Functional role of betalains in Disphyma australe under salinty stress

<p>Foliar betalainic plants are commonly found in dry and exposed environments such as deserts and sandbanks. This marginal habitat has led many researchers to hypothesise that foliar betalains provide tolerance to abiotic stressors such as strong light, drought, salinity and low temperatures. Among these abiotic stressors, soil salinity is a major problem for agriculture affecting approximately 20% of the irrigated lands worldwide. Betacyanins may provide functional significance to plants under salt stress although this has not been unequivocally demonstrated. The purpose of this thesis is to add knowledge of the various roles of foliar betacyanins in plants under salt stress. For that, a series of experiments were performed on Disphyma australe, which is a betacyanic halophyte with two distinct colour morphs in vegetative shoots. In chapter two, I aimed to find the effect of salinity stress on betacyanin pigmentation in D. australe and it was hypothesised that betacyanic morphs are physiologically more tolerant to salinity stress than acyanic morphs. Within a coastal population of red and green morphs of D. australe, betacyanin pigmentation in red morphs was a direct result of high salt and high light exposure. Betacyanic morphs were physiologically more tolerant to salt stress as they showed greater maximum CO₂ assimilation rates, water use efficiencies, photochemical quantum yields and photochemical quenching than acyanic morphs. Contrary to this, the green morphs, although possessing the ability to synthesise betalains in flower petals, did not produce betalains in vegetative shoots in response to salt stress. Moreover, green morphs, in terms of leaf photosynthesis, performed poorly under salinity stress. In chapter three I further investigated the physiological benefit of betacyanin accumulation in D. australe. I postulated that betacyanin in the leaves of D. australe can protect the salt stressed chloroplasts from harmful excessive light by absorbing significant amount of radiation. To test this, a novel experimental approach was used; the key biosynthetic step for betacyanin synthesis was identified, which was deficient in vegetative shoots of the green morphs. By supplying the product of this enzymatic reaction, L-DOPA, betacyanin synthesis could be induced in the leaves of green morphs. This model system was used to compare the photoprotective responses of red vs. green leaves. The L-DOPA induced betacyanic leaves showed similar responses (such as smaller reductions and faster recoveries of PSII and less H₂O₂ production than in the green leaves) to naturally betacyanic leaves when exposed to high light and salinity. The differences in photoinhibition between red and green leaves were attributed to the light absorbing properties of betacyanins. L-DOPA treated and naturally red leaves showed lower photoinactivation than green leaves when exposed to white or green light, although not when exposed to monochromatic (red) light. In chapter four, I used a similar experimental model to that in the third chapter and showed that other than photoprotection, betacyanins in leaves may be involved in salt tolerance by enhancing toxic ion (such as Na⁺) sequestration in betacyanic epidermal cells, storing Na⁺ away from sensitive mesophyll tissue. The Na⁺ localization between red and green leaves was compared after salinity treatment by using a sodium binding stain (SBFI-AM) and Cryo-SEM analysis. L-DOPA treated and natural red leaves sequestered Na⁺ ions to the epidermal cell layer. In contrast, green leaves retained Na⁺ in the mesophyll tissue, which suggested that red leaves were better equipped to tolerate salt-specific effects. Therefore, betacyanic plants were more tolerant to applied salinity stress and showed relatively higher growth rates than green morphs. The findings of this thesis provide a significant contribution to our understanding of the role of betacyanins in plants under salinity stress. My data suggest that the multi-faceted properties of betacyanins (such as their photoprotective function, and their involvement in sequestration of toxic ions) clearly provide a benefit to plants under salinity stress.</p>

