Evaluation of Suitable Reference Genes for Quantitative Real-Time PCR in Various Tissues of Apocynum venetum.

Apocynum venetum L. is an economically valuable plant with tolerance to drought and salinity. Its leaves are utilized in tea production and pharmaceuticals, while the stem bark serves as a high-quality fiber material. To gain insights into the gene expression patterns of A. venetum using quantitative real-time PCR (qRT-PCR), it is crucial to identify appropriate reference genes. This study selected nine candidate genes, including α-tubulin (TUA), ß-tubulin (TUB), actin (ACT), cyclophilin (CYP), elongation factor-1α (EF-1α), the B family of regulatory subunits of protein phosphatase (PPP2R2, PPP2R3, and PPP2R5), and phosphoglycerate kinase (PGK), to determine the most appropriate reference genes in the leaf, stem, and root tissues of A. venetum. A comprehensive ranking by geNorm, NormFinder, BestKeeper, and RefFinder software and Venn diagrams was used to screen more stable reference genes in different tissues. The two most stable reference genes were CYP and TUA in leaves, PGK and PPP2R3 in stems, and TUA and EF-1α in roots, respectively. The relative expression values of the four genes involved in proline metabolism under polyethylene glycol treatment were used to validate the screened reference genes, and they exhibited highly stable expression levels. These findings represent the first set of stable reference genes for future gene expression studies in A. venetum. They significantly contribute to enhancing the accuracy and reliability of gene expression analyses in this economically important plant species.

Exogenous Feeding of Fructose and Phenylalanine Further Improves Betulin Production in Suspended Betula platyphylla Cells under Nitric Oxide Treatment.

The aim of this study was to assay by NMR the metabolites which contribute to betulin production. 8-day-old suspended birch (Betula platyphylla) cells were treated by sodium nitroprusside (SNP) treatment, an NO donor, and 2-(4-carboxyphenyl)-4,4,5,5-tetramethyl-imidazoline-1-oxyl-3-oxide (cPTIO), an NO-specific scavenger. The results showed that betulin production was increased by five times after SNP treatment, similar with that of the control under cPTIO treatment. Forty one metabolites were detected after SNP treatment or cPTIO treatment. Among them, 10 were found to significantly contribute to the differences observed between controls and treated cell culture samples. To validate the contribution of the above 10 metabolites to betulin production, myo-inositol, fructose and phenylalanine based on correlation analysis between the content of 12 metabolites and betulin were used to feed birch suspension cell cultures under SNP treatment. Exogenous feeding of fructose or phenylalanine further enhanced the betulin production under SNP treatment, but myo-inositol had the opposite result.

Endogenous and Exogenous Calcium Involved in the Betulin Production from Submerged Culture of Phellinus linteus Induced by Hydrogen Sulfide.

Using pharmacological and biochemical approaches, Ca(2+) involved in the betulin production in mycelia of Phellinus linteus induced by hydrogen sulfide (H2S) were investigated. The results showed that 2 mM H2S donor NaHS or 10 mM CaCl2 was found to enhance the betulin content in the mycelia of Phellinus to the maximum, which were 112.43 and 93.24% higher than that in the control, respectively. Further, NaHS and CaCl2 co-treatment also showed positive outcome, which were 128.95 or 24.52% higher than that in the control or NaHS treatment. At the same time, NaHS also enhanced the content of Ca(2+) and CaM. But, the above positive inductive effects for Ca(2+), CaM, and betulin production can be blocked with either Ca(2+) channel blocker (LaCl3, 2-aminoethoxydiphenyl borate) or Ca(2+) chelator (ethylenediaminetetraacetic acid (EDTA)). Among of them, betulin content was reduced 35.06% by NaHS and EGTA to the minimum, and this reduction could be reversed by the application of CaCl2 (NaHS + EGTA + CaCl2). From above results, it can be concluded that endogenous and exogenous calcium involved in the betulin production from submerged culture of P. linteus induced by hydrogen sulfide.

De novo transcriptome analysis of a medicinal fungi Phellinus linteus and identification of SSR markers.

The aim of this study was to facilitate gene discovery for functional genome studies and to identify simple sequence repeat (SSR) markers for molecular-assisted selection in Phellinus linteus. The transcriptome of Phellinus linteus was sequenced using а high-throughput RNA sequencing system - the Illumina Hiseq 2000. A total of 16,383,818 clean sequencing reads, 35,532 contigs and 25,811 unigenes were postulated. Based on similarity searches with known proteins, 19,350 genes (74.97% of the unigenes) were annotated. In the present research, 19,266, 10,978 and 7831 unigenes were mapped in Nr, Swiss-Prot and clusters of orthologous groups (COG) classifications, respectively. Of all unigenes, 6845 were categorized into three functional groups, namely biological process, cellular components and molecular function and 11,088 were annotated to 108 pathways by searching the Kyoto Encyclopedia of Genes and Genomes pathway database. A total of 1129 SSRs were identified in these unigenes. In addition, 23 candidate genes, potentially involved in sterol biosynthesis, were identified and were worthy of further investigation.

