Chloroplast-localized PvBASS2 regulates salt tolerance in the C4 plant seashore paspalum.

BILE ACID: SODIUM SYMPORTER FAMILY PROTEIN 2 (BASS2) is localized within chloroplast membranes, facilitating the translocation of pyruvate and Na+ from the cytosol to the plastid, where pyruvate supports isopentenyl diphosphate (IPP) synthesis via the methylerythritol phosphate pathway in C3 plants. Nevertheless, the biological function of BASS2 in C4 plants has not been well defined. This study unveils a previously unidentified role of PvBASS2 in Na+ and pyruvate transport in seashore paspalum (Paspalum vaginatum), a halophytic C4 grass, indicating a specific cellular function within this plant species. Data showed that overexpression of PvBASS2 in seashore paspalum attenuated salt tolerance, whereas its RNAi lines exhibited enhanced salt resistance compared to wild-type plants, suggesting a negative regulatory role of PvBASS2 in seashore paspalum salt tolerance. The constitutive overexpression of PvBASS2 was also found to reduce salt tolerance in Arabidopsis. Further study revealed that PvBASS2 negatively regulates seashore paspalum salt tolerance, possibly due to elevated Na+/K+ ratio, disrupted chloroplast structure, and reduced photosynthetic efficiency following exposure to salinity. Importantly, our subsequent investigations revealed that modulation of PvBASS2 expression in seashore paspalum influenced carbon dioxide assimilation, intermediary metabolites of the tricarboxylic acid cycle, and enzymatic activities under salinity treatment, which in turn led to alterations in free amino acid concentrations. Thus, this study reveals a role for BASS2 in the C4 plant seashore paspalum and enhances our comprehension of salt stress responses in C4 plants.

Low-cost and reliable substrate-based phenotyping platform for screening salt tolerance of cutting propagation-dependent grass, paspalum vaginatum.

BACKGROUND: Salt tolerance in plants is defined as their ability to grow and complete their life cycle under saline conditions. Staple crops have limited salt tolerance, but forage grass can survive in large unexploited saline areas of costal or desert land. However, due to the restriction of self-incompatible fertilization in many grass species, vegetative propagation via stem cuttings is the dominant practice; this is incompatible with current methodologies of salt-tolerance phenotyping, which have been developed for germination-based seedling growth. Therefore, the performance of seedlings from cuttings under salt stress is still fuzzy. Moreover, the morphological traits involved in salt tolerance are still mostly unknown, especially under experimental conditions with varying levels of stress. RESULTS: To estimate the salt tolerance of cutting propagation-dependent grasses, a reliable and low-cost workflow was established with multiple saline treatments, using Paspalum vaginatum as the material and substrate as medium, where cold stratification and selection of stem segments were the two variables used to control for experimental errors. Average leaf number (ALN) was designated as the best criterion for evaluating ion-accumulated salt tolerance. The reliability of ALN was revealed by the consistent results among four P. vaginatum genotypes, and three warm-season (pearl millet, sweet sorghum, and wild maize) and four cold-season (barley, oat, rye, and ryegrass) forage cultivars. Dynamic curves simulated by sigmoidal mathematical models were well-depicted for the calculation of the key parameter, Salt50. The reliability of the integrated platform was further validated by screening 48 additional recombinants, which were previously generated from a self-fertile mutant of P. vaginatum. The genotypes displaying extreme ALN-based Salt50 also exhibited variations in biomass and ion content, which not only confirmed the reliability of our phenotyping platform but also the representativeness of the aerial ALN trait for salt tolerance. CONCLUSIONS: Our phenotyping platform is proved to be compatible with estimations in both germination-based and cutting propagation-dependent seedling tolerance under salt stresses. ALN and its derived parameters are prone to overcome the species barriers when comparing salt tolerance of different species together. The accuracy and reliability of the developed phenotyping platform is expected to benefit breeding programs in saline agriculture.

Genome-Wide Identification and Expression Analysis of Calmodulin-Like Gene Family in Paspalums vaginatium Revealed Their Role in Response to Salt and Cold Stress.

The calmodulin-like (CML) family is an important calcium (Ca2+) sensor in plants and plays a pivotal role in the response to abiotic and biotic stresses. As one of the most salt-tolerant grass species, Paspalums vaginatum is resistant to multiple abiotic stresses, such as salt, cold, and drought. However, investigations of PvCML proteins in P. vaginatum have been limited. Based on the recently published P. vaginatum genome, we identified forty-nine PvCMLs and performed a comprehensive bioinformatics analysis of PvCMLs. The main results showed that the PvCMLs were unevenly distributed on all chromosomes and that the expansion of PvCMLs was shaped by tandem and segmental duplications. In addition, cis-acting element analysis, expression profiles, and qRT-PCR analysis revealed that PvCMLs were involved in the response to salt and cold stress. Most interestingly, we found evidence of a tandem gene cluster that independently evolved in P. vaginatum and may participate in cold resistance. In summary, our work provides important insight into how grass species are resistant to abiotic stresses such as salt and cold and could be the basis of further gene function research on CMLs in P. vaginatum.

