Agrobacterium tumefaciens-mediated genetic transformation of the phytopathogenic fungus Penicillium digitatum

Agrobacterium tumefaciens-mediated transformation （ATMT） system was assessed for conducting insertional mutagenesis in Penicillium digitatum, a major fungal pathogen infecting post-harvest citrus fruits. A transformation efficiency of up to 60 transformants per 106 conidia was achieved by this system. The integration of the hph gene into the fungal genome was verified by polymerase chain reaction （PCR） amplification and sequencing. These transformants tested were also shown to be mitotically stable. Southern blot analysis of 14 randomly selected transformants showed that the hph gene was randomly integrated as single copy into the fungal genome of P. digitatum. Thus, we conclude that ATMT of P. digitatum could be used as an alternatively practical genetic tool for conducting insertional mutagenesis in P. digitatum to study functional genomics.     

毕赤酵母FLD1启动子元件的克隆与测序

为了应用毕赤酵母表达某些食用蛋白或同时表达多个异源蛋白,本文以毕赤酵母GS115基因组DNA为模板,采用prime5.9程序设计了一对不等长引物,经多轮直接多聚酶链式反应,扩增获得了大小为597bp目标DNA片段;再经限制性内切酶和双脱氧末端终止法分析,其DNA片段排列顺序与EMBL发表的FLD1启动子的序列完全一致。     

A time-space optimal parallel sorting on a hypercube

In this paper we discuss a parallel sorting algorithm on a hypercube. Its time complexity isO(n logn/p) +O(n). Here,P is the number of processors avaliable and n, the amount of items to be sorted. Take the problem of time-space optimization into consideration, whenP≤O(logn), this algorithm is both time-space optimal and cost optimization. But this means only speedup isO(p) and it is not linear speedup. Therefore, we further discuss relevant parallel efficiency problems.     

Studies of biouptake and transformation of mercury by a typical unicellular diatom Phaeodactylum tricornutum

Mercury(Hg) is a toxic heavy metal with its biogeochemical cycling in the ocean depending on the type and behavior of the oceanic microalgae.The present work aimed to evaluate bioaccumulation and transformation of Hg by Phaeodactylum tricornutum,a typical unicellular diatom,when exposed to the extremely high level of Hg in order to understand the possible mechanisms of acute stress response.P.tricornutum can accumulate Hg(its bioaccumulation factor is at 104 level),and the 96 h EC 50 was estimated to be 145μg L-1.The amounts of surface-bound Hg being about 1.2 to 4.8 times higher than those of intracellular Hg under exposure to HgCl 2(from 20 to 120μg L-1 concentrations) suggested that the cell wall of P.tricornutum is an important "fence" towards Hg.After entering the P.tricornutum cell,Hg underwent transformation in its chemical form via interactions with high molecular weight sulfur-containing proteins(accounting for 68% of the intracellular Hg),and glutathione as well as the induced phytochelatins(PCs)(24% Hg) which alleviated the toxicity of HgCl2.In addition,the existence of organic ligands greatly influenced the uptake and transformation behavior of P.tricornutum towards HgCl2,especially in the case of cysteine(Cys),which increased the uptake of Hg,but alleviated the toxicity of Hg towards P.tricornutum due to the fact that Cys is an important precursor for the synthesis of PCs inside the cell.The uptake process of Hg by P.tricornutum was in agreement with the Freundlich isotherm,suggesting a typical heterogeneous sorption process.More importantly,we observed the conversion of HgCl2 into methylmercury inside the P.tricornutum cells and its release into the culture solution using HPLC/CVG-AFS and GC-MS,although the mechanism needs to be further investigated.     

