供水及间甲酚对小麦间作蚕豆土壤微生物多样性和酶活性的影响

通过盆栽试验,探讨供水(田间持水量的45%、60%和75%)和化感物质间甲酚对小麦、蚕豆不同种植模式生长盛期土壤微生物多样性和酶活性的影响.结果表明,随灌水水平的降低,不同处理的土壤细菌、真菌和放线菌数量随之减少,间甲酚可加剧灌水减少引起的微生物数量的减少;间甲酚对不同处理土壤微生物多样性指数均具有降低作用,提高灌水水平可缓解间甲酚对间作群体土壤微生物多样性的负效应,但间甲酚在75%灌水水平下对单作微生物多样性的负效应最大,45%的供水水平和间甲酚作用下间作可维持更高的土壤微生物多样性.间甲酚对土壤过氧化氢酶的化感作用不显著,对脲酶和酸性磷酸酶活性的化感作用显著;3种土壤酶活性随供水水平的降低均显著下降,但供水与间甲酚、种植模式的互作效应对酶活性的影响不显著;间作对土壤过氧化氢酶和酸性磷酸酶活性具有极显著影响.     

The mechanism of myoblast deformation in response to cyclic strain - A cytomechanical study

Mechanical strain is one of the important epigenetic factors that cause deformation and differentiation of skeletal muscles. This research was designed to investigate how myoblast deformation occurs after cyclic strain loading. Myoblasts were passaged three times and harvested; various cyclic strains (2.5kPa, 5kPa and 10kPa) were then loaded using a pulsatile mechanical system. The adaptive response of the myoblasts was observed at different time points (0.5h, 1h, 6h and 12h) post-loading. At the early stage of cyclic strain loading (<1h), almost no visible morphological changes were observed in the myoblasts. The actin cytoskeleton showed a disordered arrangement and a weak fluorescence expression; there was little expression of talin. At 6h and 12h post-loading, the myoblasts changed their orientation to parallel (in the 2.5kPa and 5kPa groups) or perpendicular (in the 10kPa group) to the direction of strain. Fluorescence expression of both the actin cytoskeleton and talin was significantly increased. The results suggest that cyclic strain has at least two ways to regulate adaptation of myoblasts: (1) by directly affecting actin cytoskeleton at an early stage post-loading to cause depolymerization; and (2) by later chemical signals transmitted from the extracellular side to intracellular side to initiate repolymerization.     

Vesicle-associated membrane protein-2 (VAMP2) mediates cAMP-stimulated renin release in mouse juxtaglomerular cells

Renin is essential for blood pressure control. Renin is stored in granules in juxtaglomerular (JG) cells, located in the pole of the renal afferent arterioles. The second messenger cAMP stimulates renin release. However, it is unclear whether fusion and exocytosis of renin-containing granules is involved. In addition, the role of the fusion proteins, SNAREs (soluble N-ethylmaleimide-sensitive factor attachment proteins), in renin release from JG cells has not been studied. The vesicle SNARE proteins VAMP2 (vesicle associated membrane protein 2) and VAMP3 mediate cAMP-stimulated exocytosis in other endocrine cells. Thus, we hypothesized that VAMP2 and/or -3 mediate cAMP-stimulated renin release from JG cells. By fluorescence-activated cell sorting, we isolated JG cells expressing green fluorescent protein and compared the relative abundance of VAMP2/3 in JG cells versus total mouse kidney mRNA by quantitative PCR. We found that VAMP2 and VAMP3 mRNA are expressed and enriched in JG cells. Confocal imaging of primary cultures of JG cells showed that VAMP2 (but not VAMP3) co-localized with renin-containing granules. Cleavage of VAMP2 and VAMP3 with tetanus toxin blocked cAMP-stimulated renin release from JG cells by ~50% and impaired cAMP-stimulated exocytosis by ~50%, as monitored with FM1-43. Then we specifically knocked down VAMP2 or VAMP3 by adenoviral-mediated delivery of short hairpin silencing RNA. We found that silencing VAMP2 blocked cAMP-induced renin release by ~50%. In contrast, silencing VAMP3 had no effect on basal or cAMP-stimulated renin release. We conclude that VAMP2 and VAMP3 are expressed in JG cells, but only VAMP2 is targeted to renin-containing granules and mediates the stimulatory effect of cAMP on renin exocytosis.     

Molecular mechanism underlying phosphatidylinositol 4,5-bisphosphate-induced inhibition of SpIH channels

Many ion channels have been shown to be regulated by the membrane signaling phospholipid phosphatidylinositol 4,5-bisphosphate (PIP(2)). Here, we demonstrate that the binding of PIP(2) to SpIH, a sea urchin hyperpolarization-activated cyclic nucleotide-gated ion channel (HCN), has a dual effect: potentiation and inhibition. The potentiation is observed as a shift in the voltage dependence of activation to more depolarized voltages. The inhibition is observed as a reduction in the currents elicited by the partial agonist cGMP. These two effects were separable and arose from PIP(2) binding to two different regions. Deletion of the C-terminal region of SpIH removed PIP(2)-induced inhibition but not the PIP(2)-induced shift in voltage dependence. Mutating key positively charged amino acids in the C-terminal region adjacent to the membrane selectively disrupted PIP(2)-induced inhibition, suggesting a direct interaction between PIP(2) in the membrane and amino acids in the C-terminal region that stabilizes the closed state relative to the open state in HCN channels.     

