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1.
Production of transgenic maize plants and progeny by bombardment of hi-II immature embryos 总被引:4,自引:0,他引:4
D. D. Songstad C. L. Armstrong W. L. Petersen B. Hairston M. A. W. Hinchee 《In vitro cellular & developmental biology. Plant》1996,32(3):179-183
Summary Production of transgenic maize (Zea mays L.) callus, plants, and progeny from microprojectile bombardment of 2–5-d cultured Hi-II immature embryos is described. Histological
evidence indicates that these tissues are amenable to transformation due to surface layer cell division of the scutellum.
Two out of every 100 bombarded embryos produced transgenic callus and R0 transgenic plants were both male and female fertile. Expected segregation of transgenes was observed in progeny. The primary
advantage of bombarding these tissues is increased male and female fertility of transgenic plants compared with those produced
using long-term callus or suspension cultures. 相似文献
2.
Production of fertile transgenic maize by electroporation of suspension culture cells 总被引:17,自引:0,他引:17
Cheryl Montain Laursen Richard A. Krzyzek Christopher E. Flick Paul C. Anderson T. Michael Spencer 《Plant molecular biology》1994,24(1):51-61
Fertile, transgenic maize plants were generated by electroporation of suspension culture cells that were treated with a pectin-degrading enzyme. Electroporation of cells from two different suspension cultures, one derived from A188 X B73 and one derived from a B73-related inbred, with a plasmid containing the bar gene, resulted in high-frequency recovery of stably transformed callus lines. Plants were regenerated from thirteen transformed callus lines and transmission of bar to progeny was demonstrated. 相似文献
3.
Transient expression of GUS and anthocyanin constructs in intact maize immature embryos following electroporation 总被引:4,自引:0,他引:4
D. D. Songstad F. G. Halaka D. L. DeBoer C. L. Armstrong M. A. W. Hinchee C. G. Ford-Santino S. M. Brown M. E. Fromm R. B. Horsch 《Plant Cell, Tissue and Organ Culture》1993,33(2):195-201
Electroporation was used for the delivery and subsequent expression of GUS and anthocyanin reporter genes into intact maize immature embryos. The optimal conditions consisted of culturing immature embryos for 4 days on N6 1-100-25-Ag medium prior to electroporation (375 V/cm; 960 µF capacitance) in EPR buffer containing DNA and 0.07 M sodium glutamate at room temperature (22°C) after a 10 min heat shock at 37°C. Under these conditions, over 40 spots of GUS transient activity were observed per immature embryo. Transient gene expression after electroporation was further demonstrated using an anthocyanin construct, which is specific for expression in plant cells. 相似文献
4.
Electroporation of immature maize zygotic embryos and regeneration of transgenic plants 总被引:2,自引:0,他引:2
Using the pulse-discharging electroporation system HPES-3, we have transferred the neomycin phosphotransferase II (nptII) gene and -glucuronidase (gus) gene into mechanically-woulded immature zygotic embryo cells of an elite local maize cultivar Huanong Supersweet No. 42 and have produced transgenic maize plants. DNA hybridization and NPTII dot assay showed that the foreign genes were integrated into the genomes and expressed stably in the cells of the transgenic calluses and plants. 相似文献
5.
Meizhu Yang Vesna Djukanovic Jessica Stagg Brian Lenderts Dennis Bidney S. Carl Falco L. Alexander Lyznik 《Plant molecular biology》2009,70(6):669-679
We have demonstrated that targeted mutagenesis can be accomplished in maize plants by excision, activation, and subsequent
elimination of an endonuclease in the progeny of genetic crosses. The yeast FLP/FRT site-specific recombination system was used to excise and transiently activate the previously integrated yeast I-SceI homing endonuclease in maize zygotes and/or developing embryos. An artificial I-SceI recognition sequence integrated into genomic DNA was analyzed for mutations to indicate the I-SceI endonuclease activity. Targeted mutagenesis of the I-SceI site occurred in about 1% of analyzed F1 plants. Short deletions centered on the I-SceI-produced double-strand break were the predominant genetic lesions observed in the F1 plants. The I-SceI expression cassette was not detected in the mutant F1 plants and their progeny. However, the original mutations were faithfully
transmitted to the next generation indicating that the mutations occurred early during the F1 plant development. The procedure
offers simultaneous production of double-strand breaks and delivery of DNA template combined with a large number of progeny
plants for future gene targeting experiments. 相似文献
6.