Functional role of betalains in Disphyma australe under salinty stress

<p>Foliar betalainic plants are commonly found in dry and exposed environments such as deserts and sandbanks. This marginal habitat has led many researchers to hypothesise that foliar betalains provide tolerance to abiotic stressors such as strong light, drought, salinity and low temperatures. Among these abiotic stressors, soil salinity is a major problem for agriculture affecting approximately 20% of the irrigated lands worldwide. Betacyanins may provide functional significance to plants under salt stress although this has not been unequivocally demonstrated. The purpose of this thesis is to add knowledge of the various roles of foliar betacyanins in plants under salt stress. For that, a series of experiments were performed on Disphyma australe, which is a betacyanic halophyte with two distinct colour morphs in vegetative shoots. In chapter two, I aimed to find the effect of salinity stress on betacyanin pigmentation in D. australe and it was hypothesised that betacyanic morphs are physiologically more tolerant to salinity stress than acyanic morphs. Within a coastal population of red and green morphs of D. australe, betacyanin pigmentation in red morphs was a direct result of high salt and high light exposure. Betacyanic morphs were physiologically more tolerant to salt stress as they showed greater maximum CO₂ assimilation rates, water use efficiencies, photochemical quantum yields and photochemical quenching than acyanic morphs. Contrary to this, the green morphs, although possessing the ability to synthesise betalains in flower petals, did not produce betalains in vegetative shoots in response to salt stress. Moreover, green morphs, in terms of leaf photosynthesis, performed poorly under salinity stress. In chapter three I further investigated the physiological benefit of betacyanin accumulation in D. australe. I postulated that betacyanin in the leaves of D. australe can protect the salt stressed chloroplasts from harmful excessive light by absorbing significant amount of radiation. To test this, a novel experimental approach was used; the key biosynthetic step for betacyanin synthesis was identified, which was deficient in vegetative shoots of the green morphs. By supplying the product of this enzymatic reaction, L-DOPA, betacyanin synthesis could be induced in the leaves of green morphs. This model system was used to compare the photoprotective responses of red vs. green leaves. The L-DOPA induced betacyanic leaves showed similar responses (such as smaller reductions and faster recoveries of PSII and less H₂O₂ production than in the green leaves) to naturally betacyanic leaves when exposed to high light and salinity. The differences in photoinhibition between red and green leaves were attributed to the light absorbing properties of betacyanins. L-DOPA treated and naturally red leaves showed lower photoinactivation than green leaves when exposed to white or green light, although not when exposed to monochromatic (red) light. In chapter four, I used a similar experimental model to that in the third chapter and showed that other than photoprotection, betacyanins in leaves may be involved in salt tolerance by enhancing toxic ion (such as Na⁺) sequestration in betacyanic epidermal cells, storing Na⁺ away from sensitive mesophyll tissue. The Na⁺ localization between red and green leaves was compared after salinity treatment by using a sodium binding stain (SBFI-AM) and Cryo-SEM analysis. L-DOPA treated and natural red leaves sequestered Na⁺ ions to the epidermal cell layer. In contrast, green leaves retained Na⁺ in the mesophyll tissue, which suggested that red leaves were better equipped to tolerate salt-specific effects. Therefore, betacyanic plants were more tolerant to applied salinity stress and showed relatively higher growth rates than green morphs. The findings of this thesis provide a significant contribution to our understanding of the role of betacyanins in plants under salinity stress. My data suggest that the multi-faceted properties of betacyanins (such as their photoprotective function, and their involvement in sequestration of toxic ions) clearly provide a benefit to plants under salinity stress.</p>

Stomatal Responses to Light, CO2, and Mesophyll Tissue in Vicia faba and Kalanchoë fedtschenkoi

The responses of stomatal aperture to light intensity and CO2 concentration were studied in both Vicia faba (C3) and Kalanchoë fedtschenkoi (Crassulacean acid metabolism; CAM), in material sampled from both light and dark periods. Direct comparison was made between intact leaf segments, epidermises grafted onto exposed mesophyll, and isolated epidermal peels, including transplantations between species and between diel periods. We reported the stomatal opening in response to darkness in isolated CAM peels from the light period, but not from the dark. Furthermore, we showed that C3 mesophyll has stimulated CAM stomata in transplanted peels to behave as C3 in response to light and CO2. By using peels and mesophyll from plants sampled in the dark and the light period, we provided clear evidence that CAM stomata behaved differently from C3. This might be linked to stored metabolites/ions and signalling pathway components within the guard cells, and/or a mesophyll-derived signal. Overall, our results provided evidence for both the involvement of guard cell metabolism and mesophyll signals in stomatal responses in both C3 and CAM species.

