Synthetic biology is producing a paradigm shift in biotechnology based on the introduction of engineering principles in the design of new organisms by genetic modification (Check, 2005; Haseloff and Ajioka, 2009). Whereas synthetic biology has rapidly permeated microbial biotechnology, the engineering of multicelled organisms following synthetic biology principles is now emerging and is mainly driven by the so-called top-down approaches, where newly engineered genetic circuits are embedded into naturally existing organisms used as a chassis. The plant chassis offers an extraordinarily fertile ground for synthetic biology-like engineering. However, technology still faces the huge challenge of performing engineering-driven genetic designs. One of the main technological challenges of plant synthetic biology requires the construction and transfer of multigene structures to the plant genome. This is putting pressure on developing DNA assembly and transformation technologies adapted to plants. One ...

Recent studies on the root proteome of Medicago truncatula (Gaertn.) showed an induction of pathogenesis-related (PR) proteins of the class 10 after infection with the oomycete pathogen Aphanomyces euteiches (Drechs.). To get insights into the function of these proteins during the parasitic root-microbe association, a gene knockdown approach using RNAi was carried out. Agrobacterium rhizogenes-mediated transformation of M. truncatula roots led to a knockdown of the Medicago PR10-1 gene in transgenic in vitro root cultures. Proteomic analyses of the MtPr10-1i root cultures showed that MtPr10-1 was efficiently knocked down in two MtPr10-1i lines. Moreover, five additional PR-10-type proteins annotated as abscisic acid responsive proteins (ABR17s) revealed also an almost complete silencing in these two lines. Inoculation of the root cultures with the oomycete root pathogen A. euteiches resulted in a clearly reduced colonization and thus in a suppressed infection development in MtPr10-1i roots as ...

Ice plant (Mesembryanthemum crystallinum L.) is a model plant for studying salt-tolerant mechanisms in higher plants. Many salt stress-responsive ice plant genes have been identified with molecular and biochemical approaches. However, no further functional characterization of these genes in host plant due to lack of easy and effective transformation protocols. To establish efficient transformation system of ice plants, three types of ice plant materials, hypocotyl-derived callus, aseptically-grown seedlings and pot-grown juvenile plants, were used to develop Agrobacterium-mediated transformation protocols. The highest transient transformation efficiency was with 5-day-old ice plant callus co-incubated with an Agrobacterium tumefaciens at 2.5 × 109 cells mL−1 for 48 h. The 3-day-old ice plant seedlings with root tip removed were successfully infected with A. tumefaciens or A. rhizogenes, and obtained 85% and 33-100% transient transformation rates, respectively. The transient transformation assays in

Sarker, R.H. and Biswas, A. (2002) In Vitro Plantlet Regeneration and Agrobacterium-Mediated Genetic Transformation of Wheat (Triticum aestivum L.) Plant Tissue Culture, 12, 155-165.

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Read Sonication-assisted Agrobacterium-mediated transformation of the ACC gene to interfere the production of ethylene in spring Dendrobium cv.

Transformation Protocol. BY-2 (NT1) Cell Transformation with Agrobactrium. Day 1: 1. Grow up 1 ml of the Agrobacterium overnight in LB + all selective drugs. This culture may be started from frozen glycerol cultures if necessary. Day 2: 2. NT cells are used 3 days after splitting the NT cell culture. 4 ml of NT cells are required for each transformation with an additional 4 ml for the control culture which receives no bacteria. 3. 1 ul Acetosyringone (20 mM in ethanol) is added per ml of NT cells. Typically treat the whole 50 ml culture at this point and discard any that is left over when Im finished. 4. Using a 10 ml pipette, the NT cells are pipetted in and out about 20 times. This helps to induce small lesions in the cells and increases the efficiency of the transformation. 5. 75 ul of bacteria (dense growth) or 100 ul (moderate growth) are added to a petri dish containing 4 ml of NT cells (from step 4) and mixed thoroughly. REMEMBER TO INCLUDE A CONTROL HAVING NO BACTERIA. 6. Wrap plates ...

