Starch Reserves Research Articles

Characterization of the functional interactions of plastidial starch phosphorylase and starch branching enzymes from rice endosperm during reserve starch biosynthesis

Functional interactions of plastidial phosphorylase (Pho1) and starch branching enzymes (BEs) from the developing rice endosperm are the focus of this study. In the presence of both Pho1 and BE, the same branched primer molecule is elongated and further branched almost simultaneously even at very low glucan concentrations present in the purified enzyme preparations. By contrast, in the absence of any BE, glucans are not, to any significant extent, elongated by Pho1. Based on our in vitro data, in the developing rice endosperm, Pho1 appears to be weakly associated with any of the BE isozymes. By using fluorophore-labeled malto-oligosaccharides, we identified maltose as the smallest possible primer for elongation by Pho1. Linear dextrins act as carbohydrate substrates for BEs. By functionally interacting with a BE, Pho1 performs two essential functions during the initiation of starch biosynthesis in the rice endosperm: First, it elongates maltodextrins up to a degree of polymerization of at least 60. Second, by closely interacting with BEs, Pho1 is able to elongate branched glucans efficiently and thereby synthesizes branched carbohydrates essential for the initiation of amylopectin biosynthesis.

Seed dormancy and germination

Reproduction is a critical time in plant life history. Therefore, genes affecting seed dormancy and germination are among those under strongest selection in natural plant populations. Germination terminates seed dispersal and thus influences the location and timing of plant growth. After seed shedding, germination can be prevented by a property known as seed dormancy. In practise, seeds are rarely either dormant or non-dormant, but seeds whose dormancy-inducing pathways are activated to higher levels will germinate in an ever-narrower range of environments. Thus, measurements of dormancy must always be accompanied by analysis of environmental contexts in which phenotypes or behaviours are described. At its simplest, dormancy can be imposed by the formation of a simple physical barrier around the seed through which gas exchange and the passage of water are prevented. Seeds featuring this so-called 'physical dormancy' often require either scarification or passage through an animal gut (replete with its associated digestive enzymes) to disrupt the barrier and permit germination. In other types of seeds with 'morphological dormancy' the embryo remains under-developed at maturity and a dormant phase exists as the embryo continues its growth post-shedding, eventually breaking through the surrounding tissues. By far, the majority of seeds exhibit 'physiological dormancy' - a quiescence program initiated by either the embryo or the surrounding endosperm tissues. Physiological dormancy uses germination-inhibiting hormones to prevent germination in the absence of the specific environmental triggers that promote germination. During and after germination, early seedling growth is supported by catabolism of stored reserves of protein, oil or starch accumulated during seed maturation. These reserves support cell expansion, chloroplast development and root growth until photoauxotrophic growth can be resumed.

Carbon and nitrogen allocation strategy in Posidonia oceanica is altered by seawater acidification

Rising atmospheric CO2 causes ocean acidification that represents one of the major ecological threats for marine biota. We tested the hypothesis that long-term exposure to increased CO2 level and acidification in a natural CO2 vent system alters carbon (C) and nitrogen (N) metabolism in Posidonia oceanica L. (Delile), affecting its resilience, or capability to restore the physiological homeostasis, and the nutritional quality of organic matter available for grazers. Seawater acidification decreased the C to N ratio in P. oceanica tissues and increased grazing rate, shoot density, leaf proteins and asparagine accumulation in rhizomes, while the maximum photochemical efficiency of photosystem II was unaffected. The 13C-dilution in both structural and non-structural C metabolites in the acidified site indicated quali-quantitative changes of C source and/or increased isotopic fractionation during C uptake and carboxylation associated with the higher CO2 level. The decreased C:N ratio in the acidified site suggests an increased N availability, leading to a greater storage of 15N–enriched compounds in rhizomes. The amount of the more dynamic C storage form, sucrose, decreased in rhizomes of the acidified site in response to the enhanced energy demand due to higher shoot recruitment and N compound synthesis, without affecting starch reserves. The ability to modulate the balance between stable and dynamic C reserves could represent a key ecophysiological mechanism for P. oceanica resilience under environmental perturbation. Finally, alteration in C and N dynamics promoted a positive contribution of this seagrass to the local food web.

