PVY侵染后烟草营养成分的变化及其对介体烟蚜生长发育的影响

【目的】明确马铃薯Y病毒(potato virus Y, PVY)侵染后诱导的烟草营养成分的变化及其对烟蚜Myzus persicae生命特性的影响,旨在进一步解析PVY-烟草-烟蚜三者间的互作机制。【方法】通过蒽酮比色法和氨基酸自动分析仪测定了PVY不同侵染时期烟株体内的可溶性糖和游离氨基酸含量的变化;测定和比较了感病与健康烟草植株上烟蚜种群生长发育、成虫寿命、繁殖力和有翅蚜产生量的差异性。【结果】PVY侵染前、中、后期(分别为侵染后5, 12和20 d)的烟草叶片中游离氨基酸的总量均显著高于健康烟草叶片。相较于健康烟草叶片,在PVY侵染前期的烟草叶片中,谷氨酸、脯氨酸、天冬氨酸、色氨酸、缬氨酸、赖氨酸和组氨酸的含量显著增加;PVY侵染中期,感病叶片中丝氨酸含量显著下降,谷氨酸、天冬氨酸、色氨酸、缬氨酸、亮氨酸、苯丙氨酸、精氨酸和组氨酸含量显著提高;PVY侵染后期,感病叶片中甘氨酸含量显著下降,谷氨酸、脯氨酸、天冬氨酸、苏氨酸、缬氨酸、亮氨酸、丙氨酸、苯丙氨酸、组氨酸、酪氨酸和精氨酸含量显著提高。在PVY侵染的前期和中期,感病叶片中的可溶性糖含量显著高于健康烟叶,而在侵染后期感病叶片...     

Characterization of germin-like protein with polyphenol oxidase activity from Satsuma mandarine

Polyphenol oxidases (PPOs) catalyzing the oxygen dependent oxidation of phenols to quinones are ubiquitously distributed in plants and are assumed to be involved in plant defense against pests and pathogens. A protein with high PPO activity was identified in Satsuma mandarine, extracted with Tris–HCl buffer, purified by salt precipitation and column chromatography, and characterized by mass spectrometry as germin-like protein (GLP), which belongs to pathogenesis related protein (PR) family. In the present study, the structure and enzymatic properties of GLP were characterized using spectroscopy methods. Based on native PAGE analysis, the molecular weight of GLP was estimated to be 108 kDa and GLP was identified as a pentamer containing five subunits of 22 kDa. The optimum pH and temperature for PPO catalyzing activity of GLP was 6.5 and 65 °C, respectively. Kinetic constants were 0.0365 M and 0.0196 M with the substrates catechol and pyrogallol, respectively. The structural characterization of GLP provided better insights into the regions responsible for its PPO activity.     

Application of a novel thermostable NAD(P)H oxidase from hyperthermophilic archaeon for the regeneration of both NAD⁺ and NADP⁺

A novel thermostable NAD(P)H oxidase from the hyperthermophilic archaeon Thermococcus kodakarensis KOD1 (TkNOX) catalyzes oxidation of NADH and NADPH with oxygen from atmospheric air as an electron acceptor. Although the optimal temperature of TkNOX is >90°C, it also shows activity at 30°C. This enzyme was used for the regeneration of both NADP(+) and NAD(+) in alcohol dehydrogenase (ADH)-catalyzed enantioselective oxidation of racemic 1-phenylethanol. NADP(+) regeneration at 30°C was performed by TkNOX coupled with (R)-specific ADH from Lactobacillus kefir, resulting in successful acquisition of optically pure (S)-1-phenylethanol. The use of TkNOX with moderately thermostable (S)-specific ADH from Rhodococcus erythropolis enabled us to operate the enantioselective bioconversion accompanying NAD(+) regeneration at high temperatures. Optically pure (R)-1-phenylethanol was successfully obtained by this system after a shorter reaction time at 45-60°C than that at 30°C, demonstrating an advantage of the combination of thermostable enzymes. The ability of TkNOX to oxidize both NADH and NADPH with remarkable thermostability renders this enzyme a versatile tool for regeneration of the oxidized nicotinamide cofactors without the need for extra substrates other than dissolved oxygen from air.     

SOST is a ligand for LRP5/LRP6 and a Wnt signaling inhibitor

Sclerosteosis is an autosomal recessive disease that is characterized by overgrowth of bone tissue and is linked to mutations in the gene encoding the secreted protein SOST. Sclerosteosis shares remarkable similarities with "high bone mass" diseases caused by "gain-of-function" mutations in the LRP5 gene, which encodes a coreceptor for Wnt signaling proteins. We show here that SOST antagonizes Wnt signaling in Xenopus embryos and mammalian cells by binding to the extracellular domain of the Wnt coreceptors LRP5 and LRP6 and disrupting Wnt-induced Frizzled-LRP complex formation. Our findings suggest that SOST is an antagonist for Wnt signaling and that the loss of SOST function likely leads to the hyperactivation of Wnt signaling that underlies bone overgrowth seen in sclerosteosis patients.     

