Embryonal recombination and germline inheritance of recombined FRT loci mediated by constitutively expressed FLP in tobacco

FLP site-specific recombination has been shown in transgenic plants to excise DNA sequences between target FRT sites, and thereby activate transgenes in plants. In previous reports, crossing of tobacco plants expressing FLP recombinase from a CaMV-35s promoter with plants containing the target FRT sites, hybrid plants with deletion sectors were generated, which were infrequently transmitted to progeny. In this report we evaluate the occurrence of recombination in F₁ hybrid seed derived from crosses of different FLP and FRT-reporter target lines and the germinal transmission of recombined loci from these hybrids to F₂ progeny. Twenty hybrids were generated from crosses of independent five FLP-active lines and four FRT-reporter target lines. In one hybrid, FLP deletions occurred at an early stage, prior to seed maturation, and the deletions from this hybrid were more efficiently transmitted to F₂ progeny. The demonstration of FLP-mediated recombination activity and germinal inheritance of the recombined FRT loci are supported by both molecular and enzymatic evidence. This revised version was published online in August 2006 with corrections to the Cover Date.     

Ionic balance,biomass production,and organic nitrogen as affected by salinity and nitrogen source in annual ryegrass

Studies on the effects of salinity and nitrogen (N) fertilization on ionic balance, biomass, and organic N production of annual ryegrass (Lolium multiflorum Lam.) were conducted. Plants grown in sand were irrigated with nutrient solution with an electrical conductivity of 2 or 11.2 dS#lbm^‐1, and N in the form of sodium nitrate (NaNO₃), ammonium nitrate (NH₄NO₃), or ammonium sulfate [(NH₄)₂SO₄] ranging from 0.5 to 9.0 mM. Salinity increased the concentration of total inorganic cations (C) in plants and specifically sodium (Na) by more than 3‐fold higher in plants grown at high salinity as compared with plants at low salinity. Sodium (Na) concentration in roots was higher than in shoots irrespective of the salinity level, suggesting a restriction of Na transport from roots to shoots. The concentration of total inorganic anions (A) increased with salinity and when plants were supplied with nitrate (NO₃), salinity increased the concentrations of NO₃ and chloride (Cl) in plants. Increasing salinity and N concentration in the growth medium increased organic anions concentration in plants, estimated as the difference between C and A. The effect of different N sources on C‐A followed the order: NH₄NO₃ > NO₃ > ammonium (NH₄). The base of organic anions and inorganic ions with salinity contributed significantly to the osmotic potential of plants shoots and roots. Changes in C affected N and organic acids metabolism in plants, since C were highly correlated (p=0.0001) with C‐A and organic N (N_org) concentrations regardless of the salinity level or N source in the nutrient solutions. A high and positive linear dependency was found between N_org and C‐A in plants grown at high and low salinity levels and different N sources, pointing out the close relationship between N_org and organic anions on metabolism under these conditions. The amount of biomass produced was correlated positively with organic anion concentration in plants exposed to different salinity levels. Plant biomass increased with N concentration in the nutrient solution regardless of the salinity level applied. Biomass accumulation decreased while N_org concentration increased with salinity. Organic N content remained unaffected in plants exposed to salinity when grown in N less than 9.0 mM.     

Untangling the positive and negative effects of shrubs on herbaceous vegetation in drylands

