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The Application of Nicotiana benthamiana as a Transient Expression Host to Clone the Coding Sequences of Plant Genes
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作者 Jianzhong Huang Peng Jia +3 位作者 Xiaoju Zhong Xiuying Guan Hongbin Zhang Honglei Ruan 《American Journal of Molecular Biology》 CAS 2024年第2期54-65,共12页
Coding sequences (CDS) are commonly used for transient gene expression, in yeast two-hybrid screening, to verify protein interactions and in prokaryotic gene expression studies. CDS are most commonly obtained using co... Coding sequences (CDS) are commonly used for transient gene expression, in yeast two-hybrid screening, to verify protein interactions and in prokaryotic gene expression studies. CDS are most commonly obtained using complementary DNA (cDNA) derived from messenger RNA (mRNA) extracted from plant tissues and generated by reverse transcription. However, some CDS are difficult to acquire through this process as they are expressed at extremely low levels or have specific spatial and/or temporal expression patterns in vivo. These challenges require the development of alternative CDS cloning technologies. In this study, we found that the genomic intron-containing gene coding sequences (gDNA) from Arabidopsis thaliana, Oryza sativa, Brassica napus, and Glycine max can be correctly transcribed and spliced into mRNA in Nicotiana benthamiana. In contrast, gDNAs from Triticum aestivum and Sorghum bicolor did not function correctly. In transient expression experiments, the target DNA sequence is driven by a constitutive promoter. Theoretically, a sufficient amount of mRNA can be extracted from the N. benthamiana leaves, making it conducive to the cloning of CDS target genes. Our data demonstrate that N. benthamiana can be used as an effective host for the cloning CDS of plant genes. 展开更多
关键词 Coding Sequence Genomic Sequence nicotiana benthamiana Plant Genes
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Structural and Functional Insights into an Arabidopsis NBS-LRR Receptor in Nicotiana benthamiana
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作者 Jianzhong Huang Xiuying Guan +3 位作者 Xiaoju Zhong Peng Jia Hongbin Zhang Honglei Ruan 《American Journal of Molecular Biology》 CAS 2024年第2期84-96,共13页
Nucleotide-binding site leucine-rich repeat receptors (NBS-LRR/NLRs) are crucial intracellular immune proteins in plants. Previous article reported a novel NLR protein SUT1 (SUPPRESSORS OF TOPP4-1, 1), which is involv... Nucleotide-binding site leucine-rich repeat receptors (NBS-LRR/NLRs) are crucial intracellular immune proteins in plants. Previous article reported a novel NLR protein SUT1 (SUPPRESSORS OF TOPP4-1, 1), which is involved in autoimmunity initiated by type one protein phosphatase 4 mutation (topp4-1) in Arabidopsis, however, its role in planta is still unclear. This study employed Nicotiana benthamiana, a model platform, to conduct an overall structural and functional analysis of SUT1 protein. The transient expression results revealed that SUT1 is a typical CNL (CC-NBS-LRR) receptor, both fluorescence data and biochemical results showed the protein is mainly anchored on the plasma membrane due to its N-terminal acylation site. Further truncation experiments announced that its CC (coiled-coil) domain possessed cell-death-inducing activity. The outcomes of point mutations analysis revealed that not only the CC domain, but also the full-length SUT1 protein, whose function and subcellular localization are influenced by highly conserved hydrophobic residues. These research outcomes provided favorable clues for elucidating the activation mechanism of SUT1. 展开更多
