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1.
人溶菌酶基因在烟草中的表达 总被引：4，自引：0，他引：4
根据植物偏爱的密码子，改变G＋C总量和密码子第三位碱基的G＋C含量，同时避免在真核表达中影响转录，翻译及mRNA稳定性的序列如AATTAAA和ATTTA，在原氨基酸序列不变的基础上，设计并合成了适合在植物中中高效表达的人溶菌酶(human lysozyme,简单HL）基因。构建了含HL基因的植物表达载体。用根癌农杆菌（Agrobacterium tumefaciens)介导法将其转化烟草（Nicotiana tabacum).PCR和Western检测表明，HL基因在转基因烟草中得到整合和表达，提高了转基因烟草离体叶对野火病（Pseudomonas syringaepv.tabaci)的抗性。 相似文献
2.
Nhap基因编码一种质膜Na /H 逆向转运蛋白,在耐盐方面具有重要的研究价值。实验利用根癌农杆菌(Agrobac-teriumtumefacience)介导的叶盘法将Nhap基因转入烟草(Nicotianatobacum),经PCR和RT-PCR检测已获得转Nhap基因烟草的再生植株。同时从发芽率、存活率、叶绿素含量、Na 含量及耐盐性表现进行了F1代转基因烟草的耐盐性检测。结果表明,转基因烟草的发芽率、存活率、叶绿素含量均较对照高,Na 含量较对照有所下降,并且转基因烟草在200mmol/LNaCl处理下,仍能正常存活、生长,而对照烟草的生长却受到抑制。即转基因F1代烟草的耐盐性得到了显著的提高。 相似文献
3.
拟南芥液泡膜Na+/H+逆向转运蛋白基因(AtNHXl)在植物耐盐机制中有重要作用.为了鉴定AtNHX1基因功能和创制耐盐烟草新种质,本研究用农杆菌介导法将rd29A启动子驱动的AtNHX1基因导入烟草(Nicotiana tabacum L.),获得72株卡那霉素抗性再生植株,17株在MS+85.5 mmol/L NaC1培养基上生长良好.对其进行PCR、Southern杂交和Northern杂交检测,证实AtNHX1基因已整合到烟草基因组中,并进行正常转录,且其转录受盐逆境的诱导.在含盐(85.5、171、256和342 mmol/L NaCl)培养基上进行对照植株和转基因植株生长及叶片丙二醛含量的分析,结果显示,转基因株系的生长势和根发生能力明显优于对照,且叶片丙二醛含量显著低于对照,表明rd29A启动子驱动AtNHX1基因的导入和表达可显著提高转基因烟草植株的耐盐性.结果提示rd29A启动子驱动的AtNHX1基因表达单元可在异源植物中正常转录和表达,可用于植物耐盐基因工程育种. 相似文献
4.
高粱SbSKIP基因的克隆及其在烟草中的抗旱功能分析 总被引：2，自引：0，他引：2
SKIP(ski-interacting protein)是一种RNA代谢相关蛋白,在植物抵抗非生物胁迫伤害中起着很重要的调节作用,本研究克隆了高粱(Sorghum bicolor)SbSKIP基因,cDNA编码区全长1 485 bp,编码494个氨基酸;进化树分析表明,高粱SbSKIP基因编码的氨基酸序列与节节麦(Aegilops tauschii)相似性为52％;定量PCR分析表明,在模拟干旱条件下SbSKIP基因表达量上调,在干旱处理16h后,表达量显著增加了2倍以上.构建植物表达载体pSH-SbSKIP,利用农杆菌(Agrobacterium tumefaciens)介导的叶盘法遗传转化烟草(Nicotiana tabacum),获得25个转基因植株.选取3个不同株系TP4、TP9和TP13,在不同浓度甘露醇模拟干旱条件下对种子发芽率和幼苗期进行抗旱性分析,结果发现,在300 mmol/L甘露醇处理10d,种子的发芽率比野生型高60％以上,而且从苗期根的生长发现,野生型根的生长明显受到抑制.在自然条件下生长的烟草幼苗,用20％ PEG 6000处理14d,结果显示,转基因烟草正常生长,而野生型烟草大部分叶片黄化甚至出现萎蔫,其中转基因植株超氧化物歧化酶(super-oxide dismutase,SOD)的平均活性值比野生型高91.2％,H2O2和丙二醛(malondialdehyde,MDA)含量分别比野生型降低了42％和32.7％,可溶性糖(soluble sugar)含量达野生型的1.1倍.结果表明,SbSKIP基因表达提高了烟草植株的干旱耐受能力.本研究为进一步研究SbSKIP基因功能提供依据,同时该基因的克隆和功能分析可为创制转基因抗旱材料提供基础资料. 相似文献
5.
