Morphology,Carbohydrate Composition and Vernalization Response in a Genetically Diverse Collection of Asian and European Turnips (Brassica rapa subsp. rapa)

Brassica rapa displays enormous morphological diversity, with leafy vegetables, turnips and oil crops. Turnips (Brassica rapa subsp. rapa) represent one of the morphotypes, which form tubers and can be used to study the genetics underlying storage organ formation. In the present study we investigated several characteristics of an extensive turnip collection comprising 56 accessions from both Asia (mainly Japanese origin) and Europe. Population structure was calculated using data from 280 evenly distributed SNP markers over 56 turnip accessions. We studied the anatomy of turnip tubers and measured carbohydrate composition of the mature turnip tubers of a subset of the collection. The variation in 16 leaf traits, 12 tuber traits and flowering time was evaluated in five independent experiments for the entire collection. The effect of vernalization on flowering and tuber formation was also investigated. SNP marker profiling basically divided the turnip accessions into two subpopulations, with admixture, generally corresponding with geographical origin (Europe or Asia). The enlarged turnip tuber consists of both hypocotyl and root tissue, but the proportion of the two tissues differs between accessions. The ratio of sucrose to fructose and glucose differed among accessions, while generally starch content was low. The evaluated traits segregated in both subpopulations, with leaf shape, tuber colour and number of shoots per tuber explaining most variation between the two subpopulations. Vernalization resulted in reduced flowering time and smaller tubers for the Asian turnips whereas the European turnips were less affected by vernalization.     

Quantitation of oxalates in corms and shoots of <Emphasis Type="Italic">Colocasia esculenta</Emphasis> (L.) Schott under drought conditions

Oxalate (calcium oxalate) accumulation in taro plants (Colocasia esculenta (L.) Schott) impacts their nutritional quality, producing acridity, causing lips, mouth and throat tissues swelling if consumed fresh. The oxalate content is related to photosynthesis, through the glycolate–glyoxylate oxidation pathway. The plant’s photosynthetic rate usually increases in non-stressed conditions. Differences in photosynthetic rate are indirectly related to the chlorophyll content index. Protein accumulation and starch variation are also important traits to understand the taro oxalate synthesis caused by drought and how they affect corm quality. The purpose of this study was to quantitate oxalates in taro corms and shoots submitted to drought conditions and to evaluate how stress response can affect the nutritional quality of taro whole-plant. Seven taro genotypes from Madeira, Canaries and Pacific Community (SPC) collections were grown in greenhouse conditions and submitted to different watering regimes for drought tolerance screening. Corms and shoots were harvested and evaluated for oxalates (soluble, insoluble and total), chlorophyll content index (CCI), crude protein, starch, starch solubility in water and starch swelling power. All accessions had very high calcium oxalate content. Drought-tolerant genotypes showed good osmotic response by oxalate precipitation and mobilization through shoot to corm tissues, photosynthesis adaptation by increase of CCI, protein accumulation, and very low starch hydrolysis. Sensitive-drought genotypes showed less mobilization of calcium oxalate, decreased photosynthetic rate and protein synthesis, and slight increase of starch hydrolysis. Variation in taro oxalate content is consistent and significantly correlated with the photosynthetic rate, carbohydrate metabolism and protein synthesis.     

HRE-type genes are regulated by growth-related changes in internal oxygen concentrations during the normal development of potato (Solanum tuberosum) tubers

The occurrence of hypoxic conditions in plants not only represents a stress condition but is also associated with the normal development and growth of many organs, leading to adaptive changes in metabolism and growth to prevent internal anoxia. Internal oxygen concentrations decrease inside growing potato tubers, due to their active metabolism and increased resistance to gas diffusion as tubers grow. In the present work, we identified three hypoxia-responsive ERF (StHRE) genes whose expression is regulated by the gradual decrease in oxygen tensions that occur when potato tubers grow larger. Increasing the external oxygen concentration counteracted the modification of StHRE expression during tuber growth, supporting the idea that the actual oxygen levels inside the organs, rather than development itself, are responsible for the regulation of StHRE genes. We identified several sugar metabolism-related genes co-regulated with StHRE genes during tuber development and possibly involved in starch accumulation. All together, our data suggest a possible role for low oxygen in the regulation of sugar metabolism in the potato tuber, similar to what happens in storage tissues during seed development.     