The response ofAzolla pinnata R. Brown to the split application of phosphorus and the transfer of assimilated phosphorus to flooded rice plants

A field experiment was conducted at the Bangladesh Rice Research Institute, Joydebpur, Dhaka during the late wet season. Basal application of P at both 5 and 10 kg ha⁻¹ significantly increased total biomass production and nitrogen fixation byAzolla pinnata R. Brown (local strain). Addition of both 5 and 10 kg P ha⁻¹ in equal splits at inoculation and at six day intervals thereafter during growth periods of 12, 24 and 36 days increased biomass production and nitrogen fixation by Azolla over that attained with the basal application. Biomass and nitrogen fixation using a split application of 5 kg P ha⁻¹ exceeded that attained with basal application of 10 kg P ha⁻¹ and split application of 10 kg P ha⁻¹ resulted in 0.58, 11.2, and 18.3 t ha⁻¹ more biomass, and 0.47, 18.9, and 18.3 more kg fixed N ha⁻¹ at 12, 24 and 36 days, respectively, than the same amount applied as a basal application. Analyses indicated that the critical level of dry weight P in Azolla for sustained growth was in the range of 0.15–0.17%. Compared with the control, where no P was added, and additional 30 and 36 kg N ha⁻¹ were fixed after 24 and 36 days, respectively, when P was provided at 10 kg ha⁻¹ using a split application. A separate field study showed that flooded rice plants received P from incorporated Azolla with about 28% of the P present in the supplied Azolla being incorporated into the rice plants.     

转化生长因子α－绿脓杆菌外毒素融合蛋白TGFα－PE40高效表达质粒的构建和表达

利用基因工程技术,将质粒pYX382用xbaI和EcoRl切下插入的TGFa—PE40融合 基因片段-连接到可表达载体pCB604的XbaⅠ／EcoR Ⅰ位点中,构建成新的重组质粒p2x—TP1。P2x—TP1转化E.coli BL2l感受志菌后,在ⅠPTG诱导下Ⅰpp启动子转录表达TGFα—PE40融合蛋白。表达产物主要以包涵体形式沉积在细胞内。融合蛋白表达量的高低与诱导时的细胞密度,诱导的温度以及培养基有关。在一定范围内与诱导剂的剂量以及诱导时间的长短无关。P2x—TPl重组质粒在工程菌中的表达TGFα－PE40融合蛋白量约为50mg／L。     

Copepod and microzooplankton grazing in mesocosms fertilised with different Si:N ratios: no overlap between food spectra and Si:N influence on zooplankton trophic level

We hypothesized that the trophic level of marine copepods should depend on the composition of the protist community. To test this hypothesis, we manipulated the phytoplankton composition in mesocosms and measured grazing rates of copepods and mesozooplankton in those mesocosms. Twelve mesocosms with Northeast Atlantic phytoplankton were fertilised with different Si:N ratios from 0:1 to 1:1. After 1 week, ten of the mesocosms were filled with natural densities of mesozooplankton, mainly calanoid copepods, while two remained as mesozooplankton-free controls. Both before and after the addition of copepods there was a positive correlation of diatom dominance with Si:N ratios. During the second phase of the experiment, copepod and microzooplankton grazing rates on different phytoplankton species were assessed by a modification of the Landry-Hassett dilution technique, where the bottles containing the different dilution treatments were replaced by dialysis bags incubated in situ. The results indicated no overlap in the food spectrum of microzooplankton (mainly ciliates) and copepods. Ciliates fed on nanoplankton, while copepods fed on large or chain-forming diatoms, naked dinoflagellates, and ciliates. The calculated trophic level of copepods showed a significantly negative but weak correlation with Si:N ratios. The strength of this response was strongly dependent on the trophic levels assumed for ciliates and mixotrophic dinoflagellates.     

Formation and dimerization of the phosphodiesterase active site of the Pseudomonas aeruginosa MorA,a bi-functional c-di-GMP regulator

Diguanylate cyclases (DGC) and phosphodiesterases (PDE), respectively synthesise and hydrolyse the secondary messenger cyclic dimeric GMP (c-di-GMP), and both activities are often found in a single protein. Intracellular c-di-GMP levels in turn regulate bacterial motility, virulence and biofilm formation. We report the first structure of a tandem DGC–PDE fragment, in which the catalytic domains are shown to be active. Two phosphodiesterase states are distinguished by active site formation. The structures, in the presence or absence of c-di-GMP, suggest that dimerisation and binding pocket formation are linked, with dimerisation being required for catalytic activity. An understanding of PDE activation is important, as biofilm dispersal via c-di-GMP hydrolysis has therapeutic effects on chronic infections.