The Harvard iGarden is a venture into plant engineering. Our aim is to create a toolkit for the cultivation of a personalized garden containing features introduced through synthetic biology. In addition to a genetic fence designed to prevent the spread of introduced genetic material, we have developed three independent features to be included in this toolkit - inclusion of novel flavors, knockdown of plant allergens, and modification of petal color.. All parts are BioBrick compatible and introduced into plants through agrobacterium-mediated transformation, using existing plant vectors modified with the BioBrick multiple cloning site.. The Harvard iGarden, beyond being an application of the BioBrick system to plant engineering, is an effort to raise public awareness of synthetic biology, production of food, and how the two can intertwine. We envision the iGarden as a medium through which the non-scientist can see the power and potential of synthetic biology and apply it to everyday life. ...

Date Published: July 6, 2015. Publisher: Public Library of Science. Author(s): Anna Coll, Mandy L. Wilson, Kristina Gruden, Jean Peccoud, Martina Stromvik.. http://doi.org/10.1371/journal.pone.0132502. Abstract. Plant synthetic biology requires software tools to assist on the design of complex multi-genic expression plasmids. Here a vector design strategy to express genes in plants is formalized and implemented as a grammar in GenoCAD, a Computer-Aided Design software for synthetic biology. It includes a library of plant biological parts organized in structural categories and a set of rules describing how to assemble these parts into large constructs. Rules developed here are organized and divided into three main subsections according to the aim of the final construct: protein localization studies, promoter analysis and protein-protein interaction experiments. The GenoCAD plant grammar guides the user through the design while allowing users to customize vectors according to their needs. ...

Mungbean is an important pulse crop extensively cultivated in Southeast Asia for supply of easily digestible protein. Salinity severely limits the growth and productivity of mungbean, and weeding poses nutritional and disease constraints to mungbean cultivation. To pyramid both salt tolerance and protection against herbicide in mungbean, the AtNHX1 encoding tonoplast Na+/H+ antiporter from Arabidopsis, and bar gene associated with herbicide resistance were co-expressed through Agrobacterium-mediated transformation. Stress inducible expression of AtNHX1 significantly improved tolerance under salt stress to ionic, osmotic and oxidative stresses in transgenic mungbean plants compared to the wild type (WT) plants, whereas constitutive expression of bar provided resistance to herbicide. Compared to WT, transgenic mungbean plants grew better with higher plant height, foliage, dry mass and seed yield under high salt stress (200 mM NaCl) in the greenhouse. The improved performance of transgenic plants under

The study was carried out to evaluate the amenability of tropical inbred and hybrid maize lines, using Agrobacterium mediated transformation technique. Agrobacteriumtumefaciens strains EHA101 harbouri

The developments in transformation technology have enabled the scientists to incorporate, mutate or substitute gene(s) leading to a particular trait; advancing it to a point where only few technical limitations remain. Genotype...

The diguanylate cyclase from Agrobacterium vitis has been engineered to remove phosphodiesterase activity, allowing for production of cyclic-diGMP from guanosine triphosp

In an effort to engineer plants to be resistant to glyphosate, scientists starting looking for glyphosate resistant EPSP in the 1980s. Several methods were employed including selection, directed evolution, site-directed mutagenesis, and microbial screens. It was difficult to obtain a suitable enzyme because typically the resistant enzymes had an undesirable decrease in catalytic activity. Eventually, naturally occurring glyphosate-tolerant microbes were identified including Agrobacterium sp. Strain CP4, Achromobacter sp. Strain LBAA, and Pseudomonas sp. Strain PG2982. The enzymes isolate from these maintained good catalyctic ativity while being resistant to glyphosate (Funke). Agrobacterium sp. Strain CP4 was obtained from a glyphosate rich waste area at a glyphosate production facility. There is substantial sequence variation between these resistant enzymes and those of plants or E. coli. Other versions resistant versions of EPSP have been identified in Streptococcus pneumonia and ...

