Dynamics of Aerenchyma distribution in the cortex of sulfate-deprived adventitious roots of maize

BACKGROUND AND AIMS: Aerenchyma formation in maize adventitious roots is induced in nutrient solution by the deprivation of sulfate (S) under well-oxygenated conditions. The aim of this research was to examine the extent of aerenchyma formation in the cortex of sulfate-deprived adventitious roots along the root axis, in correlation with the presence of reactive oxygen species (ROS), calcium levels and pH of cortex cells and root lignification. METHODS: The morphometry of the second whorl of adventitious (W2) roots, subject to S-deprivation conditions throughout development, was recorded in terms of root length and lateral root length and distribution. W2 roots divided into sectors according to the mean length of lateral roots, and cross-sections of each were examined for aerenchyma. In-situ detection of alterations in ROS presence, calcium levels and pH were performed by means of fluorescence microscopy using H(2)DCF-DA, fluo-3AM and BCECF, respectively. Lignification was detected using the Wiesner test. KEY RESULTS: S-deprivation reduced shoot growth and enhanced root proliferation. Aerenchyma was found in the cortex of 77 % of the root length, particularly in the region of emerging or developing lateral roots. The basal and apical sectors had no aerenchyma and no aerenchyma connection was found with the shoot. S-deprivation resulted in alterations of ROS, calcium levels and pH in aerenchymatous sectors compared with the basal non-aerenchymatous region. Lignified epidermal layers were located at the basal and the proximal sectors. S-deprivation resulted in shorter lateral roots in the upper sectors and in a limited extension of the lignified layers towards the next lateral root carrying sector. CONCLUSIONS: Lateral root proliferation is accompanied by spatially localized induced cell death in the cortex of developing young maize adventitious roots during S-deprivation.     

Synergistic effects of plant growth retardants and IBA on the formation of adventitious roots in hypocotyl cuttings of mung bean

The synergistic effect of plant growth retardants, such as daminozide, paclobutrazol and triadimefon, and of indole-3-butyric acid (IBA) on the formation of adventitious roots in hypocotyl cuttings of mung bean was studied. The three retardants and IBA all stimulated adventitious root growth, but IBA was the most effective. However, mixtures of the retardants with IBA have proven generally more effective than IBA alone in promoting adventitious root formation. When IBA was applied to the hypocotyls one day after cutting preparation followed by the growth retardant on the second day, there were even more adventitious roots produced than if applied in the reverse order. The effectiveness of the treatments were in the order, IBA followed by growth retardant, IBA + growth retardant together, and IBA alone.Abbreviations IBA indole-3-butyric acid - GA gibberellin     

Inhibition by silver ions of gas space (aerenchyma) formation in adventitious roots of Zea mays L. subjected to exogenous ethylene or to oxygen deficiency

We have studied the role of ethylene in accelerating the lytic formation of gas spaces (aerenchyma) in the cortex of adventitious roots of maize (Zea mays L.) growing in poorly aerated conditions. Such roots had previously been shown to contain increased concentrations of ethylene. Ten day-old maize plants bearing seminal roots and one whorl of emerging adventitious roots were grown in nutrient solution bubbled with air, ethylene in air (0.1 to 5.0 l l^–1), or allowed to become oxygen-deficient in nonaerated (but not completely anaerobic) solution. Additions of 0.1 l l^–1 ethylene or more promoted the formation of aerenchyma, with lysis of up to 47% of the cortical cells. The effects of non-aeration were similar to those of exogenous ethylene. When silver ions, an ethylene antagonist, were present at low, non-toxic concentrations (circa 0.6 M), aerenchyma formation was prevented in ethylene treated roots and in those exposed to oxygen deficiency. Silver ions also blocked the inhibiting effect of exogenous ethylene on root extension. By contrast, the suppression of aerenchyma formation by silver ions under oxygendeficient conditions was associated with a retardation of root extension, indicating the importance of aerenchyma for root growth in poorly aerated media. Rates of production of ethylene by excised roots were stimulated by a previous non-aeration treatment. The effectiveness of Ag⁺ in inhibiting equally the action on cortical cells of exogenous ethylene and of non-aeration, supports the view that gas space (aerenchyma) formation in adventitious roots adpted to oxygendeficient environments is mediated by increased concentrations of endogenous ethylene. The possibility that extra ethylene could arise from increased biosynthesis of a precursor in root tissues with a restricted oxygen supply is discussed.     