Increasing maize seed weight by enhancing the cytoplasmic ADP-glucose pyrophosphorylase activity in transgenic maize plants 总被引:2,自引:1,他引:2
Zhangying Wang Xiaoping Chen Jianhua Wang Tingsong Liu Yan Liu Li Zhao Guoying Wang 《Plant Cell, Tissue and Organ Culture》2007,88(1):83-92
ADP-glucose pyrophosphorylase (AGPase) plays a key role in regulating starch biosynthesis in cereal seeds and is likely the
most important determinant of seed strength. The Escherichia coli mutant glgC gene (glgC16), which encodes a highly active and allosterically insensitive AGPase, was introduced into maize (Zea mays L.) under the control of an endosperm-specific promoter. Developing seeds from transgenic maize plants showed up to 2–4-fold
higher levels of AGPase activity in the presence of 5 mM inorganic phosphate (Pi). Transgenic plants with higher cytoplasmic
AGPase activity under Pi-inhibitory conditions showed increases (13–25%) in seed weight over the untransformed control. In
addition, in all transgenic maize plants, the seeds were fully filled, and the seed number of transgenic plants had no significant
difference compared with that of untransformed control. These results indicate that increasing cytoplasmic AGPase activity
has a marked effect on sink activity and, in turn, seed weight in transgenic maize plants. 相似文献
7.
K. Sukhapinda M. E. Kozuch B. Rubin-Wilson W. M. Ainley D. J. Merlo 《Plant cell reports》1993,13(2):63-68
Transgenic haploid maize (Zea mays L.) plants were obtained from protoplasts isolated from microspore-derived cell suspension cultures. Protoplasts were electroporated in the presence of plasmid DNA containing the gus A and npt II genes encoding ß-glucuronidase (GUS) and neomycin phosphotransferase II (NPT II), respectively. Transformed calli were selected and continuously maintained on kanamycin containing medium. Stable transformation was confirmed by enzyme assays and DNA. analysis. Stably transformed tissue was transferred to regeneration medium and several plants were obtained. Most plants showed NPT II activity, and some also showed GUS activity. Chromosome examinations performed on representative plants showed that they were haploid. As expected, these plants were infertile. 相似文献
8.
Production of transgenic maize from bombarded type II callus: Effect of gold particle size and callus morphology on transformation efficiency 总被引:8,自引:0,他引:8
Bronwyn R. Frame Hongyi Zhang Suzy M. Cocciolone Lyudmila V. Sidorenko Charles R. Dietrich Sue Ellen Pegg Shifu Zhen Patrick S. Schnable Kan Wang 《In vitro cellular & developmental biology. Plant》2000,36(1):21-29
Summary Here we present a routine and efficient protocol for year-round production of fertile transgenic maize plants. Type II callus
derived from maize Hi II immature zygotic embryos was transformed using the PDS 1000/He biolistic gun and selected on bialaphos.
In an effort to improve the transformation protocol, the effects of gold particle size and callus morphology on transformation
efficiency were investigated. Reducing gold particle size from 1.0 μm or 0.6 μm resulted in a significant increase in the
rate of recovery of bialaphos-resistant clones from Type II callus. The average transformation efficiency of pre-embryogenic,
early embryogenic and late embryogenic callus did not vary significantly. Rates of transformation, regeneration and fertility
achieved for Type II callus are summarized and compared to those achieved for greenhouse- and field-derived immature zygotic
embryos. 相似文献
9.
Summary Chloroplast differentiation in relation to increasing leaf age has been investigated in maize plants exposed to continuous illumination. In the young leaves the proplastids differentiate into chloroplasts containing well organized grana as well as prolamellar bodies. In the older leaves, while plastids differentiate, the prolamellar bodies are no longer detectable. Chloroplast ability to build up prolamellar bodies does not seems so much a light dependent process as it is affected by cell differentiation rate.Supported by a grant of C.N.R. 相似文献
10.
Quantitative estimation of root exudation of maize plants 总被引:6,自引:0,他引:6
Summary The rate of root exudation of maize plants was estimated by measuring the rate of denitrification in a hermetically sealed root system. While CO2 production measured in the rhizosphere results both from root respiration and microbial respiration N2O production during nitrate respiration is solely related to the amount of root exudates available for bacterial degradation. With 4 week old plants growing in quartz sand or soil root exudation amounted to 7% of the net photosynthates. Calculations revealed that about 25% of the organic matter flowing into the root system was excreted into the rhizosphere. 相似文献
11.