Comparative Transcriptome Profiling Indicated that Leaf Mesophyll and Leaf Vasculature have Different Drought Response Mechanisms in Cassava

Drought stress is one of the major environmental factors that limited crop’s growth and production. Cassava known as a tropical crop that is widely distributed in Sub-Saharan Africa. It has a strong drought tolerance and can grow well under tough environmental conditions. Therefore, understanding how cassava responds to drought stress and coordinates survival and accumulation has great theoretical significance for improving crop drought resistance breeding. Many studies on cassava drought responses mainly focused on the leaf and whole seedling. Nevertheless, how the vasculature plays an important role in plant response to water deficiency remains to be fully elucidated. Here, comparative transcriptome analysis was performed on isolated mesophyll tissue and leaf vein vascular tissue of cassava variety KU50 after mild drought treatment to determine the molecular mechanism behind drought resistance in cassava vasculature. Our results showed that KU50 leaves had increased leaf temperature, with characters of rapidly decreased net photosynthetic rate, stomatal conductance, and transpiration rate in leaves, and the intercellular CO2 concentration accumulated under drought stress. Comparative transcriptome profiling revealed that under drought stress, leaf mesophyll tissue mainly stimulated the biosynthesis of amino acids, glutamic acid metabolism, and starch and sucrose metabolism. In particular, the arginine biosynthesis pathway was significantly enhanced to adapt to the water deficiency in leaf mesophyll tissue. However, in vascular tissue, the response to drought mainly involved ion transmembrane transport, hormone signal transduction, and depolymerization of proteasome. Concretely, ABA signaling and proteasome metabolism, which are involved in ubiquitin regulation, were changed under drought stress in KU50 leaf vascular tissue. Our work highlights that the leaf vasculature and mesophyll in cassava have completely different drought response mechanisms.

Leaf anatomical adaptation of eighteen mangrove plant species from the Sundarbans in Bangladesh

Investigation on leaf anatomical adaptation of 18 mangrove plant species was carried out. Among the 18 species 13 were dorsiventral and five were isobilateral type. All the species had special stomatal structure and variable cuticle layer to minimize transpiration. Most of the species had succulent leaves with leaf thickness ranging from around 232 to 1363 μm. As an indication of salt secretion, both glandular and non-glandular trichomes were observed in several species. Although presence of single to multilayered hypodermis might effectively function as water storage tissue, several studied mangrove plant species e.g. Cynometra ramiflora L., Phoenix paludosa Roxb., Pongamea pinnata (L.) Pierre, Sonneratia apetala Buch. - Ham., S. caseolaris (L.) Engl. and Xylocarpus moluccensis (Lamk.) M. Roem. showed complete absence of hypodermis. This might be due to moderate saline condition. In addition, marked terminal tracheids in mesophyll tissue of a number of species might help with capillary water storage within the leaf. To enhance mechanical support several species were found to develop considerable amount of diverse sclereids within the mesophyll tissue and surrounding vascular bundle. Although maximum anatomical adaptations are common for plants growing in saline habitat it may be suggested that these features were differentially developed in plants specifically grown in mesohaline zone.

