极小种群濒危植物广西火桐、丹霞梧桐的叶绿体基因组特征

广西火桐(Firmiana kwangsiensis)和丹霞梧桐(F. danxiaensis)是我国南方特有物种, 其分布范围狭窄, 种群数量少。为了解其叶绿体基因组结构及系统发生关系, 本文通过高通量测序方法获得广西火桐和丹霞梧桐的浅层基因组数据, 通过生物信息学方法对叶绿体全基因组进行组装, 并对其结构特征进行分析。结果表明: 广西火桐和丹霞梧桐的叶绿体基因组大小分别为160,836 bp和161,253 bp, 具有典型被子植物叶绿体基因组环状四分体结构, 包含长度分别为89,700 bp、90,142 bp的大单拷贝区(large single copy, LSC), 长度分别为19,970 bp、20,067 bp的小单拷贝区(small single copy, SSC)及长度分别为25,583 bp、25,522 bp的2个反向重复序列区(inverted repeat sequence, IR)。两个物种的叶绿体基因组共注释得到131个基因, 包括86个蛋白编码基因、37个tRNA基因和8个rRNA基因。广西火桐的叶绿体基因组中共检测出26个正向重复序列、2个反向重复序列、21个回文重复序列、21个串联重复序列和98个简单重复序列; 丹霞梧桐叶绿体基因组中共检测出23个正向重复序列、5个反向重复序列、21个回文重复序列、30个串联重复序列和107个简单重复序列。系统发生分析结果表明5种梧桐属(Firmiana)植物构成两个强烈支持的分支(支持率100%), 一个分支为广西火桐、美丽火桐(F. pulcherrima)和火桐(F. colorata), 其中广西火桐与美丽火桐构成姐妹群; 另一分支是互为姐妹群的丹霞梧桐和云南梧桐(F. major)。综上所述, 广西火桐和丹霞梧桐的叶绿体基因组结构、基因排列及重复序列具有较高的相似性, 系统进化树将5种梧桐属物种分为两个分支, 其中广西火桐和美丽火桐最近; 而丹霞梧桐与云南梧桐关系最近。本研究鉴定的SSR位点可为梧桐属物种系统发生、进化关系的研究提供遗传信息。     

A Bayesian hierarchical model for analyzing methylated RNA immunoprecipitation sequencing data

Background: The recently emerged technology of methylated RNA immunoprecipitation sequencing (MeRIP-seq) sheds light on the study of RNA epigenetics. This new bioinformatics question calls for effective and robust peaking calling algorithms to detect mRNA methylation sites from MeRIP-seq data. Methods: We propose a Bayesian hierarchical model to detect methylation sites from MeRIP-seq data. Our modeling approach includes several important characteristics. First, it models the zero-inflated and over-dispersed counts by deploying a zero-inflated negative binomial model. Second, it incorporates a hidden Markov model (HMM) to account for the spatial dependency of neighboring read enrichment. Third, our Bayesian inference allows the proposed model to borrow strength in parameter estimation, which greatly improves the model stability when dealing with MeRIP-seq data with a small number of replicates. We use Markov chain Monte Carlo (MCMC) algorithms to simultaneously infer the model parameters in a de novo fashion. The R Shiny demo is available at the authors' website and the R/C++ code is available at https://github.com/liqiwei2000/BaySeqPeak. Results: In simulation studies, the proposed method outperformed the competing methods exomePeak and MeTPeak, especially when an excess of zeros were present in the data. In real MeRIP-seq data analysis, the proposed method identified methylation sites that were more consistent with biological knowledge, and had better spatial resolution compared to the other methods. Conclusions: In this study, we develop a Bayesian hierarchical model to identify methylation peaks in MeRIP-seq data. The proposed method has a competitive edge over existing methods in terms of accuracy, robustness and spatial resolution.     

