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1.
NBS-LRR(nucleotide-binding site and leucine-rich-repeat)是植物中最大类抗病基因家族之一。番茄基因组测序完成为全基因组水平上分析NBS-LRR抗病基因家族提供了机遇。利用生物信息学方法对番茄NBS-LRR抗病基因家族成员数目进行鉴定,并对其染色体定位和系统发育关系进行了分析。随后,将番茄和马铃薯NBS-LRR抗病基因进行了比较基因组学分析。结果表明:番茄基因组共包括252个NBS-LRR抗病基因,分布于番茄12条染色体上;63.5%的基因成簇存在,大部分为串联重复;系统发育关系分析表明番茄CC-NBS-LRR(CNL)亚家族较其他亚家族扩展程度大;同线性分析发现番茄中共79个NBS-LRR抗病基因与马铃薯基因具有同源关系。结果将为番茄NBS-LRR抗病基因家族的深入研究提供依据,同时也为利用番茄NBS-LRR基因进行基因定位以及相关抗病基因克隆等奠定基础。  相似文献   

2.
细胞色素P450 (cytochrome P450,CYP450)是高等植物中最大的酶蛋白家族之一,广泛参与植物的次生代谢和抗逆生理.本研究采用同源克隆和cDNA末端快速克隆(rapid-amplification of cDNAends,RACE)技术,获得苦荞(Fagopyrum tataricum) CYP81家族同源基因FtP450-R4 (GenBank登录号:KM271986).FtP450-R4 cDNA全长1770bp,5'-UTR为49 bp,3'-UTR为194bp,ORF为l 527 bp,其ORF序列可编码508个氨基酸残基的蛋白.生物信息学分析表明,FtP450-R4蛋白通过N-端4~24位氨基酸残基定位于内质网,与F3'H (flavonoid 3'-hydroxylase)、I2'H (isoflavone 2'-hydroxylase)和其他CYP450的同源性在44%~46%之间;多重序列比对显示,FtP450-R4具有CYP450中经典的基序和保守序列,但不含F3'H的特征基序(GGEK);系统进化树分析显示,FtP450-R4与拟南芥(Arabidopsis thaliana) CYP81家族和I2'H聚为一大簇,说明其可能参与苦荞黄酮类化合物的羟化或对逆境胁迫的应答.UV-B、寒冷和干旱均能引起FtP450-R4在苦养子叶中表达量的显著变化,而在胚轴中其表达量变化不显著.利用大肠杆菌(Escherchia coli) BL21 (DE3)菌株对FtP450-R4蛋白进行了可溶性表达,活性鉴定表明,重组FtP450-R4蛋白能以还原型烟酰胺腺嘌呤二核苷酸磷酸(reduced form of nicotinamide-adenine dinucleotide phosphate, NADPH)和山奈酚为底物进行酶促反应,具有生物学活性.本研究可为了解CYP450在黄酮合成代谢中的功能,明确苦荞黄酮代谢调控的分子机制提供了基础资料.  相似文献   

3.
诸葛菜OvCYP86MF基因的克隆及其特性分析   总被引:2,自引:1,他引:1  
为进一步阐明细胞色素P450基因CYP86MF在诸葛菜发育中的分子机理,本文根据已知同源基因保守序列设计特异引物,利用RT-PCR和RACE技术从诸葛菜中获得一个细胞色素P450基因(OvCYP86MF)全长cDNA序列,该序列全长为1876bp,含有1605bp的完整开放阅读框,可编码534个氨基酸,分子量和等电点分别为61.4kDa和6.90,具有细胞色素P450蛋白的典型特征,即保守结构域FNAGPRLCIG;原核表达显示该基因的融合蛋白在体外可以诱导表达;DANSTAR和Clustal W软件分析表明该基因的全长cDNA序列及其编码氨基酸序列与十字花科物种拟南芥相似性很高,达到80%以上,亲缘关系最近;与CYP86C亚家族成员在氨基酸水平上的相似性均高于50%,因此推断该基因属于CYP86C这个亚家族。Northern杂交分析表明该基因在花蕾中特异表达。  相似文献   

