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1.
[目的]DET2基因编码一个5α-还原酶,是油菜素内酯(BRs)合成过程中的关键限速基因。研究DET2基因在杨树生长发育中的作用,对于进一步研究油菜素内酯在木本植物中的调控机制有重要意义。[方法]从银腺杨84K(Populus alba×P. glandulosa,‘84K’)克隆得到拟南芥AtDET2同源基因PagDET2,利用生物信息学对其进行序列比对、生化特征分析、构建系统发育进化树。通过RT-PCR分析其在杨树中的表达模式。构建由CaMV 35S强启动子驱动的过表达载体,通过农杆菌介导的叶盘转化法转化84K杨,得到PagDET2-OE转基因植株。分析过表达DET2基因对于转基因植株内源BRs含量、植株生长和抗逆的影响。[结果]克隆了包含全长编码区PagDET2基因全长,可编码一个长度为257个氨基酸的蛋白质。其蛋白序列与毛果杨、拟南芥、水稻、棉花、大豆、番茄DET2蛋白同源性较高,说明该基因在进化过程中相对保守。PagDET2在84K杨不同组织中均检测到表达,其在茎中的表达较高。通过ELISA检测植物BRs含量发现,过量表达DET2可以显著提高杨树内源BRs含量。过量表达DET2基因,可以导致转基因植株的高生长,但对盐胁迫更加敏感。[结论]DET2基因作为BRs合成的关键基因,在杨树中过量表达可以显著提高内源BRs含量,促进植株增高。DET2转基因植株的获得为进一步分析BR参与木本植物生长发育的调控机制奠定了基础。  相似文献   

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Ferritin, a universal intracellular protein, can store large amounts of iron and improve plant resistance to abiotic and biotic stress. In this study, a ferritin gene(TaFer) from Tamarix androssowii Litv. was transferred into Populus tomentosa Carr. cv 'BJR01' via Agrobacterium. Six independent transgenic lines were obtained with a tolerance to kanamycin and three were randomly selected for further analysis. The PCR and RT-PCR results indicate that the TaFer gene had been integrated into the poplar genome. The effect of the gene on abiotic stress tolerance was tested, and the results show that transgenic plants improve growth, had higher chlorophyll and lower MDA contents, and higher relative electrical conductivity,fewer changes of SOD and POD activities, higher iron content, higher root ferric reductase activity and lower levels of ROS accumulation and cell death in response to drought, Fe-insufficient or Fe-excess tolerance. These results indicate that the TaFer gene can improve abiotic stress tolerance in transgenic Populus tomentosa.  相似文献   

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杨树维管组织特异启动子的克隆与启动活性分析   总被引:1,自引:0,他引:1       下载免费PDF全文
启动子在基因表达调控中起关键性作用,它在很大程度上决定所控基因表达的时间、空间和强度。依据拟南芥ATH1芯片分析杨树维管形成时期特异表达基因的结果,选取了差异表达基因NST3,通过BLAST比对在杨树EST数据库(PopulusDB)中找到同源性较高的基因NAC068。以毛白杨为材料,在其基因组中克隆得到该基因5'侧翼区901 bp长的片段,命名为pProNAC068,将该片段置换pBI121载体中的CaMV35S启动子,并在84K杨中检测报告基因GUS的表达情况。经过GUS活性检测分析发现:该启动子可以控制外源基因在次生维管组织中特异表达,从而为基因工程中有目的的控制外源基因在维管组织中的表达奠定基础。  相似文献   

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平榛ChWRKY28基因克隆及表达模式分析   总被引:1,自引:1,他引:0       下载免费PDF全文
[目的]研究平榛ChWRKY28基因序列特征及其在不同非生物胁迫下的表达规律.[方法]以平榛为试材,采用RACE-PCR方法进行基因克隆;利用实时荧光定量PCR方法检测基因在不同组织及不同非生物胁迫下的表达模式.[结果]表明:克隆得到的WRKY基因,全长1 342 bp,基因内部包含1个长963 bp的完整开放阅读框,编码320个氨基酸残基,命名为ChWRKY28.构建的系统发育树表明:该序列与拟南芥AtWRKY28及杨树PtrWRKY93的关系最近,相似性分别为49%和60%.基因表达分析表明:ChWRKY28在雄花序、雌花芽及茎中均有表达,但在茎部(皮)中的表达量高于雄花序和雌花芽中的表达量,具有组织表达特异性;低温、干旱及盐胁迫均能诱导ChWRKY28基因的表达,但受诱导程度存在差异.亚细胞定位分析结果表明:ChWRKY28蛋白分布在细胞核内,是一个核蛋白.[结论]推测ChWRKY28基因可能参与植物响应非生物胁迫的信号转导过程.  相似文献   