Relationships between soil nutrients and plant functional traits in different degradation stages of Leymus chinensis steppe in Nei Mongol,China

Aims Understanding ecological implications of plant functional traits is helpful in exploring community assembly under different environments of nature and human disturbances, and then to reveal the maintenance mechanism of the ecosystem services. By analyzing vegetation and soil data derived from field observations in Leymus chinensis steppe of Xilin River Basin in Nei Mongol, we aimed to explore the responses of plant functional traits to changing soil nutrients at different degradation stages. Methods We observed 69 plots for both plant community structure and soil attributes using quadrat and soil-drilling methods. Five plant functional traits, namely the specific leaf area (SLA), leaf dry matter content (LDMC), leaf carbon to nitrogen ratio (C:N), leaf lignin content (LLC), and maximum height (MH), were measured for each plot. We also tested soil attributes, such as total nitrogen (TN), total phosphorus (TP), available nitrogen (AN), available phosphorus (AP), and organic carbon (OC). The sixty-nine communities were classified into four groups (undegraded L. chinensis + forbs, slightly degraded L. chinensis + Stipa sp., moderately degraded L. chinensis + Cleistogenes squarrosa, and heavily degraded L. chinensis + Artemisia frigida) using TWINSPAN software. The relationships between plant functional traits and soil nutrient variables were analyzed for the four community groups using the Pearson’s correlation test with SPSS 21.0 software. Important findings (1) The soil nutrients decreased with the grassland degradation process and there were significant differences in TN and TP between the undegraded L. chinensis + forbs and heavily degraded L. chinensis + A. frigida communities; (2) plant functional traits also showed strong differences between the degradation stages. MH and C:N decreased with degradation. A significant difference was observed in MH between the undegraded L. chinensis + forbs and slightly degraded L. chinensis + Stipa sp. communities. The difference in C:N was also significant between the undegraded L. chinensis + forbs and heavily degraded L. chinensis + A. frigida communities; (3) the effects of soil nutrients on plant functional traits changed with grassland degradation. AN was negatively correlated with MH, LLC, and C:N in the slightly degraded L. chinensis + Stipa sp. community. In the moderately degraded L. chinensis + C. squarrosa community, those three traits mentioned above showed significantly positive correlations with TP; (4) while analyzing the degraded grassland, different relationships between plant functional traits were found. In the slightly degraded L. chinensis + Stipa sp. community, LLC was positively correlated with all other traits. Moreover, positive correlations also occurred between C:N and MH, C:N and LDMC, and C:N and LLC. In the heavily degraded L. chinensis + A. frigida community, all traits demonstrated the most significantly positive correlations.     

内蒙古羊草草原不同退化阶段土壤养分与植物功能性状的关系

深入认识植物功能性状的生态学含义, 对于阐明不同自然与人为干扰环境下的群落构建途径, 进一步揭示生态系统服务维持机制具有重要的理论意义。该文以内蒙古锡林河流域羊草(Leymus chinensis)草原不同退化演替阶段的群落为研究对象, 分析了土壤养分与植物功能性状的变化特征及两者之间的关系。结果表明: (1)退化导致土壤养分含量逐渐减少, 全氮和全磷在未退化的羊草+杂类草群落与严重退化的羊草+冷蒿(Artemisia frigida)群落之间差异显著; (2)随着退化演替的进程, 群落最大高度和叶片碳氮比减小, 群落最大高度在未退化的羊草+杂类草群落与轻度退化的羊草+针茅(Stipa sp.)群落之间差异显著, 碳氮比在未退化的羊草+杂类草群落与严重退化的羊草+冷蒿群落之间差异显著; (3)不同退化演替阶段的群落, 其土壤养分对植物功能性状的影响有所差异。在羊草+针茅群落, 速效氮与群落最大高度、叶片木质素含量和叶片碳氮比均呈显著负相关关系。而在羊草+糙隐子草(Cleistogenes squarrosa)群落中, 上述3种植物功能性状则表现为均与全磷含量显著正相关; (4)群落植物功能性状之间的关系也因退化阶段不同而有所不同。在轻度退化的羊草+针茅群落中, 叶片木质素含量与其他4个功能性状显著正相关, 叶片碳氮比与群落最大高度、叶干物质含量、木质素含量呈显著正相关关系。在严重退化的羊草+冷蒿群落中, 所有性状均呈极显著正相关关系。表明植物通过功能性状的协调或组合, 以适应贫瘠的土壤环境。上述结果深化了对典型草原退化演替的认识, 对退化草地的恢复与保护具有一定的指导意义。     