Stem hypertrophic lenticels and secondary aerenchyma enable oxygen transport to roots of soybean in flooded soil

Background and Aims

Aerenchyma provides a low-resistance O₂ transport pathway that enhances plant survival during soil flooding. When in flooded soil, soybean produces aerenchyma and hypertrophic stem lenticels. The aims of this study were to investigate O₂ dynamics in stem aerenchyma and evaluate O₂ supply via stem lenticels to the roots of soybean during soil flooding.

Methods

Oxygen dynamics in aerenchymatous stems were investigated using Clark-type O₂ microelectrodes, and O₂ transport to roots was evaluated using stable-isotope ¹⁸O₂ as a tracer, for plants with shoots in air and roots in flooded sand or soil. Short-term experiments also assessed venting of CO₂ via the stem lenticels.

Key Results

The radial distribution of the O₂ partial pressure (pO₂) was stable at 17 kPa in the stem aerenchyma 15 mm below the water level, but rapidly declined to 8 kPa at 200–300 µm inside the stele. Complete submergence of the hypertrophic lenticels at the stem base, with the remainder of the shoot still in air, resulted in gradual declines in pO₂ in stem aerenchyma from 17·5 to 7·6 kPa at 13 mm below the water level, and from 14·7 to 6·1 kPa at 51 mm below the water level. Subsequently, re-exposure of the lenticels to air caused pO₂ to increase again to 14–17 kPa at both positions within 10 min. After introducing ¹⁸O₂ gas via the stem lenticels, significant ¹⁸O₂ enrichment in water extracted from roots after 3 h was confirmed, suggesting that transported O₂ sustained root respiration. In contrast, slight ¹⁸O₂ enrichment was detected 3 h after treatment of stems that lacked aerenchyma and lenticels. Moreover, aerenchyma accelerated venting of CO₂ from submerged tissues to the atmosphere.

Conclusions

Hypertrophic lenticels on the stem of soybean, just above the water surface, are entry points for O₂, and these connect to aerenchyma and enable O₂ transport into roots in flooded soil. Stems that develop aerenchyma thus serve as a ‘snorkel’ that enables O₂ movement from air to the submerged roots.     

Photosynthesis in aquatic adventitious roots of the halophytic stem-succulent Tecticornia pergranulata (formerly Halosarcia pergranulata)

In flood-tolerant species, a common response to inundation is growth of adventitious roots into the water column. The capacity for these roots to become photosynthetically active has received scant attention. The experiments presented here show the aquatic adventitious roots of the flood-tolerant, halophytic stem-succulent, Tecticornia pergranulata (subfamily Salicornioideae, Chenopodiaceae) are photosynthetic and quantify for the first time the photosynthetic capacity of aquatic roots for a terrestrial species. Fluorescence microscopy was used to determine the presence of chloroplasts within cells of aquatic roots. Net O₂ production by excised aquatic roots, when underwater, was measured with varying light and CO₂ regimes; the apparent maximum capacity ( P _max) for underwater net photosynthesis in aquatic roots was 0.45  µ mol O₂ m⁻² s⁻¹. The photosynthetic potential of these roots was supported by the immunolocalization of PsbA, the major protein of photosystem II, and ribulose-1-5-bisphosphate carboxylase/oxygenase (Rubisco) in root protein extracts. Chlorophyllous aquatic roots of T. pergranulata are photosynthetically active, and such activity is a previously unrecognized source of O₂, and potentially carbohydrates, in flooded and submerged plants.     

