Diversity, distribution and roles of osmoprotective compounds accumulated in halophytes under abiotic stress.

BACKGROUND AND AIMS: Osmolytes are low-molecular-weight organic solutes, a broad group that encompasses a variety of compounds such as amino acids, tertiary sulphonium and quaternary ammonium compounds, sugars and polyhydric alcohols. Osmolytes are accumulated in the cytoplasm of halophytic species in order to balance the osmotic potential of the Na(+) and Cl(-) accumulated in the vacuole. The advantages of the accumulation of osmolytes are that they keep the main physiological functions of the cell active, the induction of their biosynthesis is controlled by environmental cues, and they can be synthesized at all developmental stages. In addition to their role in osmoregulation, osmolytes have crucial functions in protecting subcellular structures and in scavenging reactive oxygen species. SCOPE: This review discusses the diversity of osmolytes among halophytes and their distribution within taxonomic groups, the intrinsic and extrinsic factors that influence their accumulation, and their role in osmoregulation and osmoprotection. Increasing the osmolyte content in plants is an interesting strategy to improve the growth and yield of crops upon exposure to salinity. Examples of transgenic plants as well as exogenous applications of some osmolytes are also discussed. Finally, the potential use of osmolytes in protein stabilization and solvation in biotechnology, including the pharmaceutical industry and medicine, are considered.

Silicon mitigates salinity effects on sorghum-sudangrass (Sorghum bicolor × Sorghum sudanense) by enhancing growth and photosynthetic efficiency.

The aim of this study was to investigate whether silicon (Si) supply was able to alleviate the harmful effects caused by salinity stress on sorghum-sudangrass (Sorghum bicolor ×Sorghum sudanense ), a species of grass raised for forage and grain. Plants were grown in the presence or absence of 150mM NaCl, supplemented or not with Si (0.5mM Si). Biomass production, water and mineral status, photosynthetic pigment contents, and gas exchange parameters were investigated. Special focus was accorded to evaluating the PSI and PSII. Salinity stress significantly reduced plant growth and tissue hydration, and led to a significant decrease in all other studied parameters. Si supply enhanced whole plant biomass production by 50%, improved water status, decreased Na+ and Cl- accumulation, and even restored chlorophyll a , chlorophyll b , and carotenoid contents. Interestingly, both photosystem activities (PSI and PSII) were enhanced with Si addition. However, a more pronounced enhancement was noted in PSI compared with PSII, with a greater oxidation state upon Si supply. Our findings confirm that Si mitigated the adverse effects of salinity on sorghum-sudangrass throughout adverse approaches. Application of Si in sorghum appears to be an efficient key solution for managing salt-damaging effects on plants.

Mitigating Salinity Stress in Quinoa (Chenopodium quinoa Willd.) with Biochar and Superabsorber Polymer Amendments.

In agriculture, soil amendments are applied to improve soil quality by increasing the water retention capacity and regulating the pH and ion exchange. Our study was carried out to investigate the impact of a commercial biochar (Bc) and a superabsorbent polymer (SAP) on the physiological and biochemical processes and the growth performance of Chenopodium quinoa (variety ICBA-5) when exposed to high salinity. Plants were grown for 25 days under controlled greenhouse conditions in pots filled with a soil mixture with or without 3% Bc or 0.2% SAP by volume before the initiation of 27 days of growth in hypersaline conditions, following the addition of 300 mM NaCl. Without the Bc or soil amendments, multiple negative effects of hypersalinity were detected on photosynthetic CO2 assimilation (Anet minus 70%) and on the production of fresh matter from the whole plant, leaves, stems and roots (respectively, 55, 46, 64 and 66%). Moreover, increased generation of reactive oxygen species (ROS) was indicated by higher levels of MDA (plus 142%), antioxidant activities and high proline levels (plus 311%). In the pots treated with 300 mM NaCl, the amendments Bc or SAP improved the plant growth parameters, including fresh matter production (by 10 and 17%), an increased chlorophyll content by 9 and 13% and Anet in plants (by 98 and 115%). Both amendments (Bc and SAP) resulted in significant salinity mitigation effects, decreasing proline and malondialdehyde (MDA) levels whilst increasing both the activity of enzymatic antioxidants and non-enzymatic antioxidants that reduce the levels of ROS. This study confirms how soil amendments can help to improve plant performance and expand the productive range into saline areas.

