Study on pathway and characteristics of ion secretion of salt glands of Limonium bicolor

Recretohalophytes with specialized salt-secreting structures, including salt glands and salt bladders, can secrete excess salts from plant tissues and enhance salinity tolerance of plants. However, the pathway and property of salt secretion by the salt gland has not been elucidated. In the article, Limonium bicolor Kuntze was used to investigate the pathway and characteristics of salt secretion of salt gland. Scanning electron microscope micrographs showed that each of the secretory cells had a pore in the center of the cuticle, and the rice grain-like secretions were observed above the pore. The chemical composition of secretions from secretory pores was mainly NaCl using environmental scanning electron microscope technique. Non-invasive micro-test technology was used to directly measure ion secretion rate of salt gland, and secretion rates of Na⁺ and Cl^? were greatly enhanced by a 200-mmol/L NaCl treatment. However, epidermal cells and stoma showed little secretion of ions. In conclusion, our results provide evidence that the salt glands of L. bicolor have four secretory pores and that NaCl is secreted through these pores of salt gland.     

Coping with excess salt: adaptive functions of extrarenal osmoregulatory organs in vertebrates

In all organisms, changing environmental conditions require appropriate regulatory measures to physiologically adjust to the altered situation. Uptake of excess salt in non-mammalian vertebrates having limited or no access to freshwater is balanced by extrarenal salt excretion through specialized structures called ‘salt glands’. Nasal salt glands of marine birds are usually fully developed in very early stages of their lives since individuals of these species are exposed to salt soon after hatching. In individuals of other bird species, salt uptake may occur infrequently. In these animals, glands are usually quiescent and glandular cells are kept in a fairly undifferentiated state. This is the situation in ‘naive’ ducklings, Anas platyrhynchos, which have never been exposed to excess salt. When these animals become initially osmotically stressed, the nasal glands start to secrete a moderately hypertonic sodium chloride solution but secretory performance is meager. Within 48 h after the initial stimulus, however, the number of cells per gland is elevated by a factor of 2–3, the secretory cells differentiate and acquire full secretory capacity. During this differentiation process, extensive surface specializations are formed. The number of mitochondria is increased and metabolic enzymes and transporters are upregulated. These adaptive growth and differentiation processes result in a much higher efficiency of salt excretion in acclimated ducklings compared with naive animals. Receptors and signal transduction pathways in salt gland cells controling the adaptive processes seem to be the same as those controling salt secretion, namely muscarinic acetylcholine receptors and receptors for vasoactive intestinal peptide. This review focusses on signal transduction pathways activated by muscarinic receptors which seem to fine-tune salt secretion in salt-adapted ducklings and may control adaptive growth and differentiation processes in the nasal gland of naive animals.     

Dynamic secretion changes in the salt glands of the mangrove tree species Avicennia officinalis in response to a changing saline environment

The specialized salt glands on the epidermis of halophytic plants secrete excess salts from tissues by a mechanism that is poorly understood. We examined the salt glands as putative salt and water bi‐regulatory units that can respond swiftly to altering environmental cues. The tropical mangrove tree species (Avicennia officinalis) is able to grow under fluctuating salinities (0.7–50.0 dS m^?1) at intertidal zones, and its salt glands offer an excellent platform to investigate their dynamic responses under rapidly changing salinities. Utilizing a novel epidermal peel system, secretion profiles of hundreds of individual salt glands examined revealed that these glands could secrete when exposed to varying salinities. Notably, rhythmic fluctuations observed in secretion rates were reversibly inhibited by water channel (aquaporin) blocker, and two aquaporin genes (PIP and TIP) preferentially expressed in the salt gland cells were rapidly induced in response to increasing salt concentration. We propose that aquaporins are involved and contribute to the re‐absorption of water during salt removal in Avicennia officinalis salt glands. This constitutes an adaptive feature that contributes to salt balance of trees growing in saline environments where freshwater availability is limited.     

獐茅盐腺形态结构及其泌盐性

显微观察结果显示,獐茅盐腺为典型的双细胞结构,主要分布在叶脉附近,这样有利于它快速收集来自根部的盐离子。同时,X-ray微区分析结果表明,獐茅盐腺可以有效地将Na^＋从表皮细胞和叶肉细胞转运到基细胞,由帽细胞分泌到体外,从而降低植物体内盐分水平以适应盐渍环境。用不同盐类处理材料,獐茅对Na^＋、K^＋和Ca^2＋的吸收和分泌均表现为具有不同的选择性,分泌量的顺序为Na^＋〉K^＋〉Ca^2＋,并且仅仅24h内,盐分的分泌量就已经超过了叶片内的含量,结果植株体内的总离子含量水平几乎不变,说明獐茅具有较强的泌盐能力。     

Are the glandular trichomes in Jacquinia armillaris (Theophrastoideae—Primulaceae) salt glands?

