Genome-Wide Analysis of the Apple (Malus domestica) Cysteine-Rich Receptor-Like Kinase (CRK) Family: Annotation,Genomic Organization,and Expression Profiles in Response to Fungal Infection

Plant Molecular Biology Reporter - Cysteine-rich receptor-like kinases (CRKs) took crucial roles in plant cell growth and development, as well as environmental adaption. Apple (Malus domestica) had...     

yeaI基因对奶牛源大肠杆菌NJ17生物学特性的影响

c-di-GMP是细菌中广泛存在的第二信使,可通过效应蛋白参与调控细菌的生物被膜形成、运动性和毒力等生物学特性。YeaI因含有能结合c-di-GMP分子的EGEVF基序,可能作为c-di-GMP效应蛋白发挥作用。[目的] 研究yeaI基因缺失对奶牛源大肠杆菌临床分离株NJ17生物学特性的影响。[方法] 构建NJ17的yeaI缺失株（NJ17ΔyeaI）及回复株cNJ17ΔyeaI,分析yeaI对NJ17生物学特性（如生长特性、生物被膜形成能力和对小鼠乳腺上皮细胞（EpH₄-Ev）的黏附）的影响。[结果] 成功构建NJ17的yeaI缺失株（NJ17ΔyeaI）及其回复株（cNJ17ΔyeaI）;与野生株NJ17相比,缺失株NJ17ΔyeaI生长特性及耐药性无显著变化,生物被膜形成能力显著下降,运动性显著升高（P<0.05）;透射电镜检测结果表明,yeaI缺失影响NJ17菌毛和鞭毛的形成;实时定量PCR（qPCR）结果显示,yeaI基因显著抑制NJ17鞭毛基因filG和motB的转录水平（P<0.05）;血清杀菌实验表明,yeaI缺失能显著增强其抵抗血清杀菌作用（P<0.05）;对EpH₄-Ev细胞黏附实验表明,yeaI缺失对NJ17黏附性无显著影响（P>0.05）。[结论] yeaI对奶牛源大肠杆菌NJ17的生物学特性具有重要的调控作用。     

pcsk9 siRNA对oxLDL诱导的THP-1源性巨噬细胞凋亡的影响

为研究pcsk9基因沉默后对氧化型低密度脂蛋白(oxLDL)诱导THP-1源性巨噬细胞凋亡的影响,用不同浓度oxLDL 处理THP-1源性巨噬细胞48 h,Hoechst33258染色检测细胞凋亡,RT-PCR、Western blot分别检测pcsk9 mRNA、NARC-1蛋白的表达．应用Lipofectamine2000转染3对pcsk9 siRNAs进THP-1源性巨噬细胞中,筛选出最有效的siRNA再转染入THP-1源性巨噬细胞,24 h后加入oxLDL处理 48 h,Hoechst33258染色观察细胞评价细胞凋亡,流式细胞术计数检测细胞凋亡率．结果发现,75 mg/L oxLDL处理THP-1源性巨噬细胞48 h后,Hoechst33258染色可见大量凋亡细胞．同时RT-PCR、Western blot检测发现,pcsk9 mRNA和NARC-1蛋白质表达量均随oxLDL浓度的增加而增加,75 mg/L oxLDL组增加最明显．不同浓度siRNA转染THP-1源性巨噬细胞后,RT-PCR筛选出3对siRNAs的终浓度为80 nmol/L均可出现明显的沉默效应．选取此浓度在蛋白质水平检测基因抑制情况,筛选出最有效的一对siRNA．将筛选出来的siRNA转染细胞24 h后,再用oxLDL处理48 h,Hoechst33258染色及流式细胞计数结果显示,转染siRNA组凋亡明显被抑制．结果表明,在本研究的浓度范围内,随着oxLDL浓度增加pcsk9的表达随之增加,同时,THP-1源性巨噬细胞凋亡也明显增加,75 mg/L oxLDL最明显,pcsk9 mRNA和蛋白质的表达也在该浓度最高．提示pcsk9 siRNA能有效抑制pcsk9基因的表达,从而有效抑制由oxLDL诱导的THP-1源性巨噬细胞的凋亡．     

