Molecular insights into the functional role of nitric oxide (NO) as a signal for plant responses in chickpea.

The molecular mechanisms and targets of nitric oxide (NO) are not fully known in plants. Our study reports the first large-scale quantitative proteomic analysis of NO donor responsive proteins in chickpea. Dose response studies carried out using NO donors, sodium nitroprusside (SNP), diethylamine NONOate (DETA) and S-nitrosoglutathione (GSNO) in chickpea genotype ICCV1882, revealed a dose dependent positive impact on seed germination and seedling growth. SNP at 0.1mM concentration proved to be most appropriate following confirmation using four different chickpea genotypes. while SNP treatment enhanced the percentage of germination, chlorophyll and nitrogen contents in chickpea, addition of NO scavenger, cPTIO reverted its impact under abiotic stresses. Proteome profiling revealed 172 downregulated and 76 upregulated proteins, of which majority were involved in metabolic processes (118) by virtue of their catalytic (145) and binding (106) activity. A few crucial proteins such as S-adenosylmethionine synthase, dehydroascorbate reductase, pyruvate kinase fragment, 1-aminocyclopropane-1-carboxylic acid oxidase, 1-pyrroline-5-carboxylate synthetase were less abundant whereas Bowman-Birk type protease inhibitor, non-specific lipid transfer protein, chalcone synthase, ribulose-1-5-bisphosphate carboxylase oxygenase large subunit, PSII D2 protein were highly abundant in SNP treated samples. This study highlights the protein networks for a better understanding of possible NO induced regulatory mechanisms in plants.

NO to drought-multifunctional role of nitric oxide in plant drought: Do we have all the answers?

Nitric oxide (NO) is a versatile gaseous signaling molecule with increasing significance in plant research due to its association with various stress responses. Although, improved drought tolerance by NO is associated greatly with its ability to reduce stomatal opening and oxidative stress, it can immensely influence other physiological processes such as photosynthesis, proline accumulation and seed germination under water deficit. NO as a free radical can directly alter proteins, enzyme activities, gene transcription, and post-translational modifications that benefit functional recovery from drought. The present drought-mitigating strategies have focused on exogenous application of NO donors for exploring the associated physiological and molecular events, transgenic and mutant studies, but are inadequate. Considering the biphasic effects of NO, a cautious deployment is necessary along with a systematic approach for deciphering positively regulated responses to avoid any cytotoxic effects. Identification of NO target molecules and in-depth analysis of its effects under realistic field drought conditions should be an upmost priority. This detailed synthesis on the role of NO offers new insights on its functions, signaling, regulation, interactions and co-existence with different drought-related events providing future directions for exploiting this molecule towards improving drought tolerance in crop plants.

Complex and shifting interactions of phytochromes regulate fruit development in tomato.

Tomato fruit ripening is a complex metabolic process regulated by a genetical hierarchy. A subset of this process is also modulated by light signalling, as mutants encoding negative regulators of phytochrome signal transduction show higher accumulation of carotenoids. In tomato, phytochromes are encoded by a multi-gene family, namely PHYA, PHYB1, PHYB2, PHYE and PHYF; however, their contribution to fruit development and ripening has not been examined. Using single phytochrome mutants phyA, phyB1 and phyB2 and multiple mutants phyAB1, phyB1B2 and phyAB1B2, we compared the on-vine transitory phases of ripening until fruit abscission. The phyAB1B2 mutant showed accelerated transitions during ripening, with shortest time to fruit abscission. Comparison of transition intervals in mutants indicated a phase-specific influence of different phytochrome species either singly or in combination on the ripening process. Examination of off-vine ripened fruits indicated that ripening-specific carotenoid accumulation was not obligatorily dependent upon light and even dark-incubated fruits accumulated carotenoids. The accumulation of transcripts and carotenoids in off-vine and on-vine ripened mutant fruits indicated a complex and shifting phase-dependent modulation by phytochromes. Our results indicate that, in addition to regulating carotenoid levels in tomato fruits, phytochromes also regulate the time required for phase transitions during ripening.

The root as a drill: an ethylene-auxin interaction facilitates root penetration in soil.

Plant roots forage the soil for water and nutrients and overcome the soil's physical compactness. Roots are endowed with a mechanism that allows them to penetrate and grow in dense media such as soil. However, the molecular mechanisms underlying this process are still poorly understood. The nature of the media in which roots grow adds to the difficulty to in situ analyze the mechanisms underlying root penetration. Inhibition of ethylene perception by application of 1-methyl cyclopropene (1-MCP) to tomato seedlings nearly abolished the root penetration in Soilrite. The reversal of this process by auxin indicated operation of an auxin-ethylene signaling pathway in the regulation of root penetration. The tomato pct1-2 mutant that exhibits an enhanced polar transport of auxin required higher doses of 1-MCP to inhibit root penetration, indicating a pivotal role of auxin transport in this process. In this update we provide a brief review of our current understanding of molecular processes underlying root penetration in higher plants.

Tomato root penetration in soil requires a coaction between ethylene and auxin signaling.

During seed germination, emerging roots display positive gravitropism and penetrate into the soil for nutrition and anchorage. Tomato (Solanum lycopersicum) seeds germinated in the presence of 1-methylcyclopropene (1-MCP), an inhibitor of ethylene action, failed to insert roots into Soilrite and grew in the air, forming loops. Time-lapse video imaging showed that 1-MCP-grown root tips retained positive gravitropism and made contact with the surface of Soilrite but failed to penetrate into the Soilrite. Time-course studies revealed that the effect of 1-MCP was most prominent when seed imbibition and germination were carried out in the continual presence of 1-MCP. Conversely, 1-MCP was ineffective when applied postgermination after penetration of roots in the Soilrite. Furthermore, treatment with 1-MCP caused a reduction in DR5::ß-glucuronidase auxin-reporter activity and modified the expression of SlIAA3 and SlIAA9 transcripts, indicating interference with auxin signaling. The reduced ethylene perception mutant, Never-ripe, displayed decreased ability for root penetration, and the enhanced polar auxin transport mutant, polycotyledon, showed a nearly normal root penetration in the presence of 1-MCP, which could be reversed by application of auxin transport inhibitors. Our results indicate that during tomato seed germination, a coaction between ethylene and auxin is required for root penetration into the soil.

Inhibition of the ubiquitin-proteasome pathway alters cellular levels of nitric oxide in tomato seedlings.

Nitric oxide (NO) is involved in diverse plant growth processes; however, little is known about pathways regulating NO levels in plants. In this study, we isolated a NO-overproducing mutant of tomato (Solanum lycopersicum) in which hyper-accumulation of NO, associated with increase in nitric oxide synthase (NOS)-like activity, caused diminished vegetative growth of plants and showed delayed flowering. The hyper-accumulation of NO caused drastic shortening of primary root (shr) in the seedlings, while the scavenging of NO restored root elongation in shr mutant. Inhibition of NOS-like activity reduced NO levels and stimulated root elongation in the shr mutant seedlings, while inhibition of nitrate reductase (NR) activity could not rescue shr phenotype. The stimulation of NO levels in shr mutant also conferred increased resistance to pathogen Pseudomonas syringae. Application of pharmacological inhibitors regulating ubiquitin-proteasome pathway reduced NO levels and NOS-like activity and stimulated shr root elongation. Our data indicate that a signaling pathway involving regulated protein degradation likely regulates NO synthesis in tomato.