4.
十字花科植物CYP86MF同源基因的克隆及特征分析   总被引:3,自引:0,他引:3  
用保守的特异引物进行PCR扩增,并结合RACE技术,从大白菜(Brassicacampestris L.)品种黄芽14,萝卜(Raphanus sativus L.)品种圆白和诸葛菜(Orychophrogmus violaceus)3个材料中扩增到3个完整的cDNA全长,分别命名为Bc-CYP86MF,RsCYP86MF和OvCYP86MF,它们的cDNA全长分别为1854,1818和1876bp,分别编码524,526和534个氨基酸残基。将所推导的氨基酸序列与数据库中已知基因进行排序,发现它们与拟南芥CYP86家族所有成员的氨基酸相似性都大于38%,可将它们归为细胞色素CYP86基因家族。BcCYP86MF和RsCYP86MF与CYP86C4亚家族氨基酸同源性均大于55%,则可归为CYP86C4亚家族,而OvCYP86MF与CYP86C3的同源性最高,为86.5%,应归为CYP86C3亚家族。根据氨基酸序列的排序结果构建的系统树表明,CYP450基因的氨基酸序列进化程度与物种的亲缘关系的远近相吻合。对蛋白二级结构预测结果发现,它们都具有细胞色素P450典型的保守结构域FNAGPRLCIG,而且氨基酸组成也很相似。  相似文献   

5.
为分离甘薯[Ipomoea batatas(L.)Lam.]交替氧化酶(AOX)基因家族成员,分析其在甘薯应答生物和非生物胁迫中的响应模式,本试验在筛选甘薯基因组的基础上,进行基因克隆和测序,最终从心香甘薯中获得3个AOX基因家族成员.基因结构分析结果表明,3个甘薯AOX基因的cDNA全长依次为963、1 080和1 ...  相似文献   

6.
聚合酶链式反应-单链构象多态性(polymerase chain reaction-single strand conformation polymorphism,PCR-SSCP)作为一种能检测基因组间DNA碱基微小差异的技术手段,具有操作简便、快捷且灵敏度高等优点.比较基因组学近年来已经成为研究生物基因组的最主要手段之一;大量基因组序列的产生为通过比较基因组学开发新标记和定位目标基因提供了大量的DNA序列资源.本研究以李氏超短纤维突变体(Ligon lintless-1,Li1)为材料,定位Li1基因,针对PCR扩增产物片段较大的样品(>400bp)进行PCR-SSCP技术的优化,用己发表的拟南芥(Arabidopsis thaliana)和棉花(Gossypium spp.)D基因组DNA序列为参考,探索PCR-SSCP技术与比较基因组学结合开发新标记应用于棉花基因定位的可能性.结果表明,电压为150V,胶浓度为10%,电泳温度为4℃时电泳结果最好;上样缓冲液与PCR扩增目的片段比例为5∶1为最佳比例.利用上述优化的PCR-SSCP技术将根据棉花与拟南芥同线性区段开发的分子标记构建了一个由28个标记组成的长度为149.6 cM的遗传图,该区域包含4个功能上与Li1位点密切相关的基因.此外,利用D基因组序列开发的标记构建了一个由17个SSCP标记和Li1位点组成,跨越40.3cM的遗传图谱,距其最近的两端标记W058和P095分别为0.6和0.3 cM.本研究为棉花Li1基因的精细定位及其候选基因的克隆提供了理论基础.  相似文献   

7.
家蚕谷胱甘肽转移酶基因的克隆、序列分析及其表达模式   总被引:2,自引:0,他引:2  
谷胱甘肽-S-转移酶(GSTs)是昆虫体内重要的解毒酶系,在对外源和内源化合物解毒代谢中起着重要的作用。以GenBank中已知的昆虫GST的氨基酸序列在家蚕(Bombyx mori)基因组序列中作TBLASTN检索,获得了多个可能的家蚕GST同源基因。对其中的一个GST新基因进行了EST分析和克隆,结果表明,该基因编码区全长663bp。以该基因推导的氨基酸序列与其它物种的GSTs进行相似性比较和系统发育分析,发现此基因为delta家族的成员,并命名为BmGSTd3。RT-PCR结果表明,BmGSTd3基因在家蚕5龄第3天的8个组织中都有表达,中肠和表皮的表达丰度较高。在辛硫磷处理家蚕后的72h内,BmGSTd3基因的转录水平与对照组相比没有显著的变化。对BmGSTd3基因的5'侧翼区的调控序列进行了预测,共发现12个可能与内源和外源物代谢以及抗氧化相关的转录调控元件。  相似文献   