Elongation by primary lateral roots and adventitious roots during conditions of hypoxia and high ethylene concentrations

Soil flooding results in unusually low oxygen concentrations and high ethylene concentrations in the roots of plants. This gas composition had a strongly negative effect on root elongation of two Rumex species. The effect of low oxygen concentrations was less severe when roots contained aerenchymatous tissues, such as in R. palustris Sm. R. thyrsiflorus Fingerh., which has little root porosity, was much more affected. Ethylene had an even stronger effect on root elongation than hypoxia, since very small concentrations (0.1 cm³ m^?3) reduced root extension in the two species, and higher concentrations inhibited elongation more severely than did anoxia in the culture medium. Thus, ethylene contributes strongly to the negative effects of flooding on root growth. An exception may be the highly aerenchymatous, adventitious roots of R. palustris. Aerenchyma in these roots provides a low-resistance diffusion pathway for both endogenously produced ethylene and shoot-derived oxygen. This paper shows that extension by roots of R. palustris in flooded soil depends almost completely on this shoot-derived oxygen, and that aerenchyma prevents accumulation of growth-inhibiting levels of ethylene in the root.     

Waterlogging‐induced adventitious root formation in cucumber is regulated by ethylene and auxin through reactive oxygen species signalling

Development of adventitious roots (ARs) at the base of the shoot is an important adaptation of plants to waterlogging stress; however, its physiological mechanisms remain unclear. Here, we investigated the regulation of AR formation under waterlogged conditions by hormones and reactive oxygen species (ROS) in Cucumis sativus L., an agriculturally and economically important crop in China. We found that ethylene, auxin, and ROS accumulated in the waterlogged cucumber plants. On the other hand, application of the ethylene receptor inhibitor 1‐methylcyclopropene (1‐MCP), the auxin transport inhibitor 1‐naphthylphthalamic acid (NPA), or the NADPH oxidase inhibitor diphenyleneiodonium (DPI) decreased the number of ARs induced by waterlogging. Auxin enhanced the expression of ethylene biosynthesis genes, which led to ethylene entrapment in waterlogged plants. Both ethylene and auxin induced the generation of ROS. Auxin‐induced AR formation was inhibited by 1‐MCP, although ethylene‐induced AR formation was not inhibited by NPA. Both ethylene‐ and auxin‐induced AR formation were counteracted by DPI. These results indicate that auxin‐induced AR formation is dependent on ethylene, whereas ethylene‐induced AR formation is independent of auxin. They also show that ROS signals mediate both ethylene‐ and auxin‐induced AR formation in cucumber plants.     

Adaptations of Central Amazon Tree Species to Prolonged Flooding: Root Morphology and Leaf Longevity

Abstract: Várzeas are species-rich forest communities of the Central Amazon floodplains, inhabited by highly adapted tree species that can withstand long flooding periods. The leaf shedding behaviour and morphological traits that may contribute to adaptation to low oxygen levels were studied at the Ilha de Marchantaria on the lower Solimoes-Amazonas river, Brazil, and in greenhouse experiments with cuttings of six tree species typical of the Amazon floodplain. Comparison of deciduousness in situ revealed that four of the species under investigation, Salix martiana , Tabernaemontana juruana , Laetia corymbulosa and Pouteria glomerata , are able to maintain their leaf system during the aquatic period. Adventitious roots were formed by S. martiana and T. juruana , but anatomical characteristics differed between the species. Whereas S. martiana developed lysigenous aerenchyma in its roots during aerobic and anaerobic growth, only small intercellular spaces of schizogenous origin were formed in the root cortex of T. juruana . Similar to the latter species, such spaces were constitutively formed in the deciduous species Crateva benthami and Vitex cymosa . Suberin deposits were observed in tangential and radial cell walls of the hypodermis of roots from T. juruana , L. corymbulosa and P. glomerata . Suberin deposits were less pronounced in roots of S. martiana and absent in V. cymosa and C. benthami . The data show that different, almost contrasting, survival mechanisms have evolved in roots of plants with similar life forms in the same habitat. The results further suggest that the morphological traits of the investigated trees are causally linked with the in situ leaf shedding behaviour.     