Is Pteropyrum a pathway to C4 evolution in Polygonaceae? An integrative approach to the taxonomy and anatomy of Pteropyrum (C3), an immediate relative of Calligonum (C4)

Pteropyrum is a small genus of Polygonaceae with four species from the arid regions of Iran and adjacent countries. Pteropyrum spp. are not precisely delimitated and are difficult to identify because of their high plasticity in morphological characters. Pteropyrum (C3) has a close affinity to Calligonum (C4) and is therefore a suitable case for C4 evolutionary studies. We investigated the morphology and micromorphology (including pollen morphology) of Pteropyrum and elucidated the phylogenetic relationships with Atraphaxis and Calligonum using nuclear ITS sequences. Characteristics of the photosynthetic tissues such as volume and number of layers of primary carbon assimilation tissues (PCA) and photosynthetic carbon reduction tissue (PCR) were studied. In addition, the leaf and cotyledon anatomical characters of Pteropyrum (C3), Atraphaxis (C3) and Calligonum (C4), and their δ 13C values were compared to look for evolutionary changes in assimilating organs. The molecular phylogenetic tree identifies two strongly supported clades in Pteropyrum and its close relationship with Calligonum, confirming previous studies. Some morphologically similar species belong to different clades, which is probably due to convergent evolution and homoplasy. Leaf anatomical studies show that Atraphaxis has a multilayered mesophyll tissue, whereas Calligonum has one-layered mesophyll cells. The volume and layer number of mesophyll tissue cells decreases, whereas water storage tissue area significantly increases from Atraphaxis to Pteropyrum and Calligonum. This phenomenon confirms previous studies in other lineages with C4 salsoloid anatomy that have evolved through increasing of water storage tissue and succulence of assimilating organs. In the taxonomic part of the paper, a key to identification of accepted taxa of Pteropyrum, description of species and distribution maps are presented based on numerous herbarium specimens and our own rich collections from the field. Four new species are described based on a combination of morphology of seedlings and mature plants, pollen morphology and molecular data. A subspecific classification is suggested to show morpho-geographical variation of Pteropyrum aucheri s.l.

Morphological characterization of domatium development in Callicarpa saccata

Background and aims Domatia are plant structures within which organisms reside. Callicarpa saccata (Lamiaceae) is the sole myrmecophyte, or ‘ant plant’, that develops foliar (leaf-borne) myrmeco-domatia in this genus. In this work we examined domatium development in C. saccata to understand the developmental processes behind pouch-like domatia. Methods Scanning electron microscopy, sectioning and microcomputed tomography were carried out to compare the leaves of C. saccata with those of the closely related but domatia-less myrmecophyte Callicarpa subaequalis, both under cultivation without ants. Key results Callicarpa saccata domatia are formed as a result of excess cell proliferation at the blade/petiole junctions of leaf primordia. Blade/petiole junctions are important meristematic sites in simple leaf organogenesis. We also found that the mesophyll tissue of domatia does not clearly differentiate into palisade and spongy layers. Conclusions Rather than curling of the leaf margins, a perturbation of the normal functioning of the blade/petiole junction results in the formation of domatium tissue. Excess cell proliferation warps the shape of the blade and disturbs the development of the proximal–distal axis. This process leads to the generation of distinct structures that facilitate interaction between C. saccata and ants.

Pengukuran Panjang dan Lebar Pori Stomata Daun Beberapa Varietas Tanaman Kacang Tanah (Arachis hypogaea L.)