Impact of Biochar Application at Water Shortage on Biochemical and Physiological Processes in Medicago ciliaris.

The application of biochar is mostly used to improve soil fertility, water retention capacity and nutrient uptake. The present study was conducted in order to study the impact of biochar at water deficiency conditions on the physiological and biochemical processes of Medicago ciliaris seedlings. Seedlings were cultivated under greenhouse conditions in pots filled with a mixture of soil and sand mixed in the presence or absence of 2% biochar. Plants of uniform size were subjected after a pretreatment phase (72 days) either to low (36% water holding capacity, water potential low) or high soil water potential (60% water holding capacity, water potential high). Pots were weighed every day to control and maintain a stable water holding capacity. In Medicago ciliaris, drought led to a significant reduction in plant growth and an increase in the root/shoot ratio. The growth response was accompanied by a decreased stomatal conductance and a reduction of the net CO2 assimilation rate and water use efficiency. The associated higher risk of ROS production was indicated by a high level of lipid peroxidation, high antioxidant activities and high proline accumulation. Soil amendment with biochar enhanced the growth significantly and supported the photosynthetic apparatus of Medicago ciliaris species by boosting chlorophyll content and Anet both under well and insufficient watered plants and water use efficiency in case of water shortage. This increase of water use efficiency was correlated with the biochar-mediated decrease of the MDA and proline contents in the leaves buffering the impact of drought on photosynthetic apparatus by increasing the activity of enzymatic antioxidants SOD, APX, GPOX and GR and non-enzymatic antioxidants, such as AsA and DHAsA, giving the overall picture of a moderate stress response. These results confirmed the hypothesis that biochar application significantly reduces both the degree of stress and the negative impact of oxidative stress on Medicago ciliaris plants. These results implied that this species could be suitable as a cash pasture plant in the development of agriculture on dry wasteland in a future world of water shortages.

Combined effects of long-term salinity and soil drying on growth, water relations, nutrient status and proline accumulation of Sesuvium portulacastrum.

The interaction between soil drying and salinity was studied in the perennial halophyte, Sesuvium portulacastrum. Rooted cuttings were individually cultivated for three months in silty-sandy soil under two irrigation modes: 100 and 25% of field capacity (FC). The amount of the evapotranspirated water was replaced by a nutrient solution containing either 0 or 100 mM NaCl. Whole-plant growth, leaf water content, leaf water potential (Psi(w)), and Na+, K+, and proline concentrations in the tissues were measured. When individually applied, both drought and salinity significantly restricted whole-plant growth, with a more marked effect of the former stress. However, the effects of the two stresses were not additive on whole-plant biomass or on leaf expansion. Root growth was more sensitive to salt than to soil drying, the latter being even magnified by the adverse impact of salinity. Leaf water content was significantly reduced following exposure to water-deficit stress, but was less affected in salt-treated plants. When simultaneously submitted to water-deficit stress and salinity, plants displayed higher values of water and potassium use efficiencies, leaf proline and Na+ concentrations, associated with lower leaf water potential (-1.87 MPa), suggesting the ability of S. portulacastrum to use Na+ and proline for osmotic adjustment.

Effects of Cd2+ on K+, Ca2+ and N uptake in two halophytes Sesuvium portulacastrum and Mesembryanthemum crystallinum: consequences on growth.

One of the limits of Cd2+-phytoextraction is the high toxicity of this metal to plants. Growth restriction, chlorosis and necrosis are usually accompanied with a large disturbance of the uptake of essential elements. This work aims to study the effects of cadmium (Cd2+) on potassium (K+), calcium (Ca2+) and nitrogen (N) acquisition, and their consequences on growth in two halophytes species: Sesuvium portulacastrum and Mesembryanthemum crystallinum. Seedlings were grown for 30 days in split-root conditions. One half of the root system was immersed in complete nutrient solution (Basal medium (B)) supplemented with 100 microM Cd2+, and the other half was immersed in a Cd2+-free medium, containing all nutrients (B/Cd plants) or deprived of potassium ((B-K)/Cd) or calcium ((B-Ca)/Cd) or nitrogen ((B-N)/Cd). Using this approach, we demonstrated that K+ and Ca2+ uptake was impaired in roots exposed to Cd2+. Concerning N, we noticed no indication of uptake inhibition by Cd2+. However, restriction of K+ uptake by roots was compensated by an increase in the K+-use efficiency, so that growth was not inhibited. Calcium uptake was strongly limited by Cd2. This inhibition was accompanied by a reduction in growth of ((B-Ca)/Cd) plants. Thus, we conclude that Cd2+ limits growth of both halophytes through restriction imposed on Ca2+ uptake. We suggest that the increase of Ca2+ availability in soils could improve the growth of both species in the presence of Cd2+. This would be essential for improving their utility for extraction of this metal by from salty contaminated soils.

