Phenotypes and Molecular Mechanisms Underlying the Root Response to Phosphate Deprivation in Plants.

Phosphorus (P) is an essential macronutrient for plant growth. The roots are the main organ for nutrient and water absorption in plants, and they adapt to low-P soils by altering their architecture for enhancing absorption of inorganic phosphate (Pi). This review summarizes the physiological and molecular mechanisms underlying the developmental responses of roots to Pi starvation, including the primary root, lateral root, root hair, and root growth angle, in the dicot model plant Arabidopsis thaliana and the monocot model plant rice (Oryza sativa). The importance of different root traits and genes for breeding P-efficient roots in rice varieties for Pi-deficient soils are also discussed, which we hope will benefit the genetic improvement of Pi uptake, Pi-use efficiency, and crop yields.

Functional redundancy of OsPIN1 paralogous genes in regulating plant growth and development in rice.

The OsPIN1 paralogous genes (OsPIN1a-1d) are important for root and panicle development in rice (Oryza sativa L.). However, the specific role of OsPIN1 paralogous genes is still not clear. To understand the specific roles of PIN1 paralogs in rice, we generated pin1 triple and quadruple mutants by crossing the pin1a pin1b and pin1c pin1d double mutants which we previously created. Compared with the 7-day-old wild type, the pin1a pin1c pin1d and pin1b pin1c pin1d triple mutants showed no obvious phenotype variation except that the pin1a pin1c pin1d triple mutant had shorter primary root and shoot. The pin1a pin1b pin1c and pin1a pin1b pin1d triple mutants exhibited a series of developmental abnormalities, including shorter primary roots, longer root hairs, fewer crown roots and lateral roots, shorter and curved shoots. Furthermore, the pin1a pin1b pin1c pin1d quadruple mutant displayed more severe phenotypic defects which was lethal. In addition, the expression levels of some hormone signal transduction and crown root development related genes, such as OsIAAs, OsARFs, OsRRs, and OsCRLs, were significantly altered in the stem base of all examined pin1 multiple mutants. Taken together, our results demonstrated that the four OsPIN1 paralogous genes function redundantly in regulating rice growth and development.

Correction: Constitutive expression of an A-5 subgroup member in the DREB transcription factor subfamily from Ammopiptanthus mongolicus enhanced abiotic stress tolerance and anthocyanin accumulation in transgenic Arabidopsis.

[This corrects the article DOI: 10.1371/journal.pone.0224296.].

Constitutive expression of an A-5 subgroup member in the DREB transcription factor subfamily from Ammopiptanthus mongolicus enhanced abiotic stress tolerance and anthocyanin accumulation in transgenic Arabidopsis.

Dehydration-responsive element-binding (DREB) transcription factors (TFs) are key regulators of stress-inducible gene expression in plants. Anthocyanins, an important class of flavonoids, protect plants from reactive oxygen species produced under abiotic stresses. However, regulation of DREBs on anthocyanin accumulation is largely unknown. Here, an A-5 subgroup DREB gene (AmDREB3) isolated from Ammopiptanthus mongolicus, a desert broadleaf shrub with very high tolerance to harsh environments, was characterized in terms of both abiotic stress tolerance and anthocyanin accumulation. AmDREB3 does not contain the transcriptional repression motif EAR, and the protein was located in the nucleus and has transcriptional activation capacity. The transcription of AmDREB3 was differentially induced in the shoots and roots of A. mongolicus seedlings under drought, salt, heat, cold, ultraviolet B, and abscisic acid treatments. Moreover, the transcript levels in twigs, young leaves, and roots were higher than in other organs of A. mongolicus shrubs. Constitutively expressing AmDREB3 improved the tolerance of transgenic Arabidopsis to drought, high salinity and heat, likely by inducing the expression of certain stress-inducible genes. The transgenic Arabidopsis seedlings also exhibited an obvious purple coloration and significant increases in anthocyanin accumulation and/or oxidative stress tolerance under drought, salt, and heat stresses. These results suggest that the AmDREB3 TF may be an important positive regulator of both stress tolerance and anthocyanin accumulation.

