共查询到19条相似文献，搜索用时 125 毫秒
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为探究ROP基因在棉花抵御逆境胁迫中的生物学功能,利用同源克隆的方法获得一个陆地棉GhROP6基因,通过生物信息学方法分析其理化性质、结构及进化关系,利用实时荧光定量PCR(qRTPCR)技术探究GhROP6基因的组织表达特异性及不同逆境胁迫和外源激素处理下的表达模式,构建GhROP6基因的VIGS载体并转化棉花,利用qRT-PCR技术检测其沉默效率。结果显示,GhROP6基因开放阅读框（ORF）为597 bp,编码一个含198个氨基酸的I类ROP蛋白;多重序列比对结果显示,GhROP6符合ROP蛋白结构特征,且与其他物种ROP蛋白高度同源;进化树分析结果显示GhROP6蛋白与拟南芥AtROP6蛋白同源性最高;GhROP6基因在棉花根、茎、真叶及子叶中均有表达,且在真叶中表达量最高;GhROP6基因对干旱、高盐、低温、高温等胁迫和外源脱落酸（ABA）、生长素（IAA）处理均有不同程度的响应,可能在棉花抗逆反应中扮演着重要角色。GhROP6在棉花的叶片和根部均得到有效沉默,表明已获得GhROP6基因沉默植株。本研究为进一步了解GhROP6基因的分子生物学功能奠定了基础。 相似文献
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利用PCR方法扩增凋亡蛋白融合基因片段，克隆至pMD18-T载体，鉴定和测序正确的凋亡蛋白融合基因亚克隆人带有绿色荧光蛋白的真核表达载体pECFP-C1的多克隆位点EcoRⅠ和SacⅡ之间，成功地构建带有报告基因的凋亡蛋白融合基因真核重组表达载体pECFP-C1-AFG。将带有报告基因的凋亡蛋白融合基因真核表达载体应用脂质体介导使其转染中国仓鼠卵巢细胞株（CHO），于转染48h后在荧光显微镜下观察到绿色荧光蛋白的出现，对转染的CHO细胞，应用Trizol法提取RNA，经RT-PCR鉴定，转染细胞在750bp处出现目的条带。收集转染细胞上清，应用间接ELISA及Western blot检测到目的蛋白的表达，说明凋亡蛋白融合基因构建正确。转染凋亡蛋白融合基因重组质粒的细胞，长成单层后传代，经G418（Genetiein）加压筛选（800μg／mL），3周后在荧光显微镜下观察所有细胞均出现绿色荧光，说明已建立了能够稳定表达凋亡蛋白融合基因与报告基因pECFP-C1的细胞株，为进一步研究凋亡蛋白融合蛋白在肿瘤组织中的定位及其诱导肿瘤细胞凋亡打下了基础。 相似文献
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具有多选择标记的植物基因表达载体有利于转基因植物研究中的转基因植株的筛选。本研究对植物基因表达载体pCAMBIA1301进行了改造,产生了1个具有可溶性的红移绿色荧光蛋白基因（smRS-GFP）、抗除草剂Basta、葡萄糖苷酸酶（GUS）及潮霉素（Hpt）的多选择标记的新的植物基因表达载体。运用这一表达载体的多选择标记可以有效降低检测和筛选转化植株时的假阳性率。此外,如此的基因表达载体也能满足实验室的不同筛选方法的需求。 相似文献
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绿色荧光蛋白与黄瓜花叶病毒运动蛋白融合基因的构建及在大肠杆菌中的表达 总被引：1，自引：0，他引：1
采用重组－PCR方法，扩增获得绿色荧光蛋白（GFP）与黄瓜花叶病毒运动蛋白（CMV MP）融合基因，将其克隆到pKEN上，构建了该融合基因的原核表达载体pKENG-M。SDS－PAGE及免疫印迹分析显示，57kD的融合蛋白基因在大肠杆菌DH5α中正确表达，激光共聚焦显微镜分析表明，在488nm光激发下，菌体呈亮绿色，表明融合蛋白保持绿色荧光特性。实验结果为进一步研究CMV MP的功能及其与胞间连丝和其他细胞成分的作用关系奠定了基础。 相似文献
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通用型植物表达载体pCamE的构建及功能验证 总被引：2，自引：0，他引：2
为了探索通用型植物表达载体的快捷构建,本研究通过PCR克隆、酶切连接和PCR扩增获得CaMV 35S启动子序列及pUC19多克隆位点部分限制性内切酶位点和NOS终止子,构建成完整的表达组件,并将该组件插入pCAMBIA1300的多克隆位点,构建成通用型植物表达载体pCamE(GenBank登录号:JX841315)。pCamE表达组件的启动子上游、多克隆位点和终止子下游分别含有4、8和3个单拷贝限制性内切酶位点,利于外源基因的克隆和插入以及对载体启动子或终止子的修饰和更换。以绿色荧光蛋白基因(enhanced green fluorescent protein,eGFP)为报告基因,分别构建了表达载体pCam::GFP、pCam::SalGFP和pCam::DAHPS-GFP,转化洋葱(Allium cepa L.)表皮细胞和拟南芥(Arabidopsis thaliana L.),绿色荧光蛋白(GFP)和融合蛋白DAHPS-GFP在洋葱表皮细胞中均成功稳定表达;在pCam::GFP转基因拟南芥株系的根、根尖、根毛、茎表皮毛、幼叶基部、叶柄微管组织和未成熟花药等组织均能观察到强烈的绿色荧光。表明,植物表达载体pCamE是可将不同目的基因与植物染色体整合并稳定遗传的通用型植物表达载体,不受外源片段大小及插入位点的影响,具有经济实用、适用广泛、操作简便和外源基因在转基因植物中遗传稳定且正常表达等特点。该载体可用于不同基因和不同功能表达载体的快捷构建,为植物的遗传转化和基因功能研究提供了新的通用型基因表达载体工具。 相似文献
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将从胡杨中克隆的果糖-1,6-二磷酸醛缩酶基因（PeALD）构建到pGEX-4T—1载体上,经IPTG诱导成功获得融合蛋白GST：PeALD,并将蛋白进行纯化,其大小约为52kD,转化大肠杆菌的耐盐性实验表明,PeALD基因的成功表达有利于提高大肠杆菌菌株的耐盐性。为研究该基因编码蛋白在植物细胞中的定位,将基因的ORF区构建到定位表达载体pMDC85上,通过PEG介导的拟南芥瞬时转化法观察融合蛋白PeALD：GFP在细胞中的定位情况,结果显示该蛋白定位于胞质中,由此说明实验克隆得到的该胡杨果糖-1,6-二磷酸醛缩酶基因编码的是胞质蛋白。烟草种子在盐培养基上的萌发实验结果显示转基因烟草具有更高的耐盐性;烟草水培苗经200mmol／LNaCl处理一周后,可溶性糖的质谱检测结果显示转基因植株中葡萄糖的含量有很大提高。表明胡杨果糖-1,6-二磷酸醛缩酶通过促进糖酵解和有氧呼吸途径来提高植物对盐胁迫的适应性。 相似文献
