Comparative LM, SEM and EDAX study of chalk glands on leaf and stem of two species of Plumbago Linn.

Stem and leaves of two species of PlumbagoLinn.viz. P. zeylanica Linn.andP. auriculata Lam. were investigated for the structure and chemical composition of chalk glands. Light Microscopy (LM) and Scanning Electron Microscopy (SEM) revealed the presence of chalk glands on both lower as well as upper surface of leaf and stem of both species. Chalk glands are abundant on lower surface and sparse on upper surface of leaf. Chalk glands are approximately hemispherical glands with oval or almost circular outline. It is composed of 8 cells arranged in two circles – central circle of 4 secretory cells and outer circle of 4 adjoining cells. Each secretory cell has depression which corresponds to pore. Each gland is surrounded by 4 subsidiary cells. No significant difference in the structure of chalk glands in both species was noticed. Chalk glands occupy three different positions with regard to epidermal cells –at the same level of the epidermis, slightly sunken in the epidermis and slightly raised above the epidermis. Common elements found in EDAX analysis of all chalk glands are carbon, oxygen, magnesium, sulphur, potassium and calcium. Differences in the presence of elements silicon, chlorine, aluminium, sodium, phosphorus were observed. The presence of significant amount of calcium in chalk glands and their dried deposits and absence of sodium and chlorine from dried deposits and even in some chalk glands appealed to use the term ‘Chalk gland’ instead of ‘Salt gland’ in Plumbago.

Coupled Development of Salt Glands, Stomata, and Pavement Cells in Limonium bicolor

Salt-resistant plants have different mechanisms to limit the deleterious effects of high salt in soil; for example, recretohalophytes secrete salt from unique structures called salt glands. Salt glands are the first differentiated epidermal structure of the recretohalophyte sea lavender (Limonium bicolor), followed by stomata and pavement cells. While salt glands and stomata develop prior to leaf expansion, it is not clear whether these steps are connected. Here, we explored the effects of the five phytohormones salicylic acid, brassinolide, methyl jasmonate, gibberellic acid, and abscisic acid on the development of the first expanded leaf of L. bicolor and its potential connection to salt gland, stomata, and pavement cell differentiation. We calculated the total number of salt glands, stomata, and pavement cells, as well as leaf area and pavement cell area, and assessed the correlations between these parameters. We detected strong and positive correlations between salt gland number and pavement cell area, between stomatal number and pavement cell area, and between salt gland number and stomatal number. We observed evidence of coupling between the development of salt glands, stomata, and pavement cells in L. bicolor, which lays the foundation for further investigation of the mechanism behind salt gland development.

Epidermal micromorphology of floret parts in Aeluropus (Poaceae)

Aeluropus from Poaceae comprises 5 species in the world and 3 species in Iran. This halophytic perennial is distributed in salty and dry soils of Asia, Europe, and Africa. In addition to being used as fodder, it can stabilize the soil by its rhizome or stolon. These features make Aeluropus a valuable plant. In this study, lemma and palea of 10 populations of Aeluropus were studied micromorphologically by scanning electron microscope (SEM) to determine diagnostic features among species studied. Eight characters as micro-prickle, macro-hair, long cell outline, cork and silica cells, papilla, salt gland, and epicuticular wax were studied. The occurrence of salt glands and silica cells in populations/taxa studied showed the ability of Aeluropus to tolerate harsh habitats. Our result showed the taxonomic value of floret micromorphological features to separate Aeluropus species.

Salt-tolerance screening in Limonium sinuatum varieties with different flower colors

Limonium sinuatum, a member of Plumbaginaceae commonly known as sea lavender, is widely used as dried flower. Five L. sinuatum varieties with different flower colors (White, Blue, Pink, Yellow, and Purple) are found in saline regions and are widely cultivated in gardens. In the current study, we evaluated the salt tolerance of these varieties under 250 mmol/L NaCl (salt-tolerance threshold) treatment to identify the optimal variety suitable for planting in saline lands. After the measurement of the fresh weight (FW), dry weight (DW), contents of Na+, K+, Ca2+, Cl−, malondialdehyde (MDA), proline, soluble sugars, hydrogen peroxide (H2O2), relative water content, chlorophyll contents, net photosynthetic rate, and osmotic potential of whole plants, the salt-tolerance ability from strongest to weakest is identified as Pink, Yellow, Purple, White, and Blue. Photosynthetic rate was the most reliable and positive indicator of salt tolerance. The density of salt glands showed the greatest increase in Pink under NaCl treatment, indicating that Pink adapts to high-salt levels by enhancing salt gland formation. These results provide a theoretical basis for the large-scale planting of L. sinuatum in saline soils in the future.

A novel gene LbHLH from the halophyte Limonium bicolor enhances salt tolerance via reducing root hair development and enhancing osmotic resistance

Background Identifying genes involved in salt tolerance in the recretohalophyte Limonium bicolor could facilitate the breeding of crops with enhanced salt tolerance. Here we cloned the previously uncharacterized gene LbHLH and explored its role in salt tolerance. Results The 2,067-bp open reading frame of LbHLH encodes a 688-amino-acid protein with a typical helix-loop-helix (HLH) domain. In situ hybridization showed that LbHLH is expressed in salt glands of L. bicolor. LbHLH localizes to the nucleus, and LbHLH is highly expressed during salt gland development and in response to NaCl treatment. To further explore its function, we heterologously expressed LbHLH in Arabidopsis thaliana under the 35S promoter. The overexpression lines showed significantly increased trichome number and reduced root hair number. LbHLH might interact with GLABRA1 to influence trichome and root hair development, as revealed by yeast two-hybrid analysis. The transgenic lines showed higher germination percentages and longer roots than the wild type under NaCl treatment. Analysis of seedlings grown on medium containing sorbitol with the same osmotic pressure as 100 mM NaCl demonstrated that overexpressing LbHLH enhanced osmotic resistance. Conclusion These results indicate that LbHLH enhances salt tolerance by reducing root hair development and enhancing osmotic resistance under NaCl stress.

