中国野生东北虎数量监测方法有效性评估

虎数量监测是虎保护的核心内容之一。野生虎现存数量少、领域宽广,加之习性机警,很难对其数量和种群变化趋势做出准确的评估。合适的虎数量监测方法随着监测目标、监测尺度、虎密度、猎物密度、气候及其它环境因素的变化而不同。2002—2011年,用东北虎信息收集网络法,样线调查法,猎物生物量和捕食者关系法,对东北虎数量进行监测。(1)用老虎信息收集网络法研究2006年完达山东部地区东北虎的种群现状,结果显示东完达山地区2006年东北虎数量为6—9只,由1只成年雄虎,2—3只成年雌虎,2—4只亚成体虎和1只小于1岁的幼体虎组成;(2)用猎物生物量和捕食者关系法得到东完达山地区2002年东北虎的密度为0.356只/100 km2,能容纳22—27只东北虎;(3)用样线法在黑龙江的老爷岭南部和吉林省大龙岭北部面积1735.99 km2的区域内设置样线64条,总长609 km,没有发现东北虎足迹链。样线调查的结果表明,在2011年2—3月该调查区域东北虎的数量为0只。监测结果表明,用猎物生物量和捕食者关系得到东北虎数量远远超过现实数量,人们对有蹄类的盗猎和猎套对老虎的伤害可能是其主要原因;样线法调查得出的结果低于现实种群,主要原因是老虎数量极低和调查者对野生虎行为学了解甚少,较难在野外有效的发现虎信息;且样线法监测仅应用于当东北虎以一定的密度(即有定居虎)存在的情况下(多数监测样线能发现虎信息)。虽然和样线法一样存在着诸如专家估计密度和真实密度之间的关系,老虎足迹数量和老虎真实密度间关系不确定,保守估计等内在缺点,在目前中国东北地区野生东北虎种群密度极低,且多是穿越于中俄边境地区的游荡个体的现状下,信息收集网络法是一种高效,可行东北虎监测方法。因此,建议建立更广泛的监测信息收集网络,培训监测人员,严格执行信息收集程序,减少专家估计误差以完善此监测方法。此外,其他监测方法,如占有法、基于标志重捕远红外照相法、粪便DNA法、足迹数码信息法、警犬法等,应根据各种方法的理论前提、误差来源、适用范围和老虎是否定居及密度等具体情况有选择地加以应用,且有些方法可能成为未来中国野生东北虎种群的有效监测工具。     

Identification and characterization of microRNAs and their target genes in Brassica oleracea

The microRNAs are a new class of small non-coding endogenous RNAs with lengths of approximately ~21 nt. MicroRNAs perform their biological function via the degradation of the target mRNAs or by inhibiting protein translation. Until recently, only limited numbers of miRNAs were identified in Brassica oleracea, a vegetable widely cultivated around the world. In present study, 193 potential miRNA candidates were identified from 17 expressed sequence tag (ESTs) and 152 genome survey sequences (GSSs) in B. oleracea. These miRNA candidates were classified into 70 families using a well-defined comparative genome-based computational analysis. Most miRNAs belong to the miRNA169, miR5021, miR156 and miR158 families. Of these, 36 miRNA families are firstly found in Brassica species. Around 1393 B. oleracea genes were predicted as candidate targets of 175 miRNAs. The mutual relationship between miRNAs and the candidate target genes was verified by checking differentially expression levels using quantitative real-time polymerase chain reaction (qRT-PCR) and 5' RLM-RACE analyses. These target genes participate in multiple biological and metabolic processes, including signal transduction, stress response, and plant development. Gene Ontology analysis shows that the 818, 514, and 265 target genes are involved in molecular functions, biological processes, and cellular component respectively. The Kyoto Encyclopedia of Genes and Genomes (KEGG) Pathway enrichment analysis suggests that these miRNAs might regulate 186 metabolic pathways, including those of lipid, energy, starch and sucrose, fatty acid and nitrogen.     

