Quantitative conversion of phytate to inorganic phosphorus in soybean seeds expressing a bacterial phytase

Phytic acid (PA) contains the major portion of the phosphorus in the soybean (Glycine max) seed and chelates divalent cations. During germination, both minerals and phosphate are released upon phytase-catalyzed degradation of PA. We generated a soybean line (CAPPA) in which an Escherichia coli periplasmic phytase, the product of the appA gene, was expressed in the cytoplasm of developing cotyledons. CAPPA exhibited high levels of phytase expression, >or=90% reduction in seed PA, and concomitant increases in total free phosphate. These traits were stable, and, although resulted in a trend for reduced emergence and a statistically significant reduction in germination rates, had no effect on the number of seeds per plant or seed weight. Because phytate is not digested by monogastric animals, untreated soymeal does not provide monogastrics with sufficient phosphorus and minerals, and PA in the waste stream leads to phosphorus runoff. The expression of a cytoplasmic phytase in the CAPPA line therefore improves phosphorus availability and surpasses gains achieved by other reported transgenic and mutational strategies by combining in seeds both high phytase expression and significant increases in available phosphorus. Thus, in addition to its value as a high-phosphate meal source, soymeal from CAPPA could be used to convert PA of admixed meals, such as cornmeal, directly to utilizable inorganic phosphorus.     

Effect of influent substrate ratio on anammox granular sludge: performance and kinetics

Effect of influent substrate ratio on anammox process was studied in sequencing batch reactor. Operating temperature was fixed at 35 ± 1 °C. Influent pH and hydraulic retention time were 7.5 and 6 h, respectively. When influent NO₂ ^?-N/NH₄ ⁺-N was no more than 2.0, total nitrogen removal rate (TNRR) increased whereas NH₄ ⁺-N removal rate stabilized at 0.32 kg/(m³ d). ΔNO₂ ^?-N/ΔNH₄ ⁺-N increased with enhancing NO₂ ^?-N/NH₄ ⁺-N. When NO₂ ^?-N/NH₄ ⁺-N was 4.5, ΔNO₂ ^?-N/ΔNH₄ ⁺-N was 1.98, which was much higher than theoretical value (1.32). The IC₅₀ of NO₂ ^?-N was 289 mg/L and anammox activity was inhibited at high NO₂ ^?-N/NH₄ ⁺-N ratio. With regard to influent NH₄ ⁺-N/NO₂ ^?-N, the maximum NH₄ ⁺-N removal rate was 0.36 kg/(m³ d), which occurred at the ratio of 4.0. Anammox activity was inhibited when influent NH₄ ⁺-N/NO₂ ^?-N was higher than 5.0. With influent NO₃ ^?-N/NH₄ ⁺-N of 2.5–6.5, NH₄ ⁺-N removal rate and NRR were stabilized at 0.33 and 0.40 kg/(m³ d), respectively. When the ratio was higher than 6.5, nitrogen removal would be worsened. The inhibitory threshold concentration of NO₂ ^?-N was lower than NH₄ ⁺-N and NO₃ ^?-N. Anammox bacteria were more sensitive to NO₂ ^?-N than NH₄ ⁺-N and NO₃ ^?-N. TNRR would be enhanced with increasing nitrogen loading rate, but sludge floatation occurred at high nitrogen loading shock. The Han-Levenspiel could be applied to simulate nitrogen removal resulting from NO₂ ^?-N inhibition.     

文心兰25条miRNA前体序列特性及其表达分析

为了解文心兰miRNA的序列特性及其在不同组织部位中以及软腐病侵染过程中的表达规律,该研究对文心兰转录组及miRNA数据库中的25条miRNA前体(pre miRNA)序列和15条成熟体序列进行序列特性分析,并对25条pre miRNAs在文心兰不同组织部位及软腐病侵染的假鳞茎中的表达情况进行实时荧光定量分析。结果显示：（1）文心兰25条pre miRNAs序列可分为13个家族,其中包含15条成熟体序列。（2）Mfold二级结构预测显示,文心兰25条pre miRNAs的最小折叠自由能在-32.04~ -120.98 kal/mol之间,均可以形成典型、稳定的发夹结构。（3）序列比对分析发现,同一家族的前体与成熟体存在一个约20个碱基长度的保守区域,推测该区域可能为文心兰miRNA成熟体所在区域。其中,13个前体家族中,有10个家族的成熟体可以完全定位于其前体中。（4）实时荧光定量PCR结果显示,在文心兰不同组织部位中,miR159 unigene0037857、miR167 unigene0011236、miR167 unigene0002619、miR169 unigene0022341、miR172 unigene006514、miR396 unigene19032、miR398 unigene0009996、miR2950 unigene0006151、miR2950 unigene0006422等pre miRNAs在叶片中大量累积可能有利于其叶的形态建成;miR162 unigene0003615、miR162 unigene0013441、miR166 unigene0011870、miR168 unigene0009958等pre miRNAs同时在根和叶中均大量表达有利于其根和叶的发育;而miR171 unigene0045985、miR396 unigene0011179、miR396 unigene0011180在根和茎中的大量表达可能有利于其根和茎的发育。（5）在软腐病病菌侵染文心兰假鳞茎的过程中,miR159 unigene0037857、miR167 unigene0011236、miR396 unigene0011179、miR396 unigene0011180、miR396 unigene0019032、miR845 unigene0012489的表达总体呈下降趋势;miR162 unigene0040566、miR166 unigene0011870、miR168 unigene0009958、miR171 unigene0045985在软腐病菌液侵染4 h其表达量升高,之后表达量下调;miR162 unigene0003615、miR167 unigene0002619、miR168 unigene0047942、miR169 unigene0022341、miR845 unigene0012489在菌液侵染过程中其表达量呈现先下降后上升的趋势,并在侵染8 h后表达量达到最低。研究表明,文心兰pre miRNAs 13个家族不同成员在进化进程中具有高度保守性与特异性的结构特点,并可能广泛参与文心兰不同组织的生长发育及参与软腐病菌侵染的响应。     