pcsk9 siRNA对oxLDL诱导的THP-1源性巨噬细胞凋亡的影响

为研究pcsk9基因沉默后对氧化型低密度脂蛋(oxLDL)诱导THP-1源性巨噬细胞凋亡的影响,用不同浓度oxLDL处理THP-1源性巨噬细胞48 h,Hoechst33258染色检测细胞凋亡,RT-PCR、Western blot分别检测pcsk9 mRNA、NARC-1蛋白的表达.应用Lipofectamine2000转染3对pcsk9 siRNAs进THP-1源性巨噬细胞中,筛选出最有效的siRNA再转染入THP-1源性巨噬细胞,24 h后加入oxLDL处理48 h,Hoechst33258染色观察细胞评价细胞凋亡,流式细胞术计数检测细胞凋亡率.结果发现,75 mg/L oxLDL处理THP-1源性巨噬细胞48 h后,Hoechst33258染色可见大量凋亡细胞.同时RT-PCR、Western blot检测发现,pcsk9 mRNA和NARC-1蛋白质表达量均随oxLDL浓度的增加而增加,75 mg/LoxLDL组增加最明显.不同浓度siRNA转染THP-1源性巨噬细胞后,RT-PCR筛选出3对siRNAs的终浓度为80 nmol/L均可出现明显的沉默效应.选取此浓度在蛋白质水平检测基因抑制情况,筛选出最有效的一对siRNA.将筛选出来的siRNA转染细胞24 h后,再用oxLDL处理48 h,Hoechst33258染色及流式细胞计数结果显示,转染siRNA组凋亡明显被抑制.结果表明,在本研究的浓度范围内,随着oxLDL浓度增加pcsk9的表达随之增加,同时,THP-1源性巨噬细胞凋亡也明显增加,75 mg/L oxLDL最明显,pcsk9 mRNA和蛋白质的表达也在该浓度最高.提示pcsk9 siRNA能有效抑制pcsk9基因的表达,从而有效抑制由oxLDL诱导的THP-1源性巨噬细胞的凋亡.     

Reactor performance and microbial community of an EGSB reactor operated at 20 and 15°C

Aims:  To investigate the effects of low temperatures on the performance and microbial community of anaerobic wastewater treatment.
Methods and Results:  An expanded granular sludge bed (EGSB) reactor was employed to treat synthetic brewery wastewater at 20 and 15°C. Reactor performance was represented by chemical oxygen demand (COD) removal efficiency, while the microbial community was analysed using denaturing gradient gel electrophoresis (DGGE) and clone technology. When the hydraulic retention time (HRT) was maintained at 18 h, COD removal efficiencies above 85% were obtained at both 20 and 15°C, with influent COD concentrations up to 7300 and 4100 mg l⁻¹, respectively. At 15°C, the COD removal efficiency was more easily manipulated by increasing the influent COD concentration. DGGE and clone results for both temperatures revealed that Methanosaeta and Methanobacterium were two dominant methanogens, and that the majority of the eubacterial clones were represented by Firmicutes . When the temperature decreased from 20 to 15°C, both archaeal and eubacterial communities had higher diversity, and the proportion of Methanosaeta (acetate-utilizing methanogens) decreased markedly from 60·0% to 49·3%, together with an increase in proportions of hydrogen-utilizing methanogens (especially Methanospirillum ).
Conclusions:  The feasibility of psychrophilic anaerobic treatment of low and medium strength organic wastewaters was demonstrated, although lower temperature could significantly affect both reactor performance and the anaerobic microbial community.
Significance and Impact of the Study:  The findings enrich the theory involving the microbial community and the application of anaerobic treatment in a psychrophilic environment.     

