Drought induces fructan synthesis and 1-SST (sucrose: sucrose fructosyltransferase) in roots and leaves of chicory seedlings ( Cichorium intybus L.)

Roover, Joke De; Vandenbranden, Kathleen; Laere, André Van; Ende, Wim Van den

doi:10.1007/s004250050683

Cited by 128 publications

(88 citation statements)

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“…For enzyme activity measurements, representative samples of ®eld-grown sugar beet plants (roots, petioles and leaves without the major vein) were collected weekly between June 21 and October 5, and samples were stored at À808C for later analysis. Sugar beet seedlings and young plants were also grown in a controlled growth chamber as described by De Roover et al (2000). A number of 6-day-old seedlings and 1-month-old plants were harvested.…”

Section: Plant Materialsmentioning

confidence: 99%

Unexpected presence of fructan 6‐exohydrolases (6‐FEHs) in non‐fructan plants: characterization, cloning, mass mapping and functional analysis of a novel ‘cell‐wall invertase‐like’ specific 6‐FEH from sugar beet (Beta vulgaris L.)

et al. 2003

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SummaryAbout 15% of¯owering plant species synthesize fructans. Fructans serve mainly as reserve carbohydrates and are subject to breakdown by plant fructan exohydrolases (FEHs), among which 1-FEHs (inulinases) and 6-FEHs (levanases) can be differentiated. This paper describes the unexpected ®nding that 6-FEHs also occur in plants that do not synthesize fructans. The puri®cation, characterization, cloning and functional analysis of sugar beet (Beta vulgaris L.) 6-FEH are described. Enzyme activity measurements during sugar beet development suggest a constitutive expression of the gene in sugar beet roots. Classical enzyme puri®ca-tion followed by in-gel trypsin digestion and mass spectrometry (quadruple-time-of-¯ight mass spectrometry (Q-TOF) MS) led to peptide sequence information used in subsequent RT-PCR based cloning. Levantype fructans (b-2,6) are the best substrates for the enzyme, while inulin-type fructans (b-2,1) and sucrose are poorly or not degraded. Sugar beet 6-FEH is more related to cell wall invertases than to vacuolar invertases and has a low iso-electric point (pI ), clearly different from typical high pI cell wall invertases. Poor sequence homology to bacterial or fungal FEHs makes an endophytic origin highly unlikely. The functionality of the 6-FEH cDNA was further demonstrated by heterologous expression in Pichia pastoris. As fructans are absent in sugar beet, the role of 6-FEH in planta is not obvious. Like chitinases and bglucanases hydrolysing cell-surface components of fungal plant pathogens, a straightforward working hypothesis for further research might be that plant 6-FEHs participate in hydrolysis (or prevent the formation) of levan-containing slime surrounding endophytic or phytopathogenic bacteria.

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Section: Plant Materialsmentioning

confidence: 99%

Unexpected presence of fructan 6‐exohydrolases (6‐FEHs) in non‐fructan plants: characterization, cloning, mass mapping and functional analysis of a novel ‘cell‐wall invertase‐like’ specific 6‐FEH from sugar beet (Beta vulgaris L.)

et al. 2003

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“…This happens when the photosynthetic capacity of the leaves becomes higher than what is needed for leaf growth and maintenance. De Roover et al [75] demonstrated that 1-SST activity and fructan biosynthesis are initiated when growth of chicory seedlings is arrested by drought. Direct proof of the involvement of sucrose in fructan biosynthetic gene induction comes from incubation of detached leaves in sucrose solutions [36,76], resulting in fructan biosynthesis.…”

Section: Regulation Of Fructan Metabolismmentioning

confidence: 99%

Fructan Biosynthetic and Breakdown Enzymes in Dicots Evolved From Different Invertases. Expression of Fructan Genes Throughout Chicory Development