Derivation of P–T paths for Middle–Late Triassic very low-grade metapelite from Hongcan Well 1 in the Zoigê Depression and their tectonic implications

Using XRD measurements of illite crystallinity,b0values of K-rich white micas and chemical compositions of chlorites combined with homogenization temperatures of fluid inclusions in calcite,the metamorphic P–T conditions of very low-grade metapelites from different depths in the 7 km Hongcan Well 1 in the Zoige?Depression,Sichuan,China,have been obtained in this paper.Knowledge of the tectonic evolution of the studied basin allows us to derive geothermal curves for Middle and Late Triassic time(the geothermal peak)and the present day(from thermal logging of the borehole).The retrograde P–T tracks from clockwise P–T–t loops have been revealed by the plotting of individual samples on each geotherm,which can be interpreted by the lithospheric thickening due to sedimentation and folding followed by continuous uplift and erosion.A stratigraphic log indicates that the Well penetrates the steep limb of a fold which perturbed the peak geotherm and caused some P–T paths to cross.     

热敏灸治疗肛周脓肿术后尿潴留的临床观察

目的观察热敏灸治疗肛周脓肿术后尿潴留的临床效果。方法选择2016年6月~2017年6月就诊于我院的40例肛周脓肿术后尿潴留患者为研究对象,随机分为对照组和观察组各20例,对照组给予膀胱区按摩,水流声诱导及下腹部热敷治疗,观察组在此基础上给予热敏灸治疗。治疗后观察患者自行排尿时间、排尿完全通畅时间、膀胱残留尿量进行统计,计算临床治疗的有效率。结果经治疗后与对照组相比,对照组有效率为70%,观察组有效率为95%,观察组患者的临床总有效率明显优于对照组(P0.05)。结论热敏灸能有效改善肛周脓肿术后患者的尿潴留症状,利于患者排尿、缩短病程,疗效确切,值得临床推广应用。     

形成原子核群子结构的四项原理及其等腰三角形核素周期律

根据第四统计力学——JRG群子统计理论,首次提出了形成原子核结构的四项原理及相应的原子核周期律。这四项原理为:一是原子核有群子结构单元;二是形成原子核时群子通过热核聚合反应过程使核素结构有严格的排列顺序;三是原子核内群子结构间有动态共振作用;四是偶数群子稳定,而非偶数群子是引起总角动量和β⁺,β^-衰变的根源。基于上述四项原理,提出了核群子结构基本单元有(PB),(PB₂),(P₂B₃),并随着质子数Z的增加,核群子结构由(PB)_k过渡到(PB)_k(P₂B₃)_l;由(PB)_n(PB₂)_m过渡到(P₂B₃)_s(PB₂)_t。从而导出了k(-t)=2n(s)-Z,k(-t)=Z-2l(m)的关系式。基于此,可以画出等腰三角形原子核群子周期律。还发现,不管A,N,Z如何变化,有下列严格关系式:Z/N=(n+m)/(n+2m)或Z/N=(k+l)/(k+3l)并且Z=n+l。此公式高度地反映了所有原子核内质子和中子分布的整数规律。还可以通过等腰三角形周期律得知:k,l,m,n,s,t均与核素群子(PB),(PB₂), (P₂B₃),(P₃B₄)的2,3,5,7整数倍有关。     

DEX1, a plasma membrane-localized protein, functions in microspore development by affecting CalS5 expression in Arabidopsis thaliana

Microsporogenesis in flowering plants plays important roles in sexual reproduction. It has been reported that DEFECTIVE IN EXINE FORMATION1 (DEX1) is essential for exine pattern formation in Arabidopsis thaliana. However, the functions of DEX1 in regulating microspore development are largely not understood. In this study, we show that DEX1 is strongly expressed in the tapetum by using RNA in situ hybridization. dex1 microspores were degenerated and aborted after release from the tetrads. The callose wall in tetrads was thinner in the dex1 mutant than in the wild type, suggesting that DEX1 affects callose formation at the tetrad stage during anther development. RT-PCR and real-time PCR analyses showed that CalS5, which plays an important role in callose synthesis during microspore development, was greatly down-regulated in dex1 plants. DEX1 encodes a membrane protein with one transmembrane domain, one intracellular domain and one extracellular domain. Collectively, our results demonstrate that DEX1 is essential for microspore development, possibly by regulating the expression of CalS5.     