Bar gene coding Glufosinate resistance is transformed into the alfalfa variety, Caoyuan No.1, mediated by Agrobacterium tumefaciens. The leaves of transgenic plants after being screened and tested are proved to be resistant to Glufosinate. Furthermore, PCR analysis shows that bar gene has been transformed into Alfalfa.

The aim of the proposed project (in collaboration with Geneticlab S.r.l.) is to design reliable protocols to produce biomass and to induce lipid accumulation in cell cultures obtained from a plant species known as high oleic: Jatropha curcas. This species has attracted the interest of various developmental agencies in the tropics and subtropics due to its easy adaptability to semi-arid marginal sites, use of the oil as a diesel fuel substitute and its use in erosion control. J. curcas is monoecious, with male and female flowers on the same plant, and its center of origin are Mexico and Central America. In its natural area of distribution, the species is most abundant in tropical savanna and monsoon climates and in temperate climates without a dry season and with a hot summer ...

Hello researchers, , ,I am study one gene promoter and fused it to GUS in pCAMBIA1391Z, in which ,CaMV35S drives the selectable marker Hyg gene. Among the 10 independent T3 ,lines, which were confirmed by PCR, some show very strong GUS staining, ,some no, and some weak staining. , ,I knew from this paper that vectors containing 35S promoter driving ,selectable marker gene are not good for promoter study. ,The 35S promoter used in a selectable marker gene of a plant transformation ,vector affects the expression of the transgene. Planta. Volume 221, Number ,4 / June, 2005 , ,But I found that quite many published papers used vectors (like ,pCAMBIA1391, pCAMBIA1301, etc) containing 35S promoter driving selectable ,marker. , ,As the staining patterns of my promoter fusion lines are not consistent at ,al, I want to use another vector pBI121 or pCGN1547. Does anyone know where ,to get pCGN1547? ,How do you deal with those lines from 35S containing vector? CAMBIA ,suggests doing co-transformation ...

Bifunctional protein GlmU; Catalyzes the last two sequential reactions in the de novo biosynthetic pathway for UDP-N-acetylglucosamine (UDP- GlcNAc). The C-terminal domain catalyzes the transfer of acetyl group from acetyl coenzyme A to glucosamine-1-phosphate (GlcN-1-P) to produce N-acetylglucosamine-1-phosphate (GlcNAc-1-P), which is converted into UDP-GlcNAc by the transfer of uridine 5- monophosphate (from uridine 5-triphosphate), a reaction catalyzed by the N-terminal ...

Protein translocase subunit SecY; The central subunit of the protein translocation channel SecYEG. Consists of two halves formed by TMs 1-5 and 6-10. These two domains form a lateral gate at the front which open onto the bilayer between TMs 2 and 7, and are clamped together by SecE at the back. The channel is closed by both a pore ring composed of hydrophobic SecY resides and a short helix (helix 2A) on the extracellular side of the membrane which forms a plug. The plug probably moves laterally to allow the channel to open. The ring and the pore may move ...

We will carry out an empirical study on how mobility dynamics affect emigration from France. We will do so by widening the study to other population categories than simply immigrants (second generation, and expatriates). ...

is provided, focused on the role played by the different components of the virulence system. The general assessments for the establishment of efficient transformation protocols are discussed with an emphasis in the application of this methodology to monocotyledonous plants. Based on our own experience, we present the establishment of sugarcane transformation by A. tumefaciens as a model of application of this methodology to an important culture plant species, previously considered recalcitrant and inaccessible for this type of genetic manipulation ...

Agrobacterium-medieret transformation ved hjælp af en blomster-dip metode kan anvendes med succes at skabe stabile transgene linjer af...

Triplex End-Point PCR kit. The major symptoms of infection by Agrobacterium vitis are galls found on the lower trunk, near the soil line, but galls can also develop slightly below the soil surface, or extend up to 1-m height. Infected plants produce inferior shoots, and portions of the vine above the gall may die. A. vitis mainly enters the plant through wounds caused by freezing, so climatic conditions favouring freeze injury also favour infection by A. vitis. Contaminated planting material can, however, also be a source of inoculum.. Source: EPPO Bulletin 32, 367-369. ...

