Comparative expression analysis of heavy metal ATPase subfamily genes between Cd-tolerant and Cd-sensitive turnip landraces

The heavy metal ATPase(HMA)subfamily is mainly involved in heavy metal(HM)tolerance and transport in plants,but an understanding of the definite roles and mechanisms of most HMA members are still limited.In the present study,we identified 14 candidate HMA genes named BrrHMAl—BrrHMA8 from the turnip genome and analyzed the phylogeny,gene structure,chromosome distribution,and conserved domains and motifs of HMAs in turnip(Brassica rapa var.rapa).According to our phylogenetic tree,the BrrHMAs are divided into a Zn/Cd/Co/Pb subclass and Cu/Ag subclass.The BrrHMA members show similar structural characteristics within subclasses.To explore the roles of BrrHMAs in turnip,we compared the gene sequences and expression patterns of the BrrHMA genes between a Cd-tolerant landrace and a Cd-sensitive landrace.Most BrrHMA genes showed similar spatial expression patterns in both Cd-tolerant and Cd-sensitive turnip landraces;some BrrHMA genes,however,were differentially expressed in specific tissue in Cd-tolerant and Cd-sensitive turnip.Specifically,BrrHMA genes in the Zn/Cd/Co/Pb subclass shared the same coding sequence but were differentially expressed in Cd-tolerant and Cd-sensitive turnip landraces under Cd stress.Our findings suggest that the stable expression and up-regulated expression of BrrHMA Zn/Cd/Co/Pb subclass genes under Cd stress may contribute to the higher Cd tolerance of turnip landraces.     

Deciphering the global phylogeography of a coastal shrub (Scaevola taccada) reveals the influence of multiple forces on contemporary population structure

The phylogeography of coastal plant species is heavily influenced by past sealevel fluctuations, dispersal barriers, and life-history traits, such as long-distance dispersal ability of the propagules. Unlike the widely studied mangroves, phylogeographic patterns have remained mostly obscure for other coastal plant species. In this study, we sampled 42 populations of Scaevola taccada (Gaertn.) Roxb., a coastal shrub of the family Goodeniaceae, from 17 countries across its distribution range. We used five chloroplast DNA (cpDNA) and 14 nuclear microsatellite (simple sequence repeat [SSR]) markers to assess the influence of abiotic factors and population genetic processes on the phylogeographic pattern of the species. Geographical distribution of cpDNA haplotypes suggests that the species originated in Australia, followed by historical dispersal and expansion of its geographic range. Multiple abiotic factors, including the sealevel changes during the Pleistocene, the presence of landmasses like the Malay Peninsula, and contemporary oceanic circulation patterns, restricted gene flow between geographically distinct populations, thereby creating low haplotype diversity and a strong population structure. Population genetic processes acted on these isolated populations, leading to high nuclear genetic diversity and population differentiation, as revealed from analyzing the polymorphic SSR loci. Although genetic divergence was mostly concordant between cpDNA and SSR data, asymmetrical gene flow and ancestral polymorphism could explain the discordance in the detailed genetic structure. Overall, our findings indicate that abiotic factors and population genetic processes interactively influenced the evolutionary history and current phylogeographic pattern of S. taccada across its distribution range.     

虫草多糖治疗家犬夹闭肾动脉引起的急性肾衰

探讨了虫草多糖对家犬夹闭肾动脉造成的急性肾衰的治疗作用。将实验动物家犬分为假手术组、阴性模型组、阳性组、虫草多糖高、中、低三个剂量组,观察治疗后犬的肾功能、血液生化指标、尿常规、肾脏病理的变化。发现虫草多糖组肾功能受损程度明显轻于阴性模型组（P〈0．01）,肾脏病理损伤明显减轻。实验证明虫草多糖能够治疗肾缺血诱发的急性肾功能衰竭,其机理可能与改善肾功能、增加肾血流量、纠正代谢紊乱、促进肾小管的修复与再生等作用有关。     