关键词 CC-NBS-lRR Hypersensitive Response nicotiana benthamiana Plasma Membrane localization
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缺血性脑卒中铁死亡特征基因NFE2L2的鉴定与验证
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作者 王咪 马书杰 +1 位作者 刘杨 齐瑞 《中国组织工程研究》 CAS 北大核心 2025年第7期1466-1474,共9页
背景:铁死亡与缺血性脑卒中的发病密切相关,靶向铁死亡是一种治疗缺血性脑卒中有前景的方案,但具体调控靶点尚不明确。目的:通过生物信息学和机器学习方法筛选缺血性脑卒中铁死亡相关特征基因,并通过细胞实验进行验证,探讨铁死亡在缺血... 背景:铁死亡与缺血性脑卒中的发病密切相关,靶向铁死亡是一种治疗缺血性脑卒中有前景的方案,但具体调控靶点尚不明确。目的:通过生物信息学和机器学习方法筛选缺血性脑卒中铁死亡相关特征基因,并通过细胞实验进行验证,探讨铁死亡在缺血性脑卒中的作用。方法:基于GEO数据库和FerrDb数据库选取符合条件的缺血性脑卒中相关数据集和铁死亡表达数据集,通过t检验筛选铁死亡相关差异基因。对铁死亡相关差异基因进行GO功能富集分析与KEGG信号通路富集分析。通过PPI网络分析和机器学习筛选缺血性脑卒中铁死亡的特征基因,利用ROC分析和GSEA分析探究特征基因的准确性和生物功能。然后进行细胞实验,将HT22细胞分为对照组与缺血性脑卒中组,对照组不作任何干预,缺血性脑卒中组加入0.1 mol/L的H_(2)O_(2)干预24 h诱导细胞氧化应激和铁死亡,通过实时荧光定量RT-PCR和Western Blot验证铁死亡的发生和特征基因表达。结果与结论:(1)共获取45个铁死亡相关差异基因,GO和KEGG富集分析发现差异基因与氧化应激、自噬、铁死亡、脂肪细胞因子信号通路和线粒体代谢密切相关。(2)通过PPI网络中的MCODE插件和cytoHubba插件与机器学习中的LASSO算法和SVM-RFE算法共鉴定出1个铁死亡特征基因核因子E2相关因子2(nuclear factor erythroid 2-related factor 2,NFE2L2)。(3)对NFE2L2进行ROC曲线分析,发现在训练集和验证集中构建的诊断预测模型具有良好的准确性与特异性;对NFE2L2进行GSEA分析,发现特征基因通过免疫、炎症反应、氨基酸代谢及神经因子调控等方面参与缺血性脑卒中发病机制的调控。(4)细胞实验的RT-PCR和Western Blot分析表明,与对照组对比,缺血性脑卒中组中的酰基辅酶A合成酶长链家族成员4 mRNA和蛋白表达水平显著增高(P<0.05),谷胱甘肽过氧化物酶4 mRNA和蛋白表达水平显著降低(P<0.05);与对照组对比,缺血性脑卒中组中特征基因NFE2L2 mRNA和蛋白表达水平显著增高(P<0.05)。(5)上述结果证实,缺血性脑卒中与铁死亡密切相关,靶向特征基因NFE2L2可以为研究和治疗缺血性脑卒中提供一定的思路与方向。 展开更多
关键词 缺血性脑卒中 铁死亡 生物信息学 HT22细胞 机器学习 特征基因 细胞实验 NFE2l2 ASCl4 GPX4
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Effects of P301L-TAU on post-translational modifications of microtubules in human iPSC-derived cortical neurons and TAU transgenic mice
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作者 Mohamed Aghyad Al Kabbani Christoph Köhler Hans Zempel 《Neural Regeneration Research》 SCIE CAS 2025年第8期2348-2360,共13页
TAU is a microtubule-associated protein that promotes microtubule assembly and stability in the axon.TAU is missorted and aggregated in an array of diseases known as tauopathies.Microtubules are essential for neuronal... TAU is a microtubule-associated protein that promotes microtubule assembly and stability in the axon.TAU is missorted and aggregated in an array of diseases known as tauopathies.Microtubules are essential for neuronal function and regulated via a complex set of post-translational modifications,changes of which affect microtubule stability and dynamics,microtubule interaction with other proteins and cellular structures,and mediate recruitment of microtubule-severing enzymes.As impairment of microtubule dynamics causes neuronal dysfunction,we hypothesize cognitive impairment in human disease to be impacted by impairment of microtubule dynamics.We therefore aimed to study the effects of a disease-causing mutation of TAU(P301L)on the levels and localization of microtubule post-translational modifications indicative of microtubule stability and dynamics,to assess whether P301L-TAU causes stability-changing modifications to microtubules.To investigate TAU localization,phosphorylation,and effects on tubulin post-translational modifications,we expressed wild-type or P301L-TAU in human MAPT-KO induced pluripotent stem cell-derived neurons(i Neurons)and studied TAU in neurons in the hippocampus of mice transgenic for human P301L-TAU(p R5 mice).Human neurons expressing the longest TAU isoform(2N4R)with the P301L mutation