Transcriptome Analysis Reveals the Stress Tolerance to and Accumulation Mechanisms of Cadmium in Paspalum vaginatum Swartz.

Cadmium (Cd) is a non-essential heavy metal and high concentrations in plants causes toxicity of their edible parts and acts as a carcinogen to humans and animals. Paspalum vaginatum is widely cultivating as turfgrass due to its higher abiotic stress tolerance ability. However, there is no clear evidence to elucidate the mechanism for heavy metal tolerance, including Cd. In this study, an RNA sequencing technique was employed to investigate the key genes associated with Cd stress tolerance and accumulation in P. vaginatum. The results revealed that antioxidant enzyme activities catalase (CAT), peroxidase (POD), superoxide dismutase (SOD), and glutathione S-transferase GST) were significantly higher at 24 h than in other treatments. A total of 6820 (4457/2363, up-/down-regulated), 14,038 (9894/4144, up-/down-regulated) and 17,327 (7956/9371, up-/down-regulated) differentially expressed genes (DEGs) between the Cd1 vs. Cd0, Cd4 vs. Cd0, and Cd24 vs. Cd0, respectively, were identified. The GO analysis and the KEGG pathway enrichment analysis showed that DEGs participated in many significant pathways in response to Cd stress. The response to abiotic stimulus, the metal transport mechanism, glutathione metabolism, and the consistency of transcription factor activity were among the most enriched pathways. The validation of gene expression by qRT-PCR results showed that heavy metal transporters and signaling response genes were significantly enriched with increasing sampling intervals, presenting consistency to the transcriptome data. Furthermore, over-expression of PvSnRK2.7 can positively regulate Cd-tolerance in Arabidopsis. In conclusion, our results provided a novel molecular mechanism of the Cd stress tolerance of P. vaginatum and will lay the foundation for target breeding of Cd tolerance in turfgrass.

Turfgrass intercropping prevents non-point source pollution in sweet pepper production.

Greenhouse vegetable production is one of the major non-point source (NPS) pollution due to its high fertilizer input and low nutrient use efficiency. Excessive salt and nutrient accumulation in the topsoil is responsible for the NPS pollution in greenhouse vegetable production. This study was designed to investigate the effects of turfgrass intercropping on NPS pollution in greenhouse sweet pepper production. The results showed that the pollution discharges via both surface runoff and leaching were significantly reduced by turfgrass intercropping. The reduction of pollution was associated with the capacity of turfgrass species in the absorption and accumulation of salts and nitrate. Paspalum vaginatum with preferential accumulations of nitrate and salts performed best results in the alleviation of soil salinity and NPS pollution. Paspalum vaginatum-intercropping decreased 51.2% of nitrate and 23.9% of electrical conductivity in the soil, 71.8% of total nitrogen, 54.9% of ammonia-nitrogen, 67.0% of nitrate-nitrogen, 68.8% of total phosphorus, 68.2% of phosphates, and 73.2% of potassium in the surface runoff, and 64.1% of total nitrogen, 53.4% of ammonia-nitrogen, 67.0% of nitrate-nitrogen, 44.3% of total phosphorus, 54.8% of phosphates, and 32.9% of potassium in the leachate. These results indicated that Paspalum vaginatum-intercropping could be a clean and sustainable solution to prevent NPS pollution in greenhouse vegetable production.

Intraspecific variation in elemental accumulation and its association with salt tolerance in Paspalum vaginatum.

Most plant species, including most crops, perform poorly in salt-affected soils because high sodium levels are cytotoxic and can disrupt the uptake of water and important nutrients. Halophytes are species that have evolved adaptations to overcome these challenges and may be a useful source of knowledge for salt tolerance mechanisms and genes that may be transferable to crop species. The salt content of saline habitats can vary dramatically by location, providing ample opportunity for different populations of halophytic species to adapt to their local salt concentrations; however, the extent of this variation, and the physiology and polymorphisms that drive it, remain poorly understood. Differential accumulation of inorganic elements between genotypes or populations may play an important role in local salinity adaptation. To test this, we investigated the relationships between population structure, tissue ion concentrations, and salt tolerance in 17 "fine-textured" genotypes of the halophytic turfgrass seashore paspalum (Paspalum vaginatum Swartz). A high-throughput ionomics pipeline was used to quantify the shoot concentration of 18 inorganic elements across three salinity treatments. We found a significant relationship between population structure and ion accumulation, with strong correlations between principal components derived from genetic and ionomic data. Additionally, genotypes with higher salt tolerance accumulated more K and Fe and less Ca than less tolerant genotypes. Together these results indicate that differences in ion accumulation between P. vaginatum populations may reflect locally adapted salt stress responses.