Theoretical approaches for evaluation of DNA-protein interactions

The notion of the DNA-recognizing segment of a protein for three-dimensional structures of DNA-protein complexes was formalized. Algorithms for calculating two parameters, hydrogen bonds and hydrophobic contacts, that characterize the interaction of the DNA-recognizing segment of the protein with the groove DNA major were proposed. DNA-recognizing protein segments in three-dimensional structures (of complexes) were classified according to these two parameters. These data were compared with the classification of the corresponding DNA-recognizing domains according to structure. The contribution of each pair amino acid residue-base to the interaction of protein with the DNA major groove was calculated.     

太行山南麓栓皮栎和刺槐光合作用-CO2响应模拟

采用便携式光合仪(Li-6400XT)对太行山南麓栓皮栎、刺槐2个树种叶片光合作用-CO₂响应曲线进行测定,利用直角双曲线模型(RH)、非直角双曲线模型(NRH)以及直角双曲线的修正模型—叶子飘模型(YZP)进行曲线拟合,并对3种光合模型的拟合参数(最大净光合能力A_max、初始羧化速率η、光呼吸速率R_p、CO₂补偿点CCP和CO₂饱和点CSP)进行比较.结果表明: 与NRH和YZP模型相比,RH模型所得的A_max、η、R_p和CCP较高,分别高出实测值59.8%、128.6%、133.4%和19.8%.与RH模型和YZP模型相比,NRH模型拟合得出的A_max较大,高于实测值11.1%,η、R_p和CCP接近于实测值.YZP模型能较好地模拟光合作用对CO₂的饱和现象,在A_max和CSP的拟合效果上较好.栓皮栎阴叶的A_max、R_p和CCP比阳叶分别低31.3%、5.2%和14.3%.刺槐阴叶的A_max、R_p和CCP分别高出阳叶23.5%、11.0%和5.4%.栓皮栎、刺槐阴叶的η分别比阳叶高6.9%和7.0%.刺槐叶片的R_p和CCP与温度、光强均具有显著线性关系,η与气孔导度(g_s)具有显著线性关系.栓皮栎叶片的η与光强和气孔导度具有显著线性关系,CCP主要受温度和湿度影响.栓皮栎叶片的A_max与相对湿度和g_s具有显著的正线性相关关系.     

河北省塞罕坝保护区天然植物群落数量分类

运用双向指示种(TWINSPAN)和除趋势典范对应分析(DCCA)等方法定量分析了河北省塞罕坝保护区天然植物群落分布与环境因子的关系.结果表明: 塞罕坝保护区天然植物群落可划分为8种类型,即地榆+异穗苔草+金莲花群落、地榆+老芒麦+毛蕊老鹳草群落、披针叶苔草+铁丝草群落、柔毛绣线菊-铁杆蒿群落、山杏+柔毛绣线菊-铁杆蒿+披针叶苔草群落、蒙古栎+硕桦-胡枝子+柔毛绣线菊-披针叶苔草群落、白桦+山杨-毛榛+山刺玫-糙苏+披针叶苔草+唐松草群落、云杉+白桦-山刺玫+河北柳-披针叶苔草+节节草群落;DCCA分析表明,坡位和土壤养分是影响塞罕坝保护区天然植物群落空间分布的主要因子.地形、空间和土壤因子解释了物种格局变异的34.02%,其中,土壤因子占15.67%,地形因子占10.05%,空间因子占6.00%.在各类因子间的交互作用(2.30%)中,土壤和地形因子的交互作用最大(0.93%),土壤、地形和空间因子三者的交互作用最小(0.16%).65.98%未能解释的部分,可能归因于未被选取的因子如种间的协同和竞争作用或者人为干扰.     