showed increased TAU phosphorylation at the AT8,but not the p-Ser-262 epitope,and increased polyglutamylation and acetylation of microtubules compared with endogenous TAU-expressing neurons.P301L-TAU showed pronounced somatodendritic presence,but also successful axonal enrichment and a similar axodendritic distribution comparable to exogenously expressed 2N4R-wildtype-TAU.P301L-TAU-expressing hippocampal neurons in transgenic mice showed prominent missorting and tauopathy-typical AT8-phosphorylation of TAU and increased polyglutamylation,but reduced acetylation,of microtubules compared with non-transgenic littermates.In sum,P301L-TAU results in changes in microtubule PTMs,suggestive of impairment of microtubule stability.This is accompanied by missorting and aggregation of TAU in mice but not in i Neurons.Microtubule PTMs/impairment may be of key importance in tauopathies. 展开更多
关键词 human induced pluripotent stem cell MICROTUBUlES P301l pR5 mice TAU TAUOPATHY tubulin code
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载富血小板血浆水凝胶对L929细胞氧化损伤的保护作用
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作者 王自林 牟秋菊 +2 位作者 刘宏杰 申玉雪 祝丽丽 《中国组织工程研究》 CAS 北大核心 2025年第4期771-779,共9页
背景:在慢性创面愈合过程中,活性氧的过量生成可能会损害L929成纤维细胞的功能,从而延缓创面修复,因此保护成纤维细胞免受氧化应激的影响对于促进创面愈合非常重要。目的:评估羧甲基壳聚糖-氧化硫酸软骨素/富血小板血浆(CMC-OCS/PRP)水... 背景:在慢性创面愈合过程中,活性氧的过量生成可能会损害L929成纤维细胞的功能,从而延缓创面修复,因此保护成纤维细胞免受氧化应激的影响对于促进创面愈合非常重要。目的:评估羧甲基壳聚糖-氧化硫酸软骨素/富血小板血浆(CMC-OCS/PRP)水凝胶对H2O2刺激下L929细胞的保护作用。方法:制备CMC-OCS/PRP水凝胶,表征水凝胶的微观形貌、降解性能、清除H2O2与羟基自由基的能力及生物相容性。取生长状态良好的L929细胞,分5组培养:对照组常规培养,H2O2组加入H2O2,CMC-OCS组加入羧甲基壳聚糖-氧化硫酸软骨素水凝胶浸提液+H2O2,PRP组加入富血小板血浆凝胶浸提液+H2O2,CMC-OCS/PRP组加入羧甲基壳聚糖-氧化硫酸软骨素/富血小板血浆水凝胶浸提液处理+H2O2,每组先加入水凝胶浸提液处理6 h,然后再加入H2O2处理24 h。培养结束后,检测细胞活性氧及丙二醛水平,细胞凋亡、胶原纤维Ⅰ蛋白表达。在加入H2O2的情况下,将上述水凝胶浸提液分别与L929成纤维细胞直接或间接共培养36 h,通过划痕实验、Transwell小室实验检测细胞的迁移能力。结果与结论:①CMC-OCS/PRP水凝胶具有均匀相互关联的多孔结构及良好的降解能力,体外可有效清除H2O2与羟基自由基,并且具有良好的生物相容性;②与对照组比较,H2O2组细胞凋亡率、细胞活性氧与丙二醛水平升高(P<0.05),细胞铺展面积减少(P<0.05),胶原纤维Ⅰ蛋白表达无明显变化(P>0.05);与H2O2组比较,CMC-OCS组、CMC-OCS/PRP组细胞活性氧水平降低(P<0.05),PRP组、CMC-OCS组、CMCOCS/PRP组丙二醛水平降低(P<0.05)、细胞铺展面积增加(P<0.05),CMC-OCS/PRP组细胞凋亡率降低(P<0.05),PRP组、CMC-OCS组、CMCOCS/PRP组胶原纤维Ⅰ蛋白表达增加(P<0.05);③与对照组比较,H2O2组细胞迁移数量减少(P<0.05),迁移面积无明显变化(P>0.05);与H2O2组比较,PRP组、CMC-OCS组、CMC-OCS/PRP组细胞迁移数量、迁移面积均增加(P<0.05),并且以CMC-OCS/PRP组增加最显著;④在氧化应激条件下,CMC-OCS/PRP水凝胶可改善成纤维细胞的迁移能力,抗细胞凋亡并保护细胞伸展功能。 展开更多
关键词 富血小板血浆 羧甲基壳聚糖 氧化硫酸软骨素 水凝胶 l929细胞 抗氧化
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Recombinant chitinase-3-like protein 1 alleviates learning and memory impairments via M2 microglia polarization in postoperative cognitive dysfunction mice
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作者 Yujia Liu Xue Han +6 位作者 Yan Su Yiming Zhou Minhui Xu Jiyan Xu Zhengliang Ma Xiaoping Gu Tianjiao Xia 《Neural Regeneration Research》 SCIE CAS 2025年第9期2727-2736,共10页
Postoperative cognitive dysfunction is a seve re complication of the central nervous system that occurs after anesthesia and surgery,and has received attention for its high incidence and effect on the quality of life ... Postoperative cognitive dysfunction is a seve re complication of the central nervous system that occurs after anesthesia and surgery,and has received attention for its high incidence and effect on the quality of life of patients.To date,there are no viable treatment options for postoperative cognitive dysfunction.The identification of postoperative cognitive dysfunction hub genes could provide new research directions and therapeutic targets for future research.To identify the signaling mechanisms contributing