Salt stress is harmful to plants, especially for those that live under conditions of intense salt aport. For this reason, several species present alternatives to prevent or diminish the damages that high salt concentrations may cause to the cells. Salt glands are one of these alternatives once they are specialized structures that secrete salt. Here, we aimed to investigate if the glandular trichomes in the leaves of Jacquinia armillaris are salt glands. Anatomical and ultrastructural observations showed that the glandular trichomes in J. armillaris resemble the salt glands from other recretohalophytes Primulaceae, such as, their occurrence in sunken regions in the leaf epidermis, the presence of a large basal cell that acts as a collecting cell, the detachment of the cuticle from the outer periclinal walls forming a cuticular chamber, the thickness of the cuticle in the stalk portion of the trichome, and the presence of sodium and chloride ions in the secretion and in the xylem. Altogether, the gathered results support the hypothesis that the glandular trichomes in J. armillaris are adapted to salt secretion, thus characterizing as salt glands.     

Bracelet salt glands of the recretohalophyte Limonium bicolor: Distribution,morphology, and induction

Halophytes complete their life cycles in saline environments. The recretohalophyte Limonium bicolor has evolved a specialized salt secretory structure,the salt gland, which excretes Na⁺to avoid salt damage. Typical L. bicolor salt glands consist of 16 cells with four fluorescent foci and four secretory pores. Here, we describe a special type of salt gland at the base of the L. bicolor leaf petiole named bracelet salt glands due to their beaded-bracelet-like shape of blue auto-fluoresc...     

Regulation of salt gland, gut and kidney interactions

Marine birds can drink seawater because their cephalic 'salt' glands secrete a sodium chloride (NaCl) solution more concentrated than seawater. Salt gland secretion generates osmotically free water that sustains their other physiological processes. Acclimation to saline induces interstitial water and Na move into cells. When the bird drinks seawater, Na enters the plasma from the gut and plasma osmolality (Osm(pl)) increases. This induces water to move out cells expanding the extracellular fluid volume (ECFV). Both increases in Osm(pl) and ECFV stimulate salt gland secretion. The augmented intracellular fluid content should allow more rapid expansion of ECFV in response to elevated Osm(pl) and facilitate activation of salt gland secretion. To fully utilize the potential of the salt glands, intestinally absorbed NaCl must be reabsorbed by the kidneys. Thus, Na uptake at gut and renal levels may constrain extrarenal NaCl secretion. High NaCl intake elevates plasma aldosterone concentration of Pekin ducks and aldosterone stimulates intestinal and renal water and sodium uptake. High NaCl intake induces lengthening of the small intestine of adult Mallards, especially males. High NaCl intake has little effect on glomerular filtration rate or tubular sodium Na uptake of birds with competent salt glands. Relative to body mass, kidney mass and glomerular filtration rate (GFR) are greater in birds with salt glands than in birds that do not have them. Birds with salt glands do not change GFR, when they drink saline. Thus, their renal filtrate contains excess Na that is, in some species, almost completely renally reabsorbed and excreted in a more concentrated salt gland secretion. Na reabsorption by kidneys of other species, like mallards is less complete and their salt glands make less concentrated secretion. Such species may reflux urine into the hindgut, where additional Na may also be reabsorbed for extrarenal secretion. During exposure to saline, marine birds maintain elevated aldosterone levels despite high Na intake. Marine birds are excellent examples of physiological plasticity.     

The host plant as a factor in the synthesis and secretion of salivary glucose oxidase in larval Helicoverpa zea

We investigated the effect of the host plant on the synthesis and secretion of the elicitor glucose oxidase in the salivary glands of larval Helicoverpa zea. Glucose oxidase catalyses the oxidation of d-glucose to produce d-gluconic acid and hydrogen peroxide. Previous studies have found that the product hydrogen peroxide is primarily responsible for suppressing the wound-inducible defenses of the host plant. Using an antibody specific for glucose oxidase, we determined the effect of the host plant on the rate of secretion of glucose oxidase. Larval H. zea secrete microgram amounts of the enzyme glucose oxidase from their principal salivary glands, the labial glands. Larvae reared on different host plants produce varying amounts of glucose oxidase in their labial glands. We used a tissue printing procedure with our antibody to determine if larvae secrete glucose oxidase directly at the feeding or wound sites. Significant amounts of the enzyme are deposited at the feeding site, although some is deposited outside the feeding margins.     

