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Genetic Transformation of cell suspension Cultures of Medicago truncatula via Agrobacterium tumefaciens

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Genetic Transformation of Cell Suspension Cultures of Medica...

cell replacement with stem cell-derived retinal ganglion cells from different protocols

第二届植物分子育种国际会议

作者： M.A Cordeiro P.S.Fevereiro Instituto de Tecnologia Química e Biológica （ITQB） Universidade Nova de LisboaApartado 1272781-901 OeirasPortugal Instituto de Tecnologia Química e Biológica （ITQB） Universidade Nova de LisboaApartado 1272781-901 OeirasPortugal Instituto de Biologia Experimental e Tecnológica （IBET）Apartado 122781-901 OeirasPortugal Departamento de Biologia VegetalFaculdade de Ciências da Universidade de LisboaCampo Grande1749-016 LisboaPortugal

＜正＞Medicago truncatula is a forage plant. Recently, it is used as a model species for the study of plant biology unique to legume family owing to its small and diploidy genome （2N=16）, short life cycle, autogamous

关键词： Medicago truncatula Model legume cell suspension Genetic transformation Agrobacterium tumefaciens

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Neural Regeneration Research 2024年第4期19卷 807-810页

作者： Ziming Luo Kun-Che Chang Spencer Center for Vision Research Byers Eye InstituteStanford University School of Medicine Department of Ophthalmology Louis J.Fox Center for Vision RestorationUniversity of Pittsburgh School of Medicine Department of Neurobiology Center of NeuroscienceUniversity of Pittsburgh School of Medicine

Glaucoma,characterized by a degenerative loss of retinal ganglion cells,is the second leading cause of blindness *** is currently no cure for vision loss in glaucoma because retinal ganglion cells do not regenerate and are not replaced after *** stem cell-derived retinal ganglion cell transplant is a potential therapeutic strategy for retinal ganglion cell degenerative *** this review,we first discuss a 2D protocol for retinal ganglion cell differentiation from human stem cell culture,including a rapid protocol that can generate retinal ganglion cells in less than two weeks and focus on their transplantation ***,we discuss using 3D retinal organoids for retinal ganglion cell transplantation,comparing cell suspensions and *** review provides insight into current knowledge on human stem cell-derived retinal ganglion cell differentiation and transplantation,with an impact on the field of regenerative medicine and especially retinal ganglion cell degenerative diseases such as glaucoma and other optic neuropathies.

关键词： cell clumps cell suspension cell transplantation differentiation direct-induced protocol glaucoma optic neuropathy regenerative medicine retinal ganglion cell retinal organoids stem cells

In vitro Selection for Fusarium Wilt Resistance in Gladiolus

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Journal of Integrative Plant Biology 2008年第5期50卷 601-612页

作者： Idrees Ahmad Nasir Sheikh Riazuddin National Centre of Excellence in Molecular Biology University of the PunjabCanal BankThokar Niaz Baig Lahore 53700Pakistan

Cormels pieces of four Fusarium susceptible Gladiolus cultivars （Friendship, Peter Pears, Victor Borge and Novalux） formed friable calli when cultured in vitro on Murashige and Skoog basal medium containing various concentrations of auxin and cytokinin. The friable calli established cell suspensions. Plantlet regeneration was obtained from the control callus, control cell suspension derived callus and in vitro selected Fusarium oxysporum Schlecht. resistant cell-lines of Friendship. The in vitro cormlets showed 85-95% germination after breaking dormancy of 8 weeks at 4℃. cell suspensions of all four Gladiolus cultivars were found to be highly sensitive to fusaric acid. Gradual increase in fusaric acid concentrations to the cell-suspension cultures decreased cell growth considerably. One albino plant was found from the second generation of the in vitro selected cell line of Friendship. The albino plant was found to be highly susceptible to F. oxysporum. The cormlets of all in vitro selected cell lines of Friendship were inoculated with a conidial suspension of the F. oxysporum before planting and were also sprayed with the same spore suspension for further characterization when the height of plants was about 6cm. The four selected cell lines showed the same response whether or not they were inoculated with conidia of the F. oxysporum. Plantlets of all of the selected cell lines exhibited significant growth as compared with the control after application of conidia of the F. oxysporum.

关键词： cell suspension fusaric acid fusadum Gladiolus in vitro selection regeneration.