Physiological and morphological traits associated with adaptation of lucerne (Medicago sativa) to severely drought‐stressed and to irrigated environments

Adaptation to severe drought and to irrigated cropping can both contribute to increased water use efficiency of lucerne, but knowledge on the relevant adaptive traits is limited. Five cultivars featuring contrasting adaptive responses for 3‐year forage yield across 10 agricultural environments of the western Mediterranean basin were currently studied, to identify physiological and morphological traits associated with specific and wide‐adaptation responses. The landraces Mamuntanas, Demnat 203 and Erfoud 1, and the varieties SARDI 10 and Prosementi, were grown in replicated metal containers (55 cm long × 12 cm wide × 75 cm deep; 21 plants per container) under irrigation (weekly restoring soil field capacity) and under moderate and severe drought stress (implying decreased irrigation for 30 days followed by withheld irrigation for 33 and 58 days, respectively). Cultivar post‐stress survival reflected the known cultivar adaptation to drought‐prone agricultural environments. Demnat 203, specifically adapted to irrigated, frequently mown environments, displayed higher amounts of starch, soluble proteins and total nitrogen in the crown and the root under irrigation. This was due to outstanding organ size and, for starch, higher concentrations. Mamuntanas, specifically adapted to drought‐prone environments, exhibited high water‐soluble carbohydrate concentration in storage organs under severe stress, along with a water‐conservation strategy implying less water used in initial drought‐stress phases due to limited early root development that resulted in more water available under severe stress. Drought‐tolerant germplasm also displayed lower wilting under early stress, more plants with green tissues under severe stress, and more stems per plant in stress or favourable conditions. Multivariate patterns of cultivar variation for physiological and morphological traits were strictly associated with cultivar variation for adaptation pattern. Our results highlighted the difficulty to combine some traits of environment‐specific adaptive value into a unique widely adapted variety, supporting the selection of varieties specifically adapted to irrigated or severely drought‐prone environments.     

Contribution of arbuscular mycorrhizal fungi (AMF) to the adaptations exhibited by the deciduous shrub Anthyllis cytisoides L. under water deficit

Anthyllis cytisoides L. is highly colonized by arbuscular mycorrhizal fungi (AMF) and behaves as a drought-avoider species in the field. Our objectives were: (1) to study the response of A. cytisoides when exposed to moderate (acclimation) or severe (peak) drought and subsequent rewatering under nursery conditions; and (2) to verify if AMF improved the adaptation of A. cytisoides to stress. The soil compactness in drought-acclimated treatments increased four times compared with that of well-watered controls, which could reinforce the effects of water deficit on plant physiology. Photosynthetic rates decreased by around 50% and 70% and leaf conductance decreased by 40% and 50% in drought-acclimated non-mycorrhizal and mycorrhizal plants, respectively. Peak drought limited plant growth, accelerated leaf senescence and induced the conversion of starch into soluble sugars in the leaves of stressed plants. The accumulation of sugars could contribute to a decrease in water potential in order to achieve the required tension to let water move from soil to shoot. Mycorrhizal plants showed a two-fold higher chlorotic leaf biomass than non-mycorrhizal plants under severe drought. Moreover, mycorrhizal A. cytisoides showed enhanced epicuticular waxes on the surfaces of the remaining green leaves. Increased leaf senescence, together with wax deposition, could reduce whole plant transpiration, thus allowing mycorrhizal plants to maintain a higher leaf relative water content (50%) than non-mycorrhizal plants (35%). After drought recovery, leaf abscission in stressed mycorrhizal plants was 10 times greater than that in non-mycorrhizal plants. The results suggest that AMF conferred greater responsiveness of A. cytisoides to drought. Enhanced wax deposition and leaf senescence could be an ecological adaptation to cope with severe water deficit.     