Effects of exogenous nitric oxide on growth, proline accumulation and antioxidant capacity in Cakile maritima seedlings subjected to water deficit stress.

Nitric oxide (NO) - an endogenous signalling molecule in plants and animals - mediates responses to biotic and abiotic stresses. In the present study, we examined the role of exogenous application of NO in mediating stress responses in Cakile maritima Scop. seedlings under water deficit stress using sodium nitroprusside (SNP) as NO donor and as a pre-treatment before the application of stress. Water deficit stress was applied by withholding water for 14 days. Growth, leaf water content (LWC), osmotic potential (ψs), chlorophyll, malondialdehyde (MDA), electrolyte leakage (EL), proline and Δ1-pyrroline-5-carboxylate synthetase (P5CS) and proline dehydrogenase (ProDH) protein levels were determined. Enzyme activities involved in antioxidant activities (superoxide dismutase (SOD) and catalase (CAT)) were measured upon withholding water. The results showed that shoot biomass production was significantly decreased in plants subjected to water deficit stress alone. However, in water deficit stressed plants pre-treated with SNP, growth activity was improved and proline accumulation was significantly increased. Proline accumulation was concomitant with the stimulation of its biosynthesis as shown by the accumulation of P5CS proteins. Nevertheless, no significant change in ProDH protein levels was observed. Besides plants showed lower water deficit-induced lipid membrane degradation and oxidative stress after the pretreatment with 100µM SNP. This behaviour was related to the increased activity of SOD and CAT. Thus, we concluded that NO increased C. maritima drought tolerance and mitigated damage associated with water deficit stress by the regulation of proline metabolism and the reduction of oxidative damage.

Cadmium effects on growth and mineral nutrition of two halophytes: Sesuvium portulacastrum and Mesembryanthemum crystallinum.

Growth, cadmium accumulation and potassium and calcium status were studied in two halophytes from Aizoaceae family: Sesuvium portulacastrum and Mesembryanthemum crystallinum. After multiplication, the seedlings were cultivated on nutrient solution supplemented with NaCl (100mM) and CdCl2 (0, 50, 100, 200 and 300 microM). After 1 month of treatment, plants were harvested and the dry weight, as well as the Cd, K and Ca concentrations in tissues were determined. Results showed that S. portulacastrum, a perennial halophyte with slow growth, is significantly more tolerant to Cd than M. crystallinum, an annual plant. Cd severely inhibited Mesembryanthemum growth even at the lowest Cd concentration in culture medium (50 microM), and did not modify significantly that of Sesuvium. For both halophytes, Cd accumulation was significantly higher in the roots than in the shoots. However, Cd concentration reached 350-700 microg g(-1) DM in the shoots, values characteristic of Cd hyperaccumulator plants. The addition of Cd in the culture medium led to a disturbance of Ca and especially K nutrition, suggesting the possibility to improve plant growth and Cd phytoextraction of both halophytes by increasing nutrient availability in the culture medium.

Medicago sativa--Sinorhizobium meliloti Symbiosis Promotes the Bioaccumulation of Zinc in Nodulated Roots.

In this study we investigated effects of Zn supply on germination, growth, inorganic solutes (Zn, Ca, Fe, and Mg) partitioning and nodulation of Medicago sativa This plant was cultivated with and without Zn (2 mM). Treatments were plants without (control) and with Zn tolerant strain (S532), Zn intolerant strain (S112) and 2 mM urea nitrogen fertilisation. Results showed that M. sativa germinates at rates of 50% at 2 mM Zn. For plants given nitrogen fertilisation, Zn increased plant biomass production. When grown with symbionts, Zn supply had no effect on nodulation. Moreover, plants with S112 showed a decrease of shoot and roots biomasses. However, in symbiosis with S532, an increase of roots biomass was observed. Plants in symbiosis with S. meliloti accumulated more Zn in their roots than nitrogen fertilised plants. Zn supply results in an increase of Ca concentration in roots of fertilised nitrogen plants. However, under Zn supply, Fe concentration decreased in roots and increased in nodules of plants with S112. Zn supply showed contrasting effects on Mg concentrations for plants with nitrogen fertilisation (increase) and plants with S112 (decrease). The capacity of M. sativa to accumulate Zn in their nodulated roots encouraged its use in phytostabilisation processes.

