Transcriptome analysis provides new insights into plants responses under phosphate starvation in association with chilling stress

Background Chilling temperature reduces the rate of photosynthesis in plants, which is more pronounced in association with phosphate (Pi) starvation. Previous studies showed that Pi resupply improves recovery of the rate of photosynthesis in plants much better under combination of dual stresses than in non-chilled samples. However, the underlying mechanism remains poorly understood. Results In this study, RNA-seq analysis showed the expression level of 41 photosynthetic genes in plant roots increased under phosphate starvation associated with 4 °C (-P 4 °C) compared to -P 23 °C. Moreover, iron uptake increased significantly in the stem cell niche (SCN) of wild type (WT) roots in -P 4 °C. In contrast, lower iron concentrations were found in SCN of aluminum activated malate transporter 1 (almt1) and its transcription factor, sensitive to protein rhizotoxicity 1 (stop1) mutants under -P 4 °C. The Fe content examined by ICP-MS analysis in -P 4 °C treated almt1 was 98.5 ng/µg, which was only 17% of that of seedlings grown under -P 23 °C. Average plastid number in almt1 root cells under -P 4 °C was less than -P 23 °C. Furthermore, stop1 and almt1 single mutants both exhibited increased primary root elongation than WT under combined stresses. In addition, dark treatment blocked the root elongation phenotype of stop1 and almt1. Conclusions Induction of photosynthetic gene expression and increased iron accumulation in roots is required for plant adjustment to chilling in association with phosphate starvation.

Adaptive Mechanisms Make Lupin a Choice Crop for Acidic Soils Affected by Aluminum Toxicity

Almost half of the world’s agricultural soils are acidic, and most of them present significant levels of aluminum (Al) contamination, with Al3+ as the prevailing phytotoxic species. Lupin is a protein crop that is considered as an optimal alternative to soybean cultivation in cold climates. Lupins establish symbiosis with certain soil bacteria, collectively known as rhizobia, which are capable of fixing atmospheric nitrogen. Moreover, some lupin species, especially white lupin, form cluster roots, bottlebrush-like structures specialized in the mobilization and uptake of nutrients in poor soils. Cluster roots are also induced by Al toxicity. They exude phenolic compounds and organic acids that chelate Al to form non-phytotoxic complexes in the rhizosphere and inside the root cells, where Al complexes are accumulated in the vacuole. Lupins flourish in highly acidic soils where most crops, including other legumes, are unable to grow. Some lupin response mechanisms to Al toxicity are common to other plants, but lupin presents specific tolerance mechanisms, partly as a result of the formation of cluster roots. Al-induced lupin organic acid secretion differs from P-induced secretion, and organic acid transporters functions differ from those in other legumes. Additionally, symbiotic rhizobia can contribute to Al detoxification. After revising the existing knowledge on lupin distinct Al tolerance mechanisms, we conclude that further research is required to elucidate the specific organic acid secretion and Al accumulation mechanisms in this unique legume, but definitely, white lupin arises as a choice crop for cultivation in Al-rich acidic soils in temperate climate regions.

Ultrastructural and molecular implications of ecofriendly made silver nanoparticles treatments in pea (Pisum sativum L.)

Background Silver nanoparticles (AgNPs) are the most widely used nanomaterial in agricultural and environmental applications. In this study, the impact of AgNPs solutions at 20 mg/L, 40 mg/L, 80 mg/L, and 160 mg/L on cell ultrastructure have been examined in pea (Pisum sativum L) using a transmission electron microscope (TEM). The effect of AgNPs treatments on the α, β esterase (EST), and peroxidase (POX) enzymes expression as well as gain or loss of inter-simple sequence repeats (ISSRs) markers has been described. Results Different structural malformations in the cell wall and mitochondria, as well as plasmolysis and vacuolation were recorded in root cells. Damaged chloroplast and mitochondria were frequently observed in leaves and the osmiophilic plastoglobuli were more observed as AgNPs concentration increased. Starch grains increased by the treatment with 20 mg/L AgNPs. The expressions of α, β EST, and POX were slightly changed but considerable polymorphism in ISSR profiles, using 17 different primers, were scored indicating gain or loss of gene loci as a result of AgNPs treatments. This indicates considerable variations in genomic DNA and point mutations that may be induced by AgNPs as a genotoxic nanomaterial. Conclusion AgNPs may be used to induce genetic variation at low concentrations. However, considerations should be given to the uncontrolled use of nanoparticles and calls for evaluating their impact on plant growth and potential genotoxicity are justified.