Kacang tanah (Arachis hypogaea L.) merupakan salah satu tanaman pangan bernilai ekonomi tinggi, dan sebagai tanaman kacang-kacangan terpenting kedua setelah kedelai.Pori stomata merupakan tempat terjadinya pertukaran gas dan air antara atmosfer dengan sistem ruang antar sel yang berada pada jaringan mesofil di bawah epidermis.Setiap varietas tanaman kacang tanah memiliki respon terhadap faktor lingkungan yang berbeda seperti ketahanan terhadap cekaman fisilogis termasuk kemampuan membuka dan menutupnya stomata.Penelitian ini bertujuan untuk mengukur panjang dan lebar pori stomata daun beberapa varietas kacang tanah (Arachis hypogaeaL.). Penelitian dilaksanakan di rumah plastikFakultas MIPA Unsrat selama 30 hari. Hasil penelitian menunjukkan bahwa keempat varietas memiliki panjang dan lebar stomata yang lebih panjang adalah varietas Gajah 81,80 ± 28,72 µm, varietas Lokal 79,81 ± 24,85 µm, varietas Jerapah 69,28 ± 22,60 µm dan varietas Kelinci 57,22 ± 25,02 µm. Lebar pori stomata yang membuka lebih besar adalah varietas Lokal 31,13 ± 9,77 µm, varietas Gajah 29,22 ± 3,71 µm, varietas Jerapah 27,72 ± 11,65 µm dan varietas Kelinci 21,32 ± 12,78 µm.Peanut are one of the high value food plants and as the second highest bean crop after soybeans. The Pore of Stomata plays was a place of gas and water exchange between the atmosphere and the intercellular space located in the mesophyll tissue beneath the epidermis. Each variety of peanut crops has responses to different environmental factors such as resistance to physical stress, including the ability to open and close the stomata. This study aims to measure the length and width of stomata pores of several peanut varieties (Arachis hypogaea L.). The research was conducted in plastic house of Faculty of Mathematics and Natural Sciences Unsrat for 30 days. The results showed that the four varieties had length and stomatal length 81,80 ± 28,72 μm, followed by local varieties 79,81 ± 24,85 μm, giraffe varieties 69,28 ± 22,60 μm and 57,22 ± rabbit varieties 25.02 μm. Stomatal pore 31.13 ± 9.77 μm, followed by elephant varieties 29.22 ± 3.71 μm, giraffe varieties 27.72 ± 11.65 μm and rabbit varieties 21.32 ± 12.78 μm.

A comparative anatomic study of leaves of the genus Stachys L. in Iraq

The anatomic characteristics of the leaves of 22 taxa of Stachys L. representing seven of the currently recognized sections distributed in northern Iraq, were examined. The study did not found any variations in the characteristics of the stomata system of the leaves.Therewere variations with a good taxonomic value in other anatomic characteristics.Thevariations in characters of the mesophyll tissue in S.kurdica var.brevidens Bom ex Bhattacharjeewas bifacial but in S.kurdica var.kurdica Boiss. & Hohen.was unifacial .The study also found that the taxa S.benthamiana Bioss., S.lanigera (Bornm.) Rech.f. andS.kotscyi Bioss. with 3 vascular bundles in the middle vein while the taxa S.ballotiformis Vatke., S.megalodonta Hausskn. &Bornm.exP.H.Davis and S.kurdica with one only. The spescies S.lavandulifoliaVahle.had a cross section of V shape that did not seen in other studied taxa. The results showed evidence were useful in separating between species within the same section.

Microscopical evaluation of leaf of Maytenus emarginata

Microscopical investigation was carried out on leaves of Maytenus emarginata (Willd.) Ding Hou belongs to family celastraceae, is an evergreen tree that tolerates various types of stresses of the desert, locally known as “Kankero” in Hindi, “Thorny staff tree” in English. Literature survey revealed that not much work has been done on this plant, especially on leaves. So we have taken its detailed microscopical studies to prove its appropriate identification. Microscopical study provide information that the lamina is dorsiventaral and 380µm thick. The Lamina is amphistomatic, the stomata being distributed both on the adaxial and abaxial sides. Calcium oxalate crystals of druses are fairly abundant in the mesophyll tissue. The stomata are cyclocytic type. The venation is densely reticulate. The terminations are either simple or more commonly forked into deuse clusters. Powder Microscopy show that the sclereids are fiber like in shape and size. The sclereids are 500 µm long and 30 µm thick. Long or short, cylindrical mesophyll cells are common with powder. They have dense tannin contents. The cells have thin wall and the cells are up to 200 µm long and 60 µm wide. Calciumoxlate druses are quite abundant in the power. They are spherical bodies with spiny surface. The druses are 30 µm in diameters. The above studies provide useful information in regard to its correct identity, evaluation and help to differentiate from the closely related other species of Maytenus Emarginata (Willd.)Ding Hou.