Constitutive expression of chloroplast glycerol-3-phosphate acyltransferase from Ammopiptanthus mongolicus enhances unsaturation of chloroplast lipids and tolerance to chilling, freezing and oxidative stress in transgenic Arabidopsis.

Chloroplast glycerol-3-phosphate acyltransferase (GPAT) is the first key enzyme determining the unsaturation of phosphatidylglycerol (PG) in thylakoid membranes and is involved in the tolerance of plants to chilling, heat and high salinity. However, whether the GPAT affects plant tolerance to other stressors has been scarcely reported. Ammopiptanthus mongolicus is the only evergreen broadleaf shrub growing in the central Asian desert, and it has a high tolerance to harsh environments, especially extreme cold. This study aimed to characterize the physiological function of AmGPAT from A. mongolicus. The transcription of AmGPAT was markedly induced by cold and drought but differentially suppressed by heat and high salinity in the laboratory-cultured seedlings. The gene also had the highest transcription levels in the leaves of shrubs naturally growing in the wild during the late autumn and winter months throughout the year. Moreover, AmGPAT was most abundantly expressed in leaves and immature pods rather than other organs of the shrubs. Constitutive expression of AmGPAT in Arabidopsis increased the levels of cis-unsaturated fatty acids, especially that of linolenic acid (18:3), mainly in PG but also in other chloroplast lipids in transgenic lines. More importantly, the transgene significantly increased the tolerance of the transgenics not only to chilling but also to freezing and oxidative stress at both the cellular and whole-plant levels. In contrast, this gene reduced heat tolerance of the transgenic plants. This study improves the current understanding of chloroplast GPAT in plant tolerance against abiotic stressors through regulating the unsaturation of chloroplast lipids, mainly that of PG.

Ammopiptanthus mongolicus stress-responsive NAC gene enhances the tolerance of transgenic Arabidopsis thaliana to drought and cold stresses.

Drought and cold are the primary factors limiting plant growth worldwide. The Ammopiptanthus mongolicus NAC11 (AmNAC11) gene encodes a stress-responsive transcription factor. Expression of the AmNAC11 gene was induced by drought, cold and high salinity. The AmNAC11 protein was localized in the nucleus and plays an important role in tolerance to drought, cold and salt stresses. We also found that differential expression of AmNAC11 was induced in the early stages of seed germination and was related to root growth. When the AmNAC11 gene was introduced into Arabidopsis thaliana by an Agrobacterium-mediated method, the transgenic lines expressing AmNAC11 displayed significantly enhanced tolerance to drought and freezing stresses compared to wild-type Arabidopsis thaliana plants. These results indicated that over-expression of the AmNAC11 gene in Arabidopsis could significantly enhance its tolerance to drought and freezing stresses. Our study provides a promising approach to improve the tolerance of crop cultivars to abiotic stresses through genetic engineering.

AmDREB2C, from Ammopiptanthus mongolicus, enhances abiotic stress tolerance and regulates fatty acid composition in transgenic Arabidopsis.

Dehydration-responsive element-binding (DREB) transcription factors (TFs) play a vital role in plant response to abiotic stresses. However, little is known about DREB TFs in plants adapted to harsh environments and in the formation of polyunsaturated fatty acids (PUFAs), a major membrane component closely associated with plant stress tolerance. Here, we characterized AmDREB2C in Ammopiptanthus mongolicus (Maxim. ex kom.) Cheng F., a desert evergreen broadleaf shrub with a high tolerance to harsh environments. AmDREB2C encodes a canonical DREB2-type TF, and the protein was localized in the nucleus. AmDREB2C had the highest expression levels in leaves of naturally growing shrubs in the wild during the winter season of a year of sampling. The expression was also induced by cold, heat and drought stresses in laboratory-cultured seedlings. Moreover, AmDREB2C was most abundantly expressed in young leaves and immature seeds rather than other tissues of the shrubs. Constitutive expression of AmDREB2C in Arabidopsis enhanced freezing, heat and drought tolerances of the transgenic plants, likely through inducing the expression of important stress-responsive genes. The transgene also increased the level of linolenic acid (C18:3), a major PUFA in most plant species, in leaves and seeds of the transgenic plants. Correspondingly, the transcription of FAD3, FAD7 and FAD8, three genes encoding fatty acid desaturases (FADs) responsible for the production of C18:3, showed a differential up-regulation in these two organs. This study provides new insight into the underlying molecular mechanisms of A. mongolicus' ability to endure harsh environments and DREB TF regulation of fatty acid desaturation.