Antiproliferative, genotoxic and mutagenic potential of synthetic chocolate food flavoring

Flavoring additives are of great technological importance for the food industry. However, there is little information regarding the toxicological properties of these micro-ingredients, especially at the cellular level. The present study used meristematic root cells of Allium cepa L. to evaluate the toxicity of a liquid, aroma and flavor synthetic chocolate additive, manufactured and widely marketed throughout Brazil and exported to other countries in South America. The flavoring concentrations evaluated were 100.00; 50.00; 25.00; 1.00; 0.50 and 0.25 µL/L, where the highest concentration established was one-hundred times lower than that commercially suggested for use. The concentration 100 µL/L substantially reduced cell division of meristems within 24- and 48-hours exposure. Concentrations from 100.00 to 0.50 µL/L resulted in a significant number of prophases to the detriment of the other phases of cell division, indicating an aneugenic activity, and induced a significant number of cellular changes, with emphasis on micronuclei, nuclear buds and chromosomal breaks. Under the established analysis conditions, with the exception of concentration 0.25 µL/L, the flavoring of chocolate caused cytotoxicity, genotoxicity and mutagenicity to root meristems.

Comparative organization of tubulin microtubules in root cells of Zea mays (Poaceae) and Beta vulgaris (Chenopodiaceae s. str. / Amaranthaceae s. l.) under the influence of clinorotation

In order to identify the mechanism of functioning of the tubulin cytoskeleton, we have investigated the impact of clinorotation on cortical microtubules organization in the process of cell differentiation in growth zones of plant roots of Zea mays and Beta vulgaris. The similar organization of cortical and endoplasmic microtubules’ network in both species is noted. Clinorotation did not significantly change the organization of microtubules in meristem cells and the central elongation root zone. However, in the distal elongation zone of roots, both Z. mays and B. vulgaris expressed deviations of individual microtubules from the ordered transverse organization (at an angle greater than 45º). This deviation of the microtubules is likely caused by clinorotation and results in discoordination of root growth under these conditions. In addition, it has been found that the scope of destruction of the MT network by taxol in the root cells of both species is not dependent on clinorotation.

Tonoplast-Localized Theanine Transporter CsCAT2 May Mediate Theanine Storage in the Root of Tea Plants (Camellia sinensis L.)

Theanine is the component endowing tea infusion with “umami” taste and antidepression benefits. Theanine is primarily synthesized and stored in root in winter and is transported via vascular tissues to the new shoot in spring. However, the mechanism underlying theanine storage in the root of tea plants remains largely unknown. Cationic amino acid transporter 2 (CsCAT2) in tea plants is homologous to glutamine permease 1 (GNP1), the specific glutamine transporter in yeast. In this study, we identified CsCAT2 as an H+-dependent theanine transporter with medium affinity for theanine. The result of subcellular localization showed that CsCAT2 was a tonoplast-localized transporter. Importantly, CsCAT2 highly expressed in the root in winter during theanine storage and reduced its expression in the root during theanine transport from root-to-shoot in spring. In addition, CsCAT2 expression in the roots of 5 varieties at four time points during December and April was significant negatively correlated with the capacity of theanine root-to-shoot movement. Taken together, these results suggested that CsCAT2 may mediate theanine storage in the vacuole of root cells and may negatively modulate theanine transport from root to shoot.

Cytotoxic and genotoxic effects of the ethanolic extract from the rind of the fruit of Sterculia striata St. Hil. & Naudin / Efeitos citotóxicos e genotóxicos do extrato etanólico da casca do fruto de Sterculia striata St. Hil. & Naudin

The species Sterculia striata A. St. Hil. Naudin has been used by the population in food and in the treatment of skin conditions, mainly for the treatment of boils. Recently, for this species has also been attributed an antioxidant, anti-inflammatory and antibacterial action. However, little is known about its cytogenotoxic potential. The present work aimed to evaluate the cytotoxic and genotoxic effects of the ethanolic extract from the rind of the fruit of Sterculia striata A. St. Hil. Naudin. Phytochemical analysis of the ethanolic extract was performed to determine the presence of secondary metabolites. Concentrations ranging from 0.1 to 1000 µg/ml of the ethanolic extract from the rind of the fruit Sterculia striata were tested for toxicity and cytotoxicity, by the Artemia salina bioassay and the MTT test, respectively. For the genotoxicity analysis, the Allium cepa test was used, at concentrations from 9 to 1000 µg/ml of the extract. All data were analyzed and compared to controls. The statistical test of analysis of variance (ANOVA with a fixed factor) was used, followed by Tukey's multiple comparisons test, for p0.05. Phytochemical screening revealed the presence of tannins, flavonones, flavonols, saponins, alkaloids, steroids and triterpenes. The results showed a decline in the survival rate at high concentrations, in the Artemia salina and MTT tests, the latter being more sensitive for presenting a significant reduction from the concentration of 81 µg/mL. As for the results obtained for the genotoxicity parameter, an increase in the number of chromosomal alterations in root cells exposed to concentrations was observed, also from 81 µg/ml, through the Allium cepa test. The main chromosomal alterations verified were delays, bridges and breaks, in metaphase and anaphase. Taken together, it can be concluded that the ethanolic extract of the rind of the fruit of Sterculia striata A. St. Hil. Naudin exhibits cytotoxic and genotoxic effects mainly at higher concentrations.