Low source-sink ratio reduces reserve starch in grapevine woody canes and modulates sugar transport and metabolism at transcriptional and enzyme activity levels.

Severe leaf removal decreases storage starch and sucrose in grapevine cv. Cabernet Sauvignon fruiting cuttings and modulates the activity of key enzymes and the expression of sugar transporter genes. Leaf removal is an agricultural practice that has been shown to modify vineyard efficiency and grape and wine composition. In this study, we took advantage of the ability to precisely control the number of leaves to fruits in Cabernet Sauvignon fruiting cuttings to study the effect of source-sink ratios (2 (2L), 6 (6L) and 12 (12) leaves per cluster) on starch metabolism and accumulation. Starch concentration was significantly higher in canes from 6L (42.13±1.44mggDW-1) and 12L (43.50±2.85mggDW-1) than in 2L (22.72±3.10mggDW-1) plants. Moreover, carbon limitation promoted a transcriptional adjustment of genes involved in starch metabolism in grapevine woody tissues, including a decrease in the expression of the plastidic glucose-6-phosphate translocator, VvGPT1. Contrarily, the transcript levels of the gene coding the catalytic subunit VvAGPB1 of the VvAGPase complex were higher in canes from 2L plants than in 6L and 12L, which positively correlated with the biochemical activity of this enzyme. Sucrose concentration increased in canes from 2L to 6L and 12L plants, and the amount of total phenolics followed the same trend. Expression studies showed that VvSusy transcripts decreased in canes from 2L to 6L and 12L plants, which correlated with the biochemical activity of insoluble invertase, while the expression of the sugar transporters VvSUC11 and VvSUC12, together with VvSPS1, which codes an enzyme involved in sucrose synthesis, increased. Thus, sucrose seems to control starch accumulation through the adjustment of the cane sink strength.

Effect of Different Plucking Systems on Yield and Root Starch Reserve in Two Cultivars of Tea (Camellia sinensis L.)

A study was carried out to monitor the effect of different plucking systems on root starch reserve and yield of tea in two cultivars (drought tolerant TRI2025 and moderately drought tolerant TRI2026). Depending on the severity of plucking, four plucking systems were used as treatments namely, mother-leaf plucking, fish-leaf plucking, janam-leaf plucking and estate practice. The cultivar TRI2026 showed significantly higher yield than the drought tolerant cultivar TRI2025. Estate practice produced greater yield in TRI2025 than the other treatments, while in TRI2026 the highest yield was found in fish-leaf plucking system. Although cultivars had no significant effect on root starch reserves, however significantly different found in treatments. About one year after commencing the experiment, for both cultivars higher root starch observed under mother-leaf plucking and the lowest was in janam-leaf plucking.J. Environ. Sci. & Natural Resources, 9(2): 91-95 2016

Apple dwarfing rootstocks exhibit an imbalance in carbohydrate allocation and reduced cell growth and metabolism

Apple dwarfing rootstocks cause earlier shoot termination and reduced root and shoot mass. To identify physiological factors responsible for rootstock-induced growth restriction, we compared vascular-enriched gene expression between two dwarfing rootstocks (‘M27’ and ‘M9’) and the vigorous rootstock ‘M793’ using RNA sequencing and quantitative reverse transcriptase PCR. Differentially expressed genes common to both dwarfing rootstocks belonged to five main biological processes: (1) primary metabolism, (2) cell wall synthesis and modification, (3) secondary metabolism, (4) hormone signalling and response and (5) redox homeostasis. Genes promoting the biosynthesis of amino acids, lipids and cell walls were downregulated in dwarfing rootstocks, whereas genes promoting the breakdown of these compounds were upregulated. The only exception to this trend was the upregulation of starch synthesis genes in dwarfing rootstocks. Non-structural carbohydrate analysis demonstrated that starch concentrations in ‘M9’ roots, stems and grafted ‘Royal Gala’ (‘RG’) scions were double that of equivalent tissues from ‘RG’ homo-grafted trees (‘RG’/‘RG’). Fructose and glucose concentrations were much lower in all three tissues of the ‘RG’/‘M9’ trees. Together, these data indicate that dwarfing rootstocks are in a state of sugar depletion and reduced cellular activity despite having large starch reserves. Another significant finding was the over-accumulation of flavonoids and the downregulation of auxin influx transporters MdAUX1 and MdLAX2 in dwarfing rootstocks. We propose that both factors reduce polar auxin transport. The results of this study contribute novel information about the physiological state of dwarfing rootstocks.