Insights into the physiological responses of the facultative halophyte Aeluropus littoralis to the combined effects of salinity and phosphorus availability.

In this work, we investigate the physiological responses to P deficiency (5µM KH2PO4=D), salt stress (400mM NaCl=C+S), and their combination (D+S) on the facultative halophyte Aeluropus littoralis to understand how plants adapt to these combined stresses. When individually applied, both P deficiency and salinity significantly restricted whole plant growth, with a more marked effect of the latter stress. However, the effects of the two stresses were not additive in plant biomass production since the response of plants to combined salinity and P deficiency was similar to that of plants grown under salt stress alone. In addition the observed features under salinity alone are kept when plants are simultaneously subjected to the combined effects of salinity and P deficiency such as biomass partitioning; the synthesis of proline and the K(+)/Na(+) selectivity ratio. Thus, increasing P availability under saline conditions has no significant effect on salt tolerance in this species. Plants cultivated under the combined effects of salinity and P deficiency exhibited the lowest leaf water potential. This trend was associated with a high accumulation of Na(+), Cl(-) and proline in shoots of salt treated plants suggesting the involvement of these solutes in osmotic adjustment. Proline could be involved in other physiological processes such as free radical scavenging. Furthermore, salinity has no significant effect on phosphorus acquisition when combined with a low P supply and it significantly decreased this parameter when combined with a sufficient P supply. This fact was probably due to salt's effect on P transporters. In addition, shoot soluble sugars accumulation under both P deficiency treatments with and without salt likely play an important role in the adaptation of A. littoralis plants to P shortage applied alone or combined with salinity. Moreover, there was a strong correlation between shoot and root intracellular acid phosphatase activity and phosphorus use efficiency which strengthens the assumption that intracellular acid phosphatase enzymes are involved in P remobilization in this species. Finally, our results showed that P availability has no significant effect on salt excretion in A. littorlais which suggests that independently of the P status in the plant, excretion remains priority over other functions requiring energy such as growth. This result could also indicate that salt excretion is not energy-dependent in this species.

Role of the vancomycin-ristocetin heterodimerization on the enantioselectivity of D,L-tryptophan and D,L-dansyl tryptophan.

In this paper, a chromatographic system using immobilized ristocetin as chiral stationary phase and vancomycin as chiral mobile phase additive (CMPA) was described in order to investigate the role of the glycopeptide heterodimerization on the retention and enantioselectivity of D,L-tryptophan and D,L-dansyl tryptophan. A simplified interaction model was derived considering the formation of heterodimers between immobilized ristocetin and vancomycin. This theoretical approach was convenient to describe adequately the retention behavior. When the CMPA concentration increased, the solute retention factor increased for all the solute enantiomers studied indicating that the vancomycin adsorbed on the immobilized ristocetin played a preponderant role in the retention. The D,L-tryptophan enantioselectivity on the dynamically modified stationary phase was improved by a factor of 1.3, probably due to a glycopeptide conformational change upon heterodimerization. On the other hand, a decrease in the chiral discrimination of D,L-dansyl tryptophan was observed. Such a behavior seems to result from the antagonist enantioselective properties of the two glycopeptides for the dansyl amino acids.

Displacement study on a vancomycin-based stationary phase using N-acetyl-D-alanine as a competing agent.

The analysis of the binding data of D,L-dansyl amino acids on a vancomycin stationary phase is investigated in relation to the addition of N-acetyl-D-alanine in the mobile phase. This eluent additive acts as a specific competing agent for the aglycone pocket of the immobilized chiral selector. A model taking into account both stereoselective and nonstereoselective interactions between the solutes and the stationary phase is used to fit the experimental data. From the results, the theoretical approach is considered to be adequate to describe the competing agent dependence on solute retention. To the best of our knowledge, this report constitutes the first example of a displacement study on a macrocyclic antibiotic stationary phase. This work shows that dansyl amino acids bind to the active aglycone pocket of the selector and that this interaction is enantioselective. The results also demonstrate that additional enantioselective sites at the vancomycin surface are involved in the chiral discrimination of these solutes.