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利用黑斑病的高抗无性系美洲黑杨I-69及黑斑病的高感无性系欧美杨I-45为材料建立的2个cDNA文库,随机挑取cDNA克隆进行5'端EST序列测序,共获得有效序列20 023条.序列经聚类分析和拼接后,共获得10 816个Unigene,其中包括3 734个Contig,7 082个独立的Singleton.被注释的8 853个具有同源性匹配序列基因中,按照GO的分子功能、生物过程和细胞组分3个不同分类角度进行分类.在具有功能注释的8 853个Unigene中,选出330个与完成全基因组测序的毛果杨序列进行BLAST分析,结果发现有177个抗病相关候选基因出现在282个Unigene中,其中135个分布于杨树的18个不同连锁群上,其他42个基因位于还没定位的scaffolds上.所测定的这些EST序列为后期在基因组水平上研究杨树黑斑病的水平抗性遗传机制及进一步的相关基因发现奠定基础.
Abstract:
In an attempt to elucidate the molecular mechanism for resistance of black spot disease in poplar,gene expression profiles in leaves of Populus deltoides'Lus'(I-69/55)and P.euramericana'I-45/51',which were inoculated with the pathogen Marssonina brunnea f.sp.brunnea,were analyzed based on expressed sequence tags (ESTs).A total of 20 023 valid ESTs from the 5'terminal ends derived from corresponding cDNA libraries of the two poplar species were sequenced.Cluster analysis of the 20 023 sequences yielded 10 816 tentative unigenes,including 3 734 contigs and 7 082 singletons.All tentative unigenes were classified by Gene Ontology functional categories.To find resistance-associated candidate genes and locate them on poplar genome,330 unigenes was chosen from 8 853 annotated tentative unigenes,and their BLAST alignment was conducted with Populus trichocarpa assembly,1 77 related candidate resistant genes were found,and they presented in 282 unigenes.Among these genes,there were 1 35 genes located on 18 different linkage groups of poplar genome,and 42 genes located on the different scaffolds.This study supplied a resource of candidate genes for further exploring the genetic mechanism for the host horizontal resistance to the pathogen Marssonina brunnea at the whole genome range,and provided important information for further gene discovery.  相似文献   

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不同品种的杨树对重金属污染土壤的修复和耐受性存在显著差异,为了解不同品种的杨树对铅(Pb)和镉(Cd)的耐抗特性,以南林95杨和南林895杨为研究对象,研究其在不同浓度Pb胁迫和PbCd复合胁迫下的生理响应,对其根系活力、过氧化氢酶(CAT)活性、可溶性蛋白质和叶绿素含量等指标进行对比分析,并采用主成分分析和隶属函数法综合评价2种杨树对Pb和Cd胁迫的耐受性。结果表明,南林95杨和南林895杨在Pb胁迫和Pb-Cd复合胁迫下,各生理指标的变化趋势大致相同,均表现出较强的耐受性;在Pb胁迫和Pb-Cd复合胁迫下,南林895杨的根系活力和CAT活性均强于南林95杨,但其叶绿素含量少于南林95杨。Pb胁迫下,南林95杨的可溶性蛋白质含量略高于南林895杨,在Pb-Cd复合胁迫下则相反。综合评价结果表明,南林895杨对重金属Pb和Cd的耐受性强于南林95杨,具有更好的修复前景。  相似文献   

