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高羊茅DREB类转录因子基因的分离及鉴定分析 总被引：7，自引：1，他引：7
DREB转录因子是植物中特有的与低温、高盐和干旱胁迫相关的反式作用因子。为了分离和鉴定高羊茅(Festuca arundinaceaSchreb.)DREB转录因子基因,我们构建了冷诱导(4℃)的高羊茅cDNA文库,用拟南芥DREB1A基因作探针筛选该文库得到一个DREB类基因FaDREB1。序列分析表明,FaDREB1具有一个651bp的开放阅读框和229bp的3’非编码区,推测的氨基酸序列中含有一个高度保守的EREBP/AP2结构域。酵母单杂交结果表明FaDREB1蛋白能在体外特异结合DRE元件,并具有转录激活功能。Northern杂交结果发现FaDREB1基因受低温的诱导表达,对高盐、干旱和ABA没有反应,说明FaDREB1基因在高羊茅植株对低温的应答反应中起重要作用。 相似文献
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为探究大海马幼体在高温和低温胁迫条件下基因表达水平的变化规律,采用Illumina平台的PE150测序功能对大海马幼体肝脏进行了转录组测序。对照组、高温胁迫组和低温胁迫组测序数据组装后获得22 513个单基因簇(Unigene),N50为2 223 bp,平均长度为1 122.71 bp。与对照组相比,高温胁迫组共获得14 009个差异表达基因,其中5 543个差异表达基因上调,8 466个差异表达基因下调;低温胁迫组共获得20 030个差异基因,14 016个差异表达基因上调,6 014个差异表达基因下调。差异表达基因经KEGG数据库富集发现,高/低温胁迫均导致大海马体内抗氧化途径相关基因表达(HSP70、HSP90、SOD等)发生显著改变;另外,高温胁迫还造成大海马免疫系统相关基因(PIK3R、Akt、IL-10、TLR等)以及细胞凋亡相关基因(CYCS、CASP9、CASP3等)表达显著改变;低温胁迫造成DNA损伤修复(MSH、DDB2、XRCC2、RAD52、Ogg1、PMS2等)、脂肪酸代谢(StARD7、ApoA4、CYP51、Fadsd6等)和细胞凋亡(P21、P53、BAX等)相关基因显著上调。选择HSP70、Fadsd 6与IL-10等13个差异基因验证,RT-qPCR结果与RNA-Seq测序结果基本一致。本研究结果为深入探究大海马温度应激的调控机制奠定了一定的理论基础,而且有助于预防极端温度对大海马造成损伤。 相似文献
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为探究高羊茅(Festuca arundinacea)硝态氮转运蛋白基因(NRT1.1)的表达模式,本研究以黔草1号高羊茅为试验材料,采用RACE和RT-qPCR技术对高羊茅NRT1.1基因的cDNA全长序列进行扩增,并对其不同胁迫处理下的表达情况进行分析。生物信息学分析发现,高羊茅NRT1.1的理论等电点为4.81,平均亲小性为0.919,含有约32.63% α-螺旋、7.63% β-转角和53.73%不规则卷曲。结果表明,NRT1.1基因的cDNA序列全长为2 328 bp,编码606个氨基酸,预测蛋白质分子量为193.9 kDa,且高羊茅NRT1.1与黑麦草NRT1.1氨基酸序列的相似性最高。RT-qPCR表达分析发现,高羊茅叶片NRT1.1受低氮处理0.5~1 h时表达量达到峰值,显著(P< 0.05)高于对照组;在干旱和热处理下,NRT1.1表达量分别在6 h和12 h时达到峰值,且显著(P< 0.05)高于对照组;在盐处理下,仅在6 h时NRT1.1表达量高于对照组,其余时间均受显著(P< 0.05)抑制。本研究结果为解析高羊茅NRT1.1基因的表达模式提供了分子生物学基础。 相似文献
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为从分子水平揭示甲基营养型芽胞杆菌WM7低温响应机制,采用高通量测序技术对低温和常温培养的菌体进行转录组测序分析,对部分差异表达显著基因进行RT-qPCR验证。结果表明,共检测到2 029个差异表达基因,其中1 519个低温上调,510个低温下调。GO和KEGG分析显示,1 199个差异表达基因归属细胞组分、分子功能和生物过程3个部分,683个基因参与到225个代谢途径中,其中涉及基因最多的是磷酸戊糖途径、嘌呤和氨基酸合成途径。菌株低温响应主要与脂肪酸代谢、ABC转运体、转录调控、信号转导、碳氮代谢等途径相关。此外,检测到大量差异表达的未知功能基因,多个基因差异表达倍数均大于50倍。本研究从基因表达方面阐述了甲基营养型芽胞杆菌WM7低温响应机制,为全面揭示其低温生长机制奠定了理论基础,提供了基因信息。 相似文献
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高羊茅在生长季出现生殖枝,抑制新枝形成,不利于草坪质量及其持久性生长。研究春化基因的分子特征,探索抑制生殖生长的分子育种新途径,对坪用型高羊茅品种改良具有重要意义。本研究在克隆高羊茅春化基因FaVRN1的基础上,构建高羊茅春化基因FaVRN1与绿色荧光蛋白基因GFP融合的植物表达载体p-FaVRN1-hGFP,利用基因枪转化法转入洋葱表皮细胞,荧光显微镜检测融合基因的瞬时表达,并运用实时荧光定量PCR分析春化基因FaVRN1在春化与非春化条件下的表达差异。研究结果表明,FaVRN1基因编码的蛋白产物位于细胞核,符合它作为转录因子特性;春化条件下,FaVRN1基因的表达随处理时间延长逐渐增加。非春化条件下,FaVRN1基因的表达随处理时间延长而降低。FaVRN1基因在春化条件下的表达水平远高于非春化条件,FaVRN1基因的表达受春化条件正调控。 相似文献