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E8启动子是番茄果实成熟乙烯响应性基因E8的启动子区。这个区域中存在着一些响应乙烯以及在果实发育和成熟过程中起调节作用的顺式作用元件，调节E8基因在果实成熟过程中表达（Deikmaa et al．，1998）。本研究根据GenBank中E8基因的启动子区序列设计引物，从番茄基因组中克隆了该启动子，构建了番茄果实特异表达载体，并通过RT-PCR和gus基因检测了该表达载体的果实特异表达活性。 相似文献
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植物特有的NAC转录因子在植物的生长发育、胁迫应答和激素调节等方面具有重要功能。本研究从橡胶树胶乳cDNA中克隆了HbNAC33基因,生物信息学分析显示该基因的完整开放阅读框（ORF）为1 092 bp,编码363个氨基酸。HbNAC33蛋白的N-端第3～156位氨基酸之间含有一个典型的NAC结构域,酵母试验表明,HbNAC33具有转录激活活性,转录激活区在C-端,具备了NAC转录因子的基本特征。序列比对和系统进化分析表明HbNAC33蛋白属于NAC转录因子家族中的ANAC011亚族。实时荧光定量PCR分析表明,HbNAC33在叶中的表达量显著高于其他部位;割胶,乙烯利（ET）和茉莉酸（JA）处理均能诱导HbNAC33的表达上调。以上结果表明,HbNAC33可能参与植物的生长发育和胁迫应答过程。 相似文献
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磷酸蔗糖磷酸酶（SPP）是植物蔗糖生物合成最后一步催化反应的关键酶，对调控植物生长发育不同时期的蔗糖合成过程有重要作用。为探索金柑（Fortunella crassifiolia Swingle）SPP基因及其编码蛋白的序列特征及其在果实发育过程的表达特性，本研究以四倍体品种脆蜜金柑（F. crassifiolia Swingle cv. Cuimi）为试验材料，采用反转录PCR和cDNA末端快速克隆（RACE）方法从金柑果肉中克隆SPP基因，对其进行生物信息学和原核表达分析，并检测果实发育不同时期该基因在果肉中的表达量。结果表明，FcSPP基因的cDNA序列全长1 638 bp，最长开放阅读框（ORF）为1 191 bp，编码396个氨基酸，理论等电点为6.57，为稳定的亲水性蛋白，二级结构主要由α-螺旋和无规则卷曲组成。FcSPP蛋白含有S6PP和S6PP＿C保守结构域，属于植物磷酸蔗糖磷酸酶家族成员。系统进化分析结果显示，FcSPP蛋白与甜橙SPP同源性最高，在进化树上与拟南芥等双子叶植物SPP划归为一类。此外，本研究构建了FcSPP基因的原核表达载体，诱导表达获得纯化的FcSP... 相似文献
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水稻耐低锌基因型的生长发育和若干生理特性研究 总被引：11，自引：1，他引：10
在不同Zn2+活度（pZn2+9.7,pZn2+11.0和pZn2+＞11.5）的溶液培养条件下,研究了水稻耐低锌基因型的生长发育和若干生理特性。结果表明:水稻锌营养存在明显的基因型差异,降低锌离子活度会增加地下部于物质的积累,当Zn2+活度从pZn2+9.7下降到pZn2+11.0时,耐低锌品种的地上部干重虽下降,但因地下部干重显著增加,故总干重相近;锌敏感品种则地上部干重显著下降,而地下部干重增加不明显,总干重显著下降。当严重缺锌（pZn2+＞11.5）时,所有基因型水稻的干重构极显著地下降,但锌敏感品种比耐低锌品种下降得更多。降低Zn2+活度使水稻秧苗的出叶速度减慢,在极度缺锌条件下,敏感品种只能生长到3.5叶,而耐低锌品种能生长到4.5叶左右。对叶绿素和根系氧化力的测定结果表明,轻度缺锌或缺锌初期会使叶绿素含量上升和根系氧化力下降,但严重缺锌时,则使叶绿素含量显著降低,而使根系氧化力明显增加。锌敏感品种比耐缺锌品种的变化更为明显。锌离子活度对秧苗的含水量也有明显的影响。因此,耐低锌基因型在低Zn2+活度条件通过保持较低的根氧化作用,促进根系生长以维持地上部新叶生长,达到低锌适应稳态。 相似文献
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McGorum BC Fry SC Wallace G Coenen K Robb J Williamson G Aruoma OI 《Journal of agricultural and food chemistry》2000,48(6):2346-2352
To investigate the etiology of equine dysautonomia (ED), a degenerative polyneuropathy affecting grazing horses, the biochemical composition and antioxidant/prooxidant activities of aqueous extracts of plants collected from ED pastures were determined. Plants collected immediately after an outbreak of ED had reduced antioxidant and weak prooxidant activities when compared with control plants (plants collected from ED pastures out of ED season and control plants from ED pastures that were grown under favorable conditions). ED plants also had significantly increased concentrations of fructose and low molecular