Suriyan Cha-um

An Alkaline Phosphatase/Phosphodiesterase, PhoD, Induced by Salt Stress and Secreted Out of the Cells of Aphanothece halophytica, a Halotolerant Cyanobacterium

ABSTRACT Alkaline phosphatases (APases) are important enzymes in organophosphate utilization. Three prokaryotic APase gene families, PhoA, PhoX, and PhoD, are known; however, their functional characterization in cyanobacteria largely remains to be clarified. In this study, we cloned the phoD gene from a halotolerant cyanobacterium, Aphanothece halophytica (phoDAp ). The deduced protein, PhoD Ap , contains Tat consensus motifs and a peptidase cleavage site at the N terminus. The PhoD Ap enzyme was activated by Ca2+ and exhibited APase and phosphodiesterase (APDase) activities. Subcellular localization experiments revealed the secretion and processing of PhoD Ap in a transformed cyanobacterium. Expression of the phoDAp gene in A. halophytica cells was upregulated not only by phosphorus (P) starvation but also under salt stress conditions. Our results suggest that A. halophytica cells possess a PhoD that participates in the assimilation of P under salinity stress.

Comparative Effects of Salt Stress and Extreme pH Stress Combined on Glycinebetaine Accumulation, Photosynthetic Abilities and Growth Characters of Two Rice Genotypes

Glycinebetaine (Glybet) accumulation, photosynthetic efficiency and growth performance in indica rice cultivated under salt stress and extreme pH stress were investigated. Betaine aldehyde dehydrogenase (BADH) activity and Glybet accumulation in the seedlings of salt-tolerant and salt-sensitive rice varieties grown under saline and acidic conditions peaked after treatment for 72 h and 96 h, respectively, and were higher than those grown under neutral pH and alkaline salt stress. A positive correlation was found between BADH activity and Glybet content in both salt-tolerant (r^2=0.71) and salt-sensitive (r^2=0.86) genotypes. The chlorophyll a, chlorophyll b, total chlorophyll and total carotenoids contents in the stressed seedlings significantly decreased under both acidic and alkaline stresses, especially in the salt-sensitive genotype. Similarly, the maximum quantum yield of PSII (F√F(subscript m)), photon yield of PSII (Ф(subscript PSII)), non-photochemical quenching (NPQ) and net photosynthetic rate (P(subscript n)) in the stressed seedlings were inhibited, leading to overall growth reduction. The positive correlations between chlorophyll a content and F√F(subscript m), total chlorophyll content and Ф(subscript PSII), Ф(subscript PSII) and P(subscript n) as well as P(subscript n) and leaf area in both salt-tolerant and salt-sensitive genotypes were found. Saline acidic and saline alkaline soils may play a key role affecting vegetative growth prior to the reproductive stage in rice plants.

Remediation of salt-affected soil by the addition of organic matter: an investigation into improving glutinous rice productivity

Soil salinity may limit plant growth and development, and cause yield loss in crop species. This study aimed at remediating saline soil using organic matter (OM) treatment, before the cultivation of RD6 rice (Oryza sativa L. spp. indica). Physiological and morphological characters of rice plants, as well as crop yield, were evaluated from salt-affected soil with varying levels of salinity. The chlorophyll a and total chlorophyll pigments of rice plants grown in salt-affected soil (2% salt level) with the application of OM were maintained better than in plants grown without OM treatment. The degree of reduced photosynthetic pigments in rice plants was dependent on the level of salt contamination. Pigment content was positively related to maximum quantum yield of PSII (Fv/Fm) and quantum efficiency of PSII (ΦPSII), leading to reduced net photosynthetic rate (Pn) and reduced total grain weight (TGW). Photosynthetic abilities, including chlorophyll a and total chlorophyll pigments and ΦPSII, in rice plants grown with OM treatment were greater than in those cultivated in soil without the OM treatment, especially in high salt levels (1-2% salt). The remediation of salt-affected soil in paddy fields using OM should be applied further, as an effective way of enhancing food crop productivity.

Differential accumulation of glycinebetaine and choline monooxygenase in bladder hairs and lamina leaves of Atriplex gmelini under high salinity

Atriplex gmelini is a halophyte and possesses bladder hairs on the leaf surface. It is also known to accumulate the osmoprotectant glycinebetaine (GB). However, it remains unclear whether GB and its biosynthetic enzyme choline monooxygenase (CMO) accumulate in the bladder hairs. Microscopic observation of young leaves showed many bladder hairs on their surfaces, but their total number decreased along with leaf maturity. Sodium Green fluorescent approach revealed Na(+) accumulation in bladder cells of young leaves when A. gmelini was grown at high salinity (250 mM NaCl). Due to fewer bladder hairs in mature leaves, Na(+) accumulation was mostly found in mesophyll cells of mature leaves under high salinity. GB accumulation was found at significant level in both bladder- and laminae-cells without any addition of NaCl and its content increased at high salinity. CMO was not found in bladder hairs or young leaf laminae. Instead, the CMO protein expression was observed in mature leaves and that showed increased accumulation with increasing concentration of NaCl. Furthermore, in situ hybridization experiments revealed the expression of a transporter gene for GB, AgBetT, in the bladder hairs. Based on these results, the synthesis and translocation of GB in A. gmelini were discussed.