Agrobacterium vitis genes for 16S rRNA, tRNA-Ile, tRNA-Ala, 23S rRNA 150-b fragment, IVS, 23S rRNA 1.3-kb fragment A, partial and complete sequence, strain: NCPPB-1771, locus: ...

A enzima mio-inositol-1-fosfato sintase (MIPS1) (E.C.5.5.1.4) catalisa a conversão irreversível de D-Glicose-6-P para 1-L-mio-inositol-1-P. O mio-inositol desempenha papel de destaque no metabolismo vegetal, fornecendo inositol e inositídeos em processos metabólicos essenciais à formação do vegetal. Dentre as várias vias nas quais o mio-inositol está envolvido algumas são alvo do melhoramento vegetal, a exemplo a manipulação do conteúdo de ácido fítico e de oligossacarídeos em sementes e açúcares estruturais na parede celular. Até o presente momento, plantas MIPS silenciadas não foram caracterizadas em detalhes, principalmente em suas estruturas vegetativas. Com isso, o presente trabalho objetivou caracterizar tomateiros Moneymaker com diferentes níveis de silenciamento de MIPS1. Para a indução do silenciamento utilizou- se tranformação genética via Agrobacterium tumefaciens contendo plasmídeo pCambia com construção tipo intron hairpin de fragmentos do gene MIPS de ...

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Supplementary MaterialsSupplementary Shape 1. and viability assay had been performed. Outcomes: With this research, we display that BM-MSCs can induce the EMT development of CRC cells Delpazolid test, CRC shown the morphological features of epithelialCmesenchymal changeover after co-cultured with BM-MSCs for 72?h (Supplementary Shape 1A). To recognize whether MSC-CRC cell-cell adhesion was very important to this alteration further, three different co-culture versions had been founded. After Delpazolid 72?h co-cultivation in ibidi 31.9%, 11.730.9979, CRC+MSC, 603.8 MSC, 297) in cancer cells from co-cultivation organizations. Cancers cells underwent epithelial-mesenchymal changeover and MSC differentiated into adult cancer-associated fibroblasts (CAF) in the co-culture model In the MSC-CRC wound-healing assay, MSCs demonstrated greater flexibility than CRC cells (Supplementary Shape 1B). Besides, MSCs exhibited some morphological adjustments, including elongated phenotype, decreased adhesion, and ...

Co-cultivation of production cells at the limit of the in vitro age is an assay we offer to test for latent retroviruses for our clients. Essentially, your production cells are plated along with susceptible cell lines that exhibit stronger phenotypic CPE when the virus enters them. Thus, viruses that are expressed as non-infectious particles or exhibit low level of expression in production cells are detected ...

Co-cultivation of production cells at the limit of the in vitro age is an assay we offer to test for latent retroviruses for our clients. Essentially, your production cells are plated along with susceptible cell lines that exhibit stronger phenotypic CPE when the virus enters them. Thus, viruses that are expressed as non-infectious particles or exhibit low level of expression in production cells are detected ...

Graduates of Bu-Ali Sina University - the names, photos, skill, job, location. Information on the Bu-Ali Sina University - contacts, students, faculty, finances.

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一基本信息姓名：周虚出生年月：1965年8月15日生职称：吉林大学动物科学学院教授、博导联系方式：0431-87835142，Email:[email protected]招生学科:动物遗传育种与繁殖研究方向：动物生殖调控二研究特色1.牛、猪卵泡和早期胚胎发育调控的分子机制2.营养对母猪繁殖的影响及其内分泌和分子机制3.猪繁殖障碍检测与防治三学术任职教育部高校教学指导委员会委员中国奶业协会繁殖专业委员会副主任中国畜牧兽医学会动物繁殖分会常务理事、副秘书长吉林省畜牧兽医学会常务理事四学历和学位1981