Tecia solanivora infestation increases tuber starch accumulation in Pastusa Suprema potatoes

In response to infestation with larvae of the Guatemalan tuber moth(Tecia solanivora), some Solanum tuberosum(potato) varieties exhibit an overcompensation response, whereby the total dry mass of uninfested tubers is increased. Here, we describe early responses,within the first few days, of T. solanivora feeding, in the Colombian potato variety Pastusa Suprema. Nontargeted metabolite profiling showed significant secondary metabolism changes in T. solanivora-infested tubers,but not in uninfested systemic tubers. In contrast,changes in primary metabolism were greater in uninfested systemic tubers than in the infested tubers, with a notable 80% decline in systemic tuber sucrose levels within 1 d of T. solanivora infestation. This suggested either decreased sucrose transport from the leaves orincreased sink strength, i.e., more rapid sucrose to starch conversion in the tubers. Increased sucrose synthesis was indicated by higher rubisco activase and lower starch synthase gene expression in the leaves of infested plants.Elevated sink strength was demonstrated by 45% more total starch deposition in systemic tubers of T. solanivorainfested plants compared to uninfested control plants.Thus, rather than investing in increased defense of uninfested tubers, Pastusa Suprema promotes deposition of photoassimilates in the form of starch as a response to T. solanivora infestation.     

Screening of cassava (Manihot esculenta Crantz) accessions against Phytophthora palmivora induced tuber rot of cassava

Cassava (Manihot esculenta Crantz), is an important tropical tuber crop with global importance and plays a significant role in the food, nutritional and livelihood security of around 500 million people. In India, the low productivity of cassava attributes to the soil borne disease, particularly tuber rot caused by Phytophthora palmivora (Butl.) which is destructive and the attack is spreading in alarming rate in all the cassava growing regions causing heavy yield loss of more than 50%. Introduction of disease resistant varieties may alleviate the problem to a certain extent. This paper describes the screening procedures and findings on the disease resistant variety of cassava accession against tuber rot. Variety Sree Padmanabha imparted high resistance against tuber rot, while Sree Sahya was moderately resistant and all other accessions studied were found to be susceptible in in vitro and in field trials. In screening studies, a reproducible positive correlation was obtained between attached tubers in live plant with detached tubers which showed that detached tuber part can be used for the prediction of resistance in attached live plants of cassava for cultivar resistance. The procedure described here could be used as a simple, rapid and efficient method for screening of cassava accessions against tuber rot of cassava.     

Characterization of oil and starch accumulation in tubers of Cyperus esculentus var. sativus (Cyperaceae): A novel model system to study oil reserves in nonseed tissues

? Premise of the study: Storage oil (triacylglycerol) accumulates in tissues such as the embryo and endosperm of seeds and the fruit mesocarp, but seldom in underground organs. As a rare exception, cultivated variants of yellow nutsedge (Cyperus esculentus) contain high amounts of both oil and starch in the mature tubers. ? Methods: Biochemical analyses and light and electron microscopy were used to study the accumulation patterns of storage nutrients in developing nutsedge tubers. ? Key results: During the initial phase of tuber development, the conducting rhizome tissue is transformed into a storage compartment, then massive storage reserves accumulate in the tuber. At the beginning of tuber development, a large sugar load coincided with the onset of starch accumulation. Oil accumulation started later, concomitant with a substantial drop in the sugar content. Initially, oil accumulated at a lower rate compared to starch, but the rate later increased; after 6 wk, oil made up 24% of tuber dry mass, while starch made up 32%. Protein concentration changed only a small amount throughout this development. Oil and starch accumulated in the same cells throughout the tubers in a sequential fashion during tuber development. ? Conclusions: The developmental pattern in the build up of storage nutrients in the tubers highlights nutsedge as a novel model plant, having potential to significantly widen our understanding on how synthesis of storage reserves, and in particular oils, is regulated and directed in nonseed tissues such as tubers and roots.     

Leaf responses to drought stress in Mediterranean accessions of Solanum lycopersicum: anatomical adaptations in relation to gas exchange parameters

In a previous study, important acclimation to water stress was observed in the Ramellet tomato cultivar (TR) from the Balearic Islands, related to an increase in the water‐use efficiency through modifications in both stomatal (g_s) and mesophyll conductances (g_m). In the present work, the comparison of physiological and morphological traits between TR accessions grown with and without water stress confirmed that variability in the photosynthetic capacity was mostly explained by differences in the diffusion of CO₂ through stomata and leaf mesophyll. Maximization of g_m under both treatments was mainly achieved through adjustments in the mesophyll thickness and porosity and the surface area of chloroplasts exposed to intercellular airspace (S_c). In addition, the lower g_m/S_c ratio for a given porosity in drought‐acclimated plants suggests that the decrease in g_m was due to an increased cell wall thickness. Stomatal conductance was also affected by drought‐associated changes in the morphological properties of stomata, in an accession and treatment‐dependent manner. The results confirm the presence of advantageous physiological traits in the response to drought stress in Mediterranean accessions of tomato, and relate them to particular changes in the leaf anatomical properties, suggesting specific adaptive processes operating at the leaf anatomical level.     