8.
植物生长素在植物的生长发育过程中至关重要,GH3基因家族是植物生长素早期应答的成员。本研究采用比较基因组学的方法,利用已经分离的拟南芥GH3(Gretchen Hagen3)蛋白为检索序列,在全基因组水平上搜索拟南芥、水稻、葡萄、白杨和苜蓿的GH3基因的同源序列。最终确定了59个GH3候选基因,其中拟南芥19个,水稻14个,葡萄9个,白杨14个,苜蓿3个。对同源序列作进一步的多序列联配、MEME、ESTs和系统发生表达分析,结果表明:GH3基因家族的基本特征在单双子叶植物分离之前就已经形成;GH3结构域在蛋白质间较保守,可以分为3个亚家族,其中个别蛋白发生了基序丢失;59个同源蛋白中的40个成员找到了ESTs的证据,且表达部位多样,不同成员之间的表达部位存在差异。该研究结果将为植物的GH3基因家族的研究提供参考。  相似文献   

9.
PHAs合酶phaC2基因转化烟草叶绿体的研究   总被引:3,自引:0,他引:3  
中长链羟基脂肪酸聚酯 (medium-chain-length-PHAs, mcl-PHAs) 属于微生物聚酯。Ⅱ型PHA合酶是mcl-PHAs生物合成途径中的关键酶。将编码Ⅱ型PHA合酶的基因phaC2与水稻(Oryza sativa )叶绿体psbA基因的启动子和终止子构建表达盒(RpsbA-pro-phaC2-RpsbA-ter),连同壮观霉素抗性基因aadA表达盒(prrn-aadA-TpsbA-ter)一起克隆进烟草叶绿体基因组同源片段rbcL和accD中,构建了烟草叶绿体转化表达载体pTC2。基因枪法转化烟草(Nicotiana tobacum L.)叶片,获得具有壮观霉素抗性的转基因烟草。对T0代和T1代转基因植株的PCR和Southern blot鉴定结果表明, 外源基因确已整合进烟草叶绿体基因组中,T1代转基因植株已达同质化。RT-PCR分析结果证实phaC2基因已在转录水平上表达。转基因植株的正反交试验证明外源基因在转基因后代中遵循母性遗传规律,不存在转基因的花粉漂移现象。通过叶绿体遗传转化, 实现中长链羟基脂肪酸聚酯合成关键酶基因phaC2在烟草叶绿体基因组中的稳定转化。  相似文献   

10.
棉花转录因子基因(GhMYB11)的克隆与表达分析   总被引:1,自引:0,他引:1  
MYB基因家族在植物应对外界生物和非生物胁迫的过程中有重要的调控作用.本研究利用二倍体雷蒙德氏棉(Gossypium raimondii)基因组数据库,从陆地棉(G.hirsutum)品种鲁棉研32号中克隆到一个新的MYB转录因子GhMYB111(GenBank登录号:HQ234875.1),Cdna全长1 001 bp,开放阅读框828bp.通过与模式植物拟南芥(Arabidopsis thaliana)和主要作物小麦(Triticum aestivum L.)、玉米(Zea mays L.)、水稻(Oryza sativa L.) MYB蛋白的氨基酸序列比对发现,GhMYB 11蛋白与拟南芥中脱落酸(abscisic acid,ABA)合成和信号传导的重要基因AtMYB13/14编码的蛋白高度相似(E值分别为7e-83和1e-90),含两个高度保守的MYB结构域R2R3及一个转录激活结构域.实时定量PCR分析发现,GhMYB 11基因在黄萎病菌(Verticillium dahliae)侵染、干旱、盐和氧化胁迫处理后的棉花幼苗叶片中表达显著上调.GhMYB11基因可能在棉花生物和非生物胁迫反应中起重要调控作用,是陆地棉品种抗逆性遗传改良的重要候选基因.本研究为利用基因工程手段提高棉花抗逆性提供了基础材料  相似文献   