阔叶红松林6种阔叶树单个叶片叶面积和叶干质量的垂直变异及模型构建

快捷、准确地测定植物叶片尺度上的叶面积(LA)和叶干质量(LDM)对于探讨植物性状对气候变化的响应机制至关重要,但适于测定区域尺度上不同阔叶植物单个叶片LA和LDM的经验模型尚未提出.本研究以中国东北4个分布区阔叶红松林内的白桦、紫椴、山杨、枫桦、水曲柳和裂叶榆6种阔叶树种为研究对象,分别测定其不同冠层高度(上层、中层和下层)叶片的叶长、叶宽、叶厚、LA及LDM.以叶长与叶宽之比(叶长宽比)的中位数为标准将6种阔叶树分为两组,检验每组树种不同冠层高度对构建预测LA和LDM的经验模型是否存在显著影响;构建适于预测区域尺度上不同冠层单个叶片LA和LDM的经验模型,验证其预测精度;并进一步评估构建的经验模型预测其他区域相同阔叶树种LA和LDM的适用性.结果表明: 整体上6种阔叶树单个叶片的LA随冠层高度的降低呈显著增大趋势,而部分树种的LDM呈下降趋势;冠层高度对构建预测LA和LDM的经验模型具有显著影响;构建的经验模型预测两组阔叶树种不同冠层单个叶片LA和LDM的平均精度分别为95%和83%,且基于这些模型预测其他区域相应树种LA和LDM的平均精度分别为94%和80%,表明本研究构建的经验模型在我国东北区域具有普遍适用性.     

太行山南麓栓皮栎和刺槐光合作用-CO2响应模拟

采用便携式光合仪(Li-6400XT)对太行山南麓栓皮栎、刺槐2个树种叶片光合作用-CO₂响应曲线进行测定,利用直角双曲线模型(RH)、非直角双曲线模型(NRH)以及直角双曲线的修正模型—叶子飘模型(YZP)进行曲线拟合,并对3种光合模型的拟合参数(最大净光合能力A_max、初始羧化速率η、光呼吸速率R_p、CO₂补偿点CCP和CO₂饱和点CSP)进行比较.结果表明: 与NRH和YZP模型相比,RH模型所得的A_max、η、R_p和CCP较高,分别高出实测值59.8%、128.6%、133.4%和19.8%.与RH模型和YZP模型相比,NRH模型拟合得出的A_max较大,高于实测值11.1%,η、R_p和CCP接近于实测值.YZP模型能较好地模拟光合作用对CO₂的饱和现象,在A_max和CSP的拟合效果上较好.栓皮栎阴叶的A_max、R_p和CCP比阳叶分别低31.3%、5.2%和14.3%.刺槐阴叶的A_max、R_p和CCP分别高出阳叶23.5%、11.0%和5.4%.栓皮栎、刺槐阴叶的η分别比阳叶高6.9%和7.0%.刺槐叶片的R_p和CCP与温度、光强均具有显著线性关系,η与气孔导度(g_s)具有显著线性关系.栓皮栎叶片的η与光强和气孔导度具有显著线性关系,CCP主要受温度和湿度影响.栓皮栎叶片的A_max与相对湿度和g_s具有显著的正线性相关关系.     

Population structure and spatial pattern of Caragana tibetica communities in Nei Mongol shrub-encroached grassland