A NAC transcription factor OsNAC3 positively regulates ABA response and salt tolerance in rice

Background NAC (NAM, ATAF and CUC) transcription factors (TFs) play vital roles in plant development and abiotic stress tolerance. Salt stress is one of the most limiting factors for rice growth and production. However, the mechanism underlying salt tolerance in rice is still poorly understood. Results In this study, we functionally characterized a rice NAC TF OsNAC3 for its involvement in ABA response and salt tolerance. ABA and NaCl treatment induced OsNAC3 expression in roots. Immunostaining showed that OsNAC3 was localized in all root cells. OsNAC3 knockout decreased rice plants’ sensitivity to ABA but increased salt stress sensitivity, while OsNAC3 overexpression showed an opposite effect. Loss of OsNAC3 also induced Na+ accumulation in the shoots. Furthermore, qRT-PCR and transcriptomic analysis were performed to identify the key OsNAC3 regulated genes related to ABA response and salt tolerance, such as OsHKT1;4, OsHKT1;5, OsLEA3–1, OsPM-1, OsPP2C68, and OsRAB-21. Conclusions This study shows that rice OsNAC3 is an important regulatory factor in ABA signal response and salt tolerance.

Leaf nodule endosymbiotic Burkholderia confer targeted allelopathy to their Psychotria hosts

After a century of investigations, the function of the obligate betaproteobacterial endosymbionts accommodated in leaf nodules of tropical Rubiaceae remained enigmatic. We report that the α-d-glucose analogue (+)-streptol, systemically supplied by mature Ca.Burkholderia kirkii nodules to their Psychotria hosts, exhibits potent and selective root growth inhibiting activity. We provide compelling evidence that (+)-streptol specifically affects meristematic root cells transitioning to anisotropic elongation by disrupting cell wall organization in a mechanism of action that is distinct from canonical cellulose biosynthesis inhibitors. We observed no inhibitory or cytotoxic effects on organisms other than seed plants, further suggesting (+)-streptol as a bona fide allelochemical. We propose that the suppression of growth of plant competitors is a major driver of the formation and maintenance of the Psychotria–Burkholderia association. In addition to potential agricultural applications as a herbicidal agent, (+)-streptol might also prove useful to dissect plant cell and organ growth processes.

Transcriptome and Metabolome Analyses Reveal Potential Salt Tolerance Mechanisms Contributing to Maintenance of Water Balance by the Halophytic Grass Puccinellia nuttalliana

Elevated soil salinity exacerbated by human activities and global climate change poses serious threats to plant survival. Although halophytes provide many important clues concerning salt tolerance in plants, some unanswered questions remain to be addressed, including the processes of water and solute transport regulation. We performed high-throughput RNA-sequencing in roots and metabolome characterizations in roots and leaves of Puccinellia nuttalliana halophytic grass subjected to 0 (control) and 150 mM NaCl. In RNAseq, a total of 31 Gb clean bases generated were de novo assembled into 941,894 transcripts. The PIP2;2 and HKT1;5 transcript levels increased in response to the NaCl treatment implying their roles in water and ion homeostasis. Several transcription factors, including WRKY39, DEK3, HY5, and ABF2, were also overexpressed in response to NaCl. The metabolomic analysis revealed that proline and dopamine significantly increased due to the upregulation of the pathway genes under salt stress, likely contributing to salt tolerance mechanisms. Several phosphatidylcholines significantly increased in roots suggesting that the alterations of membrane lipid composition may be an important strategy in P. nuttalliana for maintaining cellular homeostasis and membrane integrity under salt stress. In leaves, the TCA cycle was enriched suggesting enhanced energy metabolism to cope with salt stress. Other features contributing to the ability of P. nuttalliana to survive under high salinity conditions include salt secretion by the salt glands and enhanced cell wall lignification of the root cells. While most of the reported transcriptomic, metabolomics, and structural alterations may have consequences to water balance maintenance by plants under salinity stress, the key processes that need to be further addressed include the role of the changes in the aquaporin gene expression profiles in the earlier reported enhancement of the aquaporin-mediated root water transport.