Applicability of hyperspectral imaging during salinity stress in rice for tracking Na+ and K+ levels in planta.

The ratio of Na+ and K+ is an important determinant of the magnitude of Na+ toxicity and osmotic stress in plant cells. Traditional analytical approaches involve destructive tissue sampling and chemical analysis, where real-time observation of spatio-temporal experiments across genetic or breeding populations is unrealistic. Such an approach can also be very inaccurate and prone to erroneous biological interpretation. Analysis by Hyperspectral Imaging (HSI) is an emerging non-destructive alternative for tracking plant nutrient status in a time-course with higher accuracy and reduced cost for chemical analysis. In this study, the feasibility and predictive power of HSI-based approach for spatio-temporal tracking of Na+ and K+ levels in tissue samples was explored using a panel recombinant inbred line (RIL) of rice (Oryza sativa L.; salt-sensitive IR29 x salt-tolerant Pokkali) with differential activities of the Na+ exclusion mechanism conferred by the SalTol QTL. In this panel of RILs the spectrum of salinity tolerance was represented by FL499 (super-sensitive), FL454 (sensitive), FL478 (tolerant), and FL510 (super-tolerant). Whole-plant image processing pipeline was optimized to generate HSI spectra during salinity stress at EC = 9 dS m-1. Spectral data was used to create models for Na+ and K+ prediction by partial least squares regression (PLSR). Three datasets, i.e., mean image pixel spectra, smoothened version of mean image pixel spectra, and wavelength bands, with wide differences in intensity between control and salinity facilitated the prediction models with high R2. The smoothened and filtered datasets showed significant improvements over the mean image pixel dataset. However, model prediction was not fully consistent with the empirical data. While the outcome of modeling-based prediction showed a great potential for improving the throughput capacity for salinity stress phenotyping, additional technical refinements including tissue-specific measurements is necessary to maximize the accuracy of prediction models.

Comparative physiological and transcriptomic analysis reveals salinity tolerance mechanisms in Sorghum bicolor (L.) Moench.

MAIN CONCLUSION: Mota Maradi is a sorghum line that exhibits holistic salinity tolerance mechanisms, making it a viable potential donor in breeding efforts for improved sorghum lines. High soil salinity is one of the global challenges for crop growth and productivity. Understanding the salinity tolerance mechanisms in crops is necessary for genetic breeding of salinity-tolerant crops. In this study, physiological and molecular mechanisms in sorghum were identified through a comparative analysis between a Nigerien salinity-tolerant sorghum landrace, Mota Maradi, and the reference sorghum line, BTx623. Significant differences on physiological performances were observed, particularly on growth and biomass gain, photosynthetic rate, and the accumulation of Na+, K+, proline, and sucrose. Transcriptome profiling of the leaves, leaf sheaths, stems, and roots revealed contrasting differentially expressed genes (DEGs) in Mota Maradi and BTx623 which supports the physiological observations from both lines. Among the DEGs, ion transporters such as HKT, NHX, AKT, HAK5, and KUP3 were likely responsible for the differences in Na+ and K+ accumulation. Meanwhile, DEGs involved in photosynthesis, cellular growth, signaling, and ROS scavenging were also identified between these two genotypes. Functional and pathway analysis of the DEGs has revealed that these processes work in concert and are crucial in elevated salinity tolerance in Mota Maradi. Our findings indicate how different complex processes work synergistically for salinity stress tolerance in sorghum. This study also highlights the unique adaptation of landraces toward their respective ecosystems, and their strong potential as genetic resources for future plant breeding endeavors.

Gain-of-function mutations of AtNHX1 suppress sos1 salt sensitivity and improve salt tolerance in Arabidopsis.

Soil salinity severely hampers agricultural productivity. Under salt stress, excess Na+ accumulation causes cellular damage and plant growth retardation, and membrane Na+ transporters play central roles in Na+ uptake and exclusion to mitigate these adverse effects. In this study, we performed sos1 suppressor mutant (named sup) screening to uncover potential genetic interactors of SOS1 and additional salt tolerance mechanisms. Map-based cloning and sequencing identified a group of mutants harboring dominant gain-of-function mutations in the vacuolar Na+/H+ antiporter gene AtNHX1. The gain-of-function variants of AtNHX1 showed enhanced transporter activities in yeast cells and increased salt tolerance in Arabidopsis wild type plants. Ion content measurements indicated that at the cellular level, these gain-of-function mutations resulted in increased cellular Na+ accumulation likely due to enhanced vacuolar Na+ sequestration. However, the gain-of-function suppressor mutants showed reduced shoot Na+ but increased root Na+ accumulation under salt stress, indicating a role of AtNHX1 in limiting Na+ translocation from root to shoot. We also identified another group of sos1 suppressors with loss-of-function mutations in the Na+ transporter gene AtHKT1. Loss-of-function mutations in AtHKT1 and gain-of-function mutations in AtNHX1 additively suppressed sos1 salt sensitivity, which indicates that the three transporters, SOS1, AtNHX1 and AtHKT1 function independently but coordinately in controlling Na+ homeostasis and salt tolerance in Arabidopsis. Our findings provide valuable information about the target amino acids in NHX1 for gene editing to improve salt tolerance in crops.

Contributions of Adaptive Plant Architecture to Transgressive Salinity Tolerance in Recombinant Inbred Lines of Rice: Molecular Mechanisms Based on Transcriptional Networks.

Genetic novelties are important nucleators of adaptive speciation. Transgressive segregation is a major mechanism that creates genetic novelties with morphological and developmental attributes that confer adaptive advantages in certain environments. This study examined the morpho-developmental and physiological profiles of recombinant inbred lines (RILs) from the salt-sensitive IR29 and salt-tolerant Pokkali rice, representing the total range of salt tolerance including the outliers at both ends of the spectrum. Morpho-developmental and physiological profiles were integrated with a hypothesis-driven interrogation of mRNA and miRNA transcriptomes to uncover the critical genetic networks that have been rewired for novel adaptive architecture. The transgressive super-tolerant FL510 had a characteristic small tiller angle and wider, more erect, sturdier, and darker green leaves. This unique morphology resulted in lower transpiration rate, which also conferred a special ability to retain water more efficiently for osmotic avoidance. The unique ability for water retention conferred by such adaptive morphology appeared to enhance the efficacy of defenses mediated by Na+ exclusion mechanism (SalTol-effects) inherited from Pokkali. The super-tolerant FL510 and super-sensitive FL499 had the smallest proportions of differentially expressed genes with little overlaps. Genes that were steadily upregulated in FL510 comprised a putative cytokinin-regulated genetic network that appeared to maintain robust growth under salt stress through well-orchestrated cell wall biogenesis and cell expansion, likely through major regulatory (OsRR23, OsHK5) and biosynthetic (OsIPT9) genes in the cytokinin signaling pathway. Meanwhile, a constitutively expressed cluster in FL510 prominently featured two transcription factors (OsIBH1, TAC3) that control tiller angle and growth habit through the brassinosteroid signaling pathway. Both the putative cytokinin-mediated and brassinosteroid-mediated clusters appeared to function as highly coordinated network synergies in FL510. In contrast, both networks appeared to be sub-optimal and inferior in the other RILs and parents as they were disjointed and highly fragmented. Transgressively expressed miRNAs (miR169, miR397, miR827) were also identified as prominent signatures of FL510, with functional implications to mechanisms that support robust growth, homeostasis, and osmotic stress avoidance. Results of this study demonstrate how genetic recombination creates novel morphology that complements inducible defenses hence transgressive adaptive phenotypes.