TY - JOUR. T1 - A novel phenolic compound, chloroxynil, improves agrobacterium-mediated transient transformation in Lotus japonicus. AU - Kimura, Mitsuhiro. AU - Cutler, Sean. AU - Isobe, Sachiko. PY - 2015/7/15. Y1 - 2015/7/15. N2 - Agrobacterium-mediated transformation is a commonly used method for plant genetic engineering. However, the limitations of Agrobacterium host-plant interactions and the complexity of plant tissue culture often make the production of transgenic plants difficult. Transformation efficiency in many legume species, including soybean and the common bean, has been reported to be quite low. To improve the transformation procedure in legumes, we screened for chemicals that increase the transformation efficiency of Lotus japonicus, a model legume species. A Chemical library was screened and chemicals that increase in transient transformation efficiency of L. japonicus accession, Miyakojima MG-20 were identified. The transient transformation efficiency was quantified by ...

6.Thaw frozen competent cells on ice. Effect of DNA incubation time on NEB Express competent E. coli transformation efficiency: 50 μl of competent cells were transformed with 100 pg of pUC19 control DNA following the provided High Efficiency Transformation Protocol except DNA incubation time varied from 0 to 40 minutes. Heat shock at exactly 42°C for exactly 30 seconds. Return to Protocols End Chemically Competent Cell Production ∅∅∅∅∅ ∅∅∅∅∅ Chemical Transformation modified from NEB transformation protocol. modification of the reported protocols used for preparation of chemical competent cells of Agrobacterium (McCormac et al., 1998) and E. coli (Green and Rogers 2013), and the freeze-thaw method for the genetic transformation of A. tumefaciens (Höfgen and Willmitzer,1988). a. Protocols BH3 Project. version 1.0 Updated:1/21/2013 Store competent cells at -80°C only! Bacterial transformation, as mentioned above, means the uptake of DNA molecules through the cell wall from ...

TY - JOUR. T1 - Expression of sweet potato asparaginyl endopeptidase caused altered phenotypic characteristics in transgenic Arabidopsis. AU - Chen, Hsien Jung. AU - Wen, I. Chia. AU - Huang, Guan Jhong. AU - Hou, Wen Chi. AU - Lin, Yaw Huei. PY - 2008/4. Y1 - 2008/4. N2 - We have previously isolated an asparaginyl endopeptidase, SPAE, from senescent leaves of sweet potato (Ipomoea batatas cv. Tainong 57). Gene expression of SPAE was activated and enhanced in natural and induced senescent leaves (Chen et al., 2004). In this report the full-length SPAE cDNA was constructed in the T-DNA portion of recombinant pBI121 vector under the control of CaMV 35S promoter and transferred to Arabidopsis with Agrobacterium-mediated floral dip transformation. Three transgenic Arabidopsis plants were isolated and confirmed by kanamycin-resistance and genomic PCR amplification of SPAE. Protein gel blot also demonstrated sweet potato SPAE expression in these transgenic plants. Phenotypic analysis showed that ...

Peanut, an important oilseed crop, is gaining priority for the development of drought tolerant genotypes in recent times, since the area under drought is constantly on the rise. To achieve this, one of the important strategies is to genetically engineer the ruling peanut varieties using transcription factor regulating the expression of several downstream, abiotic-stress responsive gene(s). In this study, eight independent transgenic peanut (cv. GG20) lines were developed using AtDREB1A gene, encoding for a transcription factor, through Agrobacterium-mediated genetic transformation. The transgene insertion was confirmed in (T0) using PCR and Dot-blot analysis, while copy-number(s) was ascertained using Southern-blot analysis. The inheritance of AtDREB1A gene in individual transgenic plants (T1 and T2) was confirmed using PCR. In homozygous transgenic plants (T2), under soil-moisture deficit stress, elevated level of AtDREB1A transgene expression was observed by RT-PCR assay. The transgenic plants at 45-d

Finger millet (Eleusine corocana subsp. coracana), an important cereal in East Africa and India, is a tetraploid species with unknown genomic components. A recent cytogenetic study confirmed the direct origin of this millet from the tetraploid E. coracana subsp. africana but questioned Eleusine indica as a genomic donor. Chloroplast (ct) DNA sequence analysis using restriction fragment pattern was used to examine the phylogenetic relationships between E. coracana subsp. coracana (domesticated finger millet), E. coracana subspecies africana (wild finger millet), and E. indica. Eleusine tristachya was included since it is the only other annual diploid species in the genus with a basic chromosome number of x = 9 like finger millet. Eight of the ten restriction endonucleases used had 16 to over 30 restriction sites per genome and were informative. E. coracana subsp. coracana and subsp. africana and E. indica were identical in all the restriction sites surveyed, while the ct genome of E, tristachya ...