Aerenchyma formation and associated oxygen movement in seminal and nodal roots of wheat

Abstract The present paper describes the effects of growth of roots of wheat (Triticum aestivum cv. Gamenya) in hypoxic nutrient solutions on acrenchyma formation and O₂ movement from shoots to roots. Two types of roots were investigated: (1) seminal roots of 4–7-d-old seedlings, and (2) seminal and nodal roots of 10–28-d-old plants. Gas-filled porosity of seminal and nodal roots increased from 3 to 12% and from 5–7 to 11–15%, respectively, when the roots emerged in stagnant or N₂-flushed solutions (0.003 mol m ^?3 O₂) compared with growth in continuously acrated solutions (0.26 mol m ^?3 O₂). However, neither root type increased in porosity when they were longer than 100–200 mm at the start of the exposure to these stagnant or N₂-flushed treatments. A vernier microscope and cylindrical platinum-electrode were used to examine the relationship between root extension and transport of O₂ from shoots to roots via the gas spaces. Measurements were made when the roots were in an anoxic medium and were dependent solely on O₂ supplied from the shoots. For seminal roots of 5–7-d-old seedlings raised in stagnant solutions (90–100 mm), internal O₂ transport was sufficient to support a rate of root elongation in the O₂-free medium of between 0.03 and 0.17 mm h^?1. When the O₂ pressure around the shoots was increased from 20 to 100 kPa O₂, the O₂ concentrations at the walls of the expanding zone (2–7 mm from the tip) of these roots increased from 0.006 mol m^?3 to between 0.04 and 0.26 mol m^?3, and the rate of root extension increased five-fold. Oxygen transport to roots grown continuously in acrated solutions was considerably less than for roots raised in stagnant solutions; this difference was greater for seminal than for nodal roots. When the acrated seminal roots were longer than 100 mm and transferred to an O₂-free root medium, O₂ concentration became zero at the root tip causing elongation to cease. After 24 h of anoxia, none of these roots were able to resume elongation following a return to acrated solutions.     

Root aeration in rice (Oryza sativa): evaluation of oxygen, carbon dioxide, and ethylene as possible regulators of root acclimatizations

Adventitious roots of rice (Oryza sativa) acclimatize to root-zone O(2) deficiency by increasing porosity, and induction of a barrier to radial O(2) loss (ROL) in basal zones, to enhance longitudinal O(2) diffusion towards the root tip. Changes in root-zone gas composition that might induce these acclimatizations, namely low O(2), elevated ethylene, ethylene-low O(2) interactions, and high CO(2), were evaluated in hydroponic experiments. Neither low O(2) (0 or 0.028 mol m(-3) O(2)), ethylene (0.2 or 2.0 microl l(-1)), or combinations of these treatments, induced the barrier to ROL. This lack of induction of the barrier to ROL was despite a positive response of aerenchyma formation to low O(2) and elevated ethylene. Carbon dioxide at 10 kPa had no effect on root porosity, the barrier to ROL, or on growth. Our findings that ethylene does not induce the barrier to ROL in roots of rice, even though it can enhance aerenchyma formation, shows that these two acclimatizations for improved root aeration are differentially regulated.     

Histological observation of secondary aerenchyma formed immediately after flooding in Sesbania cannabina and S. rostrata

The effect of up to 48 h of flooding on the development of roots of Sesbania cannabina an0d S. rostrata seedlings was examined in a pot experiment. Light microscopy revealed that the outermost cells of the phellogen of the taproot of S. cannabina expanded and elongated during the first 12 h of flooding. After 18 h, the outermost of these regions was composed of cells that had expanded radially direction to form a spongy zone inside the endodermis. These elongated cells were radially connected to each other and formed the secondary aerenchyma surrounding the stele of taproot. While those histological alterations were not observed in S. rostrata, the number of layers of cells originating in the pericycle increased slightly, but elongation of the cells was not found during the first 18 h of flooding. After 36 h of flooding, cell elongation was also detected as outer layers of the phellogen. The delayed response to flooding in aerenchyma production in S. rostrata was compensated by immediate development of adventitious roots on submerged parts of the hypocotyl.     