Agrobacterium-mediated transformation is the method of choice to engineer desirable genes into plants. Here we describe a protocol for
demonstrating T-DNA transfer from Agrobacterium into the economically important graminaceous plant maize. Expression of the T-DNA-located GUS gene was observed with high
efficiency on shoots of young maize seedlings after cocultivation with Agrobacterium. 相似文献
12.
Patricia Coello Rogelio Rodríguez Elpidio García Jorge M. Vázquez-Ramos 《Plant molecular biology》1992,20(6):1159-1168
Three different DNA polymerase activities can be resolved by passing a protein extract from 24 h imbibed maize axes through DEAE-cellulose. These activities have been numbered 1, 2 and 3, according to their elution order. One of them, DNA polymerase 2, elutes at 100–120 mM phosphates. This enzyme was further purified by passing it through Heparin-Sepharose, Sephacryl S-300 and DNA cellulose. Purification was nearly 5000-fold. The enzyme needs Mg2+, is stimulated by K+, has an optimum pH of 7.0 and its optimum temperature is 30–37 °C. Specific inhibitors for different types of polymerases, such as aphidicolin, dideoxythymidine triphosphate and N-ethyl maleimide, gave intermediate values of inhibition, making impossible the definition of the type of enzyme purified by its inhibitory pattern. SDS-PAGE indicated the presence of several bands of molecular masses of 28–40, 56 and 15 kDa. Most of these bands could be visualized when proteins from crude extracts were analyzed by western blot, using an antibody against calf thymus DNA polymerase . A high molecular mass (around 500 kDa) was calculated by western blot of native gels using the same antibody. Finally, specific activity of this enzyme increased 100-fold during maize germination whereas polymerase 3 virtually did not increase. Furthermore, immunoprecipitation experiments with the antipolymerase -antibody showed a decrease in DNA polymerase activity by 70%. The possibility that polymerase 2 is a replicative enzyme is discussed. 相似文献
13.
Biochemical features of maize tissues with different capacities to regenerate plants 总被引:4,自引:0,他引:4
Metabolic profiling using GC–MS and LC–MS analyses of soluble metabolites and cell wall bound phenolic compounds from maize calluses of different morphogenic competence revealed a number of biochemical characteristics that distinguish tissues with high plant regeneration ability from tissues that cannot efficiently regenerate plants in vitro. Maize cultures of different ages from H99 (compact type I callus) and HiII (friable type II callus) were divided into two different samples: regenerable (R) and non-regenerable (NR) based on known morphologies. Tissues from both genotypes with high morphogenic potential had higher asparagine and aspartate and indole-3-butenol concentrations, decreased sugar and DIMBOA (2,4-dihydroxy-7-methoxy-1,4-benzoxazin-3-one) concentrations, low levels of 4-aminobutyric acid (GABA) and chlorogenic acid and lower levels of feruloyl- and sinapoyl glucosides compared to NR tissues. The ether bound cell wall phenolics of tissues with high regeneration potential had higher levels of the predominant G (guaiacyl) units and lower levels of H (p-hydroxyphenyl) and S (syringyl) units and higher ferulic acid/coumaric acid and ferulic acid/diferulic acid ratios. The same trends were found with the ester-bound phenolics of HiII, however, there were only small differences between the H99 R and NR tissues. Concentrations of the major sugars, organic acids, amino acids and soluble aromatic compounds tended to increase as the time after culture initiation increased. The results show that there are differences in general metabolism, phenolic secondary compounds and cell wall composition between R and NR cell types. 相似文献
14.
Immature maize (Zea mays L.) embryos were treated with aflatoxin B1 concentrations, ranging from 0.1 g ml–1 to 25 g ml–1. Below 5 g ml–1 aflatoxin B1, root and shoot elongation was not significantly inhibited. Ultrastructurally, root tip cells showed little deterioration, except a possible diffused clearing in mitochondria and plastids. As the toxin concentration was increased above 5 gml–1, shoot, and particularly root elongation, was progressively inhibited. Associated with this, there was an apparent decrease in the ribosome population. Furthermore, membranes, particularly the vacuolar membrane, became abnormal and vacuolar distension occurred. At 20 and 25 g ml–1, these effects were exacerbated, and mitochondria and plastid structure was disrupted. At these concentrations, there was evidence of a disruption in lipid metabolism. The results are discussed in the context of known aflatoxin effects on cellular control mechanisms and ultrastructure in animal systems. 相似文献
15.