高效液相色谱法测定大黄素在Caco-2细胞中的摄取

目的:采用反相高效液相色谱法,观察大黄素在Caco-2细胞中的摄取特点。方法:将大黄素与Caco-2细胞共同孵育,收集细胞样品,液氮反复冻融。取细胞裂解液,加入甲醇提取,提取液采用HPLC进行分析。色谱分析柱为C18柱(250mm×4．6mm,5μm,Diamonsil),流动相组成为85%乙腈及15%水(含0．1%乙酸),流速1ml·min-1,进样量20μl,柱温25℃,3D模式采集数据。结果:检测Caco-2细胞中大黄素的工作曲线的回归方程为Y=0．278x 0．148(Y=0．9996,n=5),线性范围为0．037～4．8μmol·L-1,最低检测浓度为0．018μmol·L-1。当细胞中大黄素的浓度为0．05、2和8．5μg·ml-1时,回收率分别为(101．3±7．3)%、(96．7±3．0)%和(98．7±2．1)%(n=5);相应的日内标准偏差分别为0．25%、2．9%和1．4%;相应的日间标准偏差分别为2．3%、5．6%和6．3%。大黄素在Caco-2细胞中的摄取达峰时间为10分钟,峰浓度为108．56±11．57 nmol/L·mg·protein,10分钟后Caco-2细胞中大黄素的含量迅速下降。浓度处于2-50μM之间时,Caco-2细胞对大黄素的摄取量呈线性增加,浓度达50μM后,随着剂量的增加大黄素的摄取量变化不明显。结论:大黄素可被Caco-2细胞迅速摄取,随着剂量的增加,大黄素在Caco-2细胞中的摄取存在饱和现象。     

籼稻稻米蛋白质含量与外观品质性状间不同遗传体系的条件遗传相关分析

选用稻米蛋白质含量等品质差异较大的7个籼型不育系(A)及相应的保持系(B)与5个籼型恢复系(R)杂交,组成7×5不完全双列杂交组合。应用包括胚乳、细胞质和母体植株基因的遗传主效应以及基因型×环境互作效应的数量性状遗传模型及非条件和条件分析方法,研究了籼稻稻米蛋白质含量与外观品质性状间的遗传相关性,进一步揭示了籼稻糙米重、稻米直链淀粉含量对稻米蛋白质含量与外观品质性状间遗传相关性的影响。非条件分析的结果表明,除了与糙米厚的相关性未达到显著水平以外,蛋白质含量与其它稻米外观品质性状间的遗传相关性均达极显著水平,其中与糙米宽间的相关性表现为正值,其余为负向相关。条件分析的结果显示,糙米重和直链淀粉含量对稻米蛋白质含量与外观品质性状间的遗传相关性均可产生较大的影响。糙米重主要通过基因型×环境互作效应影响蛋白质含量与外观品质性状间的遗传相关性,其中对蛋白质含量与糙米长、糙米宽、糙米厚间遗传相关性的影响主要表现为负向作用,而对蛋白质含量与糙米长宽比、糙米长厚比间遗传相关性的影响则表现为正向作用。稻米直链淀粉主要通过细胞质遗传主效应和母体加性效应影响蛋白质含量与糙米长、糙米长宽比、糙米长厚比间的遗传相关性,而对蛋白质含量与糙米宽间的相关性影响主要表现为负向作用。     

Halothane binding proteome in human brain cortex

Inhaled anesthetics bind specifically to a wide variety of proteins in the brain. This set of proteins must include those that contribute to the physiological and behavioral phenotypes of anesthesia and the related side effects. To identify the anesthetic-binding targets and functional pathways associated with these targets in human brain, halothane photolabeling and two-dimensional (2D) gel electrophoresis were used. Both membrane and soluble proteins from human temporal cortex were prepared. More than 300 membrane and 400 soluble protein spots were detected on the stained blots, of which 23 membrane and 34 soluble proteins were labeled by halothane and identified by mass spectroscopy. Their functional classification reveals five groups, including carbohydrate metabolism, protein folding, oxidative phosphorylation, nucleoside triphosphatase, and dimer/kinase activity with different correlative stringency. When network analysis of the interaction between these protein molecules is used, the weighted interaction accentuates the cellular protein components important in cell growth and proliferation, cell cycle and cell death, and cell-cell signaling and interactions, although no pathway was specific. This study provides evidence for multiple anesthetic binding targets and suggests potential pathways involved in their actions.     