Expression and functional analysis of putative vacuolar Ca2+-transporters (CAXs and ACAs) in roots of salt tolerant and sensitive rice cultivars

Vacuolar Ca2+-transporters could play an important role for salt tolerance in rice (Oryza sativa L.) root. Here, we compared the expression profiles of putative vacuolar cation/H+ exchanger (CAX) and calmodulin-regulated autoinhibited Ca2+-ATPase (ACA) in rice roots of salt tolerant cv. Pokkali and salt sensitive cv. IR29. In addition to five putative vacuolar CAX genes in the rice genome, a new CAX gene (OsCAX4) has been annotated. In the present study, we isolated the OsCAX4 gene and showed that its encoded protein possesses a unique transmembrane structure and is potentially involved in transporting not only Ca2+ but also Mn2+ and Cu2+. These six OsCAX genes differed in their mRNA expression pattern in roots of tolerant versus sensitive rice cultivars exposed to salt stress. For example, OsCAX4 showed abundant expression in IR29 (sensitive) upon prolonged salt stress. The mRNA expression profile of four putative vacuolar Ca2+-ATPases (OsACA4-7) was also examined. Under control conditions, the mRNA levels of OsACA4, OsACA5, and OsACA7 were relatively high and similar among IR29 and Pokkali. Upon salt stress, only OsACA4 showed first a decrease in its expression in Pokkali (tolerant), followed by a significant increase. Based on these results, a role of vacuolar Ca2+ transporter for salt tolerance in rice root was discussed.

Water-deficit tolerant classification in mutant lines of indica rice

Water shortage is a major abiotic stress for crop production worldwide, limiting the productivity of crop species, especially in dry-land agricultural areas. This investigation aimed to classify the water-deficit tolerance in mutant rice (Oryza sativa L. spp. indica) genotypes during the reproductive stage. Proline content in the flag leaf of mutant lines increased when plants were subjected to water deficit. Relative water content (RWC) in the flag leaf of different mutant lines dropped in relation to water deficit stress. A decrease RWC was positively related to chlorophyll a degradation. Chlorophyll a , chlorophyll b , total chlorophyll , total carotenoids , maximum quantum yield of PSII , stomatal conductance , transpiration rate and water use efficiency in mutant lines grown under water deficit conditions declined in comparison to the well-watered, leading to a reduction in net-photosynthetic rate. In addition, when exposed to water deficit, panicle traits, including panicle length and fertile grains were dropped. The biochemical and physiological data were subjected to classify the water deficit tolerance. NSG19 (positive control) and DD14 were identified as water deficit tolerant, and AA11, AA12, AA16, BB13, BB16, CC12, CC15, EE12, FF15, FF17, G11 and IR20 (negative control) as water deficit sensitive, using Wards method.

Physio-Biochemical Responses of Oil Palm (Elaeis guineensis Jacq.) Seedlings to Mannitol- and Polyethylene Glycol-Induced Iso-Osmotic Stresses

References The aim of this investigation was to comparatively examine the physio-biochemical responses of oil palm seedlings to mannitol- and PEG-induced iso-osmotic stresses. The water content of osmotically stressed oil palm seedlings decreased, but the proline content and the electrolyte leakage of the seedlings increased with decreasing water potential (Ψw). However, the responses varied with the strength of osmotic stress and type of osmotic agent. Relative electrolyte leakage (REL) was negatively correlated to chlorophyll content in the osmotically stressed leaves. Chlorophyll a (Chla), chlorophyll b (Chlb), total carotenoids (Cx+c) and total chlorophyll (TC) in the seedlings were significantly reduced by osmotic stress, subsequently reducing maximum quantum yield of PSII (Fv/Fm) and photon yield of PSII (Φpsii), thereby lowering net-photosynthetic rate (Pn) and inhibiting growth. Physio-biochemical parameters, including REL, Fv/Fm, and Φpsii in oil palm seedlings were reduced more greatly by PEG-induced osmotic stress than by mannitol-induced stress. A deterioration in morphological characters, including leaf chlorosis, leaf burn, and green leaf area reduction were demonstrated in oil palm seedlings under osmotic stress induced by either mannitol or PEG. However, the toxic symptoms in oil palm seedlings under PEG-induced stress were severer than in those under mannitol-induced iso-osmotic stress, especially under severe osmotic stress.