to postoperative cognitive dysfunction,we first conducted Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses of the Gene Expression Omnibus GSE95426 dataset,which consists of mRNAs and long non-coding RNAs differentially expressed in mouse hippocampus3 days after tibial fracture.The dataset was enriched in genes associated with the biological process"regulation of immune cells,"of which Chill was identified as a hub gene.Therefore,we investigated the contribution of chitinase-3-like protein 1 protein expression changes to postoperative cognitive dysfunction in the mouse model of tibial fractu re surgery.Mice were intraperitoneally injected with vehicle or recombinant chitinase-3-like protein 124 hours post-surgery,and the injection groups were compared with untreated control mice for learning and memory capacities using the Y-maze and fear conditioning tests.In addition,protein expression levels of proinflammatory factors(interleukin-1βand inducible nitric oxide synthase),M2-type macrophage markers(CD206 and arginase-1),and cognition-related proteins(brain-derived neurotropic factor and phosphorylated NMDA receptor subunit NR2B)were measured in hippocampus by western blotting.Treatment with recombinant chitinase-3-like protein 1 prevented surgery-induced cognitive impairment,downregulated interleukin-1βand nducible nitric oxide synthase expression,and upregulated CD206,arginase-1,pNR2B,and brain-derived neurotropic factor expression compared with vehicle treatment.Intraperitoneal administration of the specific ERK inhibitor PD98059 diminished the effects of recombinant chitinase-3-like protein 1.Collectively,our findings suggest that recombinant chitinase-3-like protein 1 ameliorates surgery-induced cognitive decline by attenuating neuroinflammation via M2 microglial polarization in the hippocampus.Therefore,recombinant chitinase-3-like protein1 may have therapeutic potential fo r postoperative cognitive dysfunction. 展开更多
关键词 Chil1 hippocampus learning and memory M2 microglia NEUROINFlAMMATION postoperative cognitive dysfunction(POCD) recombinant CHI3l1
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Autism spectrum disorder:difficulties in diagnosis and microRNA biomarkers
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作者 Bridget Martinez Philip V.Peplow 《Neural Regeneration Research》 SCIE CAS 2025年第10期2776-2786,共11页
We performed a PubMed search for microRNAs in autism spectrum disorder that could serve as diagnostic biomarkers in patients and selected 17 articles published from January 2008 to December 2023,of which 4 studies wer... We performed a PubMed search for microRNAs in autism spectrum disorder that could serve as diagnostic biomarkers in patients and selected 17 articles published from January 2008 to December 2023,of which 4 studies were performed with whole blood,4 with blood plasma,5 with blood serum,1 with serum neural cell adhesion molecule L1-captured extracellular vesicles,1 with blood cells,and 2 with peripheral blood mononuclear cells.Most of the studies involved children and the study cohorts were largely males.Many of the studies had performed microRNA sequencing or quantitative polymerase chain reaction assays to measure microRNA expression.Only five studies had used real-time polymerase chain reaction assay to validate microRNA expression in autism spectrum disorder subjects compared to controls.The microRNAs that were validated in these studies may be considered as potential candidate biomarkers for autism spectrum disorder