Physiological response of halophytes to multiple stresses.

As halophytes grow vigorously in saline soils, they serve as extraordinary resources for the identification and development of new crop systems. Understanding the mechanisms of tolerance of halophytes to salinity in combination with other co-occurring constraints such as drought, flooding, heavy metals and nutrient deficiencies, would facilitate efforts to use halophytes for saline land revegetation, as well as provide new insights that might be considered in future breeding of plants for salt-affected agricultural lands. Recent results suggest that salinity may improve the response of halophytes to other stresses. Some physiological and biochemical mechanisms of tolerance to salinity are common to many halophytes when plants are subjected to salinity, whereas others are specifically amplified under a combination of stresses. Therefore, the response of halophytes to multiple stresses may not reflect an additive effect of these constraints, but rather, constitute specific response to a new situation where many constraints are operating simultaneously. Comparative studies between halophytes and glycophytes have shown that halophytes are better equipped with the mechanisms of cross-stress tolerance and are constitutively prepared for stress. Moreover, other data has shown that the pre-treatment of halophytes with salinity or other constraints in the early stages of development improves their subsequent response to salinity, which suggests the capacity of these plants to 'memorise' a previous stress allows them respond positively to subsequent stress.

Implementation of a design space approach for enantiomeric separations in polar organic solvent chromatography.

This paper focuses on implementing a design space approach and on the critical process parameters (CPPs) to consider when applying the Quality by Design (QbD) concepts outlined in ICH Q8(R2), Q9 and Q10 to analytical method development and optimization for three chiral compounds developed as modulators of small conductance calcium-activated potassium (SK) channels. In this sense, an HPLC method using a polysaccharide-based stationary phase containing a cellulose tris (4-chloro-3-methylphenylcarbamate) chiral selector in polar organic solvent chromatography mode was considered. The effects of trifluoroacetic acid (TFA) and n-hexane concentration in an acetonitrile (MeCN) mobile phase were investigated under a wide range of column temperatures. Good correlations were found between the observed data obtained after using a central composite design and the expected chromatographic behaviours predicted by applying the design of experiments-design space (DoE-DS) methodology. The critical quality attribute represented here by the separation criterion (S(crit)) allowed assessing the quality of the enantioseparation. Baseline separation for the compounds of interest in an analysis time of less than 20 min was possible due to the original and powerful tools applied which facilitated an enhanced method comprehension. Finally, the advantage of the DoE-DS approach resides in granting the possibility to concurrently assess robustness and identify the optimal conditions which are compound dependent.

Effects of water deficit stress on growth, water relations and osmolyte accumulation in Medicago truncatula and M. laciniata populations.

The effects of water stress were investigated in two Tunisian Medicago truncatula populations collected from arid (Mt-173) and sub-humid (Mt-664) climates and two Tunisian M. laciniata populations originating from arid (Ml-173) and semi-arid (Ml-345) regions. After a pre-treatment phase (24 days after sowing, DAS) of watering at 100% of field capacity (FC), the plants were either irrigated at 100% FC or at only 33% FC. After 12 days of treatment (36 DAS), one lot of dehydrated plants was rewatered at 100% FC. A final harvest was carried out after 24 days of treatment (48 DAS). Measured parameters were total dry weight (TDW), root shoot ratio (RSR), leaf relative water content (RWC), osmotic potential (OP), photosynthetic parameters (CO(2) net assimilation A, stomatal conductance g(s) and transpiration E), malondialdehyde (MDA) concentration and leaf contents in inorganic (Na(+) and K(+)) and organic solutes (proline and soluble sugars). Under water deficit conditions, compared to M. laciniata, M. truncatula populations showed a higher reduction in TDW, A, g(s) and RWC associated with a higher increase in MDA concentration. Thus, the relative tolerance of M. laciniata populations to water shortage would be related to their lower intrinsic growth rate and stomatal control of gas exchange. TDW, A, g(s), E and RWC were more decreased by water deficit in Ml-345 than in Ml-173. Drought tolerance of Ml-173 was found to be associated with a more pronounced decrease of OP and a lower reduction in RWC due to the accumulation of solutes such as proline, soluble sugars and K(+). In addition, Ml-173 showed the highest water use efficiency values (WUE) and the lowest MDA concentrations under water deficit stress.

Cd-induced growth reduction in the halophyte Sesuvium portulacastrum is significantly improved by NaCl.