Starch as a determinant of plant fitness under abiotic stress.

Contents 943 I. 943 II. 944 III. 945 IV. 945 V. 948 VI. 949 950 References 950 SUMMARY: Abiotic stresses, such as drought, high salinity and extreme temperatures, pose one of the most important constraints to plant growth and productivity in many regions of the world. A number of investigations have shown that plants, including several important crops, remobilize their starch reserve to release energy, sugars and derived metabolites to help mitigate the stress. This is an essential process for plant fitness with important implications for plant productivity under challenging environmental conditions. In this Tansley insight, we evaluate the current literature on starch metabolism in response to abiotic stresses, and discuss the key enzymes involved and how they are regulated.

Implications of the Presence of Maturing Fruit on Carbohydrate and Nitrogen Distribution in Grapevines under Postveraison Water Constraints

Grapevine (Vitis vinifera) berries are sugar and nitrogen (N) sinks between veraison and fruit maturity. Limited photoassimilation, often caused by water constraints, induces reserve total nonstructural carbohydrate (TNC) remobilization, contributing to berry sugar accumulation, while fruit N accumulation can be affected by vine water supply. Although postveraison root carbohydrate remobilization toward the fruit has been identified through 14C tracing studies, it is still unclear when this remobilization occurs during the two phases of berry sugar accumulation (rapid and slow). Similarly, although postveraison N reserve mobilization toward the fruit has been reported, the impact of water constraints during berry N accumulation on its translocation from the different grapevine organs requires clarification. Potted grapevines were grown with or without fruit from the onset of veraison. Vines were irrigated to sustain water constraints, and fortnightly root, trunk, shoot, and leaf structural biomass, starch, soluble sugar, total N, and amino N concentrations were determined. The fruit sugar and N accumulation was also assessed. Root starch depletion coincided with root sucrose and hexose accumulation during peak berry sugar accumulation. Defruiting at veraison resulted in continuous root growth, earlier starch storage, and root hexose accumulation. Leaf N depletion coincided with fruit N accumulation, while the roots of defruited vines accumulated N reserves. Root growth, starch, and N reserve accumulation were affected by maturing fruit during water constraints. Root starch is an alternative source to support fruit sugar accumulation, resulting in reserve starch depletion during rapid fruit sugar accumulation, while root starch refills during slow berry sugar accumulation. On the other hand, leaf N is a source toward postveraison fruit N accumulation, and the fruit N accumulation prevents root N storage.

Substantial resource reallocation during germination of Araucaria bidwillii (bunya pine), an Australian rainforest conifer with large seeds and cryptogeal germination