Effect of epidermal growth factor on the migration of neural stem cells

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Neural Regeneration Research 2006年第6期1卷 502-504页

作者： Faliang Duan Guoping Yang Junwu Wei Jinglei Wu Department of Neurosurgery First Hospital of Wuhan Wuhan 430022 Hubei Province China

BACKGROUND： Recently, researches on neural stem cells （NSCs） are focus on differentiation and migration ot stem cells. How to regulate and control differentiation and migration of NSCs based on human wills is still a hot topic.OBJECTIVE ： To investigate the effect of epidermal growth factor （EGF） on the migration and proliferation of NSCs and analyze duration of the effect. DESIGN ： Contrast study based on cells SETTING ： Department of Neurological Surgery, the First Hospital of Wuhan MATERIALS： Healthy SD rats aged 13-14 embryonic days. EGF （Sigma Company） METHODS： The experiment was carried out in the Animal Laboratory of Experimental Center Affiliated to the First Hospital of Wuhan from October 2004 to July 2006. NSCs selected from embryonic striatum of rats with 13-14 embryonic days were cultured; 7 days later, suspended neural sphere was used to make simple cell suspension and cultured once more. Then, DMEM-F12＋20 μg/L EGF was added into culture medium; 14 days later, the rats were divided into experimental group and control group. Rats in the experimental group were cultured with the same medium mentioned above; however, rats in the control group were cultured with only DMEM-F12. Migration of cells was observed under microscope every day. MAIN OUTCOME MEASURES： NSCs migration in both experimental group and control group RESULTS： cell spheres in primary culture were NSCs. In addition, 14 days later, proliferation of stem cells were observed remarkably in EGF culture, and size of cell sphere was about that of 100 cells. In experimental group, proliferation of cell sphere was slow down on the 14^th culture day, and apophysis was erupted to neighbor cell sphere. Moreover, NSCs migrated from big cell sphere to small cell sphere during 14-17 culture days, and then, cell migration was disappeared at 17 days after culture. In control group, cell migration was not observed. CONCLUSION： EGF can induce proliferation and migration of NSCs during a special

关键词： neural stem cells epidermal growth factor primary culture cell suspension culture medium small cell slow down hot topic

Non-CG DNA methylation-deficiency mutations enhance mutagenesis rates during salt adaptation in cultured Arabidopsis cells

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Stress Biology 2021年第1期1卷 144-155页

作者： Xiaohong Zhu Shaojun Xie Kai Tang Rajwant KKalia Na Liu Jinbiao Ma Ray ABressan Jian-Kang Zhu State Key Laboratory of Crop Stress Adaptation and Improvement School of Life SciencesHenan UniversityKaifengChina State Key Laboratory of Cotton Biology School of Life SciencesHenan UniversityKaifengChina Bioinformatics Core Purdue UniversityWest LafayetteIN 47907USA Department of Horticulture and Landscape Architecture Purdue UniversityWest LafayetteIN 47907USA Central Arid Zone Research Institute Jodhpur 342003India State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-Products Institute of VegetablesZhejiang Academy of Agricultural SciencesHangzhou 310021China State Key Laboratory of Desert and Oasis Ecology Xinjiang Institute of Ecology and GeographyChinese Academy of Sciences 830011UrumqiChina Shanghai Center for Plant Stress Biology Shanghai Institutes for Biological SciencesChinese Academy of SciencesShanghai 200032China

Much has been learned about how plants acclimate to stressful environments,but the molecular basis of stress adaptation and the potential involvement of epigenetic regulation remain poorly ***,we examined if salt stress induces mutagenesis in suspension cultured plant cells and if DNA methylation affects the mutagenesis using whole genome resequencing *** generated suspension cell cultures from two Arabidopsis DNA methylation-deficient mutants and wild-type plants,and subjected the cultured cells to stepwise increases in salt stress intensity over 40 culture *** show that ddc(drm1 drm2 cmt3)mutant cells can adapt to grow in 175 mM NaCl-containing growth medium and exhibit higher adaptability compared to wild type Col-0 and nrpe1 cells,which can adapt to grow in only 125mM NaCl-containing growth *** treated nrpe1 and ddc cells but not wild type cells accumulate more mutations compared with their respective untreated *** is no enrichment of stress responsive genes in the list of mutated genes in salt treated cells compared to the list of mutated genes in untreated *** results suggest that DNA methylation prevents the induction of mutagenesis by salt stress in plant cells during stress adaptation.

关键词： Salt stress cell suspension DNA methylation-deficient mutants Stress-induced mutagenesis Stress adaptation

Analytical model for power dissipation in cell membranes in suspensions exposed to electric fields

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Journal of Southeast University(English Edition) 2005年第2期21卷 145-148页

作者：覃玉荣江悦华赖声礼华南理工大学电子与信息信学院广州510640 广州军区广州总医院医学实验科广州510010

Due to interaction among cells, it is too complex to build an exactanalytical model for the power dissipation within the cell membrane in suspensions exposed toexternal fields. An approximate equivalence method is proposed to resolve this problem. Based on theeffective medium theory, the transmembrane voltage on cells in suspensions was investigated by theequivalence principle. Then the electric field in the cell membrane was determined. Finally,analytical solutions for the power dissipation within the cell membrane in suspensions exposed toexternal fields were derived according to the Joule principle. The equations show that theconductive power dissipation is predominant within the cell membrane in suspensions exposed todirect current or lower frequencies, and dielectric power dissipation prevails at high frequenciesexceeding the relaxation frequency of the exposed membrane.