Decreased expression of sucrose phosphate synthase strongly inhibits the water stress-induced synthesis of sucrose in growing potato tubers

Water stress stimulates sucrose synthesis and inhibits starch synthesis in wild-type tubers. Antisense and co-suppression potato transformants with decreased expression of sucrose–phosphate synthase (SPS) have been used to analyse the importance of SPS for the regulation of this water-stress induced change in partitioning. (i) In the absence of water stress, a 70–80% decrease in SPS activity led to a 30–50% inhibition of sucrose synthesis and a slight (10–20%) increase of starch synthesis in tuber discs in short-term labelling experiments with low concentrations of labelled glucose. Similar changes were seen in short-term labelling experiments with intact tubers attached to well-watered plants. Provided plants were grown with ample light and water, transformant tubers had a slightly lower water and sucrose content and a similar or even marginally higher starch content than wild-type tubers. (ii) When wild-type tuber slices were incubated with labelled glucose in the presence of mannitol to generate a moderate water deficit (between –0.12 and –0.72 MPa), there was a marked stimulation of sucrose synthesis and inhibition of starch synthesis. A similar stimulation was seen in labelling experiments with wild-type tubers that were attached to water-stressed wild-type plants. These changes were almost completely suppressed in transformants with a 70–80% reduction of SPS activity. (iii) Decreased irrigation led to an increase in the fraction of the dry-matter allocated to tubers in wild-type plants. This shift in allocation was prevented in transformants with reduced expression of SPS. (iv) The results show that operation of SPS and the sucrose cycle in growing potato tubers may lead to a marginal decrease in starch accumulation in non-stressed plants. However, SPS becomes a crucial factor in water-stressed plants because it is required for adaptive changes in tuber metabolism and whole plant allocation.     

Photosynthetic response of four fern species from different habitats to drought stress: relationship between morpho-anatomical and physiological traits

Ferns flourish in many habitats, from epiphytic to terrestrial and from sunny to shady, and such varied conditions require contrasting photosynthetic strategies to cope with drought. Four species of temperate ferns from different habitats were subjected to drought by withholding irrigation in order to investigate their photosynthetic responses. Lepisorus thunbergianus (epiphytic) had low stomatal density and showed high water-use efficiency (WUE) retaining photosynthetic activity with low relative frond water content under drought stress, which suggested their high adaptation to drought. On the other hand, low WUE with low light-saturated photosynthetic rate in Adiantum pedatum (terrestrial, shady environment) was associated with much lower photosynthesis than in the other species under drought stress, suggesting lower adaptation to drought-prone habitats. Morphological stomatal traits such as stomatal density and photosynthetic response to drought in ferns involved species-specific adaptation to survive and grow in their natural habitats with different levels of drought.     

马铃薯块茎膨大期不同程度干旱后复水的源库补偿效应

旱后复水的补偿效应在多种作物的不同生育时期都存在,是植物抵抗逆境胁迫和伤害的重要自我调节机制,也是对有限水分高效利用的体现.本研究在马铃薯块茎膨大期进行两轮干旱后复水处理,明确马铃薯补偿效应产生的干旱胁迫阈值,并从源-库角度探索马铃薯旱后复水补偿效应产生的缘由.试验选取‘大西洋’马铃薯脱毒组培苗为材料,设置充分供水(W)、轻度干旱后复水(D₁-W)、中度干旱后复水(D₂-W)和重度干旱后复水(D₃-W)4个水分处理并经过两个循环.结果表明:在经过两轮轻度干旱复水后,马铃薯产量表现出超补偿效应,水分利用效率和产量比充分供水分别提高了17.5%和6.3%;中度水分胁迫表现出近等量补偿效应,产量与充分供水差异不大,而水分利用效率提高了8.4%;而重度水分胁迫没有表现出产量补偿效应.不同程度的干旱胁迫均降低马铃薯叶片叶绿素含量、净光合速率、叶面积等源的大小和活性,而在复水后,轻度和中度胁迫出现了超补偿和补偿效应,增强了源的供应能力.同时,适度干旱后复水显著增强了块茎(库)中蔗糖-淀粉代谢途径关键酶的活性,提高了库活性,进而表现为块茎平均重量的增加.综上,马铃薯块茎膨大期适度的水分亏缺在复水后源-库均存在补偿和超补偿效应,以此来弥补干旱带来的损失,最终在产量上表现为补偿或者超补偿效应,并显著提高了水分利用效率.     