Aims The community structure and function of the shrub-encroached grassland, as well as its formation mechanism, is one of hot scientific problems in vegetation science. The spatial pattern analysis is an important means for studying plant population interactions and the relationships between plant population and environment. The shrub-encroached grassland is one major vegetation type in Nei Mongol. Knowledge on the spatial pattern of shrub can improve our understanding on adaptive characteristics of shrub to steppe environment.
Methods We chose Caragana tibetica encroached grassland distributed in Ordos City as research objects. The spatial position of each C. tibetica individual in 64 m × 64 m study plot was determined by a method of “three-distance location method”, and then the spatial pattern and intraspecific associations of C. tibetica individuals was analyzed with the point pattern analysis.
Important findings The results showed that C. tibetica population was at the rising stage. The mature individuals showed a uniform distribution at the scale of less than 3.7 m, subsequently exhibiting a random distribution with an increased scale. The juvenile individuals showed an aggregation distribution at the scale of 0.3-20.4 m, but a uniform distribution at other scales. The dead C. tibetica individuals were distributed aggregately at the scale from 0.7 to 1.4 m, but randomly at other scales. An indistinctive positive association was found for live C. tibetica individuals to dead individuals at the scale of less than 9.9 m, but no associations was found between them at other scales. The mature individuals negatively interacted with juveniles at the scale of larger than 2.2 m, but unassociated at the other scales.     

两种不同来源的绿色木霉降解木质纤维素研究

利用纤维素酶高产菌绿色木霉Trichoderma viride降解木质纤维素是实现废料资源化的重要手段。本研究选取来自不同生境的两株T. viride,分别以玉米秸秆和甘草药渣为基质,测定两者滤纸纤维素酶（filter paper cellulase,FPase）活性和还原糖产量。从时间、温度、水分、pH 4个方面比较两株T. viride的环境适应性和不同基质的差异性。结果表明,以玉米秸秆为基质,T. viride XJ最适初始料液比为1:4-1:5.5,T. viride AG最适初始料液比为1:5-1:5.5。初始料液比1:5.5时,T. viride AG产FPase活性显著高于T. viride XJ。两株T. viride最适发酵温度均为28℃,各温度处理下不同菌株间无显著差异。两株T. viride均表现为还原糖消耗。以甘草药渣为基质,T. viride XJ最适初始料液比为1:2-1:2.5,T. viride AG最适初始料液比为1:3-1:3.5。料液比高于1:3,T. viride AG产FPase活性显著高于T. viride XJ。T. viride AG最适发酵温度为28℃,T. viride XJ最适发酵温度为23-28℃。温度低于28℃,T. viride XJ产FPase活性显著高于T. viride AG。两株T. viride均表现为还原糖积累。两株T. viride最适初始pH均为6-7,最适发酵时间均为3d。最优发酵条件下FPase活性：T. viride AG>T. viride XJ。对T. viride产FPase诱导能力：甘草药渣>玉米秸秆。变差分解表明两株T. viride产FPase活性差异主要源于菌株对生境的生态适应。比较分析菌种来源、基质类型、环境条件对T. viride发酵效果的影响,将有助于该菌大规模应用性研究。     

Cloning of a full-length cDNA encoding bovine thymus poly(ADP-ribose) synthetase: evolutionarily conserved segments and their potential functions

The primary structure of bovine thymus poly(ADP-ribose) synthetase, as deduced from the nucleotide sequence of a cloned cDNA, indicated that this enzyme is composed of 1016 amino acids (aa) with an M_r of 113481. An abundance of Lys and Arg residues was in accord with the known basic nature of this protein. A comparison with reported sequences of human counterparts revealed: (1) three functional domains separated by partial proteolysis, i.e., DNA-binding (N-terminal), auto-modification (central), and NAD-binding (C-terminal) domains, have, in this order, increasing degrees of homology; (2) the DNA-binding domain is composed of two distinct regions: one, less conserved, containing zinc-binding fingers and the other, more conserved, containing helix-turn-helix motifs; (3) all Glu and Asp residues in the automodification domain are conserved; and (4) a 78-aa stretch encompassing the nucleotide-binding fold in the NAD-binding domain is completely conserved. These results are compatible with specific features of each domain, i.e., complex DNA-enzyme interactions, multiple automodification at acidic aa residues, and a stringent specificity for the substrate, NAD.     

