Twelve alfalfa genotypes that were selected for biomass under salinity, differences in Na and Cl concentrations in shoots and K/Na ratio were evaluated in this long-term salinity experiment. The selected plants were cloned to reduce genetic variability within each genotype. Salt tolerance (ST) index of the genotypes ranged from 0.39 to 1. The most salt-tolerant genotypes SISA14-1 (G03) and AZ-90ST (G10), the top performers for biomass, exhibited the least effect on shoot number and height. SISA14-1 (G03) accumulated low Na and Cl under salinity. Most genotypes exhibited a net reduction in shoot Ca, Mg, P, Fe, and Cu, while Mn and Zn increased under salinity. Salinity reduced foliar area and stomatal conductance; while net photosynthetic rate and transpiration were not affected. Interestingly, salinity increased chlorophyll and antioxidant capacity in most genotypes; however neither parameter correlated well to ST index. Salt-tolerant genotypes showed upregulation of the SOS1, SOS2, SOS3, HKT1, AKT1, NHX1, P5CS1, HSP90.7, HSP81.2, HSP71.1, HSPC025, OTS1, SGF29 and SAL1 genes. Gene expression analyses allowed us to classify genotypes based on their ability to regulate different components of the salt tolerance mechanism. Pyramiding different components of the salt tolerance mechanism may lead to superior salt-tolerant alfalfa genotypes.

Alfalfa (Medicago sativa L.) is an important crop utilized in irrigated regions that are commonly impacted by salinity. We evaluated the effect of salinity continually from emergence to mature plant growth in successive harvests. We studied emergence, biomass production, salt tolerance, and shoot ion composition as potential physiological mechanisms in four nondormant salt‐tolerant cultivars: Salado, SW 8421S, SW 9720, and SW 9215. Irrigation water salinity dominated by sodium sulfate ranging from 3.1 to 30 dS m−1 of electrical conductivity (ECiw) was imposed at planting date. Plants were grown in outdoor sand tanks in Riverside, CA for 300 d. Relative emergence (%) decreased above ECiw 12.7 dS m−1 and was reduced to 53 and 13.4% at 18.4 and 24 dS m−1, respectively. At ECiw 30 dS m−1 there were no survivor plants. Absolute and relative accumulated biomass from 6 harvests significantly decreased for ECiw above 12.7 dS m−1 (6.0 dS m−1 in the saturation extract [ECe]). Plants grown at 18.4 and 24 dS m−1 produced 68% and 30% respectively of the biomass produced at 3.1 dS m−1. Salado showed the least reduction in biomass at elevated salinity and, as with all the cultivars, exhibited yield increases in successive harvests from first through seventh. Increasing salinity increased shoot Na+, total‐S, Cl−, Mg2+, and P and decreased K+ and Ca2+. The ability of Salado to maintain low shoot Na was the mechanism most associated with salt tolerance. Saline waters with resultant ECe values of up to 6 dS m−1 did not significantly reduce total forage production of the second through the seventh harvests. This suggests that irrigation with saline waters resulting in ECe values less than 6 dS m‐1 can be used throughout the production cycle (planting to multiple harvests) without significant yield loss for the cultivars examined.

Although alfalfa is moderately tolerant of salinity, the effects of salinity on nutrient composition and forage parameters are poorly understood. In addition, there are no data on the effect of salinity on the antioxidant capacity of alfalfa. We evaluated four non-dormant, salinity-tolerant commercial cultivars, irrigated with saline water with electrical conductivities of 3.1, 7.2, 12.7, 18.4, 24.0, and 30.0 dS·m −1 , designed to simulate drainage waters from the California Central Valley. Alfalfa shoots were evaluated for nutrient composition, forage parameters, and antioxidant capacity. Salinity significantly increased shoot N, P, Mg, and S, but decreased Ca and K. Alfalfa micronutrients were also affected by salinity, but to a lesser extent. Na and Cl increased significantly with increasing salinity. Salinity slightly improved forage parameters by significantly increasing crude protein, the net energy of lactation, and the relative feed value. All cultivars maintained their antioxidant capacity regardless of salinity level. The results indicate that alfalfa can tolerate moderate to high salinity while maintaining nutrient composition, antioxidant capacity, and slightly improved forage parameters, thus meeting the standards required for dairy cattle feed. OPEN ACCESSAgriculture 2015, 5 578

Alfalfa (Medicago sativa L.) is a major forage crop in Argentina with an estimated cultivated area of 4 million ha in the 2009–2010 season, which constitutes a primary component for the animal production chain. In early summer of 2010, alfalfa plants showing virus-like symptoms were identified in 20 commercial fields in La Pampa Province with 95% disease prevalence. Diseased plants had shortened internodes, a bushy appearance, deformations, puckering, epinasty of leaflet blades, vein enations, and varying sized papillae on the adaxial leaflet surfaces. Similar symptoms were observed in alfalfa crops in Buenos Aires, Cordoba, Santa Fe, and Santiago del Estero provinces. Electron microscopy (EM) and molecular assays were performed on leaf tissue from one asymptomatic and two symptomatic plants. EM of ultrathin sections revealed membrane-bound bullet-shaped particles associated with the endoplasmic reticulum of phloem cells from symptomatic plants only. Total RNA was extracted from symptomatic and asymptomatic plants with the RNeasy Plant Mini Kit (Qiagen, Hilden, Germany) for a template in one-step reverse transcription (RT)-PCR assays with the Access RT-PCR Kit (Promega, Madison, WI). RT-PCR assays employed degenerate primers targeting conserved regions of plant rhabdovirus polymerase (L) genes (2). An amplicon of approximately 1 kilobase pairs (detected only from symptomatic plants) was gel purified with the Wizard SV Gel and PCR Clean-Up System (Promega), cloned into pGEM-T Easy Vector System (Promega), and sequenced. Three independents clones were sequenced in both directions at Macrogen Inc. (Korea Republic) to generate a consensus sequence (GenBank Accession No. HQ380230) and compared to sequences of other plant rhabdoviruses available on GenBank. The partial L gene sequence of the alfalfa-infecting rhabdovirus shared highest nucleotide (68.0%) and amino acid (76.5%) sequence identity with the cytorhabdovirus Strawberry crinkle virus (Accession No. AY331390). A phylogenetic tree based on partial amino acid sequences of the polymerase gene indicated the alfalfa-infecting virus was more closely related to cytorhabdoviruses than to nucleorhabdoviruses. Symptoms observed resembled those reported for alfalfa plants infected with a plant rhabdovirus named Alfalfa enation virus (1), which has never been fully characterized. Collectively, the data implicate the observed rhabdovirus as the etiological agent. To our knowledge, this is the first report in Argentina (and South America) of a rhabdovirus infecting alfalfa. Additional field surveys and monitoring of vector/s and yield losses need to be conducted. References: (1) B. Alliot and P. A. Signoret. Phytopathol. Z. 74:69, 1972. (2) R. L. Lamprecht et al. Eur. J. Plant Pathol. 123:105, 2009.