11.
低氮和干旱胁迫对富士和秦冠生长及氮素利用的影响   总被引:2,自引:2,他引:0  
【目的】以富士(Fuji)、 秦冠(Qinguan)嫁接在平邑甜茶(Malus hupehensis Rehd.)上的当年生盆栽苗为试验材料,采用砂培方法,研究了缺氮胁迫和干旱对富士和秦冠生长情况、 光合参数、 植株各部位氮磷钾含量及氮素利用效率的影响,分析比较了低氮干旱条件下富士和秦冠生长及氮素利用的差异,以期为果树生产高效肥水利用提供理论指导。【方法】试验共设四个处理: 正常氮正常水(ZZ)、 低氮正常水(DZ)、 正常氮干旱(ZG)、 低氮干旱(DG)。氮素和水分均设置两个水平,分别为正常氮(6 mmol/L NO-3-N)、 低氮(0.3 mmol/LNO-3-N)、 正常供水(保持盆中砂子相对含水量为饱和含水量的80%~85%)、 干旱处理(保持盆中砂子相对含水量为饱和含水量的60%~65%)。【结果】富士和秦冠的生物量(茎和叶)、 株高茎粗等生长指标以及光合速率、 气孔导度、 蒸腾速率均为正常氮正常水(ZZ)>低氮正常水(DZ)>正常氮干旱(ZG)>低氮干旱(DG),并且相对应处理下秦冠的以上指标均高于富士;正常供水下,缺氮处理使富士、 秦冠的根冠比比正常氮处理均有所增加,富士提高了2.05%,秦冠提高了22.40%。富士和秦冠的氮、 磷、 钾含量均表现出正常氮正常水(ZZ)>低氮正常水(DZ)>正常氮干旱(ZG)>低氮干旱(DG); 氮、 钾元素含量在植株各部位的分布顺序依次是叶>根>茎,磷元素则是根>叶>茎;光合氮素利用效率(PNUE)和氮素利用效率表现为秦冠处理之间差异极显著,富士处理之间差异不显著;秦冠的PNUE和NUE明显高于富士,在低氮正常水(DZ)处理下,秦冠氮肥利用率比富士高42.07%,在低氮干旱(DG)处理下高64.14%;低氮胁迫下富士和秦冠的NUE显著提高,并且秦冠提高的幅度高于富士。【结论】施用氮肥能够显著提高富士与秦冠的干物质量,同等水肥条件下,秦冠生长优于富士;水分亏缺会减少叶片对氮的吸收,干旱条件下适度增施氮肥,可提高果树的抗旱能力;低氮干旱胁迫下秦冠的生长指标、 光合指标及氮素利用效率指标均优于富士,表现出较强的抗低氮干旱胁迫的能力。  相似文献   

12.
Staff members of the Department of Botany of Palacký University in Olomouc and Gene Bank Department – Workplace Olomouc, Research Institute of Crop Production in Prague, Czech Republic, conducted an expedition in seven European countries (Austria, Czech Republic, France, Germany, Italy, the Netherlands, Switzerland) in August/September 1999 to collect wild Lactuca spp. germplasm and study its geographic distribution, ecology and biodiversity. During the mission, more than 600 locations were visited resulting in the collection of 602 seed samples (accessions) of wild Lactuca species and 13 seed samples of related genera (Chondrilla and Mycelis). Lactuca serriola f. serriola, L. serriola f. integrifolia, L. saligna and L. viminea subsp. chondrilliflora were prevalent in southern Europe (Italy, France), however, only L. serriola was common in central and western Europe (Austria, Czech Republic, Germany, Netherlands, Switzerland). The greatest diversity of Lactuca species was found in France, where also the most seed samples (165) were collected. The most characteristic habitats with a high density of Lactuca spp. populations were observed along roads and highways, grassy ditches, ruderal communities, and dust-heaps. Natural infections by powdery mildew (Erysiphe cichoracearum) and downy mildew (Bremia lactucae) on some wild Lactuca spp. were observed. Recent observations concerning the geographic distribution, population structure, habitats, and natural occurrence of diseases of Lactuca spp. are discussed. This assemblage of genetic resources of Lactuca spp. can serve as the basis of future studies of species diversification, spatial population structure, plant microevolution, domestication processes, and genetic variability of host-parasite interactions.  相似文献   