山田胶锈菌和亚洲胶锈菌吸器提取体系建立

利用锈菌吸器特异性结合伴刀豆球蛋白Concanavalin A（ConA）的特性,用ConA-sepharose 4b作为层析介质,对自然状态下感染山田胶锈菌Gymnosporangium yamadae的绚丽海棠叶片以及接种亚洲胶锈菌G. asiaticum后不同发病时期沙梨叶片中胶锈菌的吸器进行分离,构建了吸器提取体系。结果表明,在感病叶片产生大量花斑时提取到这两种胶锈菌的完整吸器,经光学显微镜与透射电子显微镜观察,发现这两种胶锈菌的吸器形态相似,呈菌丝状或肾形或不规则形状,吸器体与叶绿体含量接近1:1。研究中提取得到了G. yamadae吸器总RNA,为胶锈菌的吸器转录组中候选效应分子筛选以及病原与寄主互作分子机制研究奠定了重要基础。     

基于COⅠ基因序列的蜜蜂属系统发育分析及其舞蹈和筑巢行为进化分析

【目的】分析蜜蜂属Apis的系统发育关系,并在此基础上探讨蜜蜂属舞蹈方向、舞蹈声音、营巢环境、巢脾结构的祖先状态和演变过程。【方法】PCR扩增并测定中国分布的蜜蜂属内东方蜜蜂A. cerana、西方蜜蜂A. mellifera、大蜜蜂A. dorsata、黑大蜜蜂A. laboriosa、小蜜蜂A. florea和黑小蜜蜂A. andreniformis的线粒体COⅠ基因序列,并从NCBI数据库中下载分布于其他国家或地区的上述6种蜜蜂以及绿努蜂A. nulunsis、苏拉威西蜂A. nigrocinta、沙巴蜂A. koschevnikovi、炳式大蜜蜂A. dorsata binghami的COⅠ同源序列。分别利用最大简约法(MP)、最大似然法(ML)和贝叶斯分析(BI)依据这些序列数据构建蜜蜂属系统发育关系。对蜜蜂属上述种的舞蹈语言和筑巢行为进行编码并作为性状特征标记到系统发育树中,利用简约法对祖先状态进行追溯。【结果】系统发育分析结果表明,蜜蜂属可划分为3大类群,即穴居蜜蜂类群(东方蜜蜂、西方蜜蜂、苏拉威西蜂、绿努蜂、沙巴蜂)、大蜜蜂类群(大蜜蜂、黑大蜜蜂)和小蜜蜂类群(小蜜蜂、黑小蜜蜂);小蜜蜂类群更加接近于祖型,大蜜蜂类群和穴居蜜蜂类群是两支单系;炳式大蜜蜂是独立于大蜜蜂和黑大蜜蜂的蜂种,且与黑大蜜蜂的亲缘关系更近。祖先状态重建结果显示：蜜蜂属祖先在露天环境下筑造垂直的单脾,且在传播食物或巢址信息时跳水平方向的无声摆尾舞,有嗡嗡声的舞蹈及筑造复脾是后来形成的。【结论】COⅠ基因可作为分子标记用于分析蜜蜂属的舞蹈和筑巢行为的祖先状态及进化过程;蜜蜂筑造复脾、跳有嗡嗡声的舞蹈是后期的适应性进化行为。     

茯苓碳氮磷化学计量学特征

为探明茯苓的碳、氮、磷生态化学计量学特征,采集了云南省11个州、市42个居群的茯苓样本,分析了其菌核与表皮中碳(C)、氮(N)、磷(P)的化学计量特征.结果 表明:茯苓菌核中C、N、P的含量分别为40.24％-43.58％、0.176％-0.532％和0.020％-0.077％;C∶N、C∶P和N∶P的范围分别为93....     