RESISTENZGENE (GENETIK); PROMOTOR (MOLEKULARE GENETIK); TRANSGENE PFLANZEN + GENETISCHE TRANSFORMATION (PFLANZENGENETIK); NICOTIANA (BOTANIK); DIPLOMARBEITEN UND EXAMENSARBEITEN (DOKUMENTENTYP); RESISTANCE GENES (GENETICS); PROMOTER (MOLECULAR GENETICS); TRANSGENIC PLANTS + GENETIC TRANSFORMATION (PLANT GENETICS); NICOTIANA (BOTANY); EXAMINATION PAPERS + DEGREE PAPERS (DOCUMENT TYPES ...

Jatropha curcas is an important biofuel crop due to the presence of high amount of oil in its seeds suitable for biodiesel production. Triacylglycerols (TAGs) are the most abundant form of storage oil in plants. Diacylglycerol O-acyltransferase (DGAT1) enzyme is responsible for the last and only committed step in seed TAG biosynthesis. Direct upregulation of TAG biosynthesis in seeds and vegetative tissues through overexpression of the DGAT1 could enhance the energy density of the biomass, making significant impact on biofuel production. The enzyme diacylglycerol O-acyltransferase is the rate-limiting enzyme responsible for the TAG biosynthesis in seeds. We generated transgenic Jatropha ectopically expressing an Arabidopsis DGAT1 gene through Agrobacterium-mediated transformation. The resulting AtDGAT1 transgenic plants showed a dramatic increase in lipid content by 1.5- to 2 fold in leaves and 20-30 % in seeds, and an overall increase in TAG and DAG, and lower free fatty

Adenosine diaminase deficiency is a heritable disorder caused by the absence of adenosine deaminase (ADA), an important enzyme in purine salvage pathway. The absence of ADA results in a dysfunctional immune system due to the build-up of toxic metabolites. This led Sanjeewa Singhabahu and colleagues at University of East London to produce functional human ADA in tobacco plants. They inserted a human ADA cDNA into a plant expression vector and transformed the tobacco plants through Agrobacterium-mediated transformation. Analyses confirmed the integration of the construct into the plant nuclear genome and expression of the recombinant ADA in transgenic tobacco leaves. Further analysis have revealed that the size of the recombinant ADA is similar with the human ADA. ADA-specific activities of between 0.001 and 0.003 units per mg total soluble protein were measured in crude extracts collected from transgenic tobacco plant leaves.. Read the research article at ...

We have achieved efficient transformation system for forage-type tall fescue plants by Agrobacterium tumefaciens. Mature seed-derived embryogenic calli were infected and co-cultivated with each of three A. tumefaciens strains, all of which harbored a standard binary vector pIG121Hm encoding the neomycin phosphotransferase II (NPTII), hygromycin phosphotransferase (HPT) and intron-containing |TEX|$\beta$|/TEX|-glucuronidase (intron-GUS) genes in the T-DNA region. Transformation efficiency was influenced by the A. tumefaciens strain, addition of the phenolic compound acetosyringone and duration of vacuum treatment. Of the three A. tumefaciens strains tested, EHA101/pIG121Hm was found to be most effective followed by GV3101/pIG121Hm and LBA4404/pIG121Hm for transient GUS expression after 3 days co-cultivation. Inclusion of 100 |TEX|$\mu$|/TEX|M acetosyringone in both the inoculation and co-cultivation media lead to an improvement in transient GUS expression observed in targeted calli. Vacuum treatment