Physio-biochemical and morphological characters of halophyte legume shrub, Acacia ampliceps seedlings in response to salt stress under greenhouse

Acacia ampliceps (salt wattle), a leguminous shrub, has been introduced in salt-affected areas in the northeast of Thailand for the remediation of saline soils. However, the defense mechanisms underlying salt tolerance A. ampliceps are unknown. We investigated various physio-biochemical and morphological attributes of A. ampliceps in response to varying levels of salt treatment (200–600 mM NaCl). Seedlings of A. ampliceps (25 ± 2 cm in plant height) raised from seeds were treated with 200 mM (mild stress), 400 and 600 mM (extreme stress) of salt treatment (NaCl) under greenhouse conditions. Na+ and Ca2+ contents in the leaf tissues increased significantly under salt treatment, whereas K+ content declined in salt-stressed plants. Free proline and soluble sugar contents in plants grown under extreme salt stress (600 mM NaCl) for 9 days significantly increased by 28.7 (53.33 μmol g–1 FW) and 3.2 (42.11 mg g–1 DW) folds, respectively over the control, thereby playing a major role as osmotic adjustment. Na+ enrichment in the phyllode tissues of salt-stressed seedlings positively related to total chlorophyll (TC) degradation (R2 = 0.72). Photosynthetic pigments and chlorophyll fluorescence in salt-stressed plants increased under mild salt stress (200 mM NaCl). However, these declined under high levels of salinity (400–600 mM NaCl), consequently resulting in a reduced net photosynthetic rate (R2 = 0.81) and plant dry weight (R2 = 0.91). The study concludes that A. ampliceps has an osmotic adjustment and Na+ compartmentation as effective salt defense mechanisms, and thus it could be an excellent species to grow in salt-affected soils.

Expression of developmentally regulated plasma membrane polypeptide (DREPP2) in rice root tip and interaction with Ca 2+ /CaM complex and microtubule

The cytoplasmic free Ca2+ could play an important role for salt tolerance in rice root (Oryza sativa L.). Here, we compared the expression profiles of two putative developmentally regulated plasma membrane polypeptides (DREPP1 and DREPP2) in rice roots of salt-tolerant cv. Pokkali and salt-sensitive cv. IR29. The messenger RNA (mRNA) for OsDREPP1 could be detected in all parts of root and did not change upon salt stress, whereas the mRNA for OsDREPP2 was detected only in root tips. The transcript level of OsDREPP2 first disappeared upon salt stress, then recovered in Pokkali, but not recovered in IR29. The gene-encoding OsDREPP2 was cloned from cv. Pokkali and expressed in Escherichia coli, and its biochemical properties were studied. It was found that OsDREPP2 is a Ca2+-binding protein and binds also to calmodulin (CaM) as well as microtubules. The mutation of Trp4 and Phe16 in OsDREPP2 to Ala decreased the binding of DREPP2 to Ca2+/CaM complex, indicating the N-terminal basic domain is involved for the binding. The binding of OsDREPP2 to microtubules was inhibited by Ca2+/CaM complex, while the binding of double-mutant OsDREPP2 protein to microtubules was not inhibited by Ca2+/CaM complex. We propose that CaM inhibits the binding of DREPP2 to cortical microtubules, causes the inhibition of microtubule depolymerization, and enhances the cell elongation.

EXOGENOUS APPLICATION OF POTASSIUM NITRATE TO ALLEVIATE SALT STRESS IN RICE SEEDLINGS

□ Overall growth characteristics of many plant species cultivated in soil affected by salinity could be alleviated by the application of potassium nitrate (KNO3) to the soil. The aim of this research was to investigate salt-tolerance in a salt-sensitive rice cultivar, ‘Pathumthani 1’ (PT1), in response to the exogenous application of 11.8 mM KNO3, in comparison to ‘Homjan’ (HJ), a salt tolerant cultivar. Water potential (ψw) in both the roots and leaves of PT1 seedlings under salt stress dropped significantly, while it was maintained in PT1 pretreated with KNO3, and similarly in HJ. The reduction of leaf water potential was positively related to total chlorophyll degradation, leading to diminished chlorophyll fluorescence, directly affecting growth in plants exposed to salt stress. In salt-sensitive PT1, the application of 11.8 mM KNO3 improved salt-tolerance via the conservation of water use efficiency, the maintenance of photosynthetic pigments, enhancement of chlorophyll a fluorescence, and stimulation of growth characters.