and include miR-500a-5p,-197-5p,-424-5p,-664a-3p,-365a-3p,-619-5p,-664a-3p,-3135a,-328-3p,and-500a-5p in blood plasma and miR-151a-3p,-181b-5p,-320a,-328,-433,-489,-572,-663a,-101-3p,-106b-5p,-19b-3p,-195-5p,and-130a-3p in blood serum of children,and miR-15b-5p and-6126 in whole blood of adults.Several important limitations were identified in the studies reviewed,and need to be taken into account in future studies.Further studies are warranted with children and adults having different levels of autism spectrum disorder severity and consideration should be given to using animal models of autism spectrum disorder to investigate the effects of suppressing or overexpressing specific microRNAs as a novel therapy. 展开更多
关键词 autism spectrum disorder BIOMARKER blood cells blood plasma blood serum DIAGNOSIS MICRORNA peripheral blood mononuclear cells serum neural cell adhesion molecule l1-captured extracellular vesicles whole blood
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以单细胞压片技术观察烟草(Nicotiana tabacum L.)合子细胞器DNA的分布 被引量:1
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作者 何玉池 曲良焕 +1 位作者 孙蒙祥 杨弘远 《武汉植物学研究》 CSCD 北大核心 2005年第1期63-67,共5页
通过实验建立了单个合子的压片技术,尝试了将染色与固定分开或同时进行的两种压片方法,两种方法均可以用于不同发育阶段合子细胞器DNA分布的检测,但二者各具特色。同时也利用共聚焦激光扫描显微镜观察了烟草(NicotianatabacumL.)合子细... 通过实验建立了单个合子的压片技术,尝试了将染色与固定分开或同时进行的两种压片方法,两种方法均可以用于不同发育阶段合子细胞器DNA分布的检测,但二者各具特色。同时也利用共聚焦激光扫描显微镜观察了烟草(NicotianatabacumL.)合子细胞器的分布,并与单细胞压片技术进行了比较。实验表明它们各有其优越性。 展开更多
关键词 单细胞压片 合子 细胞器 烟草
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Using Isolated Embryo Sacs and Early Proembryos for Localization of Calmodulin mRNA Before and After Fertilization in Nicotiana 被引量:3
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作者 陈绍荣 吕应堂 +1 位作者 杨弘远 周嫦 《Acta Botanica Sinica》 CSCD 1999年第7期686-689,共4页
An in situ hybridization technique for localization of calmodulin(CaM) mRNA in isolated entire embryo sacs and proembryos in Nicotiana tabacum L.cv.W38 has been developed. This technique can be applied to smal... An in situ hybridization technique for localization of calmodulin(CaM) mRNA in isolated entire embryo sacs and proembryos in Nicotiana tabacum L.cv.W38 has been developed. This technique can be applied to small amounts of materials in which a whole view of CaM mRNA distribution can be obtained. The authors revealed that CaM mRNA expression changes dramatically before and after fertilization. Especially interesting is that a prominent CaM mRNA band appears between the egg apparatus and polar nuclei temporarily during the period of pollination and fertilization. The band disappears just prior to fertilization and expands to a fan_shaped region that occupies the micropylar portion of the embryo sac. After fertilization, CaM mRNA accumulates in the elongated zygotes with higher concentration in their chalazal portion than in the micropylar portion. Such an asymmetrical pattern continues to manifest in the early proembryos. It is supposed that CaM mRNA may be involved in the early events and signaling steps associated with double fertilization and zygote polarization in higher plants. 展开更多
关键词 Calmodulin mRNA in situ hybridization Embryo sac PROEMBRYO nicotiana tabacum
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枯草芽孢杆菌(Bacillus subtilis)cspB基因转化烟草(Nicotiana tabacuum)的研究 被引量:6
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作者 徐萍莉 陈丽萍 +1 位作者 周秀杰 何道一 《激光生物学报》 CAS CSCD 2013年第4期343-347,353,共6页