Finger millet (Eleusine corocana) is a staple of some communities living on the Jos Plateau of north-central Nigeria. Having reported in 2003 on the nutrient content of the more-common tan-colored finger millet, we were interested in knowing the content of essential amino acids, fatty acids and minerals and trace elements of a dark, rust-colored finger millet called black millet that is also cul

kezhvaragu/finger millet - the centre of attraction First in the series of Dosais would be Kezhvaragu Dosai. Kezhvaragu in Tamil, is Finger Millet in English and locally Ragi in Karnataka, the southern Indian state which is the largest producer of Finger Millet in India. It is also called Keppai in Tamil. I give…

Agrobacterium infection, which is widely used to generate transgenic plants, is often accompanied by T-DNA-linked mutations and transpositions in flowering plants. It is not known if Agrobacterium infection also affects the rates of point mutations, somatic homologous recombinations (SHR) and frame-shift mutations (FSM). We examined the effects of Agrobacterium infection on five types of somatic mutations using a set of mutation detector lines of Arabidopsis thaliana. To verify the effect of secreted factors, we exposed the plants to different Agrobacterium strains, including wild type (Ach5), its derivatives lacking vir genes, oncogenes or T-DNA, and the heat-killed form for 48 h post-infection; also, for a smaller set of strains, we examined the rates of three types of mutations at multiple time-points. The mutation detector lines carried a non-functional β-glucuronidase gene (GUS) and a reversion of mutated GUS to its functional form resulted in blue spots. Based on the

Background Agrobacterium tumefaciens has long been known to transform plant tissue in nature as part of its infection process. This natural mechanism has been utilised over the last few decades in laboratories world wide to genetically manipulate many species of plants. More recently this technology has been successfully applied to non-plant organisms in the laboratory, including fungi, where the plant wound hormone acetosyringone, an inducer of transformation, is supplied exogenously. In the natural environment it is possible that Agrobacterium and fungi may encounter each other at plant wound sites, where acetosyringone would be present, raising the possibility of natural gene transfer from bacterium to fungus. Methodology/Principal Findings We investigate this hypothesis through the development of experiments designed to replicate such a situation at a plant wound site. A. tumefaciens harbouring the plasmid pCAMDsRed was co-cultivated with the common plant pathogenic fungus Verticillium albo-atrum

Agrobacterium-mediated transformation and direct gene transfer using the gene gun (microparticle -bombardment) are the two most widely used methods for plant genetic modification. The Agrobacterium method has been successfully practiced in dicots for

Transient genetic transformation of plant organs is an indispensable way of studying gene function in plants. This study was aimed to develop an optimized system for transient Agrobacterium-mediated transformation of the Arabidopsis leaves. The beta-glucuronidase (GUS) reporter gene was employed to evaluate growth and biochemical parameters that influence the levels of transient expression. The effects of plant culture conditions, Agrobacterial genetic backgrounds, densities of Agrobacterial cell suspensions, and of several detergents were analyzed. We found that optimization of plant culture conditions is the most critical factor among the parameters analyzed. Higher levels of transient expression were observed in plants grown under short day conditions (SDs) than in plants grown under long day conditions (LDs). Furthermore, incubation of the plants under SDs at high relative humidity (85-90%) for 24 h after infiltration greatly improved the levels of transient expression. Under the optimized ...

Ragi Peas Paratha - a healthy, gluten-free Indian flatbread made from ragi flour/finger millet, stuffed with boiled peas, ginger, garlic and few other spices. Ragi / Finger millet is a diabetic friendly whole grain and is packed with essential nutrients like Vitamin C, E, B- complex, iron and calcium. This gluten-free grain is known for…

Agrobacterium-mediated transformation is the most widely used technique for generating transgenic plants. However, many crops remain recalcitrant. We found that an Arabidopsis myb family transcription factor (MTF1) inhibited plant transformation susceptibility. Mutating MTF1 increased attachment of several Agrobacterium strains to roots and increased both stable and transient transformation in both susceptible and transformation-resistant Arabidopsis ecotypes. Cytokinins from Agrobacterium tumefaciens decreased the expression of MTF1 through activation of the cytokinin response regulator ARR3. Mutating AHK3 and AHK4, genes that encode cytokinin-responsive kinases, increased the expression of MTF1 and impaired plant transformation. Mutant mtf1 plants also had increased expression of AT14A, which encodes a putative transmembrane receptor for cell adhesion molecules. Plants overexpressing AT14A exhibited increased susceptibility to transformation, whereas at14a mutant plants exhibited decreased attachment