关键词： power dissipation analytical model cell suspension external electricfields

Screening the Best Material for Ploidy Identification of Sugarcane-related Genera

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Agricultural Biotechnology 2021年第1期10卷 12-14,18页

作者： Lianying MAO Yiyun GUI Jinju WEI Haibi LI Ronghua ZHANG Hui ZHOU Xihui LIU Sugarcane Research Institute Guangxi Academy of Agricultural Sciences/Sugarcane Research CenterChinese Academy of Agricultural Sciences/Key Laboratory of Sugarcane Genetic Improvement in GuangxiNanning 530007China

[Objectives] Flow cytometry is widely used to identify plant chromosome ploidy because of its simplicity, rapidity and accuracy. Chromosome ploidy identification is an important part of sugarcane ploidy breeding and application research. It is particularly important to find out the best detection part for ploidy identification in sugarcane. [Methods] The cell suspensions of sugarcane stem tips and leaves were prepared by blade chopping method. The cell suspensions were detected by flow cytometry. The best position for ploidy identification was determined by comparing the cell suspension prepared from stem tips and cell suspension prepared from leaves. [Results] The results showed that the cell suspension dissociated from stem tips was more clear than that from leaf cell suspension;the proportion of non-adherent cells in the suspension prepared from stem tips was larger than that from the leaf cell suspension;the main peak of the stem tip cell suspension was single and the number of cells was more than that of the stem tip cell suspension by flow cytometry. Using the known ploidy ‘Badilar’ as the internal reference, the ploidy of cyathomi 87-16 was detected to be 8.37. [Conclusions] Sugarcane shoot tips are an ideal material for ploidy identification. This study provides a theoretical basis for selecting the best detection site for ploidy identification of sugarcane.

关键词： Flow cytometry Chromosome ploidy Sugarcane cell suspension

Ultrastructural and Physiological Characterization of YELP, a Novel, Yellow, Chlorophyll-Deficient cell Mutant Line That Develops Etioplast-Like Plastids in the Light

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American Journal of Plant Sciences 2016年第3期7卷 510-524页

作者： Betzaida Jiménez-Francisco Carlos Trejo Hilda Araceli Zavaleta-Mancera Blanca Moreno-Gómez Edmundo García-Moya Florentino Víctor Conde-Martínez Gerardo Armando Aguado-Santacruz Potgrado en Botánica Colegio de Postgraduados Texcoco México Campo Experimental Bajío Instituto Nacional de Investigaciones Forestales y Agropecuarias Celaya México Programa de Posgrado Instituto Tecnológico de Roque Celaya México

In this work, we present an ultrastructural and physiological description of a novel chlorophyll-deficient, yellow cell line of the grass Bouteloua gracilis that develops etioplast-like plastids in presence of light (YELP). These mutant cells were compared to the parental, wild-type, highly chlorophyllous cells from which they were isolated. Growth analysis, based on fresh and dry weights, indicated that YELP accumulates biomass at a slower rate than the parental, green cells. Besides, YELP developed very low levels of photosynthetic pigments, reaching only 9.3% and 38.4% of chlorophyll a and chlorophyll b, respectively, developed by the wild-type cells. Likewise, the accessory pigments, carotenes and xanthophylls, were only synthesized at 8.0% and 5.4%, respectively, of the levels reached by the green cells. Electron microscopy revealed remarkable differences in plastid ultrastructure between the wild-type and mutant cells. Plastids of YELP were heterogeneous and smaller than those found in wild-type cells. YELP plastids were abnormal with poorly developed membrane systems that prevented the accumulation of chlorophyll and accessory pigments in the mutant cell line. We expect this novel, mutant cell line will provide new tools for studying plastid development and differentiation.

关键词： cell suspension Chloroplast Biogenesis Prolamellar Body Ultrastructure

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大麦原生质体融合和体细胞杂种形成

上海农业学报 1996年第4期 1-4页

作者：颜昌敬黄剑华张玉华陆瑞菊孙月芳周润梅上海市农业科学院

以申麦１号和花培系Ａ１０２为材料，分别用它们的叶细胞和胚性细胞悬浮系游离出的原生质体作为融合亲本，进行了原生质体融合实验，通过改进融合液组成，采用微孔滤膜技术，改进培养方法等措施，获得了它们的体细胞杂种细胞系．并经杂... 详细信息

以申麦１号和花培系Ａ１０２为材料，分别用它们的叶细胞和胚性细胞悬浮系游离出的原生质体作为融合亲本，进行了原生质体融合实验，通过改进融合液组成，采用微孔滤膜技术，改进培养方法等措施，获得了它们的体细胞杂种细胞系．并经杂种筛选和鉴定，证明它们是由融合产生的体细胞杂种．大麦；叶细胞；细胞悬浮系；原生质体融合；体细胞杂种

关键词： Barley Leaf cell cell suspension Protoplast fusion Somatic hybrid