of a slave continuous wave (CW) laser to an optical frequency comb. The beat frequency between CW laser and the adjacent comb mode was fed to an acousto-optical frequency shifter (AOFS) to compensate the frequency dithering of the CW laser. A digital feedback loop was achieved to expand the operation bandwidth limitation of the AOFS by over an order of magnitude. The signal-to-noise ratio of the interference signal was optimized using a grating-based spectral filtering detection unit. The complete system achieved an ultrastable offset-locking of the slave CW laser to the frequency comb with a relative stability of ±3.62 × 10-14. The Allan deviations of the beat frequency were 8.01 × 10-16 and 2.19 × 10-16 for a gate time of 10 s and 1000 s, respectively. The findings of this study may further improve laser interferometry by providing a simple and robust method for ultrastable frequency control.

Suppression of B-cell development genes is key to glucocorticoid efficacy in treatment of acute lymphoblastic leukemia.

Glucocorticoids (GCs), including dexamethasone (dex), are a central component of combination chemotherapy for childhood B-cell precursor acute lymphoblastic leukemia (B-ALL). GCs work by activating the GC receptor (GR), a ligand-induced transcription factor, which in turn regulates genes that induce leukemic cell death. Which GR-regulated genes are required for GC cytotoxicity, which pathways affect their regulation, and how resistance arises are not well understood. Here, we systematically integrate the transcriptional response of B-ALL to GCs with a next-generation short hairpin RNA screen to identify GC-regulated "effector" genes that contribute to cell death, as well as genes that affect the sensitivity of B-ALL cells to dex. This analysis reveals a pervasive role for GCs in suppression of B-cell development genes that is linked to therapeutic response. Inhibition of phosphatidylinositol 3-kinase δ (PI3Kδ), a linchpin in the pre-B-cell receptor and interleukin 7 receptor signaling pathways critical to B-cell development (with CAL-101 [idelalisib]), interrupts a double-negative feedback loop, enhancing GC-regulated transcription to synergistically kill even highly resistant B-ALL with diverse genetic backgrounds. This work not only identifies numerous opportunities for enhanced lymphoid-specific combination chemotherapies that have the potential to overcome treatment resistance, but is also a valuable resource for understanding GC biology and the mechanistic details of GR-regulated transcription.

Development of polymorphic microsatellite markers by using de novo transcriptome assembly of Calanthe masuca and C. sinica (Orchidaceae).

BACKGROUND: Calanthe masuca and C. sinica are two genetically closely related species in Orchidaceae. C. masuca is widely distributed in Asia, whereas C. sinica is restricted to Yunnan and Guangxi Provinces in southwest China. Both play important roles in horticulture and are under the pressure of population decline. Understanding their genetic background can greatly help us develop effective conservation strategies for these species. Simple sequence repeats (SSRs) are useful for genetic diversity analysis, presumably providing key information for the study and preservation of the wild populations of the two species we are interested in. RESULTS: In this study, we performed RNA-seq analysis on the leaves of C. masuca and C. sinica, obtaining 40,916 and 71,618 unigenes for each species, respectively. In total, 2,019/3,865 primer pairs were successfully designed from 3,764/7,189 putative SSRs, among which 197 polymorphic SSRs were screened out according to orthologous gene pairs. After mononucleotide exclusion, a subset of 129 SSR primers were analysed, and 13 of them were found to have high polymorphism levels. Further analysis demonstrated that they were feasible and effective against C. masuca and C. sinica as well as transferable to another species in Calanthe. Molecular evolutionary analysis revealed functional pathways commonly enriched in unigenes with similar evolutionary rates in the two species, as well as pathways specific to each species, implicating species-specific adaptation. The divergence time between the two closely related species was tentatively determined to be 3.42 ± 1.86 Mya. CONCLUSIONS: We completed and analysed the transcriptomes of C. masuca and C. sinica, assembling large numbers of unigenes and generating effective polymorphic SSR markers. This is the first report of the development of expressed sequence tag (EST)-SSR markers for Calanthe. In addition, our study could enable further genetic diversity analysis and functional and comparative genomic studies on Calanthe.