细菌冷休克蛋白cspB是原核生物RNA的分子伴侣,含有原始型的冷休克结构域,具有与核酸结合功能,可以防止RNase对mRNA的降解,也能纠正mRNA的折叠错误。为了寻找作物改良的潜在基因资源,从枯草芽孢杆菌XS-01基因组中克隆出cspB基因,并与pBI... 细菌冷休克蛋白cspB是原核生物RNA的分子伴侣,含有原始型的冷休克结构域,具有与核酸结合功能,可以防止RNase对mRNA的降解,也能纠正mRNA的折叠错误。为了寻找作物改良的潜在基因资源,从枯草芽孢杆菌XS-01基因组中克隆出cspB基因,并与pBI121构建成pBI121-cspB植物表达载体。利用叶圆盘转化法转化烟草,通过卡那霉素筛选和PCR、Southern杂交鉴定5个转化体株系。除TN010外,其他转基因株系在外观形态与野生亲本的没有差别,但TN001和TN011育性降低,TN010和TN012则完全丧失育性。干旱处理结果表明,转基因植株在土壤水分恢复后10 d,其平均单株干物质质量较之野生亲本的显著增加;叶片光合速率测定结果表明,在干旱处理时,转基因植株和对照亲本叶片光合速率均显著下降,在土壤水分恢复后,转基因植株的光合速率能快速恢复,但对照亲本的恢复缓慢。实验结果说明,cspB能够促进植物细胞从逆境伤害中快速恢复功能。 展开更多
关键词 cspB基因 抗旱性 转基因 烟草
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具翼烟草(Nicotiana alata)核型、C带与rDNA染色体定位研究 被引量:1
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作者 王霖娇 任学良 +2 位作者 盛茂银 杜家颖 温培才 《中国烟草学报》 EI CAS CSCD 北大核心 2017年第1期95-101,共7页
采用去壁低渗法、BSG法和双色荧光原位杂交技术,分别开展了具翼烟草(Nicotiana alata)的核型分析、C显带与45S和5S rDNA染色体定位研究。4个具翼烟草收集系根尖细胞有丝分裂中期染色体平均核型为2n=2x=12m+6st,染色体臂比的变异幅度为1.... 采用去壁低渗法、BSG法和双色荧光原位杂交技术,分别开展了具翼烟草(Nicotiana alata)的核型分析、C显带与45S和5S rDNA染色体定位研究。4个具翼烟草收集系根尖细胞有丝分裂中期染色体平均核型为2n=2x=12m+6st,染色体臂比的变异幅度为1.14~4.22,平均臂比值为2.17,最长与最短染色体之比为2.16,属2B型。C带研究结果显示,具翼烟草中期染色体C带丰富,多态性良好,共显带25条,包括7条着丝粒带、15条端带和3条中间带。45S和5S rDNA染色体定位研究结果显示,具翼烟草根尖细胞有丝分裂中期染色体细胞相分别显示了3对45S rDNA和2对5S rDNA基因位点信号,45S rDNA基因位点分别位于I S、III S、V L上,2对5S rDNA基因位点分别位于I L、IV S上(字母L和S分别代表染色体长臂和短臂,罗马数字代表同源染色体序号)。在I号同源染色体上同时存在45S和5S rDNA基因位点。研究结果对烟草属系统起源、遗传进化以及遗传育种等领域的进一步研究具有重要的参考价值。 展开更多
关键词 烟草 核型 染色体 荧光原位杂交 C带
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烟草Nicotiana glauca遗传瘤的诱导
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作者 曲冠证 郑唐春 +2 位作者 杜恒瑞 臧丽娜 夏德安 《安徽农业大学学报》 CAS CSCD 北大核心 2012年第3期417-420,共4页
以往的研究证明一些烟草杂种能自发形成遗传瘤,而正常条件下杂交亲本各自本身并不能自发形成遗传瘤。该研究首次从烟草Nicotiana glauca品种中诱导产生出遗传瘤,这些N.glauca遗传瘤与杂种N.glauca×N.langsdorffii遗传瘤具有相同的... 以往的研究证明一些烟草杂种能自发形成遗传瘤,而正常条件下杂交亲本各自本身并不能自发形成遗传瘤。该研究首次从烟草Nicotiana glauca品种中诱导产生出遗传瘤,这些N.glauca遗传瘤与杂种N.glauca×N.langsdorffii遗传瘤具有相同的特征,即在无激素的培养基上能正常分化、对外源植物激素不敏感等特点。该研究结果同时还进一步证明种间杂交并不是诱导烟草产生遗传瘤的唯一因素。 展开更多
关键词 烟草 杂种 遗传瘤 nicotiana glauca
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粉蓝烟草(Nicotiana glauca)花柱S-糖蛋白的发达及对花粉管生长的影响 被引量:1
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作者 李润植 毛雪 李彩霞 《作物学报》 CAS CSCD 北大核心 1999年第1期120-125,共6页
本文报道了粉蓝烟草花柱S-糖蛋白的发达特性及对花粉萌发和花粉管生长的影响。开花后1~20d内,花柱S-糖蛋白在柱头下1~1.scm处大量表达,与活体上花粉管停止生长的时间和部位相吻和。在培养基中加入S-糖蛋白时,不同来源的花粉均能顺利萌... 本文报道了粉蓝烟草花柱S-糖蛋白的发达特性及对花粉萌发和花粉管生长的影响。开花后1~20d内,花柱S-糖蛋白在柱头下1~1.scm处大量表达,与活体上花粉管停止生长的时间和部位相吻和。在培养基中加入S-糖蛋白时,不同来源的花粉均能顺利萌发,但是同源花粉管生长却被特异性地抑制,而非同源花粉则能继续正常生长。在优化离体培养条件下,花粉管生长速率仍低于在活体上的生长速率。与对照相比,加入s-糖蛋白后,花粉管生长长度变异性增加。 展开更多
关键词 粉蓝烟草 自交不亲和性 花柱S糖蛋白 花粉管生长
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Cloning and Gentic Transformation of NbDAD1 Gene from Nicotiana benthamiana
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作者 徐玲玲 陈崇崇 +2 位作者 王梨嬛 潘永娟 杨莉 《Agricultural Science & Technology》 CAS 2012年第5期938-941,944,共5页
[Objective] The aim was to clone NbDAD1 gene from Nicotiana benthami- ana and study its genetic transformation. [Method] NbDAD1 gene was isolated from N. benthamiana by using RT-PCR technology and over-expression vect... [Objective] The aim was to clone NbDAD1 gene from Nicotiana benthami- ana and study its genetic transformation. [Method] NbDAD1 gene was isolated from N. benthamiana by using RT-PCR technology and over-expression vectors were con- structed to obtain NbDADl-overexpression resistant plants and NbDADl-overexpres- sion resistant plants carrying HA tag. [Result] The 351 bp long NbDAD1 gene was cloned from N. benthamiana; recombinant plasmids pCAMBIA1301-NbDAD1 and pCAMBIA1301-NbDAD1HAtag were constructed successfully; 50T0-generation N. ben- thamiana Hyg-resistant transgenic lines of three genotypes were obtained, including 23 positive transgenic plants. [Conclusion] This study laid the foundation for investi- gating the specific functions of NbDAD1 gene in N. benthamiana and exploring the possible functional mechanism of DAD1 protein in programmed cell death of plants. 展开更多