Araucaria bidwillii has cryptogeal germination, with seedlings not starting photosynthesis for several months after imbibition. Our results indicate that this complex process is metabolically relatively efficient. Araucaria bidwillii (bunya pine, Araucariaceae), an Australian rainforest conifer, produces heavy (15–20 g fresh mass) recalcitrant seeds that feature an unusual cryptogeal germination sequence. Germination features a substantial reallocation of starch from the massive megagametophyte (12–14 g fresh mass) to the hypocotyl which progressively develops into a tuberous, belowground storage organ. While germination (radicle emergence) is very rapid after imbibition, seedlings do not commence photosynthesis for several months. We were interested in the metabolic efficiency of this lengthy and complex developmental process that depends solely on stored resources. On average, the seeds had a dry mass of 10.1 g (seed coat 29 %, megagametophyte 68 %, embryo 3 %). Over 47 days the megagametophyte lost 91 % of its mass (mainly through the almost complete export of its abundant starch reserves), while the embryo/seedling increased in mass by 19 times and the hypocotyl almost 200 times. In total, a 13–14 % loss of dry mass occurred in converting a large seed at soil level into an established seedling with substantial protected belowground reserves. If only considering the living components of the seed/seedling (i.e. excluding the substantial and woody seed coat) then an 18 % loss occurred. Comparison with its closest cryptocotylar relatives from South America (A. araucana and A. angustifolia) and a range of hypocotylar rainforest angiosperms indicates that the complicated germination sequence of A. bidwillii is relatively efficient. While A. bidwillii now has a highly disjunct and contracted distribution this would appear not to be directly related to the metabolic efficiency of its unusual germination sequence.

Hydrogen photoproduction by immobilized S-deprived Chlamydomonas reinhardtii: Effect of light intensity and spectrum, and initial medium pH

Sulfur deprivation creates anaerobic conditions and thereby induces hydrogen photoproduction by the green microalga Chlamydomonas reinhardtii. In the present work we showed that immobilization of C. reinhardtii within a thin Ca-alginate film extended the duration of hydrogen evolution under sulfur depletion up to three weeks with hydrogen production limited mainly by exhaustion of intracellular starch reserves. The optimal incident light intensity for hydrogen photoproduction appeared to be about 80μmolphotonsm−2s−1 at which the total amount of evolved hydrogen reached 0.8mmol per mg chlorophyll. Illumination by monochromatic blue light peaking at 435nm or red light with maximum at 690nm, both favoring PSI excitation, improved the stability of PSII centers and enhanced hydrogen evolution. Initial medium pH in the range 6.5–7.0 was optimal for hydrogen photoproduction, likely due to enhanced accumulation of starch and elevated initial PSII activity.

The qTSN Positive Effect on Panicle and Flag Leaf Size of Rice is Associated with an Early Down-Regulation of Tillering.

The qTSN4 was identified as rice QTL (Quantitative Traits Locus) increasing total spikelet number per panicle and flag leaf area but potentially reducing panicle number depending on the environment. So far, this trade-off was mainly observed at grain maturity and not specifically studied in details, limiting the apprehension of the agronomic interest of qTSN4. This study aimed to understand the effect of qTSN4 and of the environment on panicle sizing, its trade-off with panicle number, and finally plant grain production. It compared two high yielding genotypes to their Near Isogenic Lines (NIL) carrying either QTL qTSN4 or qTSN12, two distinct QTLs contributing to the enlarged panicle size, thereafter designated as qTSN. Traits describing C sink (organ appearance rate, size, biomass) and source (leaf area, photosynthesis, sugar availability) were dynamically characterized along plant and/or panicle development within two trials (greenhouse, field), each comparing two treatments contrasting for plant access to light (with or without shading, high or low planting densities). The positive effect of qTSN on panicle size and flag leaf area of the main tiller was confirmed. More precisely, it could be shown that qTSN increased leaf area and internode cross-section, and in some cases of the photosynthetic rate and starch reserves, of the top 3–4 phytomers of the main tiller. This was accompanied by an earlier tillering cessation, that coincided with the initiation of these phytomers, and an enhanced panicle size on the main tiller. Plant leaf area at flowering was not affected by qTSN but fertile tiller number was reduced to an extent that depended on the environment. Accordingly, plant grain production was enhanced by qTSN only under shading in the greenhouse experiment, where panicle number was not affected and photosynthesis and starch storage in internodes was enhanced. The effect of qTSN on rice phenotype was thus expressed before panicle initiation (PI). Whether early tillering reduction or organ oversizing at meristem level is affected first cannot be entirely unraveled. Further studies are needed to better understand any signal involved in this early regulation and the qTSN × Environment interactions underlying its agronomic interest.