种子败育的无籽瓜果具有较强的市场竞争力和可观的经济价值.来源于解淀粉芽胞杆菌(Bacillus A myloliquefaciens)的Barnase基因编码12kD的胞外小分子核糖核酸酶(RNAase).在缺乏其天然抑制物Barstar的情况下,单独表达Barnase通常会造成宿主细胞的死亡.本研究用油菜(Brassica napus)种子特异启动子Napin调控Barnase基因,构建T-DNA表达载体,并用叶盘转化法转化烟草(Nicotiana benthamiana),获得55株转基因植株.随机抽取10株对其花器官进行形态学分析,并比较转基因植株和非转基因植株在不同发育时期花器官的形态学变化,发现转基因植株和非转基因植株在授粉前的花器官形态没有明显差异,但是受精以后转基因植株花瓣逐渐枯萎,不能正常发育形成种子,而非转基因植株能够正常形成种子.为了进一步了解转基因烟草种子败育的原因,将转基因植株和非转基因植株正反杂交,均不能正常结籽,表明转基因烟草的种子败育不是由自交不亲和性引起的.将转基因植株和非转基因植株花粉分别使用亚历山大染液染色,显微观察花粉的大小、形状和色泽,发现转基因花粉和非转基因花粉无明显差别,转基因植株和非转基因植株均能产生具有正常授粉活性的花粉.以上研究表明,使用种子特异性启动子驱动Barnase基因表达,外源基因不会通过花粉渗漏表达.本研究成功获得了Barnase基因在种子中特异表达的无籽转基因烟草,该研究为无籽植物的研制提供了一种全新的模式和思路. 相似文献
6.
向日葵种子特异性启动子Ha ds10G1的克隆及其功能验证 总被引：3，自引：0，他引：3
利用PCR技术从向日葵基因组DNA中克隆了Lea蛋白基因家族中Ha ds10 G1基因上游1414bp的调控序列，序列分析表明-1—1414bp与报道序列同源性为100%，将其与GUS基因融合构建植物表达载体后，通过农杆菌介导法转化烟草NC89，PCR扩增初步证明目的片段已整合到烟草基因组中，转基因植株的GUS活性检测表明，在茎、叶中无GUS活性，GUS活性只存在于种子中，因此，Ha ds10 G1启动子具有种子特异性的功能。 相似文献
7.
赤霉素和细胞激动素对白刺种子萌发的调控研究 总被引：2，自引：0，他引：2
为探讨白刺种子萌发特性,采用不同浓度赤霉素和细胞激动素处理方法,测定了白刺种子生活力、吸水率、发芽率、发芽势、发芽指数和萌发指数.结果表明:浸种最初的2 h是白刺种子的迅速吸水期,整个吸水过程呈快--慢--缓慢规律.赤霉素和细胞激动素处理对白刺种子萌发有明显调控作用.150 mg·kg-1的赤霉素处理24 h,种子发芽势、发芽率、发芽指数、种子生活力最高,300 mg·kg-1赤霉素处理的发芽势,发芽率、发芽指数最低.6 mg·kg-1细胞激动素处理白刺种子1 h,其发芽势、发芽率,发芽指数、种子生活力均最高,而10 mg·kg-1细胞激动素处理白刺种子发芽率、发芽势、发芽指数、活力指数均最低. 相似文献
8.
抗青枯病转群体感应猝灭基因烟草的培育 总被引：3，自引：2，他引：1
利用具有猝灭青枯菌群体感应信号分子功能的aac基因构建高效植物表达载体,通过农杆菌介导的方法转化烟草,获得了卡那霉素抗性植株57株。经PCR及RT-PCR检测,筛选得到30株阳性转基因再生苗,并证实了目的基因已经整合到烟草基因组中,且在转录水平上得到表达。转基因植株苗期抗青枯病试验表明,16天后转基因烟草较对照植株病情指数降低了51,相对病情指数为56.5％。 相似文献
9.
以玉米黄早四幼苗叶片为材料,通过RT-PCR扩增后获得玉米磷酸丙糖转移蛋白(Triose phosphate translocator．TPT)cDNA 全长片段,对其进行序列分析。结果表明：分离的目的片段核苷酸序列与报道的相比同源率为99.9％,只有1个碱基发生改变。将得到的玉米 cDNA与CaMV35S启动子和NOS终止子融合,构建了正义表达载体,通过根癌土壤杆菌介导转化烟草,转基因植株经PCR和Southern杂交检测,证明TPT基因已整合到烟草基因组上 相似文献
10.
利用PCR方法从油菜(Brassica napus)基因组中扩增得到花瓣特异性启动子XY355,构建该启动子与玉米(Zea mays)花青素调节基因Lc的植物表达载体pXY60。通过根癌农杆菌(Agrobacterium tumefaciens)介导,将pXY60分别转化烟草(Nicotiana tabacum)和矮牵牛(Petunia hybrida)。对再生植株进行PCR检测,结果表明,外源基因已经整合到转基因烟草和矮牵牛的基因组中。观察发现,转基因植株的表型发生了明显变化:转基因烟草的花色由浅红变成了红色,转基因矮牵牛的花色由白色变成了浅紫色。 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.
Aleš Lebeda Ivana Doležalová Eva Křístková Barbora Mieslerová 《Genetic Resources and Crop Evolution》2001,48(2):153-164
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.
Kyle Devey Michael Mucalo Gordon Rajendram Joseph Lane 《Communications in Soil Science and Plant Analysis》2015,46(4):72-80
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.
Thomas Larsen Jesper Luxhøi Jakob Magid Lars Stoumann Jensen Paul Henning Krogh 《Biology and Fertility of Soils》2007,44(1):59-68
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.
《中国生态农业学报》2014,(6)
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.
《中国生态农业学报》2014,(7)
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.
《中国生态农业学报》2014,(8)
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.
M.A. Callaham Jr. J.M. BlairT.C. Todd D.J. KitchenM.R. Whiles 《Soil biology & biochemistry》2003,35(8):1079-1093
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
19.
I.L. Boyd 《Biological conservation》2010,143(7):1664-1674
“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
20.
Soil-profile distribution of primary and secondary plant-available nutrients under conventional and no tillage 总被引：1，自引：0，他引：1
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