Hot pepper is one of the economically important crops in Asia. A large number of gene sequences, including expressed sequence tag (EST) and genomic sequences are publicly available. However, it is still a daunting task to determine gene function due to difficulties in genetic modification of a pepper plants. Here, we show the application of the virus-induced gene silencing (VIGS) repression for the study of 459 pepper ESTs selected as non-host pathogen-induced cell death responsive genes from pepper microarray experiments in Nicotiana benthamiana. Developmental abnormalities in N. benthamiana plants are observed in the 32 (7%) pepper ESTs-silenced plants. Aberrant morphological phenotypes largely comprised of three groups: stunted, abnormal leaf, and dead. In addition, by employing the combination of VIGS and Agrobacterium-mediated transient assays, we identified novel pepper ESTs that involved in Bax or INF1-mediated cell death responses. Silencing of seven pepper ESTs homologs suppressed Bax or INF1

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Fluorescent nanodiamonds (FNDs) are promising nanoprobes, owing to their stable and magnetosensitive fluorescence. Therefore they can probe properties as magnetic resonances, pressure, temperature or strain. The unprecedented sensitivity of diamond defects can detect the faint magnetic resonance of a single electron or even a few nuclear spins. However, these sensitivities are only achieved if the diamond probe is close to the molecules that need to be detected. In order to utilize its full potential for biological applications, the diamond particle has to enter the cell. Some model systems, like HeLa cells, readily ingest particles. However, most cells do not show this behavior. In this article we show for the first time generally applicable methods, which are able to transport fluorescent nanodiamonds into cells with a thick cell wall. Yeast cells, in particular Saccharomyces cerevisiae, are a favored model organism to study intracellular processes including aging on a cellular level. In order ...

The objectives of the study were to evaluate the yield performance of each genotype in relation to each environment, to examine the possible existence of different mega environments and to identify the winning genotype for each mega environment using GGE bi-plot method. Ten finger millet varieties which were for the experiment. The experiment was laid out using randomized complete block design with four replications in all testing sites. The plot size was four harvestable rows and spacing between rows was 40cm with the length of 5m long. All management practices such as cultivation, fertilization and weeding were done based on the recommendations for each test sites. Data for all relevant agronomic traits were collected, but only plot yield data converted to qt/ha was subjected to statistical analysis. The mean grain yield of the varieties was ranged from 18.9qt/ha to 15.3qt/ha. The highest mean was harvested from the variety Gudetu (18.9qt/ha) and the lowest from variety Meba (15.3qt/ha).Out of ten

Finger millet health benefits include weight loss, diabetes control, stronger bones, better digestion, cancer prevention, etc. Learn the recipes to include finger miller in diet.

Study was intended for the development of ready to eat chicken snacks incorporating finger millet flour, which could be stored at ambient temperature.

A novel system for detection of gene transfer between A. tumefaciens and mammalian cells was established. Using this system, Agrobacterium was found to be able to deliver DNA located on its plasmid and chromosome into human cells. Agrobacterium is actively involved in this process; such a kind of DNA transfer might occur widely between bacteria and mammalian cells. The frequency of such a gene transfer was approximately 10-4-10-5 transformants per recipient. Our data showed that this DNA transfer is dependent upon Agrobacterium and independent of its virulence genes. The polymerization of actin played an important role in Agrobacterium internalization and subsequent gene transfer into mammalian cells. In addition, an Agrobacterium sensor protein ChvG was found to be important for the gene transfer into both plant and mammalian cells. The data suggested that ChvG was involved in the regulation of acid-inducible genes and might function as a global sensor protein that can directly or indirectly ...