Similarity and diversity in adventitious root anatomy as related to root aeration among a range of wetland and dryland grass species

Assessments of the anatomy, porosity and profiles of radial O₂ loss from adventitious roots of 10 species in the Poaceae (from four subfamilies) and two species in the Cyperaceae identified a combination of features characteristic of species that inhabit wetland environments. These include a strong barrier to radial O₂ loss in the basal regions of the adventitious roots and extensive aerenchyma formation when grown not only in stagnant but also in aerated nutrient solution. Adventitious root porosity was greater for plants grown in stagnant compared with aerated solution, for all 10 species in the Poaceae. The ‘wetland root’ archetype was best developed in Oryza sativa and the two species of the Cyperaceae, in which the stele contributed less than 5% of the root cross‐sectional area, the cells of the inner cortex were packed in a cuboidal arrangement, and aerenchyma was up to 35–52%. Variations of this root structure, in which the proportional and absolute area of stele was greater, with hexagonal arrangements of cells in the inner cortex and varying in the extent of aerenchyma formation, were present in the other wetland species from the subfamilies Pooideae, Panicoideae and Arundinoideae. Of particular interest were Vetiveria zizanoides and V. filipes, wetland grass species from the tribe Andropogoneae (the same tribe as sorghum, maize and sugarcane), that had a variant of the root anatomy found in rice. The results are promising with regard to enhancing these traits in waterlogging intolerant crops.     

Influence of nicotianamine and iron supply on formation and elongation of adventitious roots in hypocotyl cuttings of the tomato mutant 'chloronerva' (Lycopersicon esculentum)

The influence of nicotianamine (NA) on formation and elongation of adventitious roots in hypocotyls of de-rooted NA-less mutant seedlings of Lycopersicon esculentum Mill, was examined in relation to the iron supply [ferric N-N'-ethylenediaminedi-(2-hydroxyphenylacetate) (FEDDHA), ferric ethylenediaminetetracetate (FeEDTA), ferric N-(2-hydroxyethyl)-ethylenediaminetriacetate (FeHEDTA, Fe-citrate and FeCl₃] in the nutrient solution. The initiation of root primordia in hypocotyl cuttings was independent of NA and occurred with about the same frequency in both, mutant and wild-type. In the mutant the development of primordia to adventitious roots was blocked at all iron sources used, except FeEDTA. Addition of NA (5x 10⁻⁶ to 2 × 10⁻⁵ M ) to the rooting medium resulted in a fast growth of adventitious roots in mutant cuttings with all iron sources tested. Rooting of wild-type cuttings was independent from NA application and iron sources. We suppose that NA is involved in the intracellular transport of iron. Its function is possibly linked with chelation of ferrous iron in the cell.     

Aerenchyma formation and radial O2 loss along adventitious roots of wheat with only the apical root portion exposed to O2 deficiency

This study investigated aerenchyma formation and function in adventitious roots of wheat (Triticum aestivum L.) when only a part of the root system was exposed to O₂ deficiency. Two experimental systems were used: (1) plants in soil waterlogged at 200 mm below the surface; or (2) a nutrient solution system with only the apical region of a single root exposed to deoxygenated stagnant agar solution with the remainder of the root system in aerated nutrient solution. Porosity increased two‐ to three‐fold along the entire length of the adventitious roots that grew into the water‐saturated zone 200 mm below the soil surface, and also increased in roots that grew in the aerobic soil above the water‐saturated zone. Likewise, adventitious roots with only the tips growing into deoxygenated stagnant agar solution developed aerenchyma along the entire main axis. Measurements of radial O₂ loss (ROL), taken using root‐sleeving O₂ electrodes, showed this aerenchyma was functional in conducting O_2. The ROL measured near tips of intact roots in deoxygenated stagnant agar solution, while the basal part of the root remained in aerated solution, was sustained when the atmosphere around the shoot was replaced by N₂. This illustrates the importance of O₂ diffusion into the basal regions of roots within an aerobic zone, and the subsequent longitudinal movement of O₂ within the aerenchyma, to supply O₂ to the tip growing in an O₂ deficient zone.     