Prevalence of visual impairment among adults in China: the Beijing Eye Study.

PURPOSE: To estimate the prevalence and distribution of blindness and low vision in Northern China. DESIGN: Population-based cohort study. METHODS: The Beijing Eye Study included 4438 subjects with an age of 40+ years. Mean age was 56.2 +/- 10.6 years (range, 40 to 101 years). RESULTS: Forty-three (1.0%) individuals had low vision (<20/60 and >/=20/400 best-corrected vision), and 17 (0.4%) individuals were blind (best-corrected visual acuity in the better-seeing eye <20/400). Low vision/blindness were significantly associated with age (P < .001), myopic refractive error (P < .001), and level of educational background (P = .035). It was not associated with gender (P = .76) and rural vs urban area (P = .88). CONCLUSIONS: Blindness or low vision affects approximately one in 100 Chinese older than 40 years. An estimated 4.1 million Chinese older than 40 years have low vision, and an estimated 1.6 million Chinese older than 40 years are blind.

Frequency of under-corrected refractive errors in elderly Chinese in Beijing.

PURPOSE: The aim of the study was to evaluate the prevalence of under-corrected refractive error among elderly Chinese in the Beijing area. METHODS: The population-based, cross-sectional, cohort study comprised 4,439 subjects out of 5,324 subjects asked to participate (response rate 83.4%) with an age of 40+ years. It was divided into a rural part [1,973 (44.4%) subjects] and an urban part [2,466 (55.6%) subjects]. Habitual and best-corrected visual acuity was measured. Under-corrected refractive error was defined as an improvement in visual acuity of the better eye of at least two lines with best possible refractive correction. RESULTS: The rate of under-corrected refractive error was 19.4% (95% confidence interval, 18.2, 20.6). In a multiple regression analysis, prevalence and size of under-corrected refractive error in the better eye was significantly associated with lower level of education (P<0.001), female gender (P<0.001), and age (P=0.001). CONCLUSIONS: Under-correction of refractive error is relatively common among elderly Chinese in the Beijing area when compared with data from other populations.

QTL for yield and its components responded to elevated CO2 in rice (Oryza sativa L.).

FACE (free air carbon dioxide enrichment) technology may provide a means by which the environment around growing plants can be modified to realistically simulate the concentration of atmospheric CO2 in the future. The plant growth and its yield of plant species can be enhanced under FACE. Identification of genomic regions influencing the response of yield and its components to elevated CO2 will be useful for understanding the genetics of active response to changed CO2 environment and developing higher yield cultivars, which will be adapted to future enriched atmospheric CO2 environment. A mapping population of 65 indica (IR24) chromosome segment substitution lines (CSSLs) in japonica (Asominori) background and their parents were used to detect QTLs for yield and its components, e. g. number of fertile tillers per plant (FT), 1000-grain weight (TGW), number of grains per panicle (GP) and grain yield per plant (GY) under FACE (200 micromol CO2/mol above current levels) and current CO2 concentration (Ambient, about 370 micromol CO2/mol) in the field experiment. The results showed that, GY, GP and FT of two parents under FACE were significant greater than that under Ambient. The transgressive segregation of the four traits was observed in the CSSLs population under both FACE and Ambient. A total of 20 QTLs for the four traits were detected on chromosome 1, 2, 4, 6, 7, 9 and 12 with LOD (Log10-likelihood ratio) of QTLs ranging from 2.5 to 5.7. Three QTLs were detected under both FACE and Ambient. However,other QTLs were detected only under one level of CO2, which indicated that these loci were sensitive to CO2 concentration. Additionally, two QTLs qFT12 and qGP4 were found for the QTL x Environment (QE) interaction effects. It is suggested that there is a high possibility to improve the yield of rice under elevated CO2 through marker-assisted selection.

Refractive error in urban and rural adult Chinese in Beijing.