Induction of human β-defensin-2 in airway epithelial cells by Streptococcus pneumoniae： involvement of IP3-dependent intracellular calcium release and NF-κB

Human β-definsin-2 （hBD-2） is mainly induced by bacterial factors and pro-inflammation mediators in epithelial cells. As the major cause of community-acquired pneumonia, whether Streptococcus pneumoniae （S. pneu-moniae） stimulation induces hBD-2 expression in airway epithelial cells is elusive. In this study, we found that S. pneumoniae stimulation induced hBD-2 expression in a time-and concentration-dependent manner in primary human airway epithelial cells. To further reveal the mechanism of S. pneumoniae inducing hBD-2, we found that S. pneumoniae stimulation activated NF-κB signaling pathway. Specific NF-κB inhibitor, PDTC, could reverse the induction of hBD-2 by S. pneumoniae. We also found that cellular inner Ca^2＋ signaling is involved in the S. pneumoniae-induced hBD-2. Taken together, our find-ings indicated that S. pneumoniae can stimulate the expression of hBD-2 in airway epithelial cells and NF-κB and inositol triphosphate-dependent intracellular calcium release is involved in this induction.     

Bioconversion of low quality lignocellulosic agricultural waste into edible protein by Pleurotus sajor-caju (Fr.) Singer

Pleurotus sajor-caju (Fr.) Singer was cultivated on selected agro wastes viz. cotton stalks, groundnut haulms, soybean straw, pigeon pea stalks and leaves and wheat straw, alone or in combinations. Cotton stalks, pigeon pea stalks and wheat straw alone or in combination were found to be more suitable than groundnut haulms and soybean straw for the cultivation. Organic supplements such as groundnut oilseed cake, gram powder and rice bran not only affected growth parameters but also increased yields. Thus bioconversion of lignocellulosic biomass by P. sajor-caju offers a promising way to convert low quality biomass into an improved human food.     

The Distribution of a CP-asymmetric quantity in the decay channelJ/Ψ→π + π − π 0

The distribution of a CP-asymmetric quantityA in the decay channelJ/ψ→π ⁺ π ^? π ⁰ is investigated. This dimensionless quantityA is constructed from the momenta of the π-meson final states, namely A=P _x ^{π +} P _y ^{π ?} -P _x ^{π ?} P _y ^{π ?}, whereP _h,P _x ^h andP _y ^h are the module of the momentum of a hadron h, the x- and y-components of that momentum respectively, theJ/ψ particle is produced by e⁺ e^? collision, and the direction of the momentum of e⁺ is taken to be the positive direction ofz axis. There would exists the violation under the combined transformation of charge conjugation and space reflection (CP violation) when the average ofA among a lot of events, 〈A〉, be examined to be nonzero clearly from data. In this way, 748 events are selected from the BES experimental data, and analyzed. The corresponding averageA-value is measured to be 〈A〉 = 0.010 39 ± 0.014 61 ± 0.015 2. Further, some discussion of our result, the size of the data sample being need for further attempt and a possible perspective are given.     

Fungal diversity on fallen leaves of Ficus in northern Thailand

Fallen leaves of Ficus altissima, F. virens, F. benjamina, F. fistulosa and F. semicordata, were collected in Chiang Mai Province in northern Thailand and examined for fungi. Eighty taxa were identified, comprising 56 anamorphic taxa, 23 ascomycetes and l basidiomycete. Common fungal species occurring on five host species with high frequency of occurrence were Beltraniella nilgirica, Lasiodiplodia theobromae, Ophioceras leptosporum, Periconia byssoides and Septonema harknessi. Colletotrichum and Stachybotrys were also common genera. The leaves of different Ficus species supported diverse fungal taxa, and the fungal assemblages on the different hosts showed varying overlap. The fungal diversity of saprobes at the host species level is discussed.     