诱导子对雷公藤不定根生长和次生代谢产物含量的影响

为探讨真菌诱导子以及硝酸银等非生物诱导子对雷公藤不定根生长及次生代谢产物含量的影响。以雷公藤不定根为材料,通过向培养基中添加不同种类的真菌诱导子以及硝酸银等非生物诱导子,采用高效液相色谱检测不定根中雷公藤甲素和生物碱含量。结果表明:各种真菌诱导子不影响不定根的生长,但对次生代谢产物含量有显著影响,其中,苹果炭疽和柿子炭疽诱导子的加入不仅使不定根中雷公藤甲素的含量分别提高了2.24和1.93倍,生物碱的含量也各提高了2.02和2.07倍。苹果炭疽诱导子浓度为50μg/m L时比较适合雷公藤不定根生长及雷公藤甲素和生物碱的积累。硝酸银抑制不定根的生长和生物碱的积累,但促进雷公藤甲素的积累。硝酸银浓度为25μmol/L时雷公藤甲素含量为对照的1.71倍。茉莉酸甲酯浓度为50μmol/L时,不定根增长量为对照的1.04倍,雷公藤甲素和生物碱含量分别为对照的1.64倍和2.12倍。酵母提取物浓度为2 g/L时,雷公藤甲素含量为对照的1.48倍。表明培养基中添加硝酸银和酵母提取物对不定根中雷公藤甲素的合成具有明显的促进作用,苹果炭疽和茉莉酸甲酯的协同作用既能促进雷公藤甲素的合成又能促进雷公藤生物碱的合成。     

Effect of nitrogen source on biomass and bioactive compound production in submerged cultures of Eleutherococcus koreanum Nakai adventitious roots

Ammonium to nitrate ratios of 0:30, 5:25, 10:20, 15:15, 20:10, 25:5, and 30:0 mM were tested to determine the optimal NH(4)(+) :NO(3)(-) ratio for improving biomass and bioactive compound production in Eleutherococcus koreanum Nakai adventitious roots using 3-L bulb-type bubble bioreactors. A high ammonium nitrogen ratio had a negative effect on root growth, and the highest fresh and dry weights were obtained when NH(4)(+):NO(3)(-) ratios were 5:25 and 10:20 (mM) after 5 weeks of culture. Although the total production of eleutherosides B and E was slightly higher at the 10:20 ratio than at the 5:25 ratio (NH(4)(+):NO(3)(-)), we proposed that the optimal NH(4)(+):NO(3)(-) ratio was 5:25 mM. This ratio achieved both the highest total production of five target bioactive compounds (eleutherosides B and E, chlorogenic acid, total phenolics, and flavonoids) and the highest root biomass. Furthermore, increasing NH(4)(+):NO(3)(-) ratios to 10:20 decreased pH in the medium, interrupted the absorption of essential minerals from the culture medium, and resulted in low biomass and increased relative oxidative stress levels, which were evaluated by determining 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity. Therefore, nitrate rather than ammonium nitrogen was more essential not for only biomass production but also for bioactive compound production in E. koreanum adventitious root cultures. The optimal nitrogen source ratio produced 5.63 g L(-1) of biomass and 24.41 mg of the five total bioactive compounds per gram of biomass (dry weight basis). The development of such in vitro culture technology will benefit the pilot-scale production of E. koreanum-based bioactive compounds for commercialization.     

Variations in the Bioactive Compounds Composition and Biological Activities of Loofah (Luffa cylindrica) Fruits in Relation to Maturation Stages

The present work aimed to determine the antioxidant and antiproliferative potential of Luffa cylindrica fruits collected at two different maturation stages and to identify and compare their functional components composition. The MeOH extracts of L. cylindrica fruits harvested at 60 – 65 days after seeding (S1) and 85 – 90 days after seeding (S2) were investigated for their antioxidant activity using various assays. Furthermore, the antiproliferative activity of the extracts against HeLa human cervical cancer cells was explored with xCELLigence real time cell analyzer, while the effect of the samples on the membrane integrity of the same cell line was assessed using LDH cytotoxicity leakage assay. Ultimately, the phytochemicals were analyzed using GC/MS and HPLC/TOF‐MS. The S1 sample had higher contents and more diversity in the phenolic compounds composition than S2. Furthermore, the S1 extract showed the highest antioxidant and antiproliferative activity, while the S2 extract had higher cytotoxicity towards HeLa cells. The findings revealed that the time of harvest has a big impact on the phytochemicals content and activity and that harvesting L. cylindrica at an early stage before the beginning of the development of the cellulose fibrous system is recommended for a rich phytochemical composition and efficient antioxidant and antiproliferative activities.