中亚滨藜盐囊泡形态结构与发育研究

应用石蜡切片技术,通过光学显微镜和扫描电镜研究了中亚滨藜(Atriplex centralasiaticaIljin)盐囊泡的形态结构与发育.结果表明:中亚滨藜的盐囊泡起源于叶原基、茎尖及幼叶处的表皮细胞,细胞质浓厚,它经过平周分裂(不均等分裂)形成泡状细胞和柄细胞,有的柄细胞继续分裂形成2个柄细胞,其结构由1~2个柄细胞和顶端1个膨大的泡状细胞构成,外面包被一层很厚的多层次的角质层.中亚滨藜的耐盐结构———盐囊泡是一种适应性结构,具有进一步研究、应用的价值.     

Ion secretion by salt glands of desert iguanas (Dipsosaurus dorsalis)

Unlike the NaCl-secreting salt glands of many birds and reptiles, the nasal salt glands of lizards can secrete potassium as well as sodium, with either chloride or bicarbonate as the accompanying anion. The factors responsible for initiating secretion by the gland and the rates of cation and anion secretion were studied in the desert iguana, Dipsosaurus dorsalis. Lizards were given combinations of ions for several days, and secreted salt was collected daily and analyzed for sodium, potassium, chloride, and bicarbonate. Maximum total cation secretion rate was 4.4+/-0.38 micromol/g/d. Cation secretion ranged from 24% to 100% potassium; even high NaCl loads did not abolish potassium secretion. Maximum bicarbonate secretion was about 0.5 micromol/g/d; chloride was the predominant anion. Secretion rate increased only in response to those treatments that included potassium and/or chloride; sodium ions and other osmotic loads (e.g., sucrose) did not increase secretion. This is in contrast to birds and some other reptiles with salt glands, which initiate NaCl secretion in response to any osmotic load. The specificity of the response of the salt gland of Dipsosaurus may be related to the ecological importance of dietary potassium and chloride for herbivorous desert lizards.     

不同类型盐生植物适应盐胁迫的生理生长机制及Na+逆向转运研究进展

盐生植物是指能在离子浓度至少200 mmol/L以上的生境中生长并完成生活史的植物。盐生植物可分为稀盐盐生植物、泌盐盐生植物、拒盐盐生植物三类。本文从生长形态、生理和分子3个方面总结三类盐生植物响应盐胁迫的不同策略及研究进展,发现盐生植物在分子水平上主要通过Na⁺转运蛋白和为其提供能量的两类基因应对体内过高Na⁺,这可能是引起盐生植物生理和生长形态异于非盐生植物的重要因素。其中稀盐盐生植物主要通过液泡离子区隔化应对盐胁迫,并表现出肉质化生长形态;泌盐盐生植物通过将体内盐分排出体外应对盐胁迫,并进化出特有的生理结构——盐腺或盐囊泡;拒盐盐生植物通过将盐离子积累在皮层细胞液泡和根部木质部薄壁细胞中减少向上运输Na⁺,同时根部多栓质化减少Na⁺吸收。本综述旨在为今后研究盐生植物及其耐盐机制提供相关依据,为植物耐盐分子育种奠定基础。     

二色补血草叶片泌盐结构的扫描电镜观察

对二色补血草盐泉结构进行比较研究,发现二色补血草叶下表皮具有花型盐腺,这些盐腺是由基细胞和帽细胞构成,植物体靠盐腺的泌盐孔和基细胞破碎释盐,表明二色补血草具有演化较高级的双重泌盐结构,是二色补血草能在盐碱地生长而不受盐害的主要原因。     

盐胁迫对獐茅生长及Na+和K+含量的影响

用含0～200mmol/L浓度的NaCl的Hoagland培养液处理獐茅幼苗,处理17 d后测定一些生理指标,结果表明獐茅的生长受NaCl抑制程度随浓度增加而增大,没观察到最适盐浓度,且叶片较根部对盐分更敏感;有机干重的比例增大表明有机物在渗透调节中的贡献随之增大.植株在加喷对盐腺向体外排盐有抑制作用的溶液-苯硫酸胆碱100mmol/L后,Na 、K 含量在叶片内增加,而在分泌物中的量降低,并表现出生长进一步受到抑制.X-ray微区分析结果表明獐茅可以将Na 区域化到盐腺细胞,以便将其分泌到体外.     