Ende

Michiels

Roover

et al. 2002

The Scientific World JOURNAL

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Fructans are fructose-based oligo-and polymers that serve as reserve carbohydrates in many plant species. The biochemistry of fructan biosynthesis in dicots has been resolved, and the respective cDNAs have been cloned. Recent progress has now succeeded in elucidating the biochemistry and molecular biology of fructan biodegradation in chicory, an economically important species used for commercial inulin extraction. Unlike fructan biosynthetic genes that originated from vacuolar-type invertase, fructan exohydrolases (FEHs) seem to have evolved from a cell-wall invertase ancestor gene that later obtained a low iso-electric point and a vacuolar targeting signal. Expression analysis reveals that fructan enzymes are controlled mainly at the transcriptional level. Using chicory as a model system, northern analysis was consistent with enzymatic activity measurements and observed carbohydrate changes throughout its development.KEY WORDS: 1-FEH, 1-FFT, 1-SST, Chicory (Cichorium intybus L.), fructan, fructosyltransferase, inulin, invertase DOMAINS: agronomy, biotechnology, plant sciences, molecular biology, gene expression INTRODUCTIONFructans occur as a reserve carbohydrate in about 15% of flowering plant species [1]. Fructans are defined as oligo-or polymeric carbohydrates in which fructosyl linkages predominate [2]. They Van den Ende et al.: Fructan Enzymes in Chicory TheScientificWorldJOURNAL (2002) 2, 1281-1295 1282 are built up by further adding fructofuranosyl units (β(2→1) or β(2→6) linkages) to three basic trisaccharides (1-kestose, 6-kestose, and neokestose), which themselves are synthesized by linking a fructose moiety to one of the three primary hydroxyl groups of sucrose [3]. Dicotyledonous species store inulin-type fructan consisting of linear β(2→1) linked fructofuranosyl units. The most extensively studied species include Jerusalem artichoke (Helianthus tuberosus) and chicory (Cichorium intybus), which belong to the Asteraceae. However, inulin-type fructans also occur in some other dicot families (Boraginaceae, Campanulaceae). Levan-type fructan consists of linear β(2→6) linked fructofuranosyl units. More complex and branched fructan types, including graminan or mixed-type fructan as well as neokestose-based fructans, are common in monocotyledonous species [4]. Fructans are used for food and non-food applications [5]. So far, chicory is the only crop that is used for commercial fructan extraction. During the last decade, annual inulin production from chicory roots increased from 1,000 to more than 100,000 tons. Therefore, this review mainly focuses on inulin-type fructans and on inulin metabolism in chicory roots.Besides their function as a reserve carbohydrate, fructans might fulfill other, perhaps more specific, roles in plants. Some data suggest a correlation with drought resistance, frost tolerance, and osmoregulation [1,6,7]. The molecular mechanism explaining the protective function of fructans is still obscure. However, it has recently been demonstrated that fructans stabilize liposomes during free...

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“…The main storage sites of fructans in grasses are the leaf sheath and, to a lesser extent, the leaf base (Housley and Volenec, 1988;MorvanBertrand et al, 2001;Morvan et al, 1997;Roth et al, 1997;Volenec, 1986). Fructans have been implicated in conferring tolerance to drought and cold stress (De Roover et al, 2000;Housley and Pollock, 1993;Pilon-Smits et al, 1995;Thomas and James, 1999;Volaire and Lelievre, 1997). Their synthesis occurs de novo in carbohydrate accumulating tissue, with sucrose as the major or sole precursor (Cairns and Pollock, 1988a;Cairns et al, 1999;Wagner et al, 1983).…”

Section: Introductionmentioning

confidence: 99%

Cloning and characterization of a putative fructosyltransferase and two putative invertase genes from the temperate grass Lolium temulentum L.

Gallagher

Cairns

Pollock

2004

Journal of Experimental Botany

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The invertases of Lolium temulentum have been characterized at the enzyme level. However, studies on the expression of the genes coding for these enzymes have been lacking. To elucidate the role of acid`invertase-like' genes in sucrose metabolism and carbon partitioning in Gramineae further, threè invertase-like' homologous clones were isolated from L. temulentum cDNA expression libraries based on leaf tissue, using maize soluble invertase probes. The effect of developmental stage and alterations in carbohydrate status on the expression and tissue distribution of these genes was investigated. The three highly homologous genes (Inv 1:2, Inv 1:4, and FT 2:2) show different patterns of expression and different tissue distribution. Inv 1:2 was predominantly expressed in root tissue. Expression increased during the dark in root and tiller base tissue. Minimal variations in gene expression were observed in leaf tissue following changes in carbohydrate status. Inv 1:4 was predominantly expressed in tiller bases, leaf sheath, and leaf base, with increased expression in tissue samples in the dark period. FT 2:2 was also predominantly expressed in tiller bases, leaf sheath, and leaf base. Higher expression was observed in leaf tissue following increases in carbohydrate content, in a manner that paralleled the regulation and spatial occurrence of fructan in the leaf tissue. Whilst invertases and fructosyltransferases are dif®cult to distinguish at the level of the whole sequence, analysis of 5¢ sequence and speci®c amino acids allows discrimination which correlates with patterns of expression within the tissue. Based on expression patterns and sequence characteristics, it is proposed that Inv 1:2 and Inv 1:4 code for soluble acid invertases, whilst FT 2:2 codes for a fructosyltransferase.

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Drought induces fructan synthesis and 1-SST (sucrose: sucrose fructosyltransferase) in roots and leaves of chicory seedlings ( Cichorium intybus L.)

Cited by 128 publications

References 35 publications

Unexpected presence of fructan 6‐exohydrolases (6‐FEHs) in non‐fructan plants: characterization, cloning, mass mapping and functional analysis of a novel ‘cell‐wall invertase‐like’ specific 6‐FEH from sugar beet (Beta vulgaris L.)

Unexpected presence of fructan 6‐exohydrolases (6‐FEHs) in non‐fructan plants: characterization, cloning, mass mapping and functional analysis of a novel ‘cell‐wall invertase‐like’ specific 6‐FEH from sugar beet (Beta vulgaris L.)

Fructan Biosynthetic and Breakdown Enzymes in Dicots Evolved From Different Invertases. Expression of Fructan Genes Throughout Chicory Development

Cloning and characterization of a putative fructosyltransferase and two putative invertase genes from the temperate grass Lolium temulentum L.

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