Different responses of two Mosla species to potassium limitation in relation to acid rain deposition

The increasingly serious problem of acid rain is leading to increased potassium (K) loss from soils, and in our field investigation, we found that even congenerically relative Mosla species show different tolerance to K-deficiency. A hydroponic study was conducted on the growth of two Mosla species and their morphological, physiological and stoichiometric traits in response to limited (0.35 mmol K/L), normal (3.25 mmol K/L) and excessive (6.50 mmol K/L) K concentrations. Mosla hangchowensis is an endangered plant, whereas Mosla dianthera a widespread weed. In the case of M. hangchowensis, in comparison with normal K concentration, K-limitation induced a significant reduction in net photosynthetic rate (P _n), soluble protein content, and superoxide dismutase (SOD) activity, but an increase in malondialdehyde (MDA) concentration. However, leaf mass ratio (LMR) and root mass ratio (RMR) were changed little by K-limitation. In contrast, for M. dianthera, K-limitation had little effect on P _n, soluble protein content, SOD activity, and MDA concentration, but increased LMR and RMR. Critical values of N (nitrogen):K and K:P (phosphorus) ratios in the shoots indicated that limitation in acquiring K occurred under K-limited conditions for M. hangchowensis but not for M. dianthera. We found that low K content in natural habitats was a restrictive factor in the growth and distribution of M. hangchowensis, and soil K-deficiency caused by acid rain worsened the situation of M. hangchowensis, while M. dianthera could well acclimate to the increasing K-deficiency. We suggest that controlling the acid rain and applying K fertilizers may be an effective way to rescue the endangered M. hangchowensis.     

Electronic structure and properties of FeS2 with the space groups of Pa3 and P1

The electronic structure and properties of FeS₂ with the space groups of Pa3 and P1 were studied by the density functional theory. The generalized-gradient approximation exchange-correlation functional was used in conjunction with a plane wave-ultrasoft pseudopotential representation. Calculation results show that differences are observed in electronic structures and properties between Pa3 and P1 crystals. The band gap and energy loss of P1 are smaller than those of Pa3 crystal, while the dielectric constant, conductivity, refractive index, extinction coefficient, and intensity of optical absorption of P1 are larger than those of Pa3. These behaviors are attributed to the differences in symmetry, atomic arrangement, and Mulliken bond population of each unit for Pa3 and P1 crystals.     

Electrochemical properties of LaFeO3-rGO composite

LaFeO_3-xwt% r GO composite(x = 8, 10, 12) was synthesized by ultraphonic stirring and lyophilization method.SEM, TEM and XRD results show that the perovskite-type LaFeO_3 was dispersed by rGO to form special porous structure due to the gauze-shaped wrinkles and folds structure of rGO. It was found that the special porous structure can effectively increase the specific surface area and suppress particle aggregation of LaFeO_3, thus improving the electrical conductivity and appreciably enhancing the electrochemical properties of LaFeO_3. As compared with LaFeO_3, the maximum discharge capacity of the composite(x=10) increased from 209.5 mAhg~(–1) to 334.6 mAhg~(–1).The High rate dischargeability at a discharge current density of 1500 mAg~(–1)(HRD1500) and the capacity retention rate after 100 charge/discharge cycles(S100) of the composite increased by 9% and 17%, respectively.     

The comparison of biomarkers released by hydropyrolysis and Soxhlet extraction from source rocks of different maturities

The Permian Dalong Formation(P2d) source rocks from the mature Guangyuan outcrop section and the overmature Wangcang outcrop section in Sichuan Basin were selected.The Soxhlet extraction and kerogen catalytic HyPy were conducted on the P2d source rocks.The biomarkers obtained by both methods were compared to discuss their difference and the influence extent of thermal maturation on covalently bound biomarkers.The results show that covalently bound biomarkers can hardly be correlated to the corresponding free biomarkers due to severe thermal alterations and/or interference of migrated hydrocarbons in both outcrop sections and thermal maturation have much lower influence on covalently bound biomarkers in kerogens than on free biomarkers in Soxhlet extracts.The application of HyPy in study of biomarker geochemistry can reduce the thermal maturation effect on biomarkers(Ro≤2.4%) to a greater extent and the interference of migrated hydrocarbons between interbedded layers.The covalently bound biomarkers released by HyPy are useful in the study of biomarker geochemistry and oil-source correlation on high-overmature source rocks.     