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苦瓜在生长季节遇到低温会影响其生长发育和产量,限制了种植分布区域。为了研究苦瓜耐寒性的分子机制,本研究以苦瓜自交系43为试验材料,在8℃低温胁迫处理后观察其表型并测定相关生理生化指标,再利用转录组测序技术从生理角度结合分子水平对苦瓜低温胁迫机制进行生物信息学分析。结果表明,在8℃低温胁迫下,苦瓜表现出耐低温适应性,在低温胁迫下丙二醛(MDA)含量、脯氨酸含量及电解质渗透率均呈现显著变化。分析了低温胁迫0和12 h后的转录组变化,结果发现低温胁迫处理后有916个基因上调,369个基因下调。对差异表达基因进行通路注释分析,发现大量与低温胁迫相关的代谢途径。进一步筛选到10个在其他物种报道过可能与冷胁迫相关的基因,包括MYB、WRKY以及NAC转录因子等。对植物耐低温相关的候选基因进行了实时荧光定量PCR(qRT-PCR)表达谱分析,结果与生理表型变化相吻合,同时进一步证实了RNA-seq数据的准确性。本研究基于全基因组水平分析苦瓜在低温胁迫下的转录组变化,以及耐低温相关基因动态变化和生理反应的分子机制,为揭示苦瓜中的耐寒性机理和筛选遗传改良候选基因提供了一定基础。 相似文献
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利用差异显示法研究水稻耐淹涝相关基因 总被引：7，自引：0，他引：7
应用mRNA差异显示法对耐淹材料籼稻(Oryzasativassp.indica)FR13A和敏感籼稻(O.sativassp.indica)IR39595-503-2-1-2在淹涝胁迫下的基因差异表达进行了分析。结果显示,从40对引物中共扩增出1428条片段,筛选到102条在耐淹涝材料和敏感材料间差异明显的基因片段,差异率为7.1%。其中有42条差异带来自耐淹涝材料,有7个差异片段已通过Northern杂交验证在耐淹涝材料FR13A中表达。进一步克隆测序并进行数据库比对分析表明,有4个与水分胁迫产生的应答反应和生理生化变化有关,其中一个与ATP结合蛋白高度同源,另3个与异柠檬酸酶脱氢酶、NADH脱氢酶和乙醛酸转移酶部分序列高度同源,其余3个为新的cDNA片段。 相似文献
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海藻糖-6-磷酸合成酶基因(trehalose-6-phosphate synthase,TPS)是海藻糖合成途径中的关键酶基因,在植物耐寒过程中发挥着重要作用。为了分析不同耐寒性的小麦品种中TPS基因在低温处理下的表达差异,明确TPS基因在抗寒中所起的关键作用,以抗寒品种东农冬麦1号(D1)和对照品种中国春(CS)为材料,设定一系列低温处理,用qRT-PCR分析7个TPS基因的表达差异。结果表明,低温驯化初期,TPS4和TPS7在D1中大量表达。低温驯化后期,D1中TPS1、TPS2、TPS4、TPS6和TPS7基因的表达量均明显高于CS中的表达量。TPS2、TPS3、TPS5和TPS6基因在小麦低温冷冻中后期相对表达量达到最高。-16℃下D1中的TPS1、TPS6和TPS7基因相对表达量分别为同时期CS的72.8倍、308.68倍和32.83倍。D1中的TPS3基因在7个冷处理中有6个为下调表达,而TPS7基因在各处理下均为上调表达。本研究结果对高抗寒性新品种的选育具有重要意义。 相似文献
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紫花苜蓿铝胁迫抑制消减文库的构建和初步分析 总被引：5，自引：0，他引：5
为了分离和鉴定铝诱导基因，以紫花苜蓿小冠品种为对象，以经过40μM铝离子胁迫的紫花苜蓿幼根为实验方，未胁迫的为驱赶方，用抑制消减杂交法（SSH）构建了一个紫花苜蓿铝胁迫消减文库。菌落PCR显示插入片段在250bp到1200bp之间，文库质量良好。通过对文库阳性克隆随机测序和BLASTX分析表明,文库中包含了许多在铝胁迫中发挥作用的基因，涉及到抗氧化作用、信号的传导、发育和能量代谢等多种生理过程，比如过氧化物酶、钙依赖蛋白激酶、蔗糖转化酶、蛋白磷酸脂酶调节因子等。而且许多EST序列与其他生物性和非生物性胁迫诱导相关。为了解紫花苜蓿的耐铝机制和抗性相关基因的克隆奠定了基础。 相似文献
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细胞色素P450属于一种含亚铁血红素单加氧酶的超基因家族,在植物多种代谢途径与防御反应中起着重要作用。为探究细胞色素P450基因在高羊茅中的表达模式,本研究采用cDNA末端快速扩增技术从高羊茅叶片中克隆了细胞色素P450基因,命名为FaP450。FaP450全长1 737 bp,含1 437 bp的开放阅读框,共编码478个氨基酸,具有P450基因家族典型的P450结构域。系统进化树分析表明,高羊茅FaP450与禾本科植物小麦、山羊草的P450蛋白亲缘关系较近。荧光定量PCR表明,高羊茅FaP450基因受干旱、高温、氮胁迫及盐胁迫的诱导表达上调,表明该基因与非生物胁迫的抗性相关。构建FaP450超量表达载体,利用农杆菌侵染法转化拟南芥,对其进行低氮胁迫,发现低氮胁迫下FaP450基因过量表达植株的鲜重与根长显著高于野生型,表明FaP450基因的超量表达可以增强拟南芥对低氮的适应性。本研究为深入探索P450基因家族在牧草生长发育与抗逆功能的研究奠定了理论基础。 相似文献