A novel gene LbHLH from the halophyte Limonium bicolor enhances salt tolerance via alleviating osmotic stress and reducing root hair development

Background Identifying genes involved in salt tolerance in the recretohalophyte Limonium bicolor could facilitate the breeding of crops with enhanced salt tolerance. Here we cloned the previously uncharacterized gene LbHLH and explored its role in salt tolerance. Results The 2,067-bp open reading frame of LbHLH encodes a 688-amino-acid protein with a typical helix-loop-helix (HLH) domain. In situ hybridization showed that LbHLH is expressed in salt glands of L. bicolor. LbHLH localizes to the nucleus, and LbHLH is highly expressed during salt gland development and in response to NaCl treatment. To further explore its function, we heterologously expressed LbHLH in Arabidopsis thaliana under the 35S promoter. The overexpression lines showed significantly increased trichome number and reduced root hair number. LbHLH might interact with GLABRA1 to influence trichome and root hair development, as revealed by yeast two-hybrid analysis. The transgenic lines showed higher germination percentages and longer roots than the wild type under NaCl treatment. Analysis of seedlings grown on medium containing mannitol with the same osmotic pressure as 100 mM NaCl or LiCl with the same ionic effect as 100 mM NaCl demonstrated that overexpressing LbHLH relieved osmotic stress. Conclusion These results indicate that LbHLH enhances salt tolerance by alleviating osmotic damage under NaCl stress.

Current Understanding of Role of Vesicular Transport in Salt Secretion by Salt Glands in Recretohalophytes

Soil salinization is a serious and growing problem around the world. Some plants, recognized as the recretohalophytes, can normally grow on saline–alkali soil without adverse effects by secreting excessive salt out of the body. The elucidation of the salt secretion process is of great significance for understanding the salt tolerance mechanism adopted by the recretohalophytes. Between the 1950s and the 1970s, three hypotheses, including the osmotic potential hypothesis, the transfer system similar to liquid flow in animals, and vesicle-mediated exocytosis, were proposed to explain the salt secretion process of plant salt glands. More recently, increasing evidence has indicated that vesicular transport plays vital roles in salt secretion of recretohalophytes. Here, we summarize recent findings, especially regarding the molecular evidence on the functional roles of vesicular trafficking in the salt secretion process of plant salt glands. A model of salt secretion in salt gland is also proposed.

Importin-β From the Recretohalophyte Limonium bicolor Enhances Salt Tolerance in Arabidopsis thaliana by Reducing Root Hair Development and Abscisic Acid Sensitivity

AimsTo elucidate the genetics underlying salt tolerance in recretohalophytes and assess its relevance to non-halophytes, we cloned the Limonium bicolor homolog of Arabidopsis thaliana (Arabidopsis) SUPER SENSITIVE TO ABA AND DROUGHT2 (AtSAD2) and named it LbSAD2, an importin-β gene associated with trichome initiation and reduced abscisic acid (ABA) sensitivity, and then we assessed the heterologously expressed LbSAD2 in Arabidopsis.MethodsWe examined LbSAD2 expression and assessed the effect of heterologous LbSAD2 expression in Arabidopsis on root hair/trichome induction; the expression levels of possible related genes in trichome/root hair development; some physiological parameters involved in salt tolerance including germination rate, root length, and contents of Na+, proline, and malondialdehyde; and the response of ABA at the germination stage.ResultsThe LbSAD2 gene is highly expressed in the salt gland development stage and salt treatment, especially located in the salt gland by in situ hybridization, and the LbSAD2 protein contains some special domains compared with AtSAD2, which may suggest the involvement of LbSAD2 in salt tolerance. Compared with the SAD2/GL1 mutant CS65878, which lacks trichomes, CS65878-35S:LbSAD2 had higher trichome abundance but lower root hair abundance. Under 100 mM NaCl treatment, CS65878-35S:LbSAD2 showed enhanced germination and root lengths; improved physiological parameters, including high proline and low contents of Na+ and malondialdehyde; higher expression of the salt-tolerance genes Δ1-PYRROLINE-5-CARBOXYLATE SYNTHETASE 1 (P5CS1) and GST CLASS TAU 5 (GSTU5); reduced ABA sensitivity; and increased expression of the ABA signaling genes RESPONSIVE TO ABA 18 (RAB18) and SNF1-RELATED PROTEIN KINASE 2 (SRK2E), but not of the ABA biosynthesis gene 9-CIS-EPOXYCAROTENOID DIOXYGENASE 3 (NCED3).ConclusionLbSAD2 enhances salt tolerance in Arabidopsis by specifically reducing root hair development, Na+ accumulation, and ABA sensitivity.

Author Correction: The earliest evidence for a supraorbital salt gland in dinosaurs in new Early Cretaceous ornithurines

An amendment to this paper has been published and can be accessed via a link at the top of the paper.