The impact of the SSIIa null mutations on grain traits and composition in durum wheat.

Starch represents a major nutrient in the human diet providing essentially a source of energy. More recently the modification of its composition has been associated with new functionalities both at the nutritional and technological level. Targeting the major starch biosynthetic enzymes has been shown to be a valuable strategy to manipulate the amylose-amylopectin ratio in reserve starch. In the present work a breeding strategy aiming to produce a set of SSIIa (starch synthases IIa) null durum wheat is described. We have characterized major traits such as seed weight, total starch, amylose, protein and β-glucan content in a set of mutant families derived from the introgression of the SSIIa null trait into Svevo, an elite Italian durum wheat cultivar. A large degree of variability was detected and used to select wheat lines with either improved quality traits or agronomic performances. Semolina of a set of two SSIIa null lines showed new rheological behavior and an increased content of all major dietary fiber components, namely arabinoxylans, β-glucans and resistant starch. Furthermore the investigation of gene expression highlighted important differences in some genes involved in starch and β-glucans biosynthesis.

Lignotubers in Mediterranean basin plants

Lignotubers are swollen woody structures located at the root-shoot transition zone and contain numerous dormant buds and starch reserves. This structure enables the plant to resprout prolifically after severe disturbances that remove the aboveground biomass. These are considered adaptive traits in ecosystems with highly frequent and severe disturbances—such as fire-prone ecosystems. In this paper, we aim to contribute to the knowledge of lignotubers in the Mediterranean basin and highlight the evolutionary implications. We first summarise existing knowledge on lignotuber species in the Mediterranean basin. We then provide a detailed morpho-anatomical description of early lignotubers in two common woody species (Arbutus unedo L. and Phillyrea angustifolia L.). Finally, we compare our anatomical results with those obtained in studies conducted with other lignotuberous species from different Mediterranean regions. Lignotubers were verified in 14 species in the Mediterranean basin; all being from lineages with origins dating to the Tertiary and thus pre-dating the Mediterranean climate. In A. unedo and P. angustifolia, lignotubers are macroscopically discernible in 4- and 2-year-old saplings, respectively. In these two species, the lignotubers have numerous buds protected by hypertrophied scales, and have a contorted xylem containing abundant starch. Our results challenge the traditional idea that pre-Mediterranean lineages suffered evolutionary inertia; instead, lignotuberous species may be considered examples of plants that adapted to the increased fire activity that occurred throughout the Tertiary and Quaternary. We also highlight the use of morpho-anatomical traits to unambiguously distinguish between lignotuberous and non-lignotuberous resprouting species.

How plants manage food reserves at night: quantitative models and open questions.

In order to cope with night-time darkness, plants during the day allocate part of their photosynthate for storage, often as starch. This stored reserve is then degraded at night to sustain metabolism and growth. However, night-time starch degradation must be tightly controlled, as over-rapid turnover results in premature depletion of starch before dawn, leading to starvation. Recent experiments in Arabidopsis have shown that starch degradation proceeds at a constant rate during the night and is set such that starch reserves are exhausted almost precisely at dawn. Intriguingly, this pattern is robust with the degradation rate being adjusted to compensate for unexpected changes in the time of darkness onset. While a fundamental role for the circadian clock is well-established, the underlying mechanisms controlling starch degradation remain poorly characterized. Here, we discuss recent quantitative models that have been proposed to explain how plants can compute the appropriate starch degradation rate, a process that requires an effective arithmetic division calculation. We review experimental confirmation of the models, and describe aspects that require further investigation. Overall, the process of night-time starch degradation necessitates a fundamental metabolic role for the circadian clock and, more generally, highlights how cells process information in order to optimally manage their resources.

Fructan and hormone connections.

