Sung Ju Ahn

Citrate Secretion Coupled with the Modulation of Soybean Root Tip under Aluminum Stress. Up-Regulation of Transcription, Translation, and Threonine-Oriented Phosphorylation of Plasma Membrane H+-ATPase

The aluminum (Al)-induced secretion of citrate has been regarded as an important mechanism for Al resistance in soybean (Glycine max). However, the mechanism of how Al induces citrate secretion remains unclear. In this study, we investigated the regulatory role of plasma membrane H+-ATPase on the Al-induced secretion of citrate from soybean roots. Experiments performed with plants grown in full nutrient solution showed that Al-induced activity of plasma membrane H+-ATPase paralleled secretion of citrate. Vanadate and fusicoccin, an inhibitor and an activator, respectively, of plasma membrane H+-ATPase, exerted inhibitory and stimulatory effects on the Al-induced secretion of citrate. Higher activity of plasma membrane H+-ATPase coincided with more citrate secretion in Al-resistant than Al-sensitive soybean cultivars. These results suggested that the effects of Al stress on citrate secretion were mediated via modulation of the activity of plasma membrane H+-ATPase. The relationship between the Al-induced secretion of citrate and the activity of plasma membrane H+-ATPase was further demonstrated by analysis of plasma membrane H+-ATPase transgenic Arabidopsis (Arabidopsis thaliana). When plants were grown on Murashige and Skoog medium containing 30 μm Al (9.1 μm Al3+ activity), transgenic plants exuded more citrate compared with wild-type Arabidopsis. Results from real-time reverse transcription-PCR and immunodetection analysis indicated that the increase of plasma membrane H+-ATPase activity by Al is caused by transcriptional and translational regulation. Furthermore, plasma membrane H+-ATPase activity and expression were higher in an Al-resistant cultivar than in an Al-sensitive cultivar. Al activated the threonine-oriented phosphorylation of plasma membrane H+-ATPase in a dose- and time-dependent manner. Taken together, our results demonstrated that up-regulation of plasma membrane H+-ATPase activity was associated with the secretion of citrate from soybean roots.

Transgenic Arabidopsis and tobacco plants overexpressing an aquaporin respond differently to various abiotic stresses

Despite the high isoform multiplicity of aquaporins in plants, with 35 homologues including 13 plasma membrane intrinsic proteins (PIPs) in Arabidosis thaliana, the individual and integrated functions of aquaporins under various physiological conditions remain unclear. To better understand aquaporin functions in plants under various stress conditions, we examined transgenic Arabidopsis and tobacco plants that constitutively overexpress Arabidopsis PIP1;4 or PIP2;5 under various abiotic stress conditions. No significant differences in growth rates and water transport were found between the transgenic and wild-type plants when grown under favorable growth conditions. The transgenic plants overexpressing PIP1;4 or PIP2;5 displayed a rapid water loss under dehydration stress, which resulted in retarded germination and seedling growth under drought stress. In contrast, the transgenic plants overexpressing PIP1;4 or PIP2;5 showed enhanced water flow and facilitated germination under cold stress. The expression of several PIPs was noticeably affected by the overexpression of PIP1;4 or PIP2;5 in Arabidopsis under dehydration stress, suggesting that the expression of one aquaporin isoform influences the expression levels of other aquaporins under stress conditions. Taken together, our results demonstrate that overexpression of an aquaporin affects the expression of endogenous aquaporin genes and thereby impacts on seed germination, seedling growth, and stress responses of the plants under various stress conditions.

Overexpression of PIP2;5 Aquaporin Alleviates Effects of Low Root Temperature on Cell Hydraulic Conductivity and Growth in Arabidopsis

The effects of low root temperature on growth and root cell water transport were compared between wild-type Arabidopsis (Arabidopsis thaliana) and plants overexpressing plasma membrane intrinsic protein 1;4 (PIP1;4) and PIP2;5. Descending root temperature from 25°C to 10°C quickly reduced cell hydraulic conductivity (Lp) in wild-type plants but did not affect Lp in plants overexpressing PIP1;4 and PIP2;5. Similarly, when the roots of wild-type plants were exposed to 10°C for 1 d, Lp was lower compared with 25°C. However, there was no effect of low root temperature on Lp in PIP1;4- and PIP2;5-overexpressing plants after 1 d of treatment. When the roots were exposed to 10°C for 5 d, Lp was reduced in wild-type plants and in plants overexpressing PIP1;4, whereas there was still no effect in PIP2;5-overexpressing plants. These results suggest that the gating mechanism in PIP1;4 may be more sensitive to prolonged low temperature compared with PIP2;5. The reduction of Lp at 10°C in roots of wild-type plants was partly restored to the preexposure level by 5 mm Ca(NO3)2 and protein phosphatase inhibitors (75 nm okadaic acid or 1 μm Na3VO4), suggesting that aquaporin phosphorylation/dephosphorylation processes were involved in this response. The temperature sensitivity of cell water transport in roots was reflected by a reduction in shoot and root growth rates in the wild-type and PIP1;4-overexpressing plants exposed to 10°C root temperature for 5 d. However, low root temperature had no effect on growth in plants overexpressing PIP2;5. These results provide strong evidence for a link between growth at low root temperature and aquaporin-mediated root water transport in Arabidopsis.