weight phenolic compounds, significantly more of one amino acid zone (probably valine), significantly less tartaric acid, and a nonsignificant decrease in ascorbic acid content when compared with control plants from ED pastures that were grown under favorable conditions. These findings suggest that ED plants may be under oxidative stress, possibly due to chilling, drought, or fungal colonization. However, experimental drought and chilling of plants did not reproduce the biochemical alterations identified in ED plants. It is possible that the altered biochemical content of ingested plants may contribute, directly or indirectly, to the development of ED in grazing horses. 相似文献
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Mycorrhizal and non-mycorrhizal onions were grown in pots containing soil at two P concentrations. Following 35S injection into the soil, both mycorrhiza) and non-mycorrhizal plants from high P treatments had significantly higher 35S concentrations in roots compared to non-mycorrhizal, low P controls. Mycorrhizal, low P plants had higher concentrations of 35S in shoots than did non-mycorrhizal, low P plants. In a second experiment detached non-mycorrhizal onion roots from plants given a nutrient solution containing P for 26 days before short-term uptake experiments absorbed at greater rates from solution than roots from plants given a complete minus-P nutrient solution. This occurred at all three concentrations of S tested. 1 mM. 10μM, and 0.1 μM. Increased S uptake by mycorrhizal plants can result from increased S absorbing power of roots with enhanced P status. 相似文献
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Metabolic response of Medicago sativa to severe nutrient imbalances and disturbances under field conditions 
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下载免费PDF全文 Our experiments were focused on the metabolic footprint of mineral‐nutrient availability under field conditions. While there are multiple factors potentially blurring such footprints, we hypothesized that physiological and metabolic adaptations of established plants are particularly important mechanisms under field conditions. To study respective differences between young and established plants and to study the impact of disturbances on the adaptive capacity of established plants, we analyzed Medicago sativa plants of different age from plots with marked differences in the levels of soil mineral nutrients established in a long‐term fertilization experiment. Gas chromatography–mass spectrometry was used to determine metabolite profiles of sink and source leaves of plants in an early state of development (“young plants”), just before the first harvest (“established plants”), and a short time after the second harvest (“regrowing