Mini-review: Microbial coaggregation: ubiquity and implications for biofilm development

Coaggregation is the specific recognition and adherence of genetically distinct microorganisms. Because most biofilms are polymicrobial communities, there is potential for coaggregation to play an integral role in spatiotemporal biofilm development and the moderation of biofilm community composition. However, understanding of the mechanisms contributing to coaggregation and the relevance of coaggregation to biofilm ecology is at a very early stage. The purpose of this review is to highlight recent advances in the understanding of microbial coaggregation within different environments and to describe the possible ecological ramifications of such interactions. Bacteria that coaggregate with many partner species within different environments will be highlighted, including oral streptococci and oral bridging organisms such as fusobacteria, as well as the freshwater sphingomonads and acinetobacters. Irrespective of environment, it is proposed that coaggregation is essential for the orchestrated development of multi-species biofilms.     

Surface display of active lipase in <Emphasis Type="Italic">Pichia pastoris</Emphasis> using Sed1 as an anchor protein

A Pichia pastoris cell-surface display system was constructed using the Sed1 anchor system that has been developed in Saccharomyces cerevisiae. Candida antarctica lipase B (CALB) was used as the model protein and was fused to an anchor that consisted of 338 amino acids of Sed1. The resulting fusion protein CALBSed1 was expressed under the control of the alcohol oxidase 1 promoter (pAOX1). Immunofluorescence microscopy of immunolabeled Pichia pastoris revealed that CALB was displayed on the cell surface. Western blot analysis showed that the fusion protein CALBSed1 was attached covalently to the cell wall and was highly glycosylated. The hydrolytic activity of the displayed CALB was more than 220 U/g dry cells after 120 h of culture. The displayed protein also exhibited a higher degree of thermostability than free CALB.     

The functional expression of calcium-sensing receptor in the differentiated THP-1 cells

The expression and function of calcium-sensing receptor (CaSR) in differentiated THP-1 (human acute monocytic leukemia cell line) cells are unknown currently. This study investigated above-mentioned issues using TRAP staining, immunofluorescence staining, Western blotting, ELISA, and Laser Confocal Scanning Microscopy techniques. We found that CaSR protein was expressed, and mainly located in the membrane and cytoplasm in differentiated THP-1 cells. Elevated extracellular calcium or GdCl₃ (an agonist of CaSR) raised intracellular calcium concentration. And this increase was inhibited or abolished by NPS2390 (an inhibitor of CaSR), U73122 (a specific inhibitor of phospholipase C, PLC) or thapsigargin (a Ca²⁺-ATPase inhibitor). The extracellular GdCl₃ elevation stimulated both of IL-1β and TNFα release, and this effect of GdCl₃ was inhibited by NPS2390. In conclusion, CaSR is functionally expressed in differentiated THP-1 cells, and the activated CaSR contributes to intracellular calcium increment through Gq-PLC- inositol triphosphate (IP3) pathway and commits to cytokine secretion. These results suggest that CaSR might be involved in a variety of pathological processes mediated by activated monocyte-macrophages.     

榄仁树的生态分布与耐盐性研究

调查了榄仁树(Terminalia catappa)在海南岛沿海地区的分布情况,并就基质盐度对温室中人工培育的榄仁幼苗的影响进行研究。结果表明,榄仁具有较强的耐盐能力,在盐度高达17.09‰条件下仍可正常生长,温室里幼苗可在10‰左右的盐度下存活。结果还表明,10‰盐度能增加幼苗叶片叶绿素含量,20‰~30‰则下降;盐胁迫下叶片光合速率呈下降趋势,而叶片可溶性糖含量则随盐度增加呈上升趋势。     

SLC17A9 Protein Functions as a Lysosomal ATP Transporter and Regulates Cell Viability

Lysosomes contain abundant ATP, which is released through lysosomal exocytosis following exposure to various stimuli. However, the molecular mechanisms underlying lysosomal ATP accumulation remain unknown. The vesicular nucleotide transporter, also known as solute carrier family 17 member 9 (SLC17A9), has been shown to function in ATP transport across secretory vesicles/granules membrane in adrenal chromaffin cells, T cells, and pancreatic cells. Here, using mammalian cell lines, we report that SLC17A9 is highly enriched in lysosomes and functions as an ATP transporter in those organelles. SLC17A9 deficiency reduced lysosome ATP accumulation and compromised lysosome function, resulting in cell death. Our data suggest that SLC17A9 activity mediates lysosomal ATP accumulation and plays an important role in lysosomal physiology and cell viability.     

CL系列甘蔗亲本的遗传力及配合力分析

为探讨CL系列甘蔗品种作杂交亲本的遗传特点,采用3×3不完全双列杂交(NCⅡ)遗传设计,估算了7个产量和品质性状的遗传方差、一般配合力(GCA)和特殊配合力(SCA)。结果表明:锤度的遗传主要受母本加性基因效应控制,株高的遗传主要受父母本加性基因控制,而锤重的遗传主要受非加性基因效应所制约;CL83-1163作为母本,糖分配合力高,且能把高糖特性传递给后代,CL88-4730为父本,产量和品质性状的配合力大,其杂交后代表现高产高糖;根据配合力总效应(TCA),综合表现好的组合有CL83-1364×CL88-4730、CL83-1900×CL84-3152、CL83-1163×CL88-4730,可用于今后的甘蔗有性育种计划。