13.
Laser-induced breakdown spectroscopy (LIBS) is a new technique for the analysis of plant material. This study investigates the application of LIBS to pasture-based plant samples. The LIBS measurements were obtained from pelletized pasture samples (100 samples) that had been also analyzed by inductively coupled plasma–optical emission spectroscopy (ICP-OES) following microwave digestion for calibration and comparison purposes. Comparisons for elements sodium (Na), potassium (K), magnesium (Mg), calcium (Ca), manganese (Mn), iron (Fe), copper (Cu), zinc (Zn), boron (B), phosphorus (P), and sulfur (S) showed that LIBS could be used for almost all the standard profile total elements with concentrations down to low mg/kg levels (observed error of Na: 0.024 percent, K: 0.18 percent, Mg: 0.016 percent, Ca: 0.073 percent, P: 0.017 percent, Mn: 31 mg/kg, Fe: 150 mg/kg, Zn: 6.6 mg/kg, and B: 1.1 mg/kg). Elemental analysis at less than mg/kg levels was not possible using LIBS. The elements S and Cu were particularly difficult to analyze with reliability using LIBS at the concentration levels found in the plant samples. Replacing microwave digestion and subsequent ICP analysis with a direct analysis of dried plant samples using LIBS has the potential to improve the productivity and reduce the cost of testing.  相似文献   

14.
We have studied the effect of anaerobically digested (ADMSW) and composted municipal solid waste (CMSW) on mineralization and foodweb dynamics to verify the hypothesis that ADMSW would immobilize N right after addition to soil in contrast to addition with CMSW. Another hypothesis was that the mesofauna (enchytraeids and microarthropods) would stimulate N release from the decomposer community. We measured excretion of -N and urea-N from the mesofauna and hypothesized that enchytraeids would release urea. ADMSW and CMSW were amended to pots with sandy loam and barley. The pots were divided into treatments with or without mesofauna. Mesofauna, plant N and biomass, soil N and ergosterol (fungal biomass) were measured over a 113-day period of four equidistant samplings. Soil respiration, N mineralization and N release by the mesofauna were modelled from concurrent studies. ADMSW- and CMSW-treated soils initially (<20 days) immobilized N. The amendments did not increase plant growth substantially, and this was probably due to N-limitation in the early stages of plant growth. Enchytraeid abundance was about three times higher in ADMSW- than CMSW-treated soils, indicating that ADMSW contained more labile compounds, bacteria, and microfauna. The mesofauna did not affect N-content, but the cumulated -N excreted by the mesofauna was estimated to be substantial and about one-fifth of total plant N in ADMSW. An explanation for this discrepancy might be that in the absence of the mesofauna, other members of the detrivore and microbivore community performed the mesofauna’s function. Neither enchytraeids nor microarthropods excreted urea.  相似文献   

15.
正The Center for Agricultural Resources Research(CARR),the Institute of Genetics and Developmental Biology(IGDB),Chinese Academy of Sciences,invites applicants for several research group leader positions.CARR is one of the research organizations in Chinese Academy of Sciences(CAS).We seek nominations and applications from individuals who have expertise and a record of accomplishment in research areas related to ecology,agro-hydrology,  相似文献   

16.
正The Center for Agricultural Resources Research(CARR),the Institute of Genetics and Developmental Biology(IGDB),Chinese Academy of Sciences,invites applicants for several research group leader positions.CARR is one of the research organizations in Chinese Academy of Sciences(CAS).We seek nominations and applications from individuals who have expertise and a record of accomplishment in research areas related to ecology,agro-hydrology,  相似文献   

17.
正The Center for Agricultural Resources Research(CARR),the Institute of Genetics and Developmental Biology(IGDB),Chinese Academy of Sciences,invites applicants for several research group leader positions.CARR is one of the research organizations in Chinese Academy of Sciences(CAS).We seek nominations and applications from individuals who have expertise and a record of accomplishment in research areas related to ecology,agro-hydrology,  相似文献   

