Unknown,Transcriptomics,Genomics,Proteomics

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Gene expression analysis of in vitro cultured tomato plantlets

ABSTRACT: Plants growth and development is highly dependent on the environment. During there lifespan, plants are submitted to changing environmental conditions that requires constant metabolic adjustments. In particular, in vitro cultured plantlets are exposed, throughout micropropagation, to unique growth conditions characterized by distinctive levels of nitrogen, exogenous sugar, light and atmospheric conditions. To resume growth upon transfer ex vitro, in vitro plantlets must acclimatize to natural conditions. In this study, the impact of the in vitro environment on the expression profile of tomato (Solanum Lycopersicum) plantlets was analysed using the microarray and real-time PCR methods. After 20 days of growth in vitro, a significant increase in expression of genes linked to photosynthesis, nitrogen metabolism and salicylic acid signaling pathways was observed. By opposition, the expression of transcripts associated to carbohydrate metabolism and jasmonic acid signaling pathway were reduced. Also, the expression of reactive oxygen scavenging (ROS) enzymes in the mitochondrias, chloroplasts and cytosol were altered in in vitro cultured tomato plantlets. More precisely, the in vitro environment led to increased photosynthetic gene expression, lower sink strength, up-regulation of nitrogen assimilation and induced a specific stress response in tomato plantlets. Our results suggest that the presence of exogenous sugar is an important environmental cue that governs the modified cell ROS pattern, defense-related genes modulation and abnormal physiology of in vitro plantlets. Keywords: environmental challenge, growth condition comparison, expression profile Three biological replicates consisting of three individual plantlets grown in different vessels (in vitro) or pots (ex vitro) were used for the competitive hybridization. To reduce possible bias due to the difference in the properties of both dyes, two technical replicates consisting of dye swap were realized. The complete experiment required six microarrays.

ORGANISM(S): Solanum lycopersicum

SUBMITTER: Yves Desjardins

PROVIDER: E-GEOD-10153 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:In Cichorium intybus, macroscopic and histological modifications were identified during in vitro cell reactivation and morphogenesis events. The relationships between these modifications and transcript profiles of genes were investigated. Two chicory genotypes derived from the Hungarian landrace Koospol, were tested: K59 is a responsive genotype (embryogenic), as it undergoes cell de- and re-differentiation that leads to somatic embryogenesis (SE). C15 is an unresponsive genotype (nonembryogenic), unable to express this morphogenetic pattern. Previously studies showed that addition of beta-D-glucosyl Yariv reagent (BGlcY), a synthetic phenyl-glycoside that specifically binds arabinogalactan-proteins (AGPs), to the culture medium blocked somatic embryogenesis in chicory root explants in a concentration-dependent manner with complete inhibition of induction occurring at 250 ÂµM of BGlcY (Chapmann et al. 2000a). A putative AGP (DT212818) encoding gene was previously found significantly over-expressed in the K59 as compared to the C15 and was suggested to be involved in chicory re-differentiation. To better understand the cell reactivation events, a treatment with BGlcY was applied in order to precipitate AGPs in muro and to block the in vitro cell re-differentiation processes. The two different genotypes together with BGlcY represented an original tool to discriminate cell reactivation events from cell fate determination. Microscopy, biochemical and transcriptome analyses showed that in vitro SE in chicory was blocked during BGlcY-treatment but cell reactivation still occurred. Consequently, AGP (DT212818) seems to be only important in cell fate determination leading to SE commitment. Gene expression profiles showed in addition to the AGP, other proteins could play a role in SE determination: 26S proteasome AAA ATPase subunit 6 (RPT6), remorin (REM), metallothionein 1 (MT1), 3-hydroxy-3-methylglutaryl-CoA reductase (HMG-CoA reductase). Cell wall events during BGlcY-treatment were observed and discussed. Gene expression of K59 explants induced (d4) was compared with gene expression of K59 explants without induction (d0). Gene expression of K59 explants induced in the presence of BGlcY (Y+; d4) was next compared with gene expression of K59 explants induced in the absence of BGlcY (Y-; d4). Similar comparisons were made for C15 leaf explants. To link up both genotypes, direct comparison was done between gene expression of K59 explants without induction (d0) and C15 explants without induction (d0). Three biological replicates were used for each analysed condition. Dye labelling for each paired sample was reversed in two subsequent individual hybridizations giving rise to a total of six hybridizations per condition except for the K59_D0_vs_C15_D0 condition.

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Gene expression analysis of in vitro cultured tomato plantlets

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