Genome Resources for Seven Fungal Isolates That Cause Dollar Spot Disease in Turfgrass, Including Clarireedia jacksonii and C. monteithiana.

Fungi in the genus Clarireedia are widespread and destructive pathogens of grasses worldwide, and are best known as the causal agents of dollar spot disease in turfgrass. Here, we report genome assemblies of seven Clarireedia isolates, including ex-types of the two most widespread species, Clarireedia jacksonii and C. monteithiana. These datasets provide a valuable resource for ongoing studies of the dollar spot pathogens that include population diversity, host-pathogen interactions, marker development, and disease control.

Comparative transcriptome profiling provides insights into plant salt tolerance in seashore paspalum (Paspalum vaginatum).

BACKGROUND: Seashore paspalum (Paspalum vaginatum), a halophytic warm-seasoned perennial grass, is tolerant of many environmental stresses, especially salt stress. To investigate molecular mechanisms underlying salinity tolerance in seashore paspalum, physiological characteristics and global transcription profiles of highly (Supreme) and moderately (Parish) salinity-tolerant cultivars under normal and salt stressed conditions were analyzed. RESULTS: Physiological characterization comparing highly (Supreme) and moderately (Parish) salinity-tolerant cultivars revealed that Supreme's higher salinity tolerance is associated with higher Na+ and Ca2+ accumulation under normal conditions and further increase of Na+ under salt-treated conditions (400 mM NaCl), possibly by vacuolar sequestration. Moreover, K+ retention under salt treatment occurs in both cultivars, suggesting that it may be a conserved mechanism for prevention of Na+ toxicity. We sequenced the transcriptome of the two cultivars under both normal and salt-treated conditions (400 mM NaCl) using RNA-seq. De novo assembly of about 153 million high-quality reads and identification of Open Reading Frames (ORFs) uncovered a total of 82,608 non-redundant unigenes, of which 3250 genes were identified as transcription factors (TFs). Gene Ontology (GO) annotation revealed the presence of genes involved in diverse cellular processes in seashore paspalum's transcriptome. Differential expression analysis identified a total of 828 and 2222 genes that are responsive to high salinity for Supreme and Parish, respectively. "Oxidation-reduction process" and "nucleic acid binding" are significantly enriched GOs among differentially expressed genes in both cultivars under salt treatment. Interestingly, compared to Parish, a number of salt stress induced transcription factors are enriched and show higher abundance in Supreme under normal conditions, possibly due to enhanced Ca2+ signaling transduction out of Na+ accumulation, which may be another contributor to Supreme's higher salinity tolerance. CONCLUSION: Physiological and transcriptome analyses of seashore paspalum reveal major molecular underpinnings contributing to plant response to salt stress in this halophytic warm-seasoned perennial grass. The data obtained provide valuable molecular resources for functional studies and developing strategies to engineer plant salinity tolerance.

Seaweed Extracts Enhance Salam Turfgrass Performance during Prolonged Irrigation Intervals and Saline Shock.

The negative effects of the ongoing climate change include unusual prolonged droughts and increased salinity pressures on the agricultural lands. Consequently, crops are facing unprecedented environmental pressure, and this calls for more research toward controlling such major stresses. The current study investigates the effects of seaweed extract sprays of Ascophyllum nodosum (5 and 7 mL·L-1; 6 day intervals) on Paspalum vaginatum Salam' during prolonged irrigation intervals (2 and 6 day) and saline growing conditions (1 and 49.7 dS·m-1) for 6 weeks in containers under greenhouse conditions. Control plants showed reduced turf quality, photochemical efficiency, root length and dry weight, total non-structural carbohydrates, and K and Ca compositions. Seaweed extracts increased turf quality, leaf photochemical efficiency, root length and dry weight, total non-structural carbohydrates, K, Ca, and proline in treated plants during prolonged irrigation intervals as well as saline shock conditions. There were also increases in the antioxidant defensive mechanisms such as catalase (CAT), superoxide dismutase (SOD) and ascorbate peroxidase (APX) activities and non-enzymatic antioxidants as well as reduced lipid peroxidation. The application of SWE at 7 mL·L-1 showed higher performance in treated plants during prolonged irrigation intervals as well as saline conditions. Our findings imply that several mechanisms including drought tolerance, osmotic adjustment and antioxidant defense system may interact to enhance the performance of plants in the face of environmental stress following SWE treatments.