Plants rely on “reserve” (stored) carbon (C) for growth and survival when newly synthesized C becomes limited. Besides a classic yet recalcitrant C reserve starch, fructans, a class of sucrose-derived soluble fructosyl-oligosaccharides, represent a major store of C in many temperate plant species including the economically important Asteraceae and Poaceae families (Hendry, 1993). Dicots typically accumulate inulin-type fructans as long-term storage (underground organs) whilst grasses and cereals accumulate fructans as short-term reserves in above-ground parts (Pollock and Cairns, 1991; Van Laere and Van den Ende, 2002). Unlike chloroplast-based water-insoluble starch, fructans are semi-soluble, possess flexible structures (Phelps, 1965; Valluru and Van den Ende, 2008), can be synthesized at low temperatures (Pollock and Cairns, 1991), and are degraded by a single type of fructan hydrolases, fructan exohydrolases (FEHs). Unlike starch that store in plastids, fructans store in vacuoles, which is physically less stressful to the active constituents of, and allows more C synthesis by, the photosynthetic cell, which may be different in dicots where fructans do not typically accumulate in green parts.

Impact of drought and flooding stresses on Gabiroba plants

The aim of this study was to evaluate the tolerance of Gabiroba plants (Campomanesia spp) to drought and flooding stresses for 0, 3, 6, 9, 12, 15, and 18 days. The contents of dry matter, water, nitrate reductase activity, total soluble sugars, sucrose, starch, insoluble and soluble protein, and soluble amino acid in leaves and roots were periodically determined. Drought and flooding induced a significant reduction in the contents of water and dry matter of leaves and roots. The contents of total soluble sugars, sucrose, and starch suggest a reduction in the photosynthetic rate, remobilization of starch reserves, and transport blocking in the phloem. Drought stress leads to decreased protein synthesis and accumulation of amino acids. The higher concentration of insoluble proteins in the leaves and roots of plants under flooding is a consequence of the synthesis of new polypeptides. For the leaves, the lowest and highest contents of amino acids were found under drought and flooding stresses, respectively; whereas for the roots, the lowest rates were found under drought stress. Gabiroba plants are tolerant to drought stress and remobilize soluble carbohydrates from the shoots to the roots.

Carbohydrate Metabolism During Wintering Period in Four Zoysiagrass Genotypes

A key barrier to widespread use of warm-season grasses in the transition zone between the temperate and subtropical climates is represented by the winter dormancy, a temporary suspension of visible growth of any plant structure including the meristem. In this environment, species with different photosynthetic carbon cycle coexist, leading to asynchrony in growth among warm- and cool-season grasses. The objectives of the present study were (i) to assess the carbohydrate content in four representative zoysiagrasses Zoysia japonica ‘El Toro’, ‘Meyer’, Z. pacifica, and Z. matrella ‘Zeon’ during three successive phases of vegetation phenology, senescence, dormancy, and green-up, and (ii) to characterize the metabolic responses to phenological variations under natural acclimation. Japanese lawn grass genotypes ‘El Toro’ and ‘Meyer’ showed longer dormancy periods than fine-leaved Z. pacifica and ‘Zeon’. In Japanese lawn grass genotypes, the progressive decline of photosynthetic pigments was similar to that observed in the color retention. Over the experimental period, a significantly greater amount of starch reserves was observed in sprigs (horizontal stems) in the Japanese lawn grasses than in the fine-leaved zoysiagrasses. In general, total soluble sugars (TSS) in leaves and sprigs did not show evidence of sugar starvation during the senescence phase. TSS increased significantly during cold acclimation, resulting in a higher metabolic activity at the onset of green-up to support spring regreening in all zoysiagrasses except ‘Meyer’ leaves.

New starch phenotypes produced by TILLING in barley.

