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Although information on the reproductive biology of the endangered plant family Gesneriaceae is well known, the pollination mechanisms of these plants in karst regions are poorly understood. This study demonstrated the flowering phenology, pollinators, and breeding system of Hemiboea ovalifolia in karst regions. Findings revealed that the anthesis of H. ovalifolia often occurred late, during sunset, or early morning, with duration of 2-4 days; there was a certain level of temporal overlap between pollen viability and stigma receptivity; the most effective pollinators were Bombus ignitus and Anthophora zonata. Controlled pollination indicated that these plants were pollen limited and exhibited late-acting inbreeding depression resulting from the seed sets; there were significant differences in fruit sets between open-pollination and self-pollination or cross pollination, and in seed sets between self-pollination and cross-pollination or open-pollination. Despite the co-existence of large numbers of fruit and seed sets, and vegetative propagation in H. ovalifolia, a failure in seedling survival, and long duration to establishing first-year seedlings in natural populations suggests that the species does not easily recover from damage.
Human activity has greatly increased the amount of biologically available nitrogen entering the natural environment. Addition of N can affect growth of and competitive interactions between native and invasive plants, thus increasing or decreasing the risk of invasion by alien species. Bidens frondosa is an invasive weed native to North America that recently has begun to spread in China. The influence of soil nutrient content on B. frondosa invasion has not yet been reported. In a common garden experiment, we compared the growth and competitive effects between B. frondosa and its co-occurring native congeners — B. tripartita and B. biternata — under three N levels (0 g m⁻², 1 g m⁻², 5 g m⁻²) to assess whether increased levels of N modifies risk of B. frondosa invasion. Our results showed that while N additions increased both the growth and competitive advantage of B. frondosa as compared with that of the native congener species, results are particularly pronounced under high N levels. While growth responses to N addition varied little among invasive populations under high N levels, the competitive effect of B. frondosa did vary among populations and was significantly greater than that of the congeners. Anthropogenic N additions are likely to increase risks of B. frondosa invasions. Thus, management efforts should focus on the reduction of N input to ecosystems to mitigate invasions by B. frondosa.
Background: Platelet derived growth factors (PDGFs) are key components of autocrine and paracrine signalling, both of which play important roles in mammalian developmental processes. PDGF expression levels also relate to oxygen levels. The characteristics of yak PDGFs, which are indigenous to hypoxic environments, have not been clearly described until the current study. Materials and methods: We amplified the open reading frame encoding yak (Bos grunniens) platelet derived growth factor-alpha (PDGFA) from a yak skin tissue cDNA library by reverse transcriptase polymerase chain reaction (PCR) using specific primers and Sanger dideoxy sequencing. Expression of PDGFA mRNA in different portions of yak brain tissue (cerebrum, cerebellum, hippocampus, and spinal cord) was detected by quantitative real-time PCR (qRT-PCR). PDGFA protein expression levels and its location in different portions of the yak brain were evaluated by western blot and immunohistochemistry. Results: We obtained a yak PDGFA 755 bp cDNA gene fragment containing a 636 bp open reading frame, encoding 211 amino acids (GenBank: KU851801). Phylogenetic analysis shows yak PDGFA to be well conserved, having 98.1% DNA sequence identity to homologous Bubalus bubalus and Bos taurus PDGFA genes. However, 8 nucleotides in the yak DNA sequence and 4 amino acids in the yak protein sequence differ from the other two species. PDGFA is widely expressed in yak brain tissue, and furthermore, PDGFA expression in the cerebrum and cerebellum are higher than in the hippocampus and spinal cord (p > 0.05). PDGFA was observed by immunohistochemistry in glial cells of the cerebrum, cerebellum, and hippocampus, as well as in pyramidal cells of the cerebrum, and Purkinje cell bodies of the hippocampus, but not in glial cells of the spinal cord. Conclusions: The PDGFA gene is well conserved in the animal kingdom; however, the yak PDGFA gene has unique characteristics and brain expression patterns specific to this high elevation species. (Folia Morphol 2017; 76, 4: 551–557)
The ETHYLENE-INSENSITIVE3 (EIN3)/EIN3- Like (EIL) EIN3/EILs, novel nuclear proteins, are located at the downstream position of the ethylene signal transduction pathway. LeEIL1, which is expressed in fruit throughout ripening, is key transcription factor in the ethylene signaling pathway in tomato. To reveal its function, the LeEIL1 gene has been transformed into and expressed in the ein2 mutant of Arabidopsis. The expression levels of the transgene in the single copy line, LeEIL1-ein2-b, were higher than those in the multiple-copy line, LeEIL1-ein2-a. The ethylene-insensitive phenotype of the ein2 mutant plants has been partially recovered by expression of LeEIL1. The florescence of LeEIL1-ein2-a and LeEIL1- ein2-b exceeded that of the ein2 mutant but was still less than that of wild type of Arabidopsis. The expression of four pathology-related genes (AtPR3, 4, AtPDF1.2 and AtGST2) has been analyzed in LeEIL1 transgenic ein2 mutant plants. The expression of AtPR3 and AtPR4, which was reduced in the ein2 mutant, was enhanced in the two transgenic Arabidopsis plants. The expression of the AtPDF1.2 gene was unaffected in the two transgenic Arabidopsis lines, the ein2 mutant and wild-type Arabidopsis plants. In addition, the expression level of AtGST2 in transgenic Arabidopsis plants was lower even than that in ein2 mutant and wild-type Arabidopsis plants.