Photosynthetic Activity and Cellular Integrity of the Andean Legume Pachyrhizus ahipa (Wedd.) Parodi under Heat and Water Stress

Pachyrhizus ahipa (Wedd.) Parodi, originally from Latin America, is an agronomy interesting legume crop due to high seed protein content and saccharides-rich tuber root. Its capacity of adaptation to Mediterranean climate, where heat and water stress are frequently associated, is being tested. Two accessions of P. ahipa (AC 102 and AC 524) differing in field production were compared as concerns the effects of water stress and high temperature on photosynthetic performance. Membrane integrity was also evaluated through electrolyte leakage (injury index, I%), lipid composition, and ultrastructure observations. Short-term heat stress (40 °C) did not affect net photosynthetic rate (P _N), stomatal conductance (g _s), and most of fluorescence parameters in both accessions, what was consistent with low electrolyte leakage. However, photosynthetic capacity (P _max) showed a significant reduction, AC 524 being more affected than AC 102. Relative water content (RWC) below 70 % caused a drastic decrease in P _N and g _s. Fluorescence parameters, P _max, and I% were affected in the two accessions, which also presented a strong reduction (42 %) in total fatty acids (TFA). Contents of galactolipids were drastically reduced, and changes in their saturation also occurred, namely a decrease in linolenic acid (C_18:3) percentage of monogalactosyl-diacylglycerol (MGDG) in both accessions. Thylakoid ultrastructure in AC 524 submitted to drought showed disorganisation of grana stacking. Mitochondria presented signs of injured cristae. When water-stressed plants were subjected to high temperature, photosynthesis and fluorescence parameters did not show significant additional changes in both accessions. The exposure of drought stressed plants to 40 °C further increased electrolyte leakage in AC 524, but not in AC 102. Chloroplasts, mitochondria, and plasmalemma showed an increased disorganisation. Vesicles appeared in the cytoplasm, which became electron-transparent, reflecting a strong reduction in the number of ribosomes. Hence AC 102 was less affected than AC 524 as regards some components of photosynthetic process, namely P _max and membrane integrity. This could account for its better yield production previously observed in field grown plants.     

Identification of a metabolic bottleneck for cold acclimation in Arabidopsis thaliana

Central carbohydrate metabolism of Arabidopsis thaliana is known to play a crucial role during cold acclimation and the acquisition of freezing tolerance. During cold exposure, many carbohydrates accumulate and a new metabolic homeostasis evolves. In the present study, we analyse the diurnal dynamics of carbohydrate homeostasis before and after cold exposure in three natural accessions showing distinct cold acclimation capacity. Diurnal dynamics of soluble carbohydrates were found to be significantly different in cold-sensitive and cold-tolerant accessions. Although experimentally determined maximum turnover rates for sucrose phosphate synthase in cold-acclimated leaves were higher for cold-tolerant accessions, model simulations of diurnal carbohydrate dynamics revealed similar fluxes. This implied a significantly higher capacity for sucrose synthesis in cold-tolerant than cold-sensitive accessions. Based on this implication resulting from mathematical model simulation, a critical temperature for sucrose synthesis was calculated using the Arrhenius equation and experimentally validated in the cold-sensitive accession C24. At the critical temperature suggested by model simulation, an imbalance in photosynthetic carbon fixation ultimately resulting in oxidative stress was observed. It is therefore concluded that metabolic capacities at least in part determine the ability of accessions of Arabidopsis thaliana to cope with changes in environmental conditions.