关键词 NbDAD1 gene Genetic transformation nicotiana benthamiana PCD
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粉蓝烟草(Nicotiana glauca)S—糖蛋白对离体花粉管生长的影响 被引量:1
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作者 李润植 毛雪 《山西农业大学学报》 CAS 1996年第3期217-221,共5页
在已论述粉蓝烟草(Nicotiana glauca)花柱S—糖蛋白分离和测序的基础之上,本文报道了离体培养条件下,纯化的S—糖蛋白对花粉萌发及花粉管生长的影响.在培养基中加入S—糖蛋白时,不同来源的花粉均能顺利萌发,但是同源花粉管生长却被特异... 在已论述粉蓝烟草(Nicotiana glauca)花柱S—糖蛋白分离和测序的基础之上,本文报道了离体培养条件下,纯化的S—糖蛋白对花粉萌发及花粉管生长的影响.在培养基中加入S—糖蛋白时,不同来源的花粉均能顺利萌发,但是同源花粉管生长却被特异性地抑制,而非同源花粉则能继续正常生长.在优化离体培养条件下,花粉管生长速率仍低于在活体上的生长速率.与对照相比,加入S—糖蛋白后,花粉管生长长度变异性增加.另外,即使绝大多数同源花粉管生长被S—糖蛋白所抑制,仍有个别花粉管继续正常生长,其原因需进一步探讨. 展开更多
关键词 粉蓝烟草 烟草 自交不亲和性 离体花粉管生长
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Effects of Ppc Gene Construction of Monocotyledon on Seedling Growth of Transgenic Nicotiana tabacum
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作者 张桂芳 丁在松 赵明 《Agricultural Science & Technology》 CAS 2015年第11期2416-2420,共5页
To compare the transformation effects of two different forms (cDNA in monocotyledonous plant Echinochloa crusgalli, DNA in monocotyledonous plant Zea mays) of phosphoenolpyruvate carboxylase (PEPC) gene (Ppc) on... To compare the transformation effects of two different forms (cDNA in monocotyledonous plant Echinochloa crusgalli, DNA in monocotyledonous plant Zea mays) of phosphoenolpyruvate carboxylase (PEPC) gene (Ppc) on the growth of transgenic dicotyledonous plant, Agrobacterium-mediated transformation of Ppc genes into Nicotiana tabacum were carried out. Transgenic leaf plates and differentiated seedling leaves were verified by GUS histochemistry, PCR, and RT-PCR. Results showed that transgenic N. tabacum with Ppc-cDNA of E. crusgalli had relatively strong differentiation ability. However, N. tabacum after transformation of complete DNA sequence of Ppc genes in Z mays had relatively poor ability of growth. The differentiated green seedlings had the phenomenon of yellowing; and photosynthesis ability of leaves was poor. This might be caused by the misidentification and wrong splicing in transcription. This indicated that the expression rate of monocotyledonous complete DNA might be reduced in the monocotyledonous cells with relatively far genetic distances. Detection results of showed that Pn in most transgenic N. tabacum with Ppc-cDNA of E. crusgalli was was higher than that in control, which preliminarily proved that PEPC of monocotyledonous plant E. crusgalli had certain regulatory effects on photosynthesis of N. tabacum. 展开更多
关键词 Echinochloa crusgalli Zea mays nicotiana tabacum Phosphoenolpyru-vate carboxylase Net photosynthetic rate Genetic distance
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Combined Toxicity Test of Complex Agents of Metalaxyl and Other Fungicides against Phytophthora parasitica var. nicotianae
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作者 刘铭 《Plant Diseases and Pests》 CAS 2011年第2期58-60,63,共4页