Acid and base secretion in the Calu-3 model of human serous cells

Submucosal glands are the primary source of airway mucus, a critical component of lung innate defenses. Airway glands are defective in cystic fibrosis (CF), showing a complete absence of secretion to vasoactive intestinal peptide or forskolin, which increase intracellular cAMP concentration. This defect is attributed to gland serous cells, which express the cystic fibrosis transmembrane conductance regulator. Calu-3 cells, which mimic many features of serous cells, secrete Cl(-) and HCO(3)(-), with HCO(3)(-) secretion predominating for forskolin stimulation and Cl(-) secretion predominating for stimuli that open basolateral K(+) channels to hyperpolarize the cells. We used pH stat and ion substitution experiments to clarify the mechanisms and consequences of these two modes of secretion. We confirm that Calu-3 cells secrete primarily HCO(3)(-) in response to forskolin. Unexpectedly, HCO(3)(-) secretion continued in response to K(+) channel openers, with Cl(-) secretion being added to it. Secretion of HCO(3)(-) from hyperpolarized cells occurs via the conversion of CO(2) to HCO(3)(-) and is reduced by approximately 50% with acetazolamide. A gap between the base equivalent current and short-circuit current was observed in all experiments and was traced to secretion of H(+) via a ouabain-sensitive, K(+)-dependent process (possibly H(+)-K(+)-ATPase), which partially neutralized the secreted HCO(3)(-). The conjoint secretion of HCO(3)(-) and H(+) may help explain the puzzling finding that mucus secreted from normal and CF glands has the same acidic pH as does mucus from glands stimulated with forskolin or ACh. It may also help explain how human airway glands produce mucus that is hypotonic.     

不同阴离子对二色补血草盐腺Na＋分泌速率的影响

本文以二色补血草（Limonium bicolor）为实验材料,用Hoagland营养液和200mmol·L—NaCl、NaBr、NaNO3溶液分别处理12h,测定二色补血草盐腺的Na＋分泌速率、叶片Na＋含量和MDA（丙二醛）含量以及质膜透性,并利用非损伤微测技术探索可能与盐腺相关的转运蛋白,以探讨不同阴离子对二色补血草盐腺分泌Na＋的作用及其可能原因。结果表明：在NaCl处理12h时二色补血草叶片Na＋分泌速率达到最大,然后逐渐下降;不同钠盐处理下叶片Na＋分泌速率为NaCl〉NaBr=NaNO3〉Hoagland,而叶片Na＋含量NaBr〉NaCl〉NaNO3〉Hoagland;不同盐处理下叶片质膜透性和MDA含量无显著性差异;利用Na—K—C1共转运体专一性抑制剂bumetanide处理发现Na＋分泌速率显著降低。这些结果表明Na—K—Cl共转运体可能参与盐腺分泌Na＋。     

Isolation of Viable Multicellular Glands from Tissue of the Carnivorous Plant,Nepenthes

Many plants possess specialized structures that are involved in the production and secretion of specific low molecular weight compounds and proteins. These structures are almost always localized on plant surfaces. Among them are nectaries or glandular trichomes. The secreted compounds are often employed in interactions with the biotic environment, for example as attractants for pollinators or deterrents against herbivores.Glands that are unique in several aspects can be found in carnivorous plants. In so-called pitcher plants of the genus Nepenthes, bifunctional glands inside the pitfall-trap on the one hand secrete the digestive fluid, including all enzymes necessary for prey digestion, and on the other hand take-up the released nutrients. Thus, these glands represent an ideal, specialized tissue predestinated to study the underlying molecular, biochemical, and physiological mechanisms of protein secretion and nutrient uptake in plants. Moreover, generally the biosynthesis of secondary compounds produced by many plants equipped with glandular structures could be investigated directly in glands.In order to work on such specialized structures, they need to be isolated efficiently, fast, metabolically active, and without contamination with other tissues. Therefore, a mechanical micropreparation technique was developed and applied for studies on Nepenthes digestion fluid. Here, a protocol is presented that was used to successfully prepare single bifunctional glands from Nepenthes traps, based on a mechanized microsampling platform. The glands could be isolated and directly used further for gene expression analysis by PCR techniques after preparation of RNA.     