低浓度SO2污染与桃蚜的生长和繁殖

用40或80ppbSO₂熏气处理过的油菜饲养桃蚜(Myzus persicae),研究低浓度SO₂污染对这种蚜虫的影响。与对照组相比,两个处理组若蚜的发育速率显著加快,5天内的平均相对生长速率(MRGR)分别增加22.3%和31.5%。成蚜的繁殖力提高40—90%,导致桃蚜种群的内禀增长率(r_m)分别升高17%和27%。比较了桃蚜和其它3种蚜虫对SO₂污染的反应强度,讨论了SO₂污染促进某些植食性昆虫生长的可能原因。     

Effects of transgenic Bt cotton on the population density, oviposition behavior, development, and reproduction of a nontarget pest, Adelphocoris suturalis (Hemiptera: Miridae)

Transgenic cotton has shown great promise for the control of target pest insects; however, frequent outbreaks of nontarget pest mirids has been recorded in recent years in northern China. To test the hypothesis that transgenic cotton contributes to nontarget pest outbreaks, we studied the impact of transgenic Bt cottons (both Bt and Bt + CpTI) on the fitness of nontarget pest Adelphocoris suturalis Jakovlev. No significant differences were detected between population densities of A. suturalis in unsprayed nontransgenic cottons and in unsprayed transgenic Bt cottons in 2007, 2008, and 2009. No difference in preferred oviposition site or egg production was detected between transgenic and nontransgenic cottons in both free choice and no choice tests. No difference in life table parameters was detected for A. suturalis between Bt cottons and nontransgenic cottons. All these results indicated that transgenic crops did not contribute to the nontarget pest outbreaks when being compared with their parental lines. The possible reasons for intensified pest status of A. suturalis, such as decrease of pesticide application, deficient natural enemies, and area-wide shift of cotton varieties, were discussed.     

High-performance liquid chromatographic analysis of bisphenol A and 4-nonylphenol in serum, liver and testis tissues after oral administration to rats and its application to toxicokinetic study

A sensitive and simple method based on solid-phase extraction (SPE) and HPLC with fluorescence detection for the determination of bisphenol A (BPA) and 4-nonylphenol (4-NP) in rat serum, liver and testis tissues has been developed. The chromatographic conditions consisted of a C18 column and mobile phase composition of acetonitrile and water with flow rate of 1.0 ml/min. The fluorescence detection was performed at excitation and emission wavelengths of 227 nm and 313 nm, respectively. Under these conditions, BPA and 4-NP were well separated and showed good linearities in the ranges of 0.01-50.0 microg/ml for BPA and 0.15-150.0 microg/ml for 4-NP with correlation coefficients greater than 0.999. The detection limits of serum and tissue samples were 2.8 ng/ml and 1.4 ng/g for BPA and 5.6 ng/ml and 2.8 ng/g for 4-NP at a signal-to-noise ratio (S/N) of 3. The intra-assay and the inter-assay precisions were better than 11.4%. Recoveries of BPA and 4-NP were 78.6-95.0% and 80.2-93.4%, respectively. The proposed method was applied to a toxicokinetic study of BPA and 4-NP including individual and combined oral administration to rats. The results showed that 4-NP remarkably altered the toxicokinetic parameters of BPA in testis, while parameters of BPA were not obviously altered in serum and liver under the experimental conditions investigated. On the other hand, there was no significant difference in the toxicokinetics of 4-NP when administered with BPA.     