Genome-based phylogeny of dsDNA viruses by a novel alignment-free method

Sequence alignment is not directly applicable to whole genome phylogeny since several events such as rearrangements make full length alignments impossible. Here, a novel alignment-free method derived from the standpoint of information theory is proposed and used to construct the whole-genome phylogeny for a population of viruses from 13 viral families comprising 218 dsDNA viruses. The method is based on information correlation (IC) and partial information correlation (PIC). We observe that (i) the IC-PIC tree segregates the population into clades, the membership of each is remarkably consistent with biologist's systematics only with little exceptions; (ii) the IC-PIC tree reveals potential evolutionary relationships among some viral families; and (iii) the IC-PIC tree predicts the taxonomic positions of certain “unclassified” viruses. Our approach provides a new way for recovering the phylogeny of viruses, and has practical applications in developing alignment-free methods for sequence classification.     

The nucleotide sequence of metallothioneins (MT) in liver of the Kafue lechwe (Kobus leche kafuensis) and their potential as biomarkers of heavy metal pollution of the Kafue River

The study determined heavy metal concentrations and MT1 nucleotide sequence [phylogeny] in liver of the Kafue lechwe. Applicability of MT1 as a biomarker of pollution was assessed. cDNA-encoding sequences for lechwe MT1 were amplified by RT-PCR to characterize the sequence of MT1 which was subjected to BLAST searching at NCBI. Phylogenetic relationships were based on pairwise matrix of sequence divergences calculated by Clustal W. Phylogenetic tree was constructed by NJ method using PHILLIP program. Metals were extracted by acid digestion and concentrations of Cr, Co, Cu, Zn, Cd, Pb, and Ni were determined using an AAS. MT1 mRNA expression levels were measured by quantitative comparative real-time RT-PCR. Lechwe MT1 has a length of 183bp, which encode for MT1 proteins of 61AA, which include 20 cysteines. Nucleotide sequence of lechwe MT1 showed identity with sheep MT (97%) and cattle MT1E (97%). Phylogenetic tree revealed that lechwe MT1 was clustered with sheep MT and cattle MT1E. Cu and Ni concentrations and MT1 mRNA expression levels of lechwe from Blue Lagoon were significantly higher than those from Lochinvar (p<0.05). Concentrations of Cd and Cu, Co and Cu, Co and Pb, Ni and Cu, and Ni and Cr were positively correlated. Spearman's rank correlations also showed positive correlations between Cu and Co concentrations and MT mRNA expression. PCA further suggested that MT mRNA expression was related to Zn and Cd concentrations. Hepatic MT1 mRNA expression in lechwe can be used as biomarker of heavy metal pollution.     

Polymorphisms of the insulin-like growth factor-binding protein 3 gene (IGFBP3) in gayal (Bos frontalis)

The gene coding for insulin-like growth factor-binding protein 3 (IGFBP3) is important for regulation of growth, development and metabolism in mammals. The present investigation was conducted to study nucleotide polymorphism of the IGFBP3 in gayal (Bos frontalis) and to compare the variations with those which occur in other ruminants. A fragment of 645 base pairs of the IGFBP3 covering a part of exon 2, the complete intron 2 and exon 3 and a part of intron 3 was amplified, sequenced (n=46) and digested (n=79) with HaeIII restriction enzyme from 125 collected gayal samples. Nine single nucleotide polymorphisms (SNPs) [C14T, A122C, C137T, G144C, C155T, G213A, C279A, G334A and G460A] were identified and located in intron 2, revealing high genetic variability. The alignment of nucleotide sequences was found to be very similar to those for other bovid species. Sequencing and HaeIII digestion showed that frequency of alleles C and A [consisting of fragments of sizes 56, 64, 228, 264, 282, 298 and 497 bp (CC genotype)] was 0.96 and 0.04 for the SNP C279A. Moreover, the genotype frequency of the SNP C279A in gayal was compared with that in other ruminants and it appears that this polymorphism may be associated with low fat content and rapid growth in this rare species.     