This study developed the hybrid partial-nitrification and anammox (HPNA) reactor to increase efficiency in ammonia nitrogen (NH₄⁺-N) removal. The HPNA reactor contained a fixed bed filled with biofilm carrier and a fluidized bed with mechanical stirrer to maximize withholding the sludge. The reactor achieved a total nitrogen (TN) removal efficiency of 70% and NH₄⁺-N removal efficiency of 84%. The maximum nitrogen loading rate (NLR) of 6.1 kg-N/m³/d and nitrogen removal rate (NRR) of 4.7 kg-N/m³/d were achieved with a short hydraulic retention time (HRT) of 3 h. After 160 days of operation, both the mixed liquid suspended solids (MLSS) and the mixed liquor volatile suspended solids (MLVSS) increased in the lower part (from 3.7 to 14.7 g/L and 1.8 to 10.9 g/L, respectively) and the upper part (from 3.7 to 11.8 g/L and 1.9 to 6.7 g/L, respectively). The proteins (PN)/ polysaccharides (PS) ratio increased from 0.96 to 1.1 in the lower part but decreased to 0.7 in the upper part. Bacterial community profiles generated from 16S rRNA sequences further showed that the ammonium-oxidizing bacteria (AOB) Nitrosomonas and the anammox bacterium Brocadiales were enriched in the HPNA reactor.
Yield loss due to insufficient potassium fertilizer supply has been well documented; however, the information about the negative effect of potassium deficiency on crop yield caused by ecophysiological determinants is not enough. A field experiment with three K treatments (severe K deficiency treatment, K₁; moderate K deficiency treatment, K₂; and sufficient K supply treatment, K₃) was conducted to (1) assess the effects of potassium deficiency on green leaf area index (GLAI) reduction; (2) quantify the contributions of single leaf area, leaf senescence, and leaf appearance to GLAI reduction under potassium deficiency; (3) reveal the changes in the contributions of accumulated radiation interception (RIacc) and radiation-use efficiency (RUE) to above-ground biomass (AM) decrease of oilseed rape under different K supplies. GLAI was restrained due to potassium deficiency, with a reduction ranging from 10.6 to 45.4%. The reduced single leaf area and accelerated leaf senescence caused by potassium starvation accounted for 5.9–23.7% and 2.4–29.0% reduction in GLAI, but delayed leaf appearance rate contributed little. The RIacc during the seedling stage in the K₁, K₂, and K₃ treatments was 101.2, 110.7, and 120.0 MJ m⁻² , respectively, and the RUE in the K₁, K₂, and K₃ treatments was 1.03, 2.22, and 2.98 g MJ⁻¹ , respectively, which caused a 61.7% and 48.2% reduction of the final harvested AM in the K₁ and K₂ treatments compared with the K₃ treatment. When AM reduction was less than 24.8%, RIacc was the main determining factor; however, it transferred to RUE when biomass decreased more. In conclusion, GLAI decreased due to potassium starvation was mainly caused by the reduced single leaf area and accelerated leaf senescence, and the relative contribution of RIacc and RUE to AM decline was related to the degree of potassium deficiency.
