Most of the native forage species, dominant in upland grassland, were able to survive and recover from extreme drought, but with various time lags. Overall the results suggest that the wide range of interspecific functional strategies for coping with drought may enhance the resilience of upland grassland plant communities under extreme drought events.

The study of carbohydrate metabolism in perennial ryegrass (Lolium perenne L. cv. Bravo) during the first 48 h of regrowth showed that fructans from elongating leaf bases were hydrolysed first whereas fructans in mature leaf sheaths were degraded only after a lag of 1.5 h. In elongating leaf bases, the decline in fructan content occurred not only in the differentiation zone (30-60 mm from the leaf base), but also in the growth zone. Unlike other soluble carbohydrates, the net deposition rate of fructose remained positive and even rose during the first day following defoliation. The activity of fructan exohydrolase (FEH; EC 3.2.1.80) was maximal in the differentiation zone before defoliation and increased in all segments, but peaked in the growth zone after defoliation. These data strongly indicate that fructans stored in the leaf growth zone were hydrolysed and recycled in that zone to sustain the refoliation immediately after defoliation. Despite the depletion of carbohydrates, leaves of defoliated plants elongated at a significantly higher rate than those of undefoliated plants, during the first 10 h of regrowth. This can be partly attributed to the transient increase in water and nitrate deposition rate. The results are discussed in relation to defoliation tolerance.

No abstract

The aim of this study was to evaluate the putative role of the sucrosyl-galactosides, loliose [␣-d-Gal (1,3) ␣-d-Glc (1,2) ␤-d-Fru] and raffinose 6) ␣-d-Glc (1,2) ␤-d-Fru], in drought tolerance of perennial ryegrass and to compare it with that of fructans. To that end, the loliose biosynthetic pathway was first established and shown to operate by a UDP-Gal: sucrose (Suc) 3-galactosyltransferase, tentatively termed loliose synthase. Drought stress increased neither the concentrations of loliose and raffinose nor the activities of loliose synthase and raffinose synthase (EC 2.4.1.82). Moreover, the concentrations of the raffinose precursors, myoinositol and galactinol, as well as the gene expressions of myoinositol 1-phosphate synthase (EC 5.5.1.4) and galactinol synthase (EC 2.4.1.123) were either decreased or unaffected by drought stress. Taken together, these data are not in favor of an obvious role of sucrosyl-galactosides in drought tolerance of perennial ryegrass at the vegetative stage. By contrast, drought stress caused fructans to accumulate in leaf tissues, mainly in leaf sheaths and elongating leaf bases. This increase was mainly due to the accumulation of long-chain fructans (degree of polymerization Ͼ 8) and was not accompanied by a Suc increase. Interestingly, Suc but not fructan concentrations greatly increased in drought-stressed roots. Putative roles of fructans and sucrosyl-galactosides are discussed in relation to the acquisition of stress tolerance.One of the strategies employed by plants to survive drought stress includes the synthesis of protective compounds, which may act by stabilizing membranes and proteins or mediating osmotic adjustment (Bohnert et al., 1995; Hare et al., 1998; Hoekstra et al., 2001). Included among these protective compounds are the water-soluble carbohydrates (WSCs), Glc, Suc, raffinose, myoinositol, and fructans.Raffinose family oligosaccharides (RFOs) such as raffinose and stachyose accumulate during seed development and are thought to play a role in the desiccation tolerance of seeds (Blackman et al., 1992; Brenac et al., 1997). Raffinose also accumulates in vegetative tissues under drought stress (Taji et al., 2002). RFO biosynthesis requires the presence of galactinol, which is formed by galactinol synthase (GolS; EC 2.4.1.123) from UDP-Gal and myoinositol. Galactinol is the galactosyl donor for the biosynthesis of raffinose from Suc by raffinose synthase (RafS; EC 2.4.1.82). Because galactinol has not been assigned any function in plants other than acting as galactosyl donor for RFOs synthesis, GolS potentially catalyzes a metabolic key step for RFO synthesis. In a recent study, two drought-responsive GolS genes were identified among seven in Arabidopsis (Taji et al., 2002). Overexpression of one of them caused an increase in endogenous galactinol and raffinose as well as an improvement in drought tolerance.In addition to GolS, myoinositol 1-phosphate synthase (INPS; EC 5.5.1.4) is another enzyme that may control the levels of galactinol and raffinose. It repr...

Summary• An analysis of fructan structures, to increase the understanding of biosynthetic pathways and enzymology of fructan synthesis in root and leaf tissues of Lolium perenne is reported.• Fructan extracted from stubble of L. perenne plants was analyzed by high performance anion exchange chromatography and pulsed amperometric detection (HPAEC-PAD) using a new desalting technique. Structures of fructan isomers, separated up to DP16 (DP, degree of polymerization), were established by chromatographic elution times or by GC-MS.• Fructans of DP8 belonged essentially to three series: inulin series, inulin neoseries and the levan neoseries, which is/are different in glucose (terminal or internal) and linked fructose residues. High DP fructans (DP > 8) comprised 75% molecules with an internal glucose residue. They had some branch points although 1 and 6 kestotetraose could not be detected and the β (2 -6) linked fructose residues were 70 times more abundant than the β (2 -1) linked fructose residues. Roots, sheaths, leaf blades and elongating leaves accumulated similar fructans although amounts of both low and high, and types of low, DP fructans, differed.• It is proposed that fructans in L. perenne are synthesized via four enzymes: 1-SST (1-sucrose-sucrosefructosyl transferase), 1-FFT (1-fructan-fructanfructosyl transferase), 6G-FT (6-glucose-fructosyl transferase) and 6-FFT (6-fructanfructanfructosyl transferase) or 6-SFT (6-sucrose-fructanfructosyl transferase).