ZmHox: a novel class of maize homeobox genes 总被引:2,自引:0,他引:2
Bettina Klinge Bärbel Überlacker Christian Korfhage Wolfgang Werr 《Plant molecular biology》1996,30(3):439-453
16.
Michael E. Compton C. M. Benton D. J. Gray D. D. Songstad 《In vitro cellular & developmental biology. Plant》1992,28(4):197-201
Summary Maize (Zea mays L.) embryogenic type-II calli were grown on medium containing 0,0.1 μM ABA or 60 g/liter sucrose or both before dehydration of solitary somatic embryos under three relative humidity regimes for
up to 6 wk. Viability of dehydrated embryos after 2 wk rehydration was assessed by their ability to produce chlorophyll (greening),
roots, coleoptiles, and/or leaves. Only embryos sequentially pretreated with ABA and high sucrose remained viable after 2
wk of dehydration at 70% RH. Up to 34% of the somatic embryos survived 2 wk dehydration at 70% RH, whereas embryos dehydrated
at 50 or 90% RH exhibited reduced viability (8.7 and 0.8%, respectively). Approximately 15% of the embryos dehydrated at 70%
RH developed into plants, whereas 0.9 and 0% of embryos dehydrated at 50 and 90% RH produced plants. Three percent of maize
somatic embryos remained viable after 6 wk of dehydration at 70% RH, and 1.7% developed into plants. Embryo size influenced
the ability of maize somatic embryos to survive dehydration. Only embryos greater than 5 mm survived 2 wk dehydration at 70%
RH. 相似文献
17.
Six media compositions and different transfer times were compared in order to improve the quality of microspore derived structures and their regeneration capacity. The production of regenerated plantlets was increased by a factor of 10 by an early transfer of anthers to medium with low sugar concentration and the use of kinetin-like growth regulators. The best transfer date was 3 weeks after initial plating, onto medium containing 25 gl–1 sucrose and 1 mg l–1 kinetin. This study demonstrates that anther transfer at an early developmental stage, before macroscopic appearance of embryo-like structures, improves embryo quality and further regeneration. This production of androgenetic structures showing normal morphological features is the first step to obtain an improvement of androgenetic plant regeneration yield. 相似文献
18.
Using a battery of methylation-sensitive restriction enzymes, cytosine methylation at 23 sites in a 7.6 kb region surrounding the Alcohol dehydrogenase-1 (Adh1) gene was measured in DNA prepared from immature maize cobs. Both the 5 upstream region and the entire coding region were hypomethylated in the two alleles examined. Methylation in Adh1 is independent of changes in Mutator transposable element methylation. The role of DNA methylation in Adh1 gene regulation is discussed. 相似文献
19.
Cleavage polyembryony in maize 总被引:1,自引:0,他引:1
Summary Two types of cleavage polyembryony are described in the inbred line VIR 17 of maize. Suspensorial embryony was observed to occur spontaneously. Typical cleavage of the zygotic proembryo occurred spontaneously, but could also be induced by treating the developing caryopses with 2,4-Dichlorophenoxyacetic acid (2,4-D) on the second day after pollination. 2,4-D was active as a decorelative factor also evoking the expression of totipotency in individual proembryonal cells. 相似文献
20.
Lubomir M. Stoilov Valeria Mirkova Jordanka Zlatanova Lalio Djondjurov 《Plant cell reports》1992,11(7):355-358
In order to elucidate some features of the topological organization of DNA within the plant nucleus, DNA fragments involved in the attachment of the DNA loops to the nuclear matrix in maize were studied. The matrix-associated DNA from dry embryo and meristematic cells after extensive digestion with DNase I and high salt treatment was about 2% of the total DNA, sized within the range of 50 and 250 bp. This DNA was found to be enriched in repetitive DNA sequences, both for nuclei from dry embryo and meristematic cells. The loop size of the DNA in cells of Zea mays appeared to be between 5 and 25 kbp.Abbreviations EDTA
Diamino-ethanetetraacetic acid
- EtBr
Ethidium bromide
- LIS
Lithium diiodosalicylate
- PMSF
Phenylmethylsulfonyl fluoride
- SDS
Sodium dodecyl sulfate 相似文献