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Qin R  Hirano Y  Brunner I 《Tree physiology》2007,27(2):313-320
We studied the effects of a 2-day exposure to aluminum (Al), copper (Cu) and zinc (Zn) stress on root exudates and root apices of aseptically grown poplar (Populus tremula L.) cuttings. Aluminum induced root exudation of oxalate and citrate, Cu induced root exudation of oxalate, malate and formate, and Zn induced root exudation of formate. The threshold treatment concentrations were 100 microM for Cu and 500 microM for Al and Zn, corresponding to about 30 microM Cu2+, 140 microM Al3+ and 290 microM Zn2+. Simultaneous with the increase in organic acid anion exudation, sulfate and the nutrient cations K+, Ca2+ and Mg2+ were released into the solutions. Significant positive correlations between the organic acid anions and the cations indicate possible co-release. Toxicity symptoms of the poplar roots included browning of the root apices, which occurred at Cu concentrations of 50 microM and above, at Zn concentrations of 500 microM and above, and at an Al concentration of 1000 microM, and callose formation, which was observed solely in response to Al concentrations of 500 microM or higher. The results indicate that the composition of the exuded organic acid anions from poplar roots and the toxicity symptoms are specific to each of the applied heavy metals.  相似文献   

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In recent years, a considerable number of studies have harnessed the power of genomics to decipher the role of pathogenesis‐related (PR) proteins in plant defence against various biotic and abiotic stresses. Chitinases are PR antifungal proteins expressed constitutively at low levels in plants and induced during biotic pressures and are demonstrated to be involved in the plant defence responses. Remarkable induction of chitinase enzymes by various abiotic agents (salicylic acid, jasmonic acid, ethylene and ozone) and biotic components (pathogens, insect pest, fungal cell wall components and oligosaccharides) is well demonstrated in plants. Several reviews on plant chitinase expression during host–pathogen interaction are available for annual species, whilst reports of their expression in tree species are limited to a few woody perennials: Populus, Pinus, Picea, Eucalyptus, Castanea and Pseudotsuga. The aim of this paper is to review the induction of chitinase during various stresses and developmental processes in forest tree species.  相似文献   

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【目的】杨树是我国人工林的重要组成部分,也是公认的模式木本植物。植物BAG蛋白作为保守的分子伴侣,在生长发育和抗逆性中起到关键作用。研究杨树BAG蛋白家族的进化与表达模式,鉴定BAG基因的生物学功能对于林木遗传改良具有重要的指导意义。【方法】以拟南芥7个BAG蛋白为诱饵,搜索毛果杨、水稻和小立碗藓在线数据库(https://phytozome.jgi.doe.gov)获得这3个物种同源的BAG蛋白。利用生物信息学构建毛果杨、拟南芥、水稻和小立碗藓BAG蛋白的系统进化树,分析这些蛋白的生化特性及进化关系。结合杨树芯片(http://bar.utoronto.ca)数据,利用qRT-PCR检测杨树BAG基因的组织和逆境(旱和热胁迫)表达模式。【结果】毛果杨包含14个BAG基因,按照国际命名法(以染色体位置进行排序),将其命名为PtrBAG1—PtrBAG14。系统进化树表明杨树、拟南芥、水稻和小立碗藓BAG蛋白相对保守,大体可以分为2个亚家族。第1个亚家族中,大部分BAG蛋白N端含有UBL结构域,可能作为分子桥梁参与降解某些蛋白;第2个亚家族的大部分BAG蛋白含有IQ结构域,意味着这些蛋白可能与Ca2+信号相关。杨树BAG基因在不同逆境处理(热胁迫和干旱胁迫)条件下有不同的表达模式。热处理诱导所有BAGs基因表达发生变化,其中BAG1、BAG4、BAG6、BAG7和BAG8表达被激活,而另9个BAG基因的表达被抑制;特别是,热诱导BAG4和BAG6表达上调超过90倍。干旱胁迫条件下,大部分BAG表达变化幅度很小。组织表达模式分析表明,大部分杨树BAG基因在根、叶、芽、小苗、雌花序、雄花序和木质部中表达量很低,只有BAG2和BAG12在木质部中大量表达。【结论】杨树BAG家族共有14个成员,大体分为2个亚家族,在进化上与本文选用的陆生植物拟南芥和水稻BAG蛋白有更高的相似性。杨树14个BAG基因可能都参与了热胁迫调控,但不同成员在抗热过程中起不同作用。相比而言,大部分杨树BAG基因可能并不参与抗旱。值得指出的是,杨树BAG4和BAG6可能在热胁迫中作用最明显,而BAG2和BAG12可能参与木材形成。  相似文献   