The relative significance of the use of stored or currently absorbed C for the growth of leaves or roots of Lolium perenne L. after defoliation was assessed by steady‐state labelling of atmospheric CO2. Leaf growth for the first two days after defoliation was to a large extent dependent on the use of C reserves. The basal part of the elongating leaves was mainly new tissue and 91% of the C in this part of the leaf was derived from reserves assimilated prior to defoliation. However, half of the sucrose in the growth zone was produced from photosynthesis by the emerged leaves. Fructans that were initially present in elongating leaf bases were hydrolysed (loss of 93 to 100%) and the resulting fructose was found in the new leaf bases, suggesting that this pool may be used to support cell division and elongation. Despite a negative C balance at the whole‐plant level, fructans were synthesized from sucrose that was translocated to the new leaf bases. After a regrowth period of 28 d, 45% of the C fixed before defoliation was still present in the root and leaf tissue and only 1% was incorporated in entirely new tissue.

Fructans, which are beta-(2,1) and/or beta-(2,6) linked polymers of fructose, are important storage carbohydrates in many plants. They are mobilized via fructan exohydrolases (FEHs). The cloning, mapping, and functional analysis of the first 1-FEH (EC 3.2.1.153) from Lolium perenne L. var. Bravo is described here. By screening a perennial ryegrass cDNA library, a 1-FEH cDNA named Lp1-FEHa was cloned. The Lp1-FEHa deduced protein has a low iso-electric point (5.22) and it groups together with plant FEHs and cell-wall type invertases. The deduced amino acid sequence shows 75% identity to wheat 1-FEH w2. The Lp1-FEHa gene was mapped at a distal position on the linkage group 3 (LG3). Functional characterization of the recombinant protein in Pichia pastoris demonstrated that it had high FEH activity towards 1-kestotriose, 1,1-kestotetraose, and inulin, but low activity against 6-kestotriose and levan. Like other fructan-plant FEHs, no hydrolase activity could be detected towards sucrose, convincingly demonstrating that the enzyme is not a classic invertase. The expression pattern analysis of Lp1-FEHa revealed transcript accumulation in leaf tissues accumulating fructans while transcript level was low in the photosynthetic tissues. The high expression level of this 1-FEH in conditions of active fructan synthesis, together with its low expression level when fructan contents are low, suggest that it might play a role as a beta-(2,1) trimming enzyme acting during fructan synthesis in concert with fructan synthesis enzymes.

Smith, K. F., Simpson, R. J., Culvenor, R. A., Oram, R. N., Humphreys, M. O., Prud'homme, M. P. (2001). The effects of ploidy and a phenotype conferring a high water-soluble carbohydrate concentration on carbohydrate accumulation, nutritive value and morphology of perennial ryegrass (Lolium perenne L.). Journal of Agricultural Science, 136, 65-74.Tetraploidy or the use of diploid genotypes with genes conferring high water-soluble carbohydrate concentrations are two mechanisms to increase the nutritive value of perennial ryegrass. This experiment compared the morphology, nutritive value and diurnal variation in water-soluble carbohydrate (WSC) concentrations of 56-day-old plants from six perennial ryegrass cultivars grown under controlled environment conditions. Three of these cultivars were diploid (Melle, Aurora and Cariad) and three were tetraploids (Meltra, Prospero and AberOnyx) which had been derived from the respective diploid cultivars. Two of the diploid cultivars (Cariad and Aurora) had previously been selected for high concentrations of water-soluble carbohydrates. The tetraploid cultivars had fewer (mean 59), larger tillers than the diploids (mean 83). However, with the exception of Melle and Meltra the dry matter yield of the diploid cultivars was not significantly different from their tetraploid derivatives. The effect of tetraploidy on WSC concentrations was dependent on the genetic background of the cultivars. Melle, which had not been previously selected for increased WSC, had a significantly lower WSC concentration than its tetraploid derivative, Meltra. However, tetraploidy did not further increase the WSC concentration in those cultivars previously selected for high WSC concentrations. WSC concentrations in the leaf of both Aurora and Melle rose by 65?70 g/kg throughout the photoperiod, suggesting that differences in the total WSC concentration of these cultivars were not due to any increase in the amount of carbon fixed by Aurora but rather due to differences in the allocation of carbon during growth and development. This experiment demonstrated that tetraploidy was not beneficial in improving the WSC concentration of perennial ryegrass when imposed on two diploid cultivars which had the genetic potential for increased WSC accumulation. However, tetraploidy significantly increased the WSC concentration and by implication the nutritive value of a cultivar derived from a perennial ryegrass cultivar with standard WSC concentrations.Peer reviewe