plants”). Metabolite profiles from young plants were markedly responsive to soil mineral nutrients and resembled respective profiles from controlled conditions, demonstrating that overall variability of growth and sampling conditions had relatively little importance for the metabolite profiles recorded. In the case of established plants, however, we observed only little impact of availability of mineral nutrients on metabolite profiles. This low metabolic responsiveness of plants was partially lost after severe disturbances (removal of the plant shoot). Metabolite profiling, in summary, is able to detect a metabolic footprint of mineral‐nutrient availability in young plants under field conditions and may provide information about the ability of older plants to partially uncouple their metabolism from the environment. In addition, it is also possible to determine the impact of disturbances on this ability of the plant organism. 相似文献
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Toshio Sugimoto Ryoichi Masuda Makoto Kito Naomasa Shiraishi Yoshikiyo Oji 《Soil Science and Plant Nutrition》2013,59(2):273-279
We compared the concentrations and contents of protein and oil in mature seeds from nodulated and non-nodulated soybean plants grown on soils with four different N levels during maturation. We observed a positive correlation between the contents of protein and oil in seeds from nodulated plants. Seeds from nodulated plants grown on urea-treated soil showed higher protein and lower oil contents than those from plants grown on soil treated with coated slow release N fertilizer (LP-100). Contents of these compounds in seeds from nodulated plants grown on LP-100 soil were almost the same as those from non-nodulated plants on the same soil. These observations indicated that N economy in roots during seed maturation affects the contents of storage compounds. We suggested that the control of the N2 fixation activity of soybean plants and management of soil N level during seed maturation are important to determine the contents of protein and oil in seeds. 相似文献
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Terry J. Rose Juan Pariasca‐Tanaka Michael T. Rose Asako Mori Matthias Wissuwa 《植物养料与土壤学杂志》2012,175(6):799-804
Interest in developing crop varieties with low grain phosphorus (P) in order to minimize the removal of P from fields in harvested grain has been limited due to the view that a low‐P grain trait may impair subsequent seedling vigor. This perception is based on relatively few studies, which typically investigated seedling growth on infertile soils, and used seed that may have differed in attributes other than P concentration. To investigate whether these anomalies cast sufficient doubt to warrant renewed research in