 OmpF plays very important roles in the influx of antibiotics and bacterial survival in the presence of antibiotics. However, high-grade mutant OmpF and its function in decreasing bacterial survival rate have not been reported to date. In the present study, we cloned a high-grade mutant OmpF (mOmpF) and sequence analysis suggested that over 45 percent of the DNA sequence was significantly mutated, leading to dramatic changes in over 55 percent of the amino acid sequence. mOmpF protein was successfully expressed. When grown in the presence of antibiotic, the bacterial survival rate decreased and the antibiotic inhibition zone became larger with the increase of the mOmpF. It was concluded that concentration of high-grade mutant mOmpF dramatically influenced the bacterial survival rate. The study presented here may provide insights into better understanding of the relationships between structure and function of OmpF.
The present study showed the toxicity caused by heavy metal and its detoxification responses in two desert plants: perennial Peganum harmala and annual Halogeton glomeratus. In pot experiments, 1-month-old seedlings were grown under control and three levels of combined heavy metal stress. Seedling growth as well as heavy metal accumulation, antioxidative enzymes [superoxide dismutase (SOD), catalase (CAT) and ascorbate peroxidase (APX)] activities and the contents of malondialdehyde (MDA), and hydrogen peroxide (H₂O₂) in leaves was examined after 2 months of heavy metal exposure. Compared with H. glomeratus, growth of P. harmala was more severely inhibited. In leaves, the heavy metal accumulation pattern in both the plants was dose-dependent, being more in H. glomeratus. H. glomeratus exhibited a typical antioxidative defense mechanism, as evidenced by the elevated activities of all the three enzymes tested. P. harmala exhibited a different enzyme response pattern, with a significant reduction in CAT activity, and elevated SOD and APX activities, but significantly elevated APX activity was only at the lowest heavy metal concentration. MDA and H₂O₂ contents were significantly enhanced in leaves of heavy metal-treated P. harmala, but in H. glomeratus were elevated only at the highest heavy metal treatment. These results indicated that H. glomeratus had a greater capacity than P. harmala to adapt to oxidative stress caused by heavy metal stress, and antioxidative defense in H. glomeratus might play an important role in heavy metal tolerance.
Cold-inducible RNA binding protein (CIRP) is over-expressed during cold and many other stresses, and could regulate the adaptation to hypothermia. In the present investigation, the objective was to determine the expression of CIRP in adult yak heart, liver, spleen, lung, kidney, brain, ovary, testis and skin by relative quantitative real time polymerase chain reaction (RT-PCR), Western blot and immunohistochemistry from mRNA and protein levels. The CIRP open reading frame encoding was cloned from the domestic yak brain. Results of RT-PCR and Western blot showed the high expression level of CIRP in the heart, kidney, brain, testis and skin, and the lower expression level of CIRP in the lung. Immunohistochemical staining showed CIRP was expressed in the nucleus of neuronal cells, spermatogonia, primary spermatocytes and epidermal cells, and in the cytoplasm of the residual tissues. These observations may provide new data to understand and further study the important role of CIRP protein in the plateau adaptation of the domestic yak on long-term evolution. (Folia Morphol 2016; 75, 4: 460–466)
Serpins are a broadly distributed family of protease inhibitors. In this study, the gene encoding Bombyx mori serpin-2 (Bmserpin-2) was cloned and expressed in E. coli. The Bmserpin-2 cDNA contains a 1125 bp open reading frame (ORF). The deduced protein has 374 amino-acid residues, contains a conserved SERPIN domain and shares extensive homology with other invertebrate serpins. RT-PCR analysis showed that Bmserpin-2 was expressed in all developmental stages of B. mori larvae and various larval tissues. Subcellular localization analysis indicated that Bmserpin-2 protein was located in the cytoplasm. Interestingly, real-time quantitative PCR revealed that the expression of Bmserpin-2 in the midgut of susceptible B. mori strain 306 significantly increased at 72 hours post inoculation (hpi) when infected with BmNPV. However, there was no significant increase of the Bmserpin-2 expression in resistant strain NB infected with BmNPV. Thus, our data indicates that Bmserpin-2 may be involved in B. mori antiviral response.
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