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Purpose
Two plant species (tall fescue and alfalfa) grown alone and in combination was investigated to evaluate phytoremediation of polychlorinated biphenyl (PCB)-contaminated soil and the impact on the catabolic genes and soil enzyme activities in the rhizosphere.Materials and methods
Surface soil was collected from Xiaoshan (a PCB-containing capacitors and transformers storage site). The phytoremediation experiment was carried out in an agricultural greenhouse experiment station at Huajiachi campus, Zhejiang University. Soil dressing method and fertilizers were applied to improve soil quality for the pot experiment. Soil dehydrogenase activity was measured spectrophotometrically by the reduction of 2, 3, 5-triphenylterazolium chloride (TTC) to triphenyl formazane (TPF) and catalase activity was determined by back-titrating residual H2O2 with KMnO4. Quantitative real-time PCR using SYBR green I was employed with the individual primer sets to determine the relative abundance of the biphenyl dioxygenase genes. Total bacterial numbers were determined by CFU counting and amplification of 16S rDNA.Results and discussion
Planting treatment significantly enhanced bacterial numbers and PCB removal. The copy numbers of the bphA, bphD.1.B, bphD.2.A, and bphD.2.A/B genes, total bacteria counts, and dehydrogenase activity were the highest in mixed cropping soil, which indicated that tall fescue (forage grass) and alfalfa (legume) mixed cropping was most beneficial to soil bacteria, the potential PCB degraders, and enzyme activity. However, the highest removal of PCBs was found in tall fescue single plant cultivation, followed by combined plant cultivation, probably because tall fescue had greater biomass and could extract more PCBs from soil. Compared with nutrients amended unplanted control, the removals of tri-, tetra-, and penta-CBs in tall fescue single-planted and tri-CBs in combined planted soils were significantly enhanced (p?<?0.01).Conclusions
The presence of vegetation significantly promoted the dissipation of PCBs and growth of total bacteria and the potential PCBs degraders in soils. Tall fescue and alfalfa mixed cropping was most beneficial to soil bacteria and enzyme activity. Tall fescue showed highest ability for remediation of PCBs in a poor quality soil. 相似文献14.