Non-invasive methods for identifying oocysts of Sarcocystis spp. from definitive hosts

Because the excreted sporocysts and/or oocysts of various species of Sarcocystis may not be discriminated morphologically, we sought to validate a diagnostic technique based on variation in the 18S rDNA sequence. Oocysts and/or sporocysts from three taxa of Sarcocystis were collected from human, feline, and canine definitive hosts that had fed upon meats infected with the muscle cysts of Sarcocystis hominis, Sarcocystis fusiformis and a species of Sarcocystis from water buffalo that could not be distinguished from Sarcocystis cruzi. Using a new collection method employing filter paper, these excreted oocysts and sporocysts were subjected to DNA extraction, as were the corresponding muscle cysts. Methods employing PCR–RFLP and DNA sequencing of a partial 18S rDNA gene (ssrRNA) sequence were then used to successfully distinguish among the three taxa. The same, unique restriction digestion pattern characterizes the tissue cysts and oocysts and/or sporocysts of each parasite taxon. The technique makes possible amplification and identification of species specific gene sequences based on DNA extracted from as few as 7 excreted sporocysts (the equivalent of 3 and 1/2 oocysts) from freshly prepared material, or as few as 50 sporocysts from feces samples that had been stored in potassium dichromate (K₂Cr₂O₇) for as long as 6 years. This represents the first report using molecular diagnostic procedures to diagnose oocysts of Sarcocystis in faecal samples, describing a valuable new tool for studying the epidemiology of various Sarcocystis species.     

蓝藻挥发物α-紫罗酮诱导莱茵衣藻细胞程序性死亡

【目的】蓝藻挥发性有机化合物(VOCs)对其他藻类的化感作用可促进蓝藻成为富营养化水体优势种群,本研究旨在以VOCs主要成分α-紫罗酮为例揭示其化感致死机制。【方法】采用α-紫罗酮处理莱茵衣藻,测定藻细胞生长以及致死浓度下藻细胞光合性能、caspase-likes活性和DNA ladders。【结果】采用0.05和0.1mmol/Lα-紫罗酮处理24h后,莱茵衣藻细胞生长均受到明显抑制,其中0.1 mmol/L处理时部分藻细胞发生死亡,死亡率为38.3%。采用0.2 mmol/Lα-紫罗酮处理时,藻细胞全部死亡,同时光合色素逐渐降解、Fv/Fm逐渐降低并消失,这表明藻细胞死亡并非坏死。在藻细胞死亡过程中,caspase-9-like和caspase-3-like活性明显增强;DNA在处理1h时出现ladders,并逐渐降解为100–250 bp片段。【结论】这表明蓝藻VOCs可通过诱导细胞程序性死亡以发挥化感作用。     

Narrow H3K4me2 is required for chicken PGC formation

The development of primordial germ cells (PGCs) undergoes epigenetic modifications. The study of histone methylation in regulating PGCs is beneficial to understand the development and differentiation mechanism of germ stem cells. Notably, it provides a theoretical basis for directed induction and mass acquisition in vitro. However, little is known about the regulation of PGC formation by histone methylation. Here, we found the high enrichment of H3K4me2 in the blastoderm, genital ridges, and testis. Chromatin immunoprecipitation sequencing was performed and the results revealed that genomic H3K4me2 is dynamic in embryonic stem cells, PGCs, and spermatogonial stem cells. This trend was consistent with the H3K4me2 enrichment in the gene promoter region. Additionally, narrow region triggered PGC‐related genes (Bmp4, Wnt5a, and Tcf7l2) and signaling pathways (Wnt and transforming growth factor‐β). After knocking down histone methylase Mll2 in vitro and vivo, the level of H3K4me2 decreased, inhibiting Cvh and Blimp1 expression, then repressing the formation of PGCs. Taken together, our study revealed the whole genome map of H3K4me2 in the formation of PGCs, contributing to improve the epigenetic study in PGC formation and providing materials for bird gene editing and rescue of endangered birds.