AtLTP1 luciferase expression during carrot somatic embryogenesis

Toonen, M.A.J.; Verhees, John; Schmidt, E.D.L.; Kammen, A. van; Vries, S.C. de

doi:10.1046/j.1365-313x.1997.12051213.x

Cited by 38 publications

(23 citation statements)

References 15 publications

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“…The Arabidopsis LTP1 promoter was isolated as a 1,150-bp HinDIII/BamHI fragment from pMT121 (Toonen et al, 1997) and used to replace the CaMV 35S promoter in pBIN 35S::mgal4:vp16 (J. Haseloff, unpublished data; here named ACT 35S; pSDM1600). The resulting construct was named ACT LTP1 (pSDM7019).…”

Section: Dna Cloningmentioning

confidence: 99%

Diphtheria Toxin-Mediated Cell Ablation Reveals Interregional Communication during Arabidopsis Seed Development

et al. 2003

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Fertilization of the female gametophyte in angiosperm plants initiates a process of coordinated development of embryo, endosperm, and seed coat that ensures the production of a viable seed. Mutant analysis has suggested that communication between the endosperm and the seed coat is an important determinant in this process. In addition, cell groups within the embryo, derived from the apical and from the basal cell, respectively, after zygote division, concertedly establish a functional root meristem, and cells in the apical region of the embryo are hypothesized to repress cell divisions in the basal cell-derived suspensor. The available evidence for these interregional communication events mostly relies on the analysis of mutant phenotypes in Arabidopsis. To provide independent and direct evidence for communication events, we used conditional domain-specific expression of the diphtheria toxin A chain (DTA) in developing Arabidopsis seeds. By using a collection of cell-or tissue-type-specific promoters, we show that the mGAL4:VP16/UAS two-component gene expression allows reliable spatiotemporal and conditional expression of the GFP:GUS reporter and the DTA gene in the developing embryo and endosperm. Expression of DTA in the protoderm of the embryo proper led to excessive proliferation of suspensor cells, sometimes resulting in the formation of secondary embryos. Endosperm-specific expression of DTA caused complete cessation of seed growth, followed by pattern defects in the embryo and embryo arrest. Taken together, the results presented here substantiate the evidence for and underline the importance of interregional communication in embryo and seed development and demonstrate the usefulness of conditional toxin expression as a method complementary to phenotypic analysis of developmental mutants.Seed development in higher plants is characterized by the coordinated development of distinct tissues. Seed tissues mainly arise from cells and tissues of the female gametophyte that are formed before fertilization. The multinucleate female gametophyte is enclosed by several sporophytic maternal cell layers that constitute the ovule. In angiosperms, double fertilization by fusion of the egg cell and the central cell of the female gametophyte with the two sperm cells delivered by the pollen tube generates the diploid embryo and triploid endosperm, respectively. The embryo and endosperm both develop within the confines of the maternal tissue, now referred to as the seed coat, but each follows a different developmental program (for review on embryo and endosperm development, see Jü rgens and Mayer, 1994; Berger, 2003, respectively). Within the embryo, a complex but precise pattern of organs and cell types is laid down from a single cell, the zygote (Jü rgens and Mayer, 1994). Genetic controls are required to establish the embryo pattern and to ensure the proper initiation and relative positioning of distinct cell groups and organs, such as the meristems and vasculature. The endosperm first undergoes a series of synchronized ...

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Section: Dna Cloningmentioning

confidence: 99%

Diphtheria Toxin-Mediated Cell Ablation Reveals Interregional Communication during Arabidopsis Seed Development

et al. 2003

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“…The AtLTP1 promoter region of 1.2 kb was released from the vectors pMT121 (Toonen et al 1997) by digestion with HindIII/BamHI, filled in with Klenow and replaced the HincII/SmaI digested 35S promoter in the pRT105 vector (Hecht et al 2001). The AtLTP::AtSERK1::pA fusion was PCR amplified with primers RT1(TCCCCCGGG-GGAAGCTTGCATGCCTG) and RT2 (TCCCCCGG-GGGACTGGATTTTGGTT) containing SmaI restriction sites.…”