Salinity stress is a major factor limiting the growth of turfgrass irrigated with recycled wastewater. The change in lipid peroxidation in terms of malondialdehyde (MDA) content and the activities of superoxide dismutase (SOD), catalase (CAT), ascorbate peroxide (APX) and glutathione reductase (GR) in the shoots and roots of Kentucky bluegrass and tall fescue were investigated under salinity stress. Plants were subjected to 0, 50, 100, 150 and 200 mmol L 1 NaCl for 40 d. The MDA content under salinity stress was lower in tall fescue than in Kentucky bluegrass in both shoots and roots. Activities of SOD in the shoots of both species increased with salinity stress. The activities of CAT and APX decreased in Kentucky bluegrass, but no significant difference in the activities of CAT and APX was observed in tall fescue. The activities of SOD, CAT and APX in the shoots of tall fescue were higher than those in Kentucky bluegrass. In the roots of Kentucky bluegrass, SOD and GR activities increased and CAT and APX activities decreased in comparison with the control. In the roots of tall fescue, salinity increased the activities of SOD, CAT, and APX. These results suggested that tall fescue exhibited a more effective protection mechanism and mitigated oxidative stress and lipid peroxidation by maintaining higher SOD, CAT and APX activities than Kentucky bluegrass. 相似文献
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高羊茅CBF转录激活因子基因片段的克隆 总被引：4，自引：0，他引：4
根据拟南芥CBF基因序列的保守区设计 1对特异引物 ,采用PCR方法从高羊茅基因组中扩增出 1个DNA片段并克隆到pUCm T载体中。经序列测定和分析 ,该片段长 60 8bp ,与 3个拟南芥CBF基因的核苷酸及其推导氨基酸序列分别具有 81～ 84%和 75～ 79%的同源性 ,而且推导氨基酸序列中包含有同源性更高的AP2DNA结合域以及CBF蛋白的两段特征序列PKK RPAGRxKFxETRHP和DSAWR。这些结果表明 ,本研究克隆的片段为高羊茅CBF基因片段 相似文献
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本试验构建了PEG6000渗透胁迫下白花柽柳的抑制性消减杂交(suppression subtractive hybridization, SSH)文库。文库克隆的重组率为95 % ，插入片段大部分集中在100～1000 bp之间。通过对文库阳性克隆的随机测序，并对表达序列标签（expressed sequence tag, EST）分析得到了100个与干旱胁迫相关的EST片断，它们涉及了植物的渗透调节、信号传递、转录调控、活性氧清除、蛋白代谢、跨膜转运和离子平衡及光合作用等方面。此结果对今后进一步研究这些基因的功能奠定了基础。 相似文献
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营养匮乏和伴生杂草是水稻种植过程中的常见影响因素。本研究应用抑制消减杂交(SSH)技术,分别研究低氮、稗草条件下,非化感水稻品种"Lemont"的上调表达基因。结果显示,低氮条件下,"Lemont"水稻中参与生长调控的生长素响应蛋白,参与抗逆防御的类NBS-LRR蛋白、过氧化氢酶、金属硫因蛋白,以及参与蛋白质代谢相关蛋白的编码基因上调表达。稗草共培下,编码NADH脱氢酶、细胞色素氧化酶、ATP依赖性RNA解旋酶等与植物生长相关的基因,与抗逆防御相关的几丁质酶和糖基水解酶基因,以及与信号转导相关的锌指蛋白及组氨酸激酶基因增强表达。研究结果表明,非化感水稻"Lemont"能够通过调节抗逆以及生长调节相关基因的表达,从而响应不同的胁迫。 相似文献