The activity of the RGA5 sensor NLR from rice requires binding of its integrated HMA domain to effectors but not HMA domain self-interaction

The rice nucleotide-binding (NB) and leucine-rich repeat (LRR) domain immune receptors (NLRs) RGA4 and RGA5 form a helper NLR/sensor NLR (hNLR/sNLR) pair that specifically recognizes the effectors AVR-Pia and AVR1-CO39 from the blast fungus Magnaporthe oryzae. While RGA4 contains only canonical NLR domains, RGA5 has an additional unconventional heavy metal-associated (HMA) domain integrated after its LRR domain. This RGA5_HMA domain binds the effectors and is crucial for their recognition. Investigation of the three-dimensional structure of the AVR1-CO39/RGA5_HMA complex by X-ray crystallography identified a candidate surface for effector binding in the HMA domain and showed that the HMA domain self-interacts in the absence of effector through the same surface. Here, we investigated the relevance of this HMA homodimerization for RGA5 function and the role of the RGA5_HMA effector-binding and self-interaction surface in effector recognition. By analysing structure-informed point mutations in the RGA5_HMA-binding surface in protein interaction studies and in Nicotiana benthamiana cell death assays, we found that HMA self-interaction does not contribute to RGA5 function. However, the effector-binding surface of RGA5_HMA identified by X-ray crystallography is crucial for both in vitro and in vivo effector binding as well as effector recognition. These results support the current hypothesis that noncanonical integrated domains of NLRs act primarily as effector traps and deepen our understanding of the sNLRs' function within NLR pairs.     

四川卧龙国家级自然保护区三种高山同域鸡形目鸟类的时空生态位比较

高海拔山区气候条件恶劣, 资源匮乏, 探究同域分布的近缘物种如何利用有限的资源以实现稳定共存, 对于了解高山生态系统生物多样性格局的形成和维持机制具有重要意义。鸡形目鸟类飞行能力弱, 属于典型的地栖物种, 生态位空间相对狭窄, 可能面临更高的种间竞争压力。本研究旨在比较几种同域分布的鸡形目鸟类的时空生态位, 为了解高山生态系统同域物种的共存机制提供新的研究案例。2020年4-9月, 研究人员在四川卧龙国家级自然保护区海拔3,300-4,200 m的高山区域进行了野外调查, 通过样线法和样方法对鸡形目鸟类群落优势物种绿尾虹雉(Lophophorus lhuysii)、雉鹑(Tetraophasis obscurus)和雪鹑(Lerwa lerwa)繁殖期的微生境进行调查, 使用红外相机对其活动节律进行监测, 并运用核密度估计法从微生境利用和日活动节律两个生态维度进行了种间生态位比较。结果显示, 雪鹑在微生境利用和日活动节律上均与其他两个物种存在显著差异。绿尾虹雉与雉鹑在微生境的利用上具有相似偏好; 但绿尾虹雉的早活动高峰晚于雉鹑, 晚活动高峰早于雉鹑, 表现出显著的种间日活动节律差异; 然而, 整合两个维度后, 绿尾虹雉和雉鹑的整体生态位仍然高度重叠, 没有显著分化。本研究表明高山鸡形目物种间的生态位分化体现于多个不同的生态维度, 并且不同物种之间的分化方式有所差异。在空间和时间生态位上的显著分化使雪鹑与同域物种间的竞争压力相对较小, 有利于其实现稳定共存。而绿尾虹雉与雉鹑的整体生态位高度重叠, 建议进一步对其食性开展研究, 探讨营养生态位上的潜在种间分化。