Barley grain starch is formed by amylose and amylopectin in a 1∶3 ratio, and is packed into granules of different dimensions. The distribution of granule dimension is bimodal, with a majority of small spherical B-granules and a smaller amount of large discoidal A-granules containing the majority of the starch. Starch granules are semi-crystalline structures with characteristic X-ray diffraction patterns. Distinct features of starch granules are controlled by different enzymes and are relevant for nutritional value or industrial applications. Here, the Targeting-Induced Local Lesions IN Genomes (TILLING) approach was applied on the barley TILLMore TILLING population to identify 29 new alleles in five genes related to starch metabolism known to be expressed in the endosperm during grain filling: BMY1 (Beta-amylase 1), GBSSI (Granule Bound Starch Synthase I), LDA1 (Limit Dextrinase 1), SSI (Starch Synthase I), SSIIa (Starch Synthase IIa). Reserve starch of nine M3 mutant lines carrying missense or nonsense mutations was analysed for granule size, crystallinity and amylose/amylopectin content. Seven mutant lines presented starches with different features in respect to the wild-type: (i) a mutant line with a missense mutation in GBSSI showed a 4-fold reduced amylose/amylopectin ratio; (ii) a missense mutations in SSI resulted in 2-fold increase in A:B granule ratio; (iii) a nonsense mutation in SSIIa was associated with shrunken seeds with a 2-fold increased amylose/amylopectin ratio and different type of crystal packing in the granule; (iv) the remaining four missense mutations suggested a role of LDA1 in granule initiation, and of SSIIa in determining the size of A-granules. We demonstrate the feasibility of the TILLING approach to identify new alleles in genes related to starch metabolism in barley. Based on their novel physicochemical properties, some of the identified new mutations may have nutritional and/or industrial applications.

Amylolytic activities and starch reserve mobilization during the germination of quinoa

Quinoa (Chenopodium quinoa) is a dicotyledonous plant producing perispermic seeds that is increasingly popular in the Western world due to its nutritional value. Germination can be used in food processing to alter the sensory, nutritional and functional properties of grains, and the increased enzyme activities may have beneficial influences in food applications. Knowledge about the germination of perispermic seeds is scarce. In this study, the development of amylolytic activities and subsequent changes in sugar profiles and starch content were followed in quinoa over a period of 72 h. The seeds germinated rapidly with radicle protrusion occurring 8 h after imbibition, when the seeds had reached a moisture content of 44 %. A low level of α-amylase activity (determined at pH 5.2 using blocked ρ-nitrophenyl-maltoheptaoside as substrate) was present in the embryo of non-germinating seeds, but emerged in the perisperm only after 24 h, followed by a sharp increase in activity in both tissues. The accumulation of glucose and fructose (determined with high pressure liquid chromatography) and the decrease in starch content (determined with an enzymatic assay) were observed after 24 and 36 h, respectively, indicating the onset of starch reserve mobilization. Overall, the levels of amylolytic activities remained very low compared with traditional malting cereals, suggesting the unsuitability of quinoa as a source of amylases in food applications. Scanning electron microscopy, used to visualize changes in starch granule morphology, showed mainly exocorrosion, suggesting a homogenous structure of the outer layers of quinoa starch.

Root Starch Reserves Are Necessary for Vigorous Re-Growth following Cutting Back in Lotus japonicus

Perenniality and vegetative re-growth vigour represent key agronomic traits in forage legume (Fabaceae) species. The known determinants of perenniality include the conservation of the vegetative meristem during and after the flowering phase, and the separation of flowering from senescence. The ability of the plants to store nutrient resources in perennial organs and remobilize them may also play an important role in the perennial growth habit, and in determining the capacity of the plant to re-grow following grazing or from one season to the next. To examine the importance of stored starch, we examined the vegetative re-growth vigour following cutting back of a unique collection of Lotus japonicus mutants impaired in their ability to synthesize or degrade starch. Our results establish that starch stored in the roots is important for re-growth vigour in Lotus japonicus. We extended this analysis to a collection of Lotus (trefoil) species and two ecotypes of Lotus japonicus displaying a large variation in their carbohydrate resource allocation. There was a positive correlation between root starch content and re-growth vigour in these natural variants, and a good general correlation between high re-growth vigour and the perennial life-form. We discuss the relationship between perenniality and the availability of root carbohydrates for re-growth.