Hormone-dependent dissociation of blood flow and secretion rate in the lingual salt glands of the estuarine crocodile, Crocodylus porosus

Salt and water balance in the estuarine crocodile, Crocodylus porosus, involves the coordinated action of both renal and extra-renal tissues. The highly vascularised, lingual salt glands of C. porosus excrete a concentrated sodium chloride solution. In the present study, we examined the in vivo actions of vasoactive intestinal peptide (VIP), B-type natriuretic peptide (BNP) and angiotensin II (ANG II) on the secretion rate and blood perfusion of the lingual salt glands. These peptides were selected for their vasoactive properties in addition to their reported actions on salt gland activity in birds and turtles and rectal gland activity in elasmobranchs. The femoral artery was cannulated in seven juvenile crocodiles for delivery of peptides and measurement of mean blood pressure and heart rate. In addition, secretion rate of, and blood flow to, the salt glands were recorded simultaneously using laser Doppler flowmetry. VIP stimulated salt secretion was coupled to an increase in blood flow and vascular conductance of the lingual salt glands. BNP was a potent stimulant of salt gland secretion, resulting in a maximal secretion rate of more than 15-fold higher than baseline; however, this was not coupled to an increase in perfusion rate, which remained unchanged. ANG II failed to stimulate salt gland secretion and there was a transient decrease in salt gland blood flow and vascular conductance. It is evident from this study that blood flow to, and secretion rate from, the lingual salt glands of C. porosus are regulated independently; indeed, it is apparent that maximal secretion from the salt glands may not require maximal blood flow.     

Salicin from host plant as precursor of salicylaldehyde in defensive secretion of Chrysomeline larvae

ABSTRACT. Phratora vitellinae L. and Chrysomela tremulae F. (Chrysomelinae, Coleoptera) feed on Salix or Populus spp. (Salicaceae). Their larvae, as well as the larvae of other chrysomelines feeding on Salicaceae, secrete salicylaldehyde. In this study, we demonstrate that salicylaldehyde is derived from salicin, a phenylglucoside present in the leaves of the host plant. The concentration of salicylaldehyde in the secretion is positively correlated with the amount of salicin in the food of the larvae. The transformation of salicin into salicylaldehyde occurs in the defence glands since the β-glucosidase activity is 4 times higher in their glands than in the gut. The larvae recover most of the glucose that results from the hydrolysis of salicin. For generalist predators, such as ants, salicylaldehyde is a more potent deterrent than saligenin or salicin.     

关于野大豆盐腺问题的探讨

以中国3个省的盐生野大豆(Glycine soja Sieb. et Zucc.)为材料,在沙基培养、溶液培养和大田种植3种种植条件下用不同浓度的盐处理,观测了茎叶表面附着物的形态分布和腺毛的超微结构,测定了叶片腺毛分泌物中和叶片组织内部Na^ 和Cl^-的含量变化,并对腺毛的3个细胞以及表皮细胞和叶肉细胞内的Na^ 相对含量变化进行了X射线微区分析。结果发现：盐生野大豆茎叶表皮上生长的附着物中只有表皮毛和腺毛,腺毛的形态类似于禾本科植物中的一些盐腺,叶片上的腺毛均生长在叶脉上;腺毛细胞内部结构具有一般盐腺的特点,如有大液泡,稠密的细胞质,大量的线粒体、叶绿体、胞间连丝以及较厚的细胞壁等。通过测定在无盐对照、盐处理和盐处理加盐腺泌盐抑制剂条件下盐生野大豆叶片腺毛分泌物中和叶片组织内部的Na^ 和Cl^-含量,结果显示,盐生野大豆腺毛具有泌盐功能,加入泌盐抑制剂后,其泌盐作用停止;腺毛的3个细胞以及表皮细胞和叶肉细胞内的Na^ 在不同的盐浓度下的微区定位分析结果表明,盐生野大豆叶片的腺毛细胞有较强的积累Na^ 的能力。综合分析认为,盐生野生大豆的腺毛就是具有泌盐功能的盐腺,没有发现其他类型的盐腺。     

Comparative transcriptome analysis of developmental stages of the Limonium bicolor leaf generates insights into salt gland differentiation

With the expansion of saline land worldwide, it is essential to establish a model halophyte to study the salt‐tolerance mechanism. The salt glands in the epidermis of Limonium bicolor (a recretohalophyte) play a pivotal role in salt tolerance by secreting excess salts from tissues. Despite the importance of salt secretion, nothing is known about the molecular mechanisms of salt gland development. In this study, we applied RNA sequencing to profile early leaf development using five distinct developmental stages, which were quantified by successive collections of the first true leaves of L. bicolor with precise spatial and temporal resolution. Specific gene expression patterns were identified for each developmental stage. In particular, we found that genes controlling salt gland differentiation in L. bicolor may evolve in a trichome formation, which was also confirmed by mutants with increased salt gland densities. Genes involved in the special ultrastructure of salt glands were also elucidated. Twenty‐six genes were proposed to participate in salt gland differentiation. Our dataset sheds light on the molecular processes underpinning salt gland development and thus represents a first step towards the bioengineering of active salt‐secretion capacity in crops.