[ Objective] To overcome the resistance of Phytophthora parasitica var. nicotianae against metalaxyl and effectively control its damage, new efficient complex agent of metalaxyl was studied and developed. [ Method] Th... [ Objective] To overcome the resistance of Phytophthora parasitica var. nicotianae against metalaxyl and effectively control its damage, new efficient complex agent of metalaxyl was studied and developed. [ Method] The toxieities of nine fungicides against P. parasitica were measured using growth rate method. On this basis, the fungicides with good effects were selected to compound with metalaxyl, and the optimum complex ratio was confirmed. [Result] The toxicity of metalaxyl was the strongest with EC50 of 2. 130 5μg/ml; followed by carbendazim, mancezeb and dimethomorph with EC50 of 2.357 9, 2.639 8 and 2. 778 8 μg/ml. The effect of cyazofamid was the poorest with EC50 of 6. 278 8 μg/ml. The optimum complex ratios of dimethomorph, carbendazim and mancezeb with metalaxyl were 40: 60, 30:70 and 20: 80, and their co-toxicity coefficients were 138.80,124.25 and 115.00, respectively. [ Conclusion] The complex agents had application and promotion value, which could be used to carry out further field trials. 展开更多
关键词 Phytophthora parasitica vat. nicotianae METAlAXYl COMPlEX TOXICITY
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基于SSR分子标记的Nicotiana tobacum–N.plumbaginifolia异源染色体植株的鉴定与筛选 被引量:5
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作者 尚维 赵申清玉 +5 位作者 党江波 郭启高 梁国鲁 杨超 张艳 陈益银 《作物学报》 CAS CSCD 北大核心 2018年第11期1640-1649,共10页
以烟草(Nicotiana tabacum)品种云烟87的八倍体(2n=8x=96)和野生烟草N. plumbaginifolia (2n=2x=20)的基因组DNA为模板,对340对烟草SSR引物进行筛选以获得能扩增多态性条带的引物。利用多态性引物对种间杂交后代及190株回交后代的基因组... 以烟草(Nicotiana tabacum)品种云烟87的八倍体(2n=8x=96)和野生烟草N. plumbaginifolia (2n=2x=20)的基因组DNA为模板,对340对烟草SSR引物进行筛选以获得能扩增多态性条带的引物。利用多态性引物对种间杂交后代及190株回交后代的基因组DNA进行扩增,并对N.plumbaginifolia中的SSR标记的连锁情况进行简要分析。经筛选获得了多态性引物29对。结果显示,在190株后代中, 159株的基因组DNA能扩增出N. plumbaginifolia的特异SSR位点,可以判定该159株为N. tabacum的N. plumbaginifolia异源染色体植株,其余31株植株可能不含有N. plumbaginifolia的染色体。经UPGMA聚类分析,本群体中植株的遗传多样性较为丰富,部分分子标记在后代中的出现具有完全相关性。29个标记中14个可确定来源于5条不同染色体,N.plumbaginifolia的29个位点在回交后代中的扩增效率并不相同,且效率均较低(低于31.00%),说明该杂种中N. plumbaginifolia基因组的垂直传递效率较低。利用SSR分子标记可以判定云烟87八倍体与N.plumbaginifolia杂交获得的后代为真杂种,且自该远缘杂种回交后代中筛选获得大量异源染色体植株。这些结果和筛选获得异源染色体植株为进一步创制N.tabacum-N.plumbaginifolia抗黑胫病单体附加系以及易位系奠定了基础。 展开更多
关键词 nicotiana tabacum N.plumbaginifolia SSR分子标记 异源染色体 黑胫病
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L型阵水下目标方位和距离联合最大似然估计 被引量:1
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作者 宋海岩 迟凤阳 唐弢 《黑龙江工程学院学报》 CAS 2024年第2期1-6,共6页
空间目标定位是阵列信号处理领域中的研究热点,广泛应用于雷达、声纳、通信、射电天文、医学诊断、地震遥感等众多领域。传统的水下声源定位仅获取目标声源的深度和距离信息,却无法估计目标的方位信息。针对这一问题,在充分考虑水下声... 空间目标定位是阵列信号处理领域中的研究热点,广泛应用于雷达、声纳、通信、射电天文、医学诊断、地震遥感等众多领域。传统的水下声源定位仅获取目标声源的深度和距离信息,却无法估计目标的方位信息。针对这一问题,在充分考虑水下声传播模型的基础上,利用L型阵列固有的结构特征,将最大似然估计技术应用到水下目标定位,提出一种基于L型水听器阵列的水下目标方位与距离联合最大似然估计算法。研究表明:最大似然估计法可有效突破传统波束形成分辨力“瑞利限”的限制,提高水下目标定位的分辨能力和精度。计算机仿真分析显示,在单目标和相干双目标的场景中,文中所提出的方法均具有更为尖锐的谱峰及更低的旁瓣水平,可对水下目标进行有效的方位角和距离联合估计。 展开更多
关键词 l型阵列 水下声传播 方位角及距离联合估计 最大似然估计法
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L波段差分干涉SAR卫星严格回归轨道优化设计方法
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作者 李楠 温俊健 +3 位作者 刘艳阳 凌惠祥 魏春 陈筠力 《测绘学报》 EI CSCD 北大核心 2024年第10期1873-1880,共8页
相比于传统SAR卫星应用方向,我国第一代L波段差分干涉SAR卫星主要用于实现全球精准地表形变测量。为了达到重轨差分干涉优于5 cm的形变测量指标,首先需要解决卫星空间基准轨道高精度回归的工程难题。对此,本文提出一种严格回归轨道优化... 相比于传统SAR卫星应用方向,我国第一代L波段差分干涉SAR卫星主要用于实现全球精准地表形变测量。为了达到重轨差分干涉优于5 cm的形变测量指标,首先需要解决卫星空间基准轨道高精度回归的工程难题。对此,本文提出一种严格回归轨道优化设计方法,通过地球重力场阶数需求分析建立卫星高阶动力学模型,将J4摄动下修正的太阳同步回归轨道参数作为优化算法初值,以卫星WGS-84坐标系位置、速度回归精度满足收敛阈值为目标,利用启发式多目标进化算法开展轨道参数寻优。仿真试验表明,寻优结果回归精度可达厘米级,优于既定工程指标,并且本文方法有效性和正确性已经过L波段差分干涉SAR卫星在轨验证。 展开更多
关键词 严格回归轨道 合成孔径雷达差分干涉 高阶重力场 优化轨道设计 l波段差分干涉SAR卫星
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