Transcriptomic landscape of tomato traditional long shelf-life landraces under low water regimes.

'Corbarino' (COR) and 'Lucariello' (LUC) belong to the family of Mediterranean long shelf-life tomato landraces, producing high quality fruits under low water input cultivation regime in their traditional cultivation area. Understanding the morpho-physiological and molecular details of the peculiar drought stress tolerance of these two genotypes may be key to their valorization as breeding material. RNA sequencing of leaf samples of COR and LUC subjected to drought stress by water withholding in a semi-controlled greenhouse identified 3089 and 2135 differentially expressed genes respectively. These included COR- and LUC-specific annotated genes, as well as genes containing single nucleotide polymorphisms as compared to reference genome. Enriched Gene Ontology categories showed that categories such as response to water, oxidoreductase activity, nucleotide salvation and lipid biosynthesis-related processes were enriched among up-regulated DEGs. By contrast, growth and photosynthesis related genes were down-regulated after drought stress, consistent with leaf gas exchange and biomass accumulation measurements. Genes encoding cell wall degrading enzymes of the pectinase family were also down-regulated in drought stress conditions and upregulated in rewatering, indicating that cell wall composition/hardness is important for drought stress responses. Globally our results contribute to understanding the transcriptomic and physiological responses of representative tomato genotypes from Southern Italy, highlighting a promising set of genes to be investigated to improve tomato tolerance to drought.

Polyphasic OKJIP Chlorophyll a Fluorescence Transient in a Landrace and a Commercial Cultivar of Sweet Pepper (Capsicum annuum, L.) under Long-Term Salt Stress.

In a soilless long-term salt-stress experiment, we tested the differences between the commercial sweet pepper cultivar "Quadrato d'Asti" and the landrace "Cazzone Giallo" in the structure and function of PSII through the JIP test analysis of the fast chlorophyll fluorescence transients (OKJIP). Salt stress inactivated the oxygen-evolving complex. Performance index detected the stress earlier than the maximum quantum yield of PSII, which remarkably decreased in the long term. The detrimental effects of salinity on the oxygen evolving-complex, the trapping of light energy in PSII, and delivering in the electron transport chain occurred earlier and more in the landrace than the cultivar. Performance indexes decreased earlier than the maximum quantum yield of PSII. Stress-induced inactivation of PSII reaction centers reached 22% in the cultivar and 45% in the landrace. The resulted heat dissipation had the trade-off of a correspondent reduced energy flow per sample leaf area, thus an impaired potential carbon fixation. These results corroborate the reported higher tolerance to salt stress of the commercial cultivar than the landrace in terms of yield. PSII was more affected than PSI, which functionality recovered in the late of trial, especially in the cultivar, possibly due to heat dissipation mechanisms. This study gives valuable information for breeding programs aiming to improve tolerance in salt stress sensitive sweet pepper genotypes.

Physiological Basis of Salt Stress Tolerance in a Landrace and a Commercial Variety of Sweet Pepper (Capsicum annuum L.).

Salt stress is one of the most impactful abiotic stresses that plants must cope with. Plants' ability to tolerate salt stress relies on multiple mechanisms, which are associated with biomass and yield reductions. Sweet pepper is a salt-sensitive crop that in Mediterranean regions can be exposed to salt build-up in the root zone due to irrigation. Understanding the physiological mechanisms that plants activate to adapt to soil salinization is essential to develop breeding programs and agricultural practices that counteract this phenomenon and ultimately minimize yield reductions. With this aim, the physiological and productive performances of Quadrato D'Asti, a common commercial sweet pepper cultivar in Italy, and Cazzone Giallo, a landrace of the Campania region (Italy), were compared under different salt stress treatments. Quadrato D'Asti had higher tolerance to salt stress when compared to Cazzone Giallo in terms of yield, which was associated with higher leaf biomass vs. fruit ratio in the former. Ion accumulation and profiling between the two genoptypes revealed that Quadrato D'Asti was more efficient at excluding chloride from green tissues, allowing the maintenance of photosystem functionality under stress. In contrast, Cazzone Giallo seemed to compartmentalize most sodium in the stem. While sodium accumulation in the stems has been shown to protect shoots from sodium toxicity, in pepper and/or in the specific experimental conditions imposed, this strategy was less efficient than chloride exclusion for salt stress tolerance.

