The M43 domain-containing metalloprotease RcMEP1 in Rhizoctonia cerealis is a pathogenicity factor during the fungus infection to wheat

Wheat(Triticum aestivum L.) is an important staple crop for global human. The necrotrophic fungus Rhizoctonia cerealis is the causal pathogen of sharp eyespot, a devastating disease of wheat. Herein, we identified RcMEP1, a zinc metalloproteaseencoding gene from R. cerealis genomic sequences, and characterized its pathogenesis function. RcMEP1 expressed at markedly-high levels during R. cerealis infection process to wheat. The predicted protein RcMEP1 comprises of 287 amino acid residues and contains a signal peptide and a M43 metalloprotease domain harboring the active site motif(HEVGHWLGLYH). The assays of Agrobacterium tumefaciens-mediated transient expression in Nicotiana benthamiana leaves indicated that RcMEP1 is an apoplastic elicitor of cell death, and that the predicted signal peptide functions and is required for secretion and cell death-induction. The purified RcMEP1 protein and its M43 domain peptide were individually able to induce plant cell death and H2 O2 accumulation, and to inhibit expression of host chitinases when infiltrated into wheat and N. benthamiana leaves, while the M43 domain-deleting peptide and negative control lacked the capacity. Moreover, compared with the control pretreatment, the purified RcMEP1 protein or its M43-domain peptide resulted in enhanced pathogenesis in the inoculated wheat, whereas the M43 domain-deleting peptide failed. These results suggest that RcMEP1 acted as an important pathogenicity factor during R. cerealis infection to wheat and that its signal peptide and M43 domain are required for the secretion and pathogenesis of RcMEP1. This study provides insights into pathogenesis role of M43 domain-containing metalloproteases during R. cerealis infection to wheat.     

勐海古茶树死亡原因及管养措施研究

勐海县种茶、制茶、用茶历史悠久,古茶树资源丰富,但近年来随着古茶树茶叶价格不断攀升,古茶树死亡和树势衰退现象变得多发。笔者认为造成勐海县古茶树的死亡原因主要有古茶树生境遭受破坏,管理不规范,私下流转现象严重、造成林权管理和服务不到位,没有规范操作规程、导致采摘过度,茶园土壤营养缺乏等;提出修复古茶树生境,加强古茶树科学研究,细化古茶园管养技术措施、制定管理规程,运用测土配方施肥技术、改善土壤养分结构,加大水肥管理和病虫害防控知识宣传力度等实现古茶树资源可持续经营的对策。     

程序性细胞死亡因子10抑制Ⅰ型干扰素表达并促进FMDV复制

旨在探究宿主蛋白程序性细胞死亡因子10（programmed cell death factor 10，PDCD10）通过抑制Ⅰ型干扰素表达进而促进口蹄疫病毒（foot-and-mouth disease virus，FMDV）的复制。首先，本研究验证了过表达和沉默PDCD10对FMDV复制的影响，接着利用双荧光素酶报告系统探究PDCD10对Ⅰ型干扰素信号通路活化的影响，最后，利用实时荧光定量PCR探究PDCD10对Ⅰ型干扰素通路下游刺激基因（IFN-stimulated genes，ISGs）转录的影响。结果表明，过表达PDCD10显著促进FMDV的复制，沉默PDCD10显著抑制FMDV的复制。与对照相比，过表达PDCD10后感染仙台病毒（Sendai virus，SeV）的细胞培养液上清液显著促进FMDV复制，进一步，PDCD10显著抑制SeV诱导的IFN-β启动子以及NF-κB的激活且呈剂量依赖性，并且PDCD10负调控Ⅰ型干扰素通路信号分子转录，最后还发现PDCD10负调控Ⅰ型干扰素下游ISGs转录。本研究结果为深入探究PDCD10在抗病毒天然免疫中的作用积累了资料。     

Reactive oxygen species are involved in cell death in wheat roots against powdery mildew

Inoculation of wheat(Triticum aestivum L.) leaves with wheat powdery mildew fungus(Blumeria graminis f. sp. tritici) induces the cell death in adventitious roots. Reactive oxygen species(ROS) play a key role in respond to biotic stress in plants. To study the involvement of ROS and the degree of cell death in the wheat roots following inoculation, ROS levels and microstructure of root cells were analyzed in two wheat cultivars that are susceptible(Huamai 8) and resistant(Shenmai 8) to powdery mildew fungus. At 18 d after powdery mildew fungus inoculation, only Huamai 8 displayed the leaf lesions, while root cell death occurred in both varieties. Huamai 8 had a high level of ROS accumulation, which is associated with increased root cell degradation, while in Shenmai 8, there was little ROS accumulation correlating with slight root cell degradation. The molecular study about the expression levels of ROS scavenging genes(MnSOD and CAT) in wheat roots showed that these genes expression decreased after the leaves of wheat was inoculated. The difference between Huamai 8 and Shenmai 8 on subcellular localization of H2 O2 and O2–· was corresponded with the different down-regulation of the genes encoding for superoxide dismutase and catalase in two wheat cultivars. These results suggested that ROS were involved in the process by which powdery mildew fungus induced cell death in wheat roots.     