黑龙江省红松人工林枝条分布数量模拟

基于黑龙江省佳木斯市孟家岗林场的12块样地65株人工红松解析木的955个枝解析数据,以Poisson回归模型和负二项回归模型作为备选模型,构建了人工红松二级枝条数量分布模型,并采用AIC、Pseudo-R²、均方根误差(RMSE)和Vuong检验对模型的拟合优度进行比较.结果表明: 每轮一级枝条分布数量集中在3~5个,均值为4个,一级枝条分布数量与人工红松自身的枝条属性相关.一级标准枝上二级枝条分布的离散程度较大,利用全部子回归技术构建二级枝条分布数量模型,最终选择以负二项回归模型为基础的E(Y)=exp(β₀+β₁lnRDINC+β₂RDINC²+β₃HT/DBH+β₄CL+β₅DBH)作为二级枝条分布数量最优预测模型(β为参数;RDINC为相对着枝深度;HT为树高;DBH为胸径;CL为冠长).最优模型的Pseudo-R²为0.79,平均偏差接近于0,平均绝对偏差<7.对于所建立的模型,lnRDINC、CL和DBH的参数为正值,RDINC²和HT/DBH的为负值,随着RDINC增大,在树冠内二级枝条分布数量存在最大值.总的来说,所建立的人工红松二级枝条分布数量模型的预测精度为96.4%,可以很好地预估该研究区域人工红松二级枝条分布数量,为以后枝条的光合作用和生物量的研究提供了理论基础.     

利用InDel标记分析中国香菇菌株的遗传多样性与群体结构

通过对5个香菇菌株重测序,以香菇L808-1菌株的全基因组序列为参考基因组,分析了这些菌株中插入/缺失（InDel）标记位点在香菇基因组10条染色体上的分布,并筛选了插入/缺失碱基数≥15的位点,合成了449对InDel标记引物。经过PCR和电泳检测,其中237对引物条带清晰。最终筛选出107个PIC≥0.3的标记作为核心InDel标记对来自国内的44份香菇菌株进行遗传多样性分析。聚类分析显示栽培菌株和野生菌株各自聚为一支,所选香菇菌株间存在明显的群体分层。群体结构分析显示香菇种质资源可分为4个亚群,主成分分析显示香菇菌株之间的位置及距离与聚类分析和群体结构分析结果相符。香菇InDel分子标记的开发与应用,为香菇核心种质的构建和种质资源育种引用提供了基础。     

东方蜜蜂微孢子虫侵染意大利蜜蜂工蜂过程中nce-miR-12220及其靶基因的表达谱

东方蜜蜂微孢子虫Nosema ceranae侵染成年蜜蜂导致蜜蜂微孢子虫病。本研究旨在验证东方蜜蜂微孢子虫nce-miR-12220的存在和表达,并检测nce-miR-12220及其靶基因在病原侵染意大利蜜蜂Apis mellifera ligustica (简称意蜂)工蜂过程的表达谱。Stem-loop RT-PCR和Sanger测序结果显示nce-miR-12220真实存在和表达。靶向预测结果显示nce-miR-12220共靶向KRAB-A和γ tubulin等15个基因。上述靶基因可注释到19个GO条目和3条KEGG通路。RT-qPCR结果显示,相较于接种后1 d (1 day post infection,1 dpi),nce-miR-12220在2 dpi上调表达,而在3-12 dpi阶段总体表现出显著下调表达的趋势。类似地,与1 dpi相比,靶基因KRAB-A在2 dpi上调表达,而在3-12 dpi阶段总体呈下调表达的趋势。另外,与1 dpi相比,靶基因γ tubulin在2-12 dpi阶段总体表现出显著下调表达的趋势。上述结果表明nce-miR-12220与KRAB-A和γ tubulin之间存在潜在的靶向结合和正向调控关系;东方蜜蜂微孢子虫通过下调表达nce-miR-12220抑制KRAB-A的表达进而促进增殖;意蜂工蜂可能通过抑制东方蜜蜂微孢子虫的γ tubulin表达抵御病原侵染。研究结果明确了nce-miR-12220及其靶基因KRAB-A和γ tubulin在东方蜜蜂微孢子虫侵染意蜂工蜂过程中的动态表达规律,为深入探究nce-miR-12220在病原侵染中的功能及调控机制提供了理论和实验依据。