兰州市SO2时空动态分布特征及其与城市生态系统的关系

强化城市大气污染物多尺度监测手段,辨识污染物时空演变规律及其与城市生态系统各要素间的相互关系,可为城市大气环境质量综合管控提供科学依据。利用OMI、EOS/MODIS卫星影像及其他统计类数据,在分析了兰州市2006—2016年对流层SO_2柱浓度的时空分布特征基础上,进一步探讨了NDVI、工业总产值、汽车保有量等城市自然经济社会生态要素的结构变化与SO_2浓度变化的关系。结果表明:1)兰州市对流层SO_2空间分布不均,呈现出以河谷低海拔区域分布为中心,向西北、东南山地两侧递减的"倒U"形格局;2)近10年内,兰州市大气SO_2柱浓度值在0.033—0.25(多布森单位)之间,总体呈下降趋势,年均降幅为14%,降速为0.02(多布森单位)/a;3)兰州市大气SO_2柱浓度变化特征与城市工业生产和交通因素关系密切,又受地形、季节风向等气象气候条件制约;4)兰州市NDVI指数与大气SO_2柱浓度值相关性明显,兰州市植被覆盖度较低,植被指数为0.09—0.66,对SO_2净化能力有限。综合以上结果,本研究认为,通过调控城市社会、经济、自然生态系统结构,并适当采用基因工程等科学技术来增加地表植被覆盖量,是下一步兰州市大气环境质量管理的重要举措之一。     

食物链长度理论研究进展

食物链长度（Food chain length,FCL）是生态系统中最重要的特点之一,它通过改变生物间的营养关系,影响着生物多样性,群落的结构以及稳定性;它是反映食物网物质转换与能量传递的综合指数,食物链及其动态特征是生态学许多重要理论的基础,食物链长度理论的研究进展,推动了人们对水域生态系统中生物和非生物相互作用的理解。回顾了食物链长度的3种度量方法及其详细的计算方法,在此基础上简述了各方法的特点。综述了食物链长度的决定因素的4种假说（资源可利用性假说、生产力空间假说、生态系统大小假说、动态稳定性假说）及其交互作用,重点总结了湖泊食物链长度的空间格局与决定因素的研究进展。最后,食物链长度研究展望,包括食物链长度决定因子研究存在的问题及发展方向的总结,以及在在水域生态学中的应用的研究进展,例如食物链长度在指示污染物的生物富集中的研究进展、食物链食物链长度在指导生物操作、以及在食物链长度在对气候变化响应方面的研究进展等等。     

Role of melatonin in the dynamics of acute spinal cord injury in rats

Melatonin is well-documented to have the ability of reducing nerve inflammation and scavenging free radicals. However, the therapeutic effect of melatonin on spinal cord injury has not been fully described. In this study, we assessed the effect of melatonin on T9 spinal cord injury established by Allen method in rats. Melatonin deficiency significantly delayed the recovery of sensory and motor functions in SCI rats. Treatment with melatonin significantly alleviated neuronal apoptosis and accelerated the recovery of spinal cord function. These results suggest that melatonin is effective to ameliorate spinal cord injury through inhibition of neuronal apoptosis and promotion of neuronal repair.     

Cancer-associated fibroblasts–derived exosomes-mediated transfer of LINC00355 regulates bladder cancer cell proliferation and invasion

The aim of this study was to investigate the regulatory mechanism of cancer-associated fibroblasts (CAFs) exosome in bladder cancer (BC) cell proliferation and invasion. CAFs and normal fibroblasts (NFs) were isolated from tumor tissues and adjacent normal tissues of BC patients, and examined by immunocytochemistry for the expression of fibroblast activation protein alpha (FAP) and α-smooth muscle actin (α-SMA). Exosomes were extracted from CAFs and NFs and observed under a transmission electron microscope, and expression of the exosome markers CD9 and CD63 was confirmed by western blotting. The distribution and intensity of fluorescence were observed by confocal laser microscopy to analyze exosomes uptake by BC cell lines T24 or 5367. BC cell proliferation and invasion were detected by MTT and Transwell assays, respectively. LINC00355 levels in CAFs, NFs, CAFs exosome, NFs exosome, and BC cells were detected by quantitative real-time polymerase chain reaction (qRT-PCR). Results showed that CAFs exosome significantly promoted BC cell proliferation and invasion relative to NFs exosome. LINC00355 expression was significantly elevated in CAFs exosome when compared with that in NFs-exosome. Up-regulated LINC00355 expression was observed both in T24 and 5367 cells co-incubated with CAFs exosome. Exosomes derived from LINC00355 siRNA-transfected CAFs observably repressed BC cell proliferation and invasion when compared with control siRNA-CAFs exosome. In conclusion, CAFs exosome–mediated transfer of LINC00355 regulates BC cell proliferation and invasion. Significance of the study. In this study, our data suggest that the exosomes released from CAFs promote BC cell proliferation and invasion. The mechanism of this effect is, at least in part, related to the increased LINC00355. Regulation of LINC00355 expression in exosomes released from CAFs might be a putative therapeutic strategy against the pathogenesis of BC.