Section: Atserk1 Ectopic Expression Constructs and Transformation Of mentioning

confidence: 99%

“…The AtLTP::AtSERK1::pA fusion was PCR amplified with primers RT1(TCCCCCGGG-GGAAGCTTGCATGCCTG) and RT2 (TCCCCCGG-GGGACTGGATTTTGGTT) containing SmaI restriction sites. The PCR fragments were then subcloned into an SmaI site of the pMOG800 binary vector (Toonen et al 1997). The construct was verified by sequencing and used to transform Arabidopsis ecotype WS plants.…”

Section: Atserk1 Ectopic Expression Constructs and Transformation Of mentioning

confidence: 99%

Use of the SSLP-based method for detection of rare apomictic events in a sexual AtSERK1 transgenic Arabidopsis population

Kantama

Lambert

et al. 2006

Sex Plant Reprod

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Here we present a screening method to evaluate the potential of genes to transfer aspects of apomixis into sexual crop plants. Based on the assumption that an apomictic progeny is an exact genetic replica of the mother plant we employed a set of single sequence length polymorphism (SSLP) markers to identify individuals displaying heterozygosity fixation in segregating sexual populations as an indication of rare apomictic events. Here we present the results of such a study using the Arabidopsis thaliana SOMATIC EMBRYOGENE-SIS RECEPTOR KINASE 1 (AtSERK1) gene expressed under the control of the AtLTP1 promoter in sexual Arabidopsis plants. In one of the three tested F2 transgenic populations expressing the AtLTP1::At-SERK1 construct we observed two plants with heterozygosity maintenance for the full set of SSLP markers indicating a possible clonal inheritance. However, as their offspring revealed a close to binomial segregation for a number of heterozygous loci, it was concluded that these two putative apomictic plants either lost their clonal ability in the next generation or resulted from incidental recombination events displaying the genotype of the parent.

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“…Expression of LTPs is a wellknown marker of somatic embryo induction (Sterk et al, 1991;Sabala et al, 2000). It is also a marker for embryo differentiation, as it has been detected in the protoderm of developing somatic and zygotic embryos (Vroemen et al, 1996;Toonen et al, 1997). Taken together, a correct expression of ltp genes is required for normal embryo development.…”

Section: Introductionmentioning

confidence: 98%

Generation of Recombinant Antibodies against Orchardgrass Acidic nsLTP-Like Proteins

Rakleova

Tsacheva

Petkova

et al. 2008

Zeitschrift Für Naturforschung C

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Embryogenic and non-embryogenic suspension cultures of orchardgrass (Dactylis glomerata L.) secreted into the culture medium a set of proteins, among which low molecular mass (11/12 kDa) proteins were found. However, only the 11/12 kDa proteins from the embryogenic suspension cultures reacted specifically with an antiserum raised against the carrot EP2 non-specific lipid transfer protein (nsLTP). Two-dimensional (2-D) electrophoretic analysis revealed that the extracellular nsLTP-like proteins from the embryogenic lines were acidic proteins, with pI values ranging between 4.3 and 6.4, and the 11/12 kDa proteins of the nonembryogenic lines were basic ones (pI 8Ð9.3). This is only the second case to report on the accumulation of extracellular acidic nsLTP-like proteins in the culture medium during somatic embryogenesis. A naïve phage display Griffin1. library was used to select single-chain phage antibodies, which specifically bind to acidic nsLTP-like proteins. Nine phage clones were selected after four rounds of biopanning of the target proteins blotted on a nitrocellulose membrane. Three soluble monoclonal single-chain phage antibodies, expressed in the non-suppressor E. coli strain HB2151, were purified by metal affinity chromatography and found to be highly specific for the acidic nsLTP-like proteins from the embryogenic suspension cultures. The application of the selected monoclonal antibodies for localization and elucidation of the role of the acidic nsLTP-like proteins in vivo is discussed.

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AtLTP1 luciferase expression during carrot somatic embryogenesis

Cited by 38 publications

References 15 publications

Diphtheria Toxin-Mediated Cell Ablation Reveals Interregional Communication during Arabidopsis Seed Development

Diphtheria Toxin-Mediated Cell Ablation Reveals Interregional Communication during Arabidopsis Seed Development

Use of the SSLP-based method for detection of rare apomictic events in a sexual AtSERK1 transgenic Arabidopsis population

Generation of Recombinant Antibodies against Orchardgrass Acidic nsLTP-Like Proteins

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