Project description:Arabidopsis HMA4 transformation facilitates Zn root-to-shoot translocation, but in Zn-supply dependent manner. It implies that distinct effects of HMA4 expression on Zn root:shoot distribution in transgenics grown under different Zn supply regimes results from the interaction between the transgene activity and the molecular background of the host plant. Microarray analysis was performed to compare expression profiles in leaves of transgenic and WT-plants grown in the presence of 5 µM Zn to identify the molecular mechanism underlying the difference in Zn root:shoot distribution between 35S:AtHMA4-expressing and wild-type tomato. To recognize tissue specific alterations in gene transcription in tomato leaves invoked by AtHMA4 expression, spongy paerenchyma + lower epidermis and palisade paerenchyma + upper epidermis were isolated using LCM technique. Understanding the interplay between the transgene and the endogens is crucial for planning genetic modification for Zn-biofortification purpose.
Project description:Ectopic expression of AtHMA4 in tomato modified Zn accumulation in a Zn-supply dependent manner. It suggests that HMA4-expression under a range of Zn-supply differentially modifies the expression of endogens, which contributes to different Zn-related phenotypes. To identify genes differentially regulated in 35S:AtHMA4-expressing and wild-type tomato, the transcription profiles were compared between roots of transgenic and WT-plants grown in the presence of 5 µM Zn. An effort was undertaken to recognize tissue specific alterations underlying the difference in Zn root:shoot distribution between tested plant lines and LCM technique was used to isolate epidermis+cortex and stele from roots.Understanding the interplay between the transgene and the endogens is crucial for planning genetic modification for Zn-biofortification purpose.
Project description:Ectopic expression of AtHMA4 in tomato modified Zn accumulation in a Zn-supply dependent manner. It suggests that HMA4-expression under a range of Zn-supply differentially modifies the expression of endogens, which contributes to different Zn-related phenotypes. To identify genes differentially regulated in 35S:AtHMA4-expressing and wild-type tomato, the transcription profiles were compared between roots of transgenic and WT-plants grown in the presence of 5 µM Zn. An effort was undertaken to recognize tissue specific alterations underlying the difference in Zn root:shoot distribution between tested plant lines and LCM technique was used to isolate epidermis+cortex and stele from roots.Understanding the interplay between the transgene and the endogens is crucial for planning genetic modification for Zn-biofortification purpose. 10-day old tomato plant (35S:AtHMA4 expressing and WT) were subjected to 5 µM Zn (added as ZnSo4) for 15 days. Three independent experiments were performed. At the end of each experiment 2.0 cm long root fragments from six plants (excluding 1.5 cm long apical fragments) were pooled, embedded in NEG-50 Frozen Section Medium (Thermo Scientific), and frozen in liquid nitrogen. Total RNA isolated from three batches of roots was amplified and used for three independent microarray analysis.
Project description:Arabidopsis HMA4 transformation facilitates Zn root-to-shoot translocation, but in Zn-supply dependent manner. It implies that distinct effects of HMA4 expression on Zn root:shoot distribution in transgenics grown under different Zn supply regimes results from the interaction between the transgene activity and the molecular background of the host plant. Microarray analysis was performed to compare expression profiles in leaves of transgenic and WT-plants grown in the presence of 5 µM Zn to identify the molecular mechanism underlying the difference in Zn root:shoot distribution between 35S:AtHMA4-expressing and wild-type tomato. To recognize tissue specific alterations in gene transcription in tomato leaves invoked by AtHMA4 expression, spongy paerenchyma + lower epidermis and palisade paerenchyma + upper epidermis were isolated using LCM technique. Understanding the interplay between the transgene and the endogens is crucial for planning genetic modification for Zn-biofortification purpose. 10-day old tomato plant (35S:AtHMA4 expressing and WT) were subjected to 5 µM Zn (added as ZnSo4) for 15 days. Three independent experiments were performed. At the end of each experiment the 3rd and 4th leaf (counting upwards) were collected, 1-2 mm long and 2-3 mm wide fragments were cut out from between the vascular bundles in the middle part of each leaf, embedded in NEG-50 Frozen Section Medium (Thermo Scientific), and frozen in liquid nitrogen. Total RNA isolated from three batches of leaves was amplified and used for three independent microarray analysis.
Project description:The tomato SlWRKY3 transcription factor was overexpressed in cultivated tomato (Solanum lycopersicum)and transgenic plants transcriptome was compared to that of wild-type plants.
Project description:The tomato SlWRKY3 transcription factor was overexpressed in cultivated tomato (Solanum lycopersicum)and transgenic plants transcriptome was compared to that of wild-type plants. At least 4 plants were collected for RNA extraction. The aim of the experiment was to compare transcriptomes of 35::SlWRKY3 plants and wild-type plants grown together and on MS (Murashige and Skoog) medium in vitro for 4 weeks. A technical replicate (dye swap) was conducted.
Project description:We report that Moniliophthora perniciosa biotype-S, which infects Solanaceae, manipulates the tomato Micro-Tom (MT) cytokinin (CK) metabolism. We evaluated the transcriptome of wild-type MT plants and of the transgenic line that overexpresses the Arabidopsis CYTOKININ OXIDASE-2 gene (35S::AtCKX2) inoculated or not with M. perniciosa at 12, 24, and 48 hours after inoculation (hai), and 5, 10, 20, and 30 days after inoculation (dai).