Emerging role of NF-κB signaling in the induction of senescence-associated secretory phenotype (SASP)

The major hallmark of cellular senescence is an irreversible cell cycle arrest and thus it is a potent tumor suppressor mechanism. Genotoxic insults, e.g. oxidative stress, are important inducers of the senescent phenotype which is characterized by an accumulation of senescence-associated heterochromatic foci (SAHF) and DNA segments with chromatin alterations reinforcing senescence (DNA-SCARS). Interestingly, senescent cells secrete pro-inflammatory factors and thus the condition has been called the senescence-associated secretory phenotype (SASP). Emerging data has revealed that NF-κB signaling is the major signaling pathway which stimulates the appearance of SASP. It is known that DNA damage provokes NF-κB signaling via a variety of signaling complexes containing NEMO protein, an NF-κB essential modifier, as well as via the activation of signaling pathways of p38MAPK and RIG-1, retinoic acid inducible gene-1. Genomic instability evoked by cellular stress triggers epigenetic changes, e.g. release of HMGB1 proteins which are also potent enhancers of inflammatory responses. Moreover, environmental stress and chronic inflammation can stimulate p38MAPK and ceramide signaling and induce cellular senescence with pro-inflammatory responses. On the other hand, two cyclin-dependent kinase inhibitors, p16INK4a and p14ARF, are effective inhibitors of NF-κB signaling. We will review in detail the signaling pathways which activate NF-κB signaling and trigger SASP in senescent cells.     

Acetylation of αA-crystallin in the human lens: Effects on structure and chaperone function

α-Crystallin is a major protein in the human lens that is perceived to help to maintain the transparency of the lens through its chaperone function. In this study, we demonstrate that many lens proteins including αA-crystallin are acetylated in vivo. We found that K70 and K99 in αA-crystallin and, K92 and K166 in αB-crystallin are acetylated in the human lens. To determine the effect of acetylation on the chaperone function and structural changes, αA-crystallin was acetylated using acetic anhydride. The resulting protein showed strong immunoreactivity against a N^ε-acetyllysine antibody, which was directly related to the degree of acetylation. When compared to the unmodified protein, the chaperone function of the in vitro acetylated αA-crystallin was higher against three of the four different client proteins tested. Because a lysine (residue 70; K70) in αA-crystallin is acetylated in vivo, we generated a protein with an acetylation mimic, replacing Lys70 with glutamine (K70Q). The K70Q mutant protein showed increased chaperone function against three client proteins compared to the Wt protein but decreased chaperone function against γ-crystallin. The acetylated protein displayed higher surface hydrophobicity and tryptophan fluorescence, had altered secondary and tertiary structures and displayed decreased thermodynamic stability. Together, our data suggest that acetylation of αA-crystallin occurs in the human lens and that it affects the chaperone function of the protein.     

中国地鼠山医群体近交系E繁殖性状的遗传力估计

目的对中国地鼠山医群体近交系E的一些繁殖性状进行遗传力估计分析。方法采用平均信息约束最大似然法（AIREML）,利用WOMBAT软件进行处理。结果产仔数、离乳数、胎间隔2-1和胎间隔3-2的遗传力均较低,分别为0.05、0.096、0.182和0.116。结论中国地鼠山医群体近交系E繁殖性状的遗传力均属于低遗传力。     

空间信息技术与深远海渔业资源开发

深远海渔业资源作为人类食用的优质蛋白来源,其开发受到各渔业国家的重视。以卫星遥感、地理信息系统等为代表的空间信息技术已经在海洋渔业中得到较为广泛的应用,主要包括卫星遥感渔场环境监测及渔情分析预报、鱼类洄游路径的监测、渔船作业位置的分布与监测等。针对深远海渔业资源的捕捞开发,重点综述了空间信息技术在深远海渔业资源开发中应用的现状以及应用前景分析。     

脓肿分枝杆菌非溶血性磷脂酶C蛋白MAB_0555基因克隆测序及生物信息学分析

目的克隆脓肿分枝杆菌（Mycobacterium abscessus,M.abscessus）非溶血性磷脂酶C基因MAB_0555,并对其测序以及生物信息学分析,为进一步研究其生物学功能及其在脓肿分枝杆菌致病机制中的作用提供基础。方法以脓肿分枝杆菌基因组DNA为模板,PCR扩增非溶血性磷脂酶C的基因片段,克隆入pQE-30质粒,构建重组原核表达载体pQE-30-MAB_0555,将其转化入大肠埃希菌DH5α,并进行序列测定及利用生物信息学软件DNAstar 5.0分析其生物学特性。结果 PCR扩增获得长1440 bp的MAB_0555基因片段,编码479个氨基酸,DNAstar等软件预测其蛋白质相对分子量（Mr）约为53 kD,并显示出具有良好的抗原性。结论成功获得序列正确的脓肿分枝杆菌MAB-0555基因,为其重组蛋白的表达及其相关研究奠定基础。