Construction of Streptomyces lydicus A01 transformant with the chit33 gene from Trichoderma harzianum CECT2413 and its biocontrol effect on Fusaria

Streptomyces lydicus A01 resists many plant pathogens (including Fusarium spp.) by producing the antifungal agent natamycin, which binds to the ergosterol of fungal cell membranes and inhibits the growth of pathogens. Trichoderma harzianum CECT2413 is a widely-distributed soil fungus that antagonizes several plant fungal pathogens (including Fusarium spp.) by producing chi-tinase and degrading chitin, a major component of the fungal cell wall. This study attempted to enhance the biocontrol effect of S. lydicus A01 on Fusarium spp. by transforming the chitinase gene of Trichoderma. Chitinase and natamycin could act synergisti-cally on both the cell walls and cell membranes of pathogens. The 33-kD chitinase-encoding gene (chit33) was cloned and conju-gal-transformed from T. harzianum CECT2413 to S. lydicus A01, and then confirmed via polymerase chain reaction (PCR) assays. Subsequent analyses using the 3,5-dinitrosalicylic acid (DNS) method and ultraviolet spectrophotometry showed that compared with its wild type strain (WT), the S. lydicus A01 conjugal transformant (CT) with chit33 gene exhibited substantially higher chi-tinase activity and natamycin production. The resistance of S. lydicus A01-chit33 CT and WT to four Fusaria in crops and vegetables was tested via the cup-plate method. Compared with the WT, the conjugal transformant of S. lydicus A01 with chit33 gene from T. harzianum CECT2413 showed greatly increased biocontrol effect on fusarium disease. This study would be beneficial to the development of high-quality antifungal bio-agents for agricultural applications via the synergy between the previously non-existent and pre-existing functions achieved through heterogeneous gene transformation.     

内置发热圆的方腔内双扩散混合对流数值模拟研究

利用计算流体动力学CFD技术,对内置发热圆的方腔内双扩散混合对流情况进行了数值模拟.发热圆位于方腔中心,其表面高温高浓度,方腔各壁面绝热,低温低浓度流体从方腔左侧底部流进,从右侧顶部流出.分析了理查德数Ri,路易斯数Le及浮升力比Br对方腔内双扩散混合对流的影响,给出了流线、等浓度线以及等温线的分布情况,得出平均努赛尔数Nuav和平均舍伍德数Shav随着Ri,Le及Br的变化规律.     

Studies on optimization of nitrogen sources for astaxanthin production by Phaffia rhodozyma

Fermentation of Phaffia rhodozyma is a major method for producing astaxanthin, an important pigment with industrial and pharmaceutical application. To improve astaxanthin productivity, single factor and mixture design experiments were used to investigate the effects of nitrogen source on Phaffia rhodozyma cultivation and astaxanthin production. Results of single factor experiments showed nitrogen source could significantly affect P. rhodozyma cultivation with respect to carbon source utilization, yeast growth and astaxanthin accumulation. Further studies of mixture design experiments using (NH₄)₂SO₄, KNO₃ and beef extract as nitrogen sources indicated that the proportion of three nitrogen sources was very important to astaxanthin production. Validation experiments showed that the optimal nitrogen source was composed of 0.28 g/L (NH₄)₂SO₄, 0.49 g/L KNO₃ and 1.19 g/L beef extract. The kinetic characteristics of batch cultivation were investigated in a 5-L pH-stat fermentor. The maximum amount of biomass and highest astaxanthin yield in terms of volume and in terms of biomass were 7.71 mg/L and 1.00 mg/g, respectively.     

1型糖尿病小鼠钙磷代谢研究

通过对1型糖尿病小鼠模型的研究,检测其尿液、血清、骨骼中钙、磷含量.腹腔注射链脲佐菌素(40mg/kg),连续5d,诱导糖尿病小鼠模型.使用钙、磷试剂盒检测血清、尿液、骨骼中的钙、磷含量.糖尿病小鼠血磷升高(P0.05),尿磷降低(P0.05),尿钙升高(P0.05).股骨中钙、磷(P0.05)含量较正常组降低,胫骨中钙含量降低(P0.05).研究表明,高血糖引起尿钙流失增加,继而导致骨骼钙溶出增加,骨钙含量降低.另外,高血糖引起尿磷排泄减少,从而导致高血磷状态.