PURPOSE: To evaluate refractive error and its demographic associations in an urban and rural population in northern China. DESIGN: Epidemiological study. PARTICIPANTS: The Beijing Eye Study is a population-based cohort study in northern China including 4439 subjects. Excluding pseudophakic and aphakic patients, the present study involved 4319 subjects. It was divided into a rural part (1905 [44.1%] subjects) and an urban part (2414 [55.9%] subjects). Mean age was 55.85+/-10.33 years (range, 40-90). METHODS: Standardized ophthalmologic examination. For statistical analysis, the spherical equivalent was converted to binary variables, and logistic regression was used to investigate the association with continuous or categorical independent variables. MAIN OUTCOME MEASURE: Refractive error. RESULTS: Mean refractive error measured -0.33+/-2.22 diopters (D) (range, -20.88 to +7.88). Myopia of >-0.50 D, -1.0 D, >-6.0 D, and >-8 D, respectively, occurred in 22.9% (95% confidence interval [CI], 21.7-24.2), 16.9% (95% CI, 15.8-18.0), 2.6% (95% CI, 2.2-3.1), and 1.5% (95% CI, 1.1-1.9) of the subjects, respectively. Myopic refractive error was associated significantly with younger age (P<0.001), urban region (vs. rural region) (P<0.001), higher educational background (P<0.001), higher degree of nuclear cataract (P<0.001), decreasing uncorrected visual acuity (UCVA) (P<0.001), decreasing best-corrected visual acuity (BCVA) (P<0.001), and female gender (P<0.001). Prevalence of high myopia (myopic refractive error >-8 D) was associated with age (P<0.001), female gender (P = 0.020), urban region (P = 0.023), and lower BCVA (P<0.001). Mean anisometropia was 1.09+/-2.03 D (median, 0.38; range, 0-22.0). Prevalence of anisometropia of > or =1 D was associated significantly with age (P<0.001), refractive error (P<0.001), BCVA (P<0.001), and region (P<0.001). Mean astigmatic error measured 0.62+/-0.90 D (median, 0.25; range, 0-7.50). Astigmatism of > or =1 D was associated significantly with age (P<0.001), lower UCVA (P = 0.003), lower BCVA (P<0.001), urban area (P<0.001), and degree of cortical cataract (P = 0.027). CONCLUSIONS: As in other population-based studies on Chinese, myopia was more prevalent in younger subjects. Myopia was associated with urban region, educational background, female gender, decreasing visual acuity, and nuclear cataract. If longitudinal studies confirm the association of refractive error with age, refractive surgery may achieve emmetropia only for a limited time.

[The prevalence and its effective factors of primary angle-closure glaucoma in defined populations of rural and urban in Beijing].

OBJECTIVE: To investigate the prevalence of primary angle-closure glaucoma (PACG) of urban and rural residents in Beijing, China. A quantitative comparison was made based on the data collected from this epidemiological survey. We also identify some of the most typical risk factors associated with PACG. METHODS: Glaucomatous screening examination was applied to specific age group populations (aged 40 and older) in the defined district of Beijing and its remote rural county, from June to October, 2001. There are 4451 subjects in all, 1980 rural subjects and 2471 urban subjects, 1939 males and 2512 females. The screen and diagnostic methods used in this survey included van Herick methods and gonioscopy examination to estimate the peripheral depth of the anterior chamber, visual acuity, intraocular pressure, refraction status, stereoscopic fundus photography, and threshold-related visual field tests. Subjects regarded as suspected glaucoma and glaucoma patients are reexamined with standard glaucomatous examination. RESULTS: The response rate of rural and urban population is 79.6% (1980/2488), 87.1% (2471/2836), respectively. The prevalence of PACG (in aged 40 years or older population) resulted from this survey was 1.2% (95% CI 0.9% - 1.5%). However, the prevalence was different between urban and rural residents, 1.1% (95% CI 0.8% - 1.4%) vs. 1.6% (95% CI 1.2% - 2.0%). The prevalence of PACG in female was more than that in male, 1.7% (95% CI 1.3% - 2.1%) vs.0.8% (95% CI 0.5% - 1.1%). A drastic increase in prevalence of PACG with age increase was identified in both survey sites, however, this increase in rural subjects (aged 60-69 years group) occurred ten years earlier than those from urban subjects (aged more than 70 years group). Compared to urban residents, rural residents showed higher prevalence of unilateral low vision (39.3% vs. 20.6%) and blindness (28.6% vs. 14.7%). This survey also confirmed that, as people aging, refraction status became hyperopia, the depth of peripheral anterior chamber became narrow. In the different age groups, female and male groups, the changes of refraction status and the depth of peripheral anterior chamber paralleled the prevalence of PACG. CONCLUSIONS: The prevalence of PACG was obviously different in different groups. This could due to several factors including gender, age, change of refraction status and chamber angle as well.