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Roots of endophyte‐infected (E+) tall fescue (Festuca arundinacea Schreb.) exude more phenolic‐like reductants than roots of endophyte‐free (E‐) plants when mineral stressed. Phenolic compounds are efficient chelators of aluminum (Al) and may influence Al tolerance in many plant species. The objective of our study was to determine if enhanced release of phenolic compounds by roots of E+ plants contributes to Al tolerance in tall fescue. Two cloned genotypes (DN2 and DN11) of tall fescue infected with their naturally occurring fungal endophyte Neotyphodium coenophialum (Morgan‐Jones and Gams) Glenn, Bacon and Hanlin and their noninfected isolines were grown in nutrient solutions at 0 μM Al (Al‐) and at 640 μM Al (Al+) under controlled environment conditions. Root and shoot dry matter (DM) of endophyte‐infected tall fescue was greater in E+ than E‐ plants by 57% and 40%, respectively, when plants were grown without Al. Endophyte infection did not affect root and shoot DM of tall fescue grown with Al but relative (to Al‐treatment) reduction in root and shoot DM was greater in E+ than E‐ plants. In response to Al stress, more Al (47%) and P (49%) could be desorbed from root surfaces of E+ than E‐ plants. Aluminum concentrations in roots of E+ plants were 35% greater and P concentrations were 10% less than those determined in roots of E‐plants. No differences in mineral concentrations were observed in shoots, regardless of endophyte status, or Al level in nutrient solution. Roots of E+ plants increased pH of both Al‐ and Al+ nutrient solutions to a greater extent than roots of E‐ plants in a 48 h interval. Our results show that more Al can be sequestered on root surfaces and in root tissues of endophyte‐infected tall fescue than in plants devoid of endophyte. Aluminum sequestration was greater on root surfaces and in root tissues of E+ than E‐ plants of a given tall fescue genotype. Our results suggest that increased exudation of phenolic‐like compounds from roots of endophyte‐infected tall fescue may be directly involved in Al tolerance and serves as a mechanism for widespread adaptability and success of endophyte‐tall fescue associations. 相似文献