this field, we compared the growth of rice seedlings from seed low in P obtained from P‐starved plants (P‐starved seed) vs. high‐P seeds (obtained from P‐fertilized plants) in P‐deficient and P‐replete soils. While plants from high‐P seed were superior in the P‐deficient soil, plants grown from P‐starved seed overcame an initial lull in early vigor to obtain similar biomass at maturity as plants grown from high‐P seed. Subsequent experiments were undertaken using high‐P seed vs. seed low in P from a range of rice genotypes that was not obtained from P‐stressed plants (low‐P seed): There was no reduction in seedling vigor or biomass and grain yields at maturity of plants from low‐P seeds in low‐P soil compared to plants from high‐P seed, though responses were genotype‐specific. The results suggest that multiple factors can confound the results of seed P × seedling vigor studies, and that a renewed research effort to define the minimum P levels in seeds required for adequate seedling growth across a range of environments is warranted. 相似文献
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河西走廊东部不同类型植物物候对气候变化的响应 总被引：3，自引：0，他引：3
基于甘肃省武威市气象局1980-2004年对多年生植物旱柳和中国槐物候期、1981-2008年对一年生植物春小麦和玉米发育期的观测资料,运用线性倾向估计法、膨化相关、SPSS统计软件进行逐步回归和方差分析,建立了关键物候期的气候预测模型。结果表明：（1）在分析期内,多年生木本植物相同物候期最晚与最早出现日期相差达20～38d,一年生草本植物达11～30d,反映出物候期对气象条件年际变化非常敏感。（2）分析期内多年生植物春季多数物候期提前（线性倾向率-2.008～-3.246d/10a,P〈0.05）,秋季物候期推迟（线性倾向率6.631～7.108d/10a,P〈0.01）。一年生植物的春、夏季物候期均有提前的趋势（线性倾向率-1.494～-4.122d/10a,P〈0.05）,秋季物候期推后不显著。（3）从各物候间隔期变化看,多年生植物绿叶期延长速率为1.046～7.738d/10a,营养生长期延长速率为0.877～8.454d/10a,干物质积累期延长速率为4.392～7.738d/10a,秋季生长期延长速率为0.477～3.015 d/10a。一年生植物春季营养生长期缩短速率为2.17～3.41d/10a,秋季生殖生长期延长速率为3.322d/10a。（4）对多年生植物而言,平均气温、平均最低气温、平均最高气温和20cm、80cm、160cm地温是影响春、夏、秋季物候期的主要气象因子,温度越高春季物候期越提前,夏、秋季物候期越推迟。对一年生植物而言,平均气温、平均最低气温、平均最高气温和20cm、80cm、160cm地温也是主要影响因子,气候变暖,春、夏、秋季物候期均提前。 相似文献
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In a greenhouse pot experiment, maize was grown inoculated with the spores of the VAM fungi Glomus mosseae and Glomas multicaulis or non‐inoculated. Low soluble ferrous phosphate (FePO4.4H2O) was added to the mycorrhized and non‐mycorrhized maize. The fresh and dry weights of mycorrhized plants with added phosphate (P) were higher than in mycorrhized plants without added P or non‐mycorrhized plants with added P. The amount of P in the soil samples from pots with mycorrhizal plants fertilized with P was evidently smaller than those in samples also fertilized non‐mycorrhizal plants. The percentage of P was higher in tissues of fertilized mycorrhizal plants than in those mycorrhized plants without or non‐mycorrhized plants with added low‐ soluble P. These results indicated that plants in VAM symbiosis mobilize P better from low‐soluble P than non‐mycorrhized plants. 相似文献