Analysis of sinking death using video images of the swimming performance of Pacific Bluefin tuna (Thunnus orientalis) larvae

In this study, we aimed to clarify the mechanism of sinking death during the larval stage of Pacific Bluefin tuna Thunnus orientalis by investigating the effects of swimming performance on sinking death, using a behavioral approach. Swimming performance was examined 3–9 days after hatching (DAH) under day and night light conditions in cuboid experimental tanks. Swimming behavior variables such as swimming speed and swimming angle were measured under both light conditions. Larvae in the daytime experiment and larvae with inflated swim bladders at night were distributed on the surface layer of the water column. In contrast, larvae with uninflated swim bladders at night were frequently observed swimming vertically or sinking to the bottom of the tank. Larvae with inflated swim bladders at night were always distributed beneath the surface until the next morning (survival rates were 100 %). However, larvae with uninflated swim bladders at night swam upward repeatedly and later sank to the bottom of the tank (survival rates were 60 % and 38 % at 5 and 9 DAH, respectively). Larvae with uninflated swim bladders were not always able to maintain their swimming depth by swimming until the next morning. Additionally, their swimming speed and vertical swimming frequency (ratio) depended on the illumination and swim bladder conditions. Our findings show that larvae with uninflated swim bladders at night were associated with a higher risk of sinking death. The swimming energy capacity of Pacific Bluefin tuna larvae, which indicates the total amount of the energy that enables individuals to swim throughout the night without feeding, was found to be linked to sinking death.     

Prevalence of Electrolyte Disturbances and Perianesthetic Death Risk Factors in 120 Horses Undergoing Colic Surgery

The objective of this study was to determine the prevalence of intraoperative electrolyte disturbances and risk factors associated with perianesthetic death in horses undergoing colic surgery. The files of 120 horses meeting the inclusion criteria were reviewed. Data retrieved from the medical records, including demographic data and the occurrence of electrolyte disturbances and other intraoperative complications, were analyzed with a two step-approach by univariate and multivariate logistic regression models. Hypocalcemia was the most represented electrolyte disturbance (52.5%), followed by hypokalemia (30.0%) that was associated with intraoperative administration of salbutamol (P = .045). Perianesthetic death occurred in 46 horses, accounting for an overall mortality rate of 38.3%. Risk factors associated with death were anesthetic duration (P = .001), body weight (P = .020), presence of gastric reflux before anesthesia (P = .021), and intraoperative tachycardia (P = .043) and acidosis (P = .025). The mortality in the study population was comparable to previously reported findings. Based on the study findings, it is advisable to optimize hemodynamics prior to anesthesia, in order to prevent intraoperative tachycardia that is associated with increased risk of death. Heavier horses and those with gastric reflux may have a higher risk of fatalities, and intraoperative salbutamol administration may contribute to hypokalemia.     

体细胞克隆荷斯坦奶牛出生后死亡犊牛内脏器官的组织病理学观察

为探究体细胞克隆荷斯坦奶牛出生后死亡的原因,对新生后死亡的克隆荷斯坦公牛和自然繁殖的荷斯坦公犊的主要组织器官进行比较。通过解剖和石蜡切片-HE(hematoxylin-eosin staining)染色技术,对主要的组织器官结构进行观察和分析。结果表明,新生后死亡的克隆荷斯坦公牛肺部结构清晰;肝脏的肝细胞明显肿大,出现脂肪轻度变性;肾小管上皮细胞出现变性;心肌的肌纤维间空隙增大;骨骼肌纤维间隙明显,空泡变性,这可能导致该克隆牛犊肌肉无力并功能不全;淋巴结皮髓质界限不分明,淋巴小结细胞较稀疏,生发中心不明显,淋巴窦细胞较少;脾脏内红细胞较少,这说明该克隆牛的造血功能可能不完善;胸腺的皮质部分和髓质部分界限不明显,嗜酸性胸腺小体不易辨认,可能发育不全。该克隆公牛免疫器官出现不同程度的发育不全现象较严重,这有可能是其出生后死亡率高的主要原因。     

miR-140-3p inhibits viability,invasion and migration of non-small-cell lung cancer A549 cells by targeting PD-L1

AIM: To explore the target relationship between microRNA-140-3p (miR-140-3p) and programmed cell death ligand 1 (PD-L1) and their effect on the viability, migration and invasion of non-small-cell lung cancer A549 cells.METHODS: RT-qPCR was used to detect the miR-140-3p expression in HLF-1, A549 and H1299 cells, and then the A549 cells with the most significant difference were selected as the subsequent research object. TargetScan software and dual-luciferase reporter assay were performed to predict and confirm the target relationship between miR-140-3p and PD-L1. RT-qPCR and Western blot were used to determine the effects of miR-140-3p mimic and inhibitor on PD-L1 expression level. MTT assay was used to detect the viability of A549 cells. Transwell assay was performed to detect the migration and invasion abilities of the A549 cells.RESULTS: miR-140-3p was significantly down-regulated in the A549 cells and H1299 cells (P<0.05). Transfection with miR-140-3p mimic decreased the expression of PD-L1 and inhibited the viability, migration and invasion of the A549 cells. Transfection with pcDNA3.0-PD-L1 reversed the inhibitory effect of miR-140-3p on the viability, migration and invasion of the A549 cells.CONCLUSION: miR-140-3p inhibits the viability, migration and invasion of A549 cells by targeting PD-L1.     

鸭梨果实石细胞分化特性研究

以鸭梨为试材,利用透射电镜显微技术和组织化学技术对梨果实石细胞分化及次生壁形成过程进行研究。结果表明,石细胞分化始于开花第7天,原生质体收缩于细胞一侧,随后胞内出现颗粒状凝聚物,次生壁逐渐增厚,之后胞质继续收缩消失,早期形成的石细胞多以群体形式同时出现。石细胞次生壁形成是逐步进行的,即先形成纤维素或半纤维素网架结构,再进行木质化填充。透射电镜下石细胞分化过程中依次呈现出细胞核染色质凝聚、细胞核变形、核膜界限不清晰的现象;胞内出现同心层次的自噬泡,形成大量囊泡;细胞核消失,内质网断裂并大量增生,质膜向内突起形成泡状结构;次生壁增厚,胞内细胞器仅有线粒体和内质网,最终细胞质收缩消失。