MYB46-Mediated Transcriptional Regulation of Secondary Wall Biosynthesis

Formation of secondary wall requires coordinated transcrip-tional regulation of the genes involved in the biosynthesis of major secondary wall components (e.g. cellulose, hemicellu-lose, and lignin). even though many aspects of plant biology have been extensively elucidated using various model spe-cies, our current understanding of secondary wall formation is limited.complex transcriptional networks appear to be involved in the coordinated regulation of secondary wall biosynthesis (Ko et al., 2009; Demura and Ye, 2010; Zhu et al., 2010; lee et al., 2011; for a recent review, see Wang and Dixon, 2012). Recently,

Differences in root-to-shoot Cd and Zn translocation and by HMA3 and 4 could influence chlorophyll and anthocyanin content in Arabidopsis Ws and Col-0 ecotypes under excess metals

Judging from the ecotypic variability in Arabidopsis thaliana L., Columbia-0 (Col-0) appears to be less tolerant to cadmium (Cd) than the Wassilewskija (Ws) ecotype that possesses the full-length Heavy Metal ATPase3 (HMA3) cDNA. In this study, the Ws and Col-0 were tested to determine toxic metal response between Ws and Col-0 due to AtHMA3 point mutation and/or other factors. The growth inhibition of Col-0 mediated by Cd and zinc (Zn) was more serious than the inhibition of Ws, while no significant difference was evident by lead (Pb) and cobalt (Co). In the presence of Cd stress, chlorosis in leaves of Col-0 was more serious than the Ws ecotype. When grown under hydroponic culture containing 500 µM Zn, leaves of Col-0 showed a remarkable increase in the anthocyanin content in a dose-dependent manner and the expression of genes encoding enzymes involved in anthocyanin synthesis in the leaves. The rate of root-to-shoot translocation of Cd and Zn in the Col-0 was 2 times higher when compared with the Ws, whereas roots of the Col-0 accumulated 2 times lower Cd and Zn concentrations than those of the Ws. Real-time polymerase chain reaction (PCR) analyses indicated that not only the alteration of the expression of HMA3 but also of the HMA4 was responsible for the root-to-shoot translocation of toxic metals. The results demonstrate that the Col-0 is readily translocating Cd and Zn to the aerial parts but not the Ws, thereby induce the alteration of phenotype in leaf color.

The Minor Spliceosomal Protein U11/U12-31K Is an RNA Chaperone Crucial for U12 Intron Splicing and the Development of Dicot and Monocot Plants

U12 intron-specific spliceosomes contain U11 and U12 small nuclear ribonucleoproteins and mediate the removal of U12 introns from precursor-mRNAs. Among the several proteins unique to the U12-type spliceosomes, an Arabidopsis thaliana AtU11/U12-31K protein has been shown to be indispensible for proper U12 intron splicing and for normal growth and development of Arabidopsis plants. Here, we assessed the functional roles of the rice (Oryza sativa) OsU11/U12-31K protein in U12 intron splicing and development of plants. The U11/U12-31K transcripts were abundantly expressed in the shoot apical meristems (SAMs) of Arabidopsis and rice. Ectopic expression of OsU11/U12-31K in AtU11/U12-31K-defecient Arabidopsis mutant complemented the incorrect U12 intron splicing and abnormal development phenotypes of the Arabidopsis mutant plants. Impaired cell division activity in the SAMs and inflorescence stems observed in the AtU11/U12-31K-deficient mutant was completely recovered to normal by the expression of OsU11/U12-31K. Similar to Arabidopsis AtU11/U12-31K, rice OsU11/U12-31K was determined to harbor RNA chaperone activity. Collectively, the present findings provide evidence for the emerging idea that the U11/U12-31K protein is an indispensible RNA chaperone that functions in U12 intron splicing and is necessary for normal development of monocotyledonous plants as well as dicotyledonous plants.

Aluminum tolerance associated with enhancement of plasma membrane H+-ATPase in the root apex of soybean

Abstract Seventeen soybean cultivars were screened to discern differences in aluminum (Al) sensitivity. The Sowon (Al-tolerant) and Poongsan (Al-sensitive) cultivars were selected for further study by simple growth measurement. Aluminum-induced root growth inhibition was significantly higher in the Poongsan cultivar than in the Sowon cultivar, although the differences depended on the Al concentration (0, 25, 50, 75 or 100 μmol L–1) and the amount of exposure (0, 3, 6, 12 or 24 h). Damage occurred preferentially in the root apex. High-sensitivity growth measurements using India ink implicated the central elongation zone located 2–3 mm from the root apex. The Al content was lower 0–5 mm from the root apices in the Sowon cultivar than in the apices of the Poongsan cultivar when exposed to 50 μmol L–1 Al for 12 h. Furthermore, the citric acid exudation rate was more than twofold higher in the Sowon cultivar. Protein production of plasma membrane (PM) H+-ATPase from the root apices (0–5 mm) was upregulated in the presence of Al for 24 h in both cultivars. This activity, however, decreased in both cultivars treated with Al and the Poongsan cultivar was more severely affected. We propose that Al-induced growth inhibition is correlated with changes in PM H+-ATPase activity, which is linked to the exudation of citric acid in the root apex.

Effect of nutrient deficiencies on the water transport properties in figleaf gourd plants

Effects of nitrogen, phosphorus, and potassium deficiencies on water transport properties in figleaf gourd plants were studied. Plants were treated for different period of deficiency and physiological parameters such as stomatal conductance, photosynthesis and transpiration were measured. Cell and root pressure probes were utilized to measure turgor and root pressures, half-times of water exchange and hydraulic conductivities to analyze water transport properties. When plants were grown in nitrogen or phosphorus deficient nutrient solutions, they became insensitive to mercury, suggesting that aquaporin was closed resulting in reduced hydraulic conductivity. Inclusion of tungstate, however, restored the sensitivity of cells to mercury, indicating the importance of internal nutrient concentration, not the incoming nutrient supply. The hydrostatic hydraulic conductivity of roots grown in nitrogen deficient solution, representing apoplastic pathway of water transport, was reduced but this reduction was dramatically recovered by the application of tungstate, indicating the importance of nutrient availability from storage pools in relation to water status of plants.