The effects of Cd(2+) and NaCl, applied together or separately, on growth and uptake of Cd(2+) were determined for the halophyte Sesuvium portulacastrum L. Seedlings were cultivated in the presence of 50 or 100 micromol L(-1) Cd(2+) alone or combined with 100 or 400 mmol L(-1) NaCl. Data showed that alone, Cd(2+) induced chlorosis, necrosis, and inhibited growth. Addition of NaCl to Cd(2+)-containing medium restored growth and alleviated the toxicity, however. NaCl also enhanced the amounts of Cd(2+) accumulated in the shoots. All Cd(2+) treatment reduced K(+) and Ca(2+) uptake and transport to the shoots. Accumulation of Na(+) in the shoots was not affected by Cd(2+), however. Thus S. portulacastrum maintained its halophytic characteristics in the presence of Cd(2+). We suggest this halophyte could be used for phytoextraction of Cd(2+) from salt-contaminated sites.

Effect of sodium chloride on the response of the halophyte species Sesuvium portulacastrum grown in mannitol-induced water stress.

Sesuvium portulacastrum is a halophytic species well adapted to salinity and drought. In order to evaluate the physiological impact of salt on water deficit-induced stress response, we cultivated seedlings for 12 days, in the presence or absence of 100 mmol l(-1) NaCl, on a nutrient solution containing either 0 mmol l(-1) or 25 mmol l(-1) mannitol. Mannitol-induced water stress reduced growth, increased the root/shoot ratio, and led to a significant decrease in water potential and leaf relative water content, whereas leaf Na(+) and K(+) concentrations remained unchanged. The addition of 100 mmol l(-1) NaCl to 25 mmol l(-1) mannitol-containing medium mitigated the deleterious impact of water stress on growth of S. portulacastrum, improved the relative water content, induced a significant decrease in leaf water potential and, concomitantly, resulted in enhancement of overall plant photosynthetic activity (i.e. CO(2) net assimilation rate, stomatal conductance). Presence of NaCl in the culture medium, together with mannitol, significantly increased the level of Na(+) and proline in the leaves, but it had no effect on leaf soluble sugar content. These findings suggest that the ability of NaCl to improve plant performance under mannitol-induced water stress may be due to its effect on osmotic adjustment through Na(+) and proline accumulation, which is coupled with an improvement in photosynthetic activity. A striking recovery in relative water content and growth of the seedlings was also recorded in the presence of NaCl on release of the water stress induced by mannitol.

A framework based on the extended Wyman concept for analyzing the salt effects on the solute retention in high-performance affinity chromatography.

The analysis of binding data of a ligand to a macromolecule in the presence of an additive can be classically formulated in terms of the linked functions of Wyman. In the case of a salt, this approach has been extended by Tanford such that the contributions of both salt and water are taken into account. In this paper, the extended Wyman theory was applied to high-performance affinity chromatography (HPAC) in order to define a general model describing the effects of the mobile-phase salts on the ligand binding. Various HPAC literature data, as well as our data concerning dansyl amino acid retention on a vancomycin stationary phase, were examined in relation to this model. From the results, this theoretical approach was considered to be adequate to describe accurately the salt dependence on solute retention. This work shows the importance of taking into account the effects of both ionic species and water in the investigation of relative contributions of the interactions involved in the ligand binding to immobilized receptor.

Vancomycin dimerization and chiral recognition studied by high-performance liquid chromatography.

The retention and separation of D,L-dansylvaline enantiomers (used as test solutes) were investigated using silica gel as stationary phase and vancomycin as chiral mobile-phase additive. A retention model was developed to describe the mechanistic aspects of the interaction between solute and vancomycin in the chromatographic system. It considered the formation of vancomycin dimers both "free" in the mobile phase and adsorbed on silica. By fitting the model equation to experimental data, it appeared clearly that the approach taking into account the vancomycin dimerization described accurately the retention behavior of the compounds. The examination of the model equation parameters showed that the glycopeptide dimerization increased the enantioselectivity by a factor of approximately 3.7. This study demonstrated the preponderant role of the vancomycin dimerization on the chiral recognition process of D,L-dansylvaline. Also, an additional analysis on a vancomycin chiral stationary phase indicated that the addition of vancomycin in the mobile phase promoted a greater enantioselectivity mediated by the formation of dimers in the stationary phase.