18.
“Wicked” problems are those that are complex and that change when solutions are applied. Many conflicts in conservation fall into this category. The study approached the problem of how to constrain the apparent wickedness of a problem in the conservation management of a species by using simple empirical indicators to carry out iterative assessment of the risk to a population and to document how this risk evolves in relation to the addition of new data and the implementation of management actions. Effects of high levels of uncertainty within data and also concerning population structure were examined through stochastic simulation and by exploration of scenarios. Historical trends in the example used, the Steller sea lion, showed rapid declines in abundance in some regions during the 1980s. The current total population is 130,000-150,000 Steller sea lions through Alaska and British Columbia and this number has been stable since about 1990 in spite of regional differences in population dynamics. Regional differences in the sequence of changes in the number of pups and non-pups, suggested that an internal re-distribution of juveniles could have happened between 1980 and 1990. Current productivity also appears close to the long-term mean. Stochastic population projection using various scenarios showed that, based upon this history, the risk of extinction for the population has declined and is below reasonable thresholds for considering the population to be endangered. The trends in risk suggest that management actions taken since 1990 have probably been effective. Consequently, the conservation management objectives for the Steller sea lion are probably being met. The approach provides a mechanism, based upon experience and scenario analysis, for exploring future policy options and may help to constrain the debate amongst stakeholders about the cost-benefit trade-offs associated with different options.  相似文献   

19.
Nutrient distributions under no tillage (NT) compared with conventional disk-and-bed tillage (CT) management in the warm, humid region of the southeastern USA need to be assessed so that future placement, quantity, and type of fertilizers can be altered, if necessary, to efficiently match crop demands. We determined soil-profile distributions of pH, N, P, S, K, Ca, Mg, Na, Zn, Fe, Mn, and Cu to a depth of 0.9 m at the end of 8.5 years of continuous CT and NT management on a Weswood silty clay loam (fine, mixed, thermic Fluventic Ustochrept) in southcentral Texas. Most dramatic changes occurred within the 0–0.05 m depth, where soil under NT had lower pH, Fe, and Cu than under CT, but greater P, K, Zn, and Mn. Greater P and K under NT than under CT also occurred below the till-zone (0.15–0.3 m). At a depth of 0–0.3 m, soil under NT contained greater amounts of extractable P, K, Zn, Fe, Mn, and Cu than under CT. Nitrogen fertilization had little effect on nutrient distributions, except resulting in greater extractable K at 0–0.05 m and greater nitrate at 0–0.15 m. Few changes in soil-profile distributions were observed for extractable S, Ca, Mg, and Na. Long-term continuous use of NT on this fine-textured, high-fertility (except for N) soil had no apparent adverse effects on nutrient distributions relative to CT, but enhanced conservation and availability of P, K, Zn, Fe, Mn, and Cu near the soil surface where crop roots proliferate.  相似文献   

20.
The responses of tallgrass prairie plant communities and ecosystem processes to fire and grazing are well characterized. However, responses of invertebrate consumer groups, and particularly soil-dwelling organisms, to these disturbances are not well known. At Konza Prairie Biological Station, we sampled soil macroinvertebrates in 1994 and 1999 as part of a long-term experiment designed to examine the effects and interactions of annual fire, mowing, and fertilization (N and P) on prairie soil communities and processes. For nearly all taxa, in both years, responses were characterized by significant treatment interactions, but some general patterns were evident. Introduced European earthworms (Aporrectodea spp. and Octolasion spp.) were most abundant in plots where fire was excluded, and the proportion of the total earthworm community consisting of introduced earthworms was greater in unburned, unmowed, and fertilized plots. Nymphs of two Cicada genera were collected (Cicadetta spp. and Tibicen spp.). Cicadetta nymphs were more abundant in burned plots, but mowing reduced their abundance. Tibicen nymphs were collected almost exclusively from unburned plots. Treatment effects on herbivorous beetle larvae (Scarabaeidae, Elateridae, and Curculionidae) were variable, but nutrient additions (N or P) usually resulted in greater densities, whereas mowing usually resulted in lower densities. Our results suggest that departures from historical disturbance regimes (i.e. frequent fire and grazing) may render soils more susceptible to increased numbers of European earthworms, and that interactions between fire, aboveground biomass removal, and vegetation responses affect the structure and composition of invertebrate communities in tallgrass prairie soils.  相似文献   

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