Transcriptomic Changes Drive Physiological Responses to Progressive Drought Stress and Rehydration in Tomato.

Tomato is a major crop in the Mediterranean basin, where the cultivation in the open field is often vulnerable to drought. In order to adapt and survive to naturally occurring cycles of drought stress and recovery, plants employ a coordinated array of physiological, biochemical, and molecular responses. Transcriptomic studies on tomato responses to drought and subsequent recovery are few in number. As the search for novel traits to improve the genetic tolerance to drought increases, a better understanding of these responses is required. To address this need we designed a study in which we induced two cycles of prolonged drought stress and a single recovery by rewatering in tomato. In order to dissect the complexity of plant responses to drought, we analyzed the physiological responses (stomatal conductance, CO2 assimilation, and chlorophyll fluorescence), abscisic acid (ABA), and proline contents. In addition to the physiological and metabolite assays, we generated transcriptomes for multiple points during the stress and recovery cycles. Cluster analysis of differentially expressed genes (DEGs) between the conditions has revealed potential novel components in stress response. The observed reduction in leaf gas exchanges and efficiency of the photosystem PSII was concomitant with a general down-regulation of genes belonging to the photosynthesis, light harvesting, and photosystem I and II category induced by drought stress. Gene ontology (GO) categories such as cell proliferation and cell cycle were also significantly enriched in the down-regulated fraction of genes upon drought stress, which may contribute to explain the observed growth reduction. Several histone variants were also repressed during drought stress, indicating that chromatin associated processes are also affected by drought. As expected, ABA accumulated after prolonged water deficit, driving the observed enrichment of stress related GOs in the up-regulated gene fractions, which included transcripts putatively involved in stomatal movements. This transcriptomic study has yielded promising candidate genes that merit further functional studies to confirm their involvement in drought tolerance and recovery. Together, our results contribute to a better understanding of the coordinated responses taking place under drought stress and recovery in adult plants of tomato.

Open-system chamber for measurements of gas exchanges at plant level.

Gas exchanges of whole canopy can be studied by covering entire plants with a chamber and using portable infrared gas analyzers (IRGAs) to measure CO2 and H2O exchanged with the air blown through the chamber enclosure. The control of temperature rise inside the chamber, which should be kept low, and the accurate measurement of the air flow are two crucial aspects for realistic and precise estimation of photosynthesis and transpiration. An automated open-system plant chamber (clear flexible balloon enclosure) for small plants was developed to ameliorate such a technique. The temperature rise is here predicted by heat balance analysis inside the chamber. The analysis shows that when as much as 500 W m2 of solar radiation is converted to sensible heat, a flow rate of 0.98 mol s(-1) (approximately = 20 L s(-1)) of air blown into a cylinder-shaped enclosure (0.8 m high, 0.5 m wide) is adequate to limit temperature increase to 2 K. An improved calibration for the measurement of the chamber airflow was obtained by combining the use of a Pitot tube anemometer with the classical CO2 injection approach. The concentration increase due to the injection of CO2 at a known rate into the chamber was predicted by the air flow calculated from the "Pitot" air velocity. The turbulent regime of air assured that a single-point Pitot measurement was enough for a good estimation (slope = 0.99; R2 = 0.999) of the actual air flow. The open-system chamber was tested on potted sunflower (Helianthus annuus, L.) and maize (Zea mays, L.) plants under variable solar radiation, temperature, and air humidity during the daytime. As expected, similar rates of maximal leaf-area based photosynthesis (about 40 micromol m(-2) s(-1)) were observed in the two species confirming the reliability of our system. The consistency of data also resulted from the typical relationships observed between photosynthetic rate and light.