Root Cell-Specific Regulators of Phosphate-Dependent Growth.

Cellular specialization in abiotic stress responses is an important regulatory feature driving plant acclimation. Our in silico approach of iterative coexpression, interaction, and enrichment analyses predicted root cell-specific regulators of phosphate starvation response networks in Arabidopsis (Arabidopsis thaliana). This included three uncharacterized genes termed Phosphate starvation-induced gene interacting Root Cell Enriched (PRCE1, PRCE2, and PRCE3). Root cell-specific enrichment of 12 candidates was confirmed in promoter-GFP lines. T-DNA insertion lines of 11 genes showed changes in phosphate status and growth responses to phosphate availability compared with the wild type. Some mutants (cbl1, cipk2, prce3, and wdd1) displayed strong biomass gain irrespective of phosphate supply, while others (cipk14, mfs1, prce1, prce2, and s6k2) were able to sustain growth under low phosphate supply better than the wild type. Notably, root or shoot phosphate accumulation did not strictly correlate with organ growth. Mutant response patterns markedly differed from those of master regulators of phosphate homeostasis, PHOSPHATE STARVATION RESPONSE1 (PHR1) and PHOSPHATE2 (PHO2), demonstrating that negative growth responses in the latter can be overcome when cell-specific regulators are targeted. RNA sequencing analysis highlighted the transcriptomic plasticity in these mutants and revealed PHR1-dependent and -independent regulatory circuits with gene coexpression profiles that were highly correlated to the quantified physiological traits. The results demonstrate how in silico prediction of cell-specific, stress-responsive genes uncovers key regulators and how their manipulation can have positive impacts on plant growth under abiotic stress.

Interactions among rice ORC subunits.

The origin recognition complex (ORC) is composed of six subunits and plays an important role in DNA replication in all eukaryotes. The ORC subunits OsORC6 as well as the other five ORC subunits in rice were experimentally isolated and sequenced. It indicated that there also exist six ORC subunits in rice. Results of RT-PCR indicated that expression of all the rice ORC genes are no significant difference under 26°C and 34°C. Yeast two hybridization indicated that OsORC2, -3, -5 interact with each other. OsORC5 can then bind OsORC4 to form the OsORC2, -3,-4,-5 core complex. It suggested that the basic interactions have been conserved through evolution. No binding of OsORC1 and OsORC6 with the other subunits were observed. A model of ORC complex in rice is proposed.

Non-destructive determination of metronidazole powder by using artificial neural networks on short-wavelength NIR spectroscopy.

The present study aimed at providing a new method in sight into short-wavelength near-infrared (NIR) spectroscopy of in pharmaceutical quantitative analysis. To do that, 124 experimental samples of metronidazole powder were analyzed using artificial neural networks (ANNs) in the 780-1100 nm region of short-wavelength NIR spectra. In this paper, metronidazole was as active component and other two components (magnesium stearate and starch) were as excipients. Different preprocessing spectral data (first-derivative, second-derivative, standard normal variate (SNV) and multiplicative scatter correction (MSC)) were applied to establish the ANNs models of metronidazole powder. The degree of approximation, a new evaluation criterion of the networks was employed to prove the accuracy of the predicted results. The results presented here demonstrate that the short-wavelength NIR region is promising for the fast and reliable determination of major component in pharmaceutical analysis.