Interaction Between Belonolaimus longicaudatus and Helicotylenchus pseudorobustus on Bermudagrass and Seashore Paspalum Hosts.

Belonolaimus longicaudatus and Helicotylenchus pseudorobustus are among the most common nematode parasites of turfgrasses in Florida. Bermudagrass (Cynodon dactylon × C. transvaalensis) and seashore paspalum (Paspalum vaginatum) are the two turf species most commonly used on Florida golf courses. This paper explores the interactions between B. longicaudatus and H. pseudorobustus on bermudagrass and seashore paspalum hosts. Data collected from thousands of nematode samples submitted to the Florida Nematode Assay Lab over a 8-yr period revealed a negative relationship between B. longicaudatus and H. pseudorobustus on bermudagrass, but not seashore paspalum. In a multi-year field plot experiment using multiple cultivars of bermudagrass, and seashore paspalum B. longicaudatus and H. pseudorobustus were negatively related on both turf species. Greenhouse trials where multiple cultivars of both turf species were inoculated with different combinations of B. longicaudatus and H. pseudorobustus found that each nematode species was inhibitory to the other on both host species. Belonolaimus longicaudatus and H. pseudorobustus clearly impact each other on turfgrass hosts, although the mechanism of the nematode-nematode interactions is unknown.

Field Responses of Bermudagrass and Seashore paspalum Cultivars to Sting and Spiral Nematodes.

Belonolaimus longicaudatus and Helicotylenchus spp. are damaging nematode species on bermudagrass (Cynodon spp.) and seashore paspalum (Paspalum vaginatum) in sandy soils of the southeastern United States. Eight bermudagrass and three seashore paspalum cultivars were tested for responses to both nematode species in field plots for two years in Florida. Soil samples were taken every three months and nematode population densities in soil were quantified. Turfgrass aboveground health was evaluated throughout the growing season. Results showed that all bermudagrass cultivars, except TifSport, were good hosts for B. longicaudatus, and all seashore paspalum cultivars were good hosts for H. pseudorobustus. Overall, bermudagrass was a better host for B. longicaudatus while seashore paspalum was a better host for H. pseudorobustus. TifSport bermudagrass and SeaDwarf seashore paspalum cultivars supported the lowest population densities of B. longicaudatus. Seashore paspalum had a higher percent green cover than bermudagrass in the nematode-infested field. Nematode intolerant cultivars were identified.

Saline Irrigation Affects Belonolaimus longicaudatus and Hoplolaimus galeatus on Seashore Paspalum.

Seashore paspalum (Paspalum vaginatum) has great potential for use in salt-affected turfgrass sites. Use of this grass on golf courses, athletic fields, and lawns in subtropical coastal areas may aid in conservation of freshwater resources. Belonolaimus longicaudatus and Hoplolaimus galeatus are considered among the most damaging root pathogens of turfgrasses in Florida. Glasshouse experiments were performed in 2002 and 2003 to examine the effects of increasing levels of irrigation salinity on B. longicaudatus and H. galeatus. Irrigation treatments were formulated by concentrating deionized water to six salinity levels (0, 5, 10, 15, 20, and 25 dS/m). Final population densities of H. galeatus followed a negative linear regression (r(2) = 0.92 and 0.83; P <= 0.01) with increasing salinity levels. Final population densities of B. longicaudatus were quadratically (r(2) = 0.72 and 0.78; P <= 0.01) related to increasing salinity levels from 0 to 25 dS/m. An increase in population densities of B. longicaudatus was observed at moderate salinity levels (10 and 15 dS/m) compared to 0 dS/m. Root-length comparisons revealed that B. longicaudatus caused root stunting at low salinity levels, 0 to 10 dS/m, but roots were not affected at 15 to 25 dS/m. These results indicate that the ability of B. longicaudatus to feed and stunt root growth was negatively affected at salinity levels of 15 dS/m and above.

Host Status of 'SeaIsle 1' Seashore Paspalum (Paspalum vaginatum) to Belonolaimus longicaudatus and Hoplolaimus galeatus.

Belonolaimus longicaudatus and Hoplolaimus galeatus are considered among the most damaging pathogens of turfgrasses in Florida. However, the host status of seashore paspalum (Paspalum vaginatum) is unknown. Glasshouse experiments were performed in 2002 and 2003 to determine the tolerance of 'SeaIsle 1' seashore paspalum to a population of B. longicaudatus and a population of H. galeatus, and to compare to 'Tifdwarf' bermudagrass for differences. Both nematode species reproduced well on either grass, but only B. longicaudatus consistently reduced root growth as measured by root length. Belonolaimus longicaudatus reduced root growth (P