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1IntroductionAdversitystudyisoneofthefocusesinplantphysiology.However,thepriorityhadbeenputontheselectionofpoplarcloneswithfastgrowthrateandbetterwoodqualityonpoplarbreedinginthepastfewdecades.Therewerelimitedresearchestargetedontheresistancebreeding.Moreover,mostoftheresearchonadversityresistancewasconductedonPopulustomentosa(Peietal.1994,Duanetal.1998)andPopulusussuriensis(Jiangetal.1999).LittlewasknownabouttheadversityresistanceofpoplarclonesinSectionAigeiros(Wang1982,Zhu1990,Liuetal.1…  相似文献   

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Reforestation in the humid tropics and arid zones, where trees are often subject to stresses, is an effective approach for mitigating global warming. Forestation with Populus species that are tolerant to the stresses in such regions has been conducted. The selection of poplar trees with higher stress tolerance leads to more efficient reforestation. The genome-wide bioinformatics approaches of gene function have been used for revealing the mechanisms of biological processes, including such stress tolerance. The decoding of the poplar genome has been followed by the genome-wide identification of genes and then the inference of gene function for systematic understanding of biological processes. To predict gene function in poplar, we analyzed poplar gene expression data using DNA microarray datasets obtained from the Gene Expression Omnibus database and developed a database for poplar gene co-expression analysis. Using the database, we illustrate the steps to retrieve two groups of co-expressed genes that are specifically expressed in experiments of hypoxic stress response in gray poplar, a flooding-tolerant tree species. Our database allows users to extract genes involved in biological processes, such as stress reaction, and then is useful for understanding such mechanisms in tree species.  相似文献   

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以毛白杨为转基因受体材料,利用根癌农杆菌介导法转化来源于益母草的阳离子抗菌肽基因LJAMP2。经卡那霉素筛选,共获得50株抗性植株。GUS组织染色和PCR检测显示有30株抗性植株呈阳性,初步证明外源目的基因已整合到毛白杨基因组中。RT-PCR证实抗菌肽基因LJAMP2在转基因植株中能大量表达。离体抗病性试验表明:转基因毛白杨细胞粗提液的抑菌能力明显强于非转基因植株。进一步将溃疡病菌接种在转基因和野生型毛白杨茎段上培养30天,转基因植株的病级指数均低于非转化植株。上述抗性试验结果表明:在毛白杨中超量表达益母草抗菌肽基因LJAMP2能显著提高其溃疡病抗病性。  相似文献   

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转双抗虫基因741杨特征及其繁殖技术   总被引:7,自引:1,他引:7  
简述了当今植物抗虫基因工程的研究进展 ,介绍了Bt基因和蛋白酶抑制剂基因 ,农杆菌介导的遗传转化以及防治害虫产生抗性的策略等。双抗虫基因是用部分改造BtCry1Ac基因和慈菇蛋白酶抑制剂基因 (API)构建的植物表达载体。被转化体 74 1杨 (Populus×aldatomen tosaCl.74 1)是以银白杨× (山杨 小叶杨 )为母本 ,毛白杨为父本 ,1994年杂交育成 ,为一优良白杨无性系 ,形态酷似毛白杨。在转双抗虫基因的研究中 ,选出高抗虫、中抗虫无性系。高抗虫无性系杀死昆虫幼虫的总死亡率为 83%~ 90 %。从苗期高生长和形态观察 ,转双抗虫基因 74 1杨生长发育正常。比较详细地介绍了 74 1杨营养繁殖技术及基本原理。介绍了硬枝扦插容器育苗、组培微繁技术、容器育苗的移栽和苗圃管理以及采穗圃的建立和经营等  相似文献   

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中国和外来杨属种质资源   总被引:1,自引:0,他引:1  
介绍了中国天然分布和从国外引进的杨属(PopulusL.)种质资源。总结了中国天然分布的杨属种质资源的派别、种类、分布范围和分布特点。在中国杨属5派中,以青杨派(Tacamahaca)种类最多,白杨派(Populus,以前称Leuce)分布最为广泛,黑杨派(Aigeiros)最具经济价值。将中国杨属树种引种历史划分为解放前的零星引种时期、50年代至80年代的以引进国外培育成功的优良品种为主时期和90年代后注重种质资源引进时期三个阶段,介绍了每个阶段的引种特点和引进的种质资源。在过去50年种,中国天然分布的白杨派和引进的黑杨派(Aigeiros)种质资源在中国的杨树栽培和育种中发挥了巨大作用。  相似文献   

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