Woody vegetation, as an ecosystem engineer, can modulate the landscape such that the levels of resources in its vicinity undergo positive and negative changes as far as the herbaceous vegetation is concerned. To better understand how these processes play out in a semi-arid ecosystem, we examined resource modulation by woody vegetation, and the response of herbaceous vegetation to that modulation, at a fine spatial scale. Experimental manipulations were employed to separate the positive and negative effects of water, light and seed dispersal in determining herbaceous species density and biomass in three patch types within and adjacent to the shrub (core, periphery and open). We synthesized our results into a multilayered landscape diversity (MLLD) model. Woody vegetation creates distinct multilayered resource patches at its core and periphery which do not correspond to the dichotomous structural pattern of shrub canopy versus intershrub background. The combined effect of these multilayered resource patches had higher herbaceous species density (8.2 vs. 4.0 species 400?cm^?2) and herbaceous biomass (5.4 vs. 1.0?g 400?cm^?2) in the periphery than in the core (3-yr averages). The periphery??s net positive effects are due to enhancement of soil properties (water infiltration depth of 11.1?cm at periphery vs. 8.1?cm at core), while the core??s net negative effects are due to modulation of seed (seed abundance per seed trap of 44.2 at periphery vs. 3.0 at core) and light availability (PAR transmittance of 41.9?% at periphery vs. 16.5?% at core) by the shrub canopy. Thus, when examined at this fine spatial resolution, woody vegetation has both net positive and net negative effects on herbaceous vegetation. Analysis of our results by means of the MLLD model emphasizes the importance of examining the landscape at the spatial scale of the modulated resources and of recognizing different patch types and their differing effects on herbaceous vegetation.     

Understory woody vegetation in manmade Mediterranean pine forests: variation in community structure along a rainfall gradient

Natural regeneration that occurs in the understory of Mediterranean pine monocultures provides the basis for the transition of these simply structured systems toward a more complex and sustainable state. However, the course and consequences of this process, and its relationships with environmental and silvicultural variables, are still inadequately understood. We investigated the relationship between rainfall amount and understory woody vegetation (UWV) structure in mature (40–50 year) Pinus halepensis plantations in the Mediterranean zone of Israel, where rainfall ranges from 280 mm/year in the south to 900 mm/year in the north. We measured abundance, diversity and species composition on south- and north-facing slopes, in forest sites distributed along the rainfall gradient. UWV abundance, as measured by cover percentage and height, increased with rainfall amount along the entire gradient (2–113% and 0.1–3.4 m, respectively), more rapidly on north-facing slopes. Species composition varied along the rainfall gradient, with ranges of species occurrences corresponding to those in unforested habitats. The relationship between rainfall and UWV species richness was positive throughout most of the rainfall gradient, possibly with a shift in pattern at the highest rainfall levels. UWV richness increased sharply with increasing abundance, up to a certain point with no further increase in richness as abundance increased further. We concluded that UWV structure in the studied forest environment and climatic range is strongly determined by rainfall and suggested that the design and management of Mediterranean forests should focus more on optimizing water availability for the various components.     

A foliar spray of micronutrient solutions induces local and systemic protection against powdery mildew (Sphaerotheca fuliginia) in cucumber plants

A single spray of solutions of 0.005M H₃BO₃, 0.0025M CuSO₄, and 0.0025 MnCl₂, on the upper surface of the first true leaf of cucumber plants 2 h before inoculation with a conidial suspension of Sphaerotheca fuliginea, induced systemic protection against powdery mildew in leaves 2 and 3 without causing any damage on the induced leaf (first leaf). A similar level of systemic protection was observed when plants were induced by micronutrients, 2, 24 and 72 h before challenge with S. fuliginea. The level of protection induced by various concentrations varied from solution to solution. In general, the systemic protection induced by K₂HPO₄ was similar to that by the microelements. Spraying of a 1:1 mixture of phosphate and micronutrient solutions did not improve the systemic protection over that obtained with each of the solutions alone. Increasing the inoculum concentration of S. fuliginea increased the number of powdery mildew colonies produced on both induced and non-induced plants and has relatively affected the systemic protection on induced plants. A single foliar spray of micronutrient solutions, as a prophylactic treatment, on the upper surface of all the leaves of 3-leaf stage cucumber plants significantly inhibited powdery mildew development. A single spray of MnCl₂ on leaf 1 elevated peroxidase activity in the soluble fraction and caused an enhancement of -1,3-glucanase content in the ionically bound fractions of leaf 2 of non-inoculated plants. Forty-eight hours after inoculation, the level of both fractions of the enzymes increased in non-treated plants and decreased (-1,3-glucanase) or remained unchanged (peroxidase) in treated (induced) plants as compared to non-treated plants. The possible mechanism for this protection, and the use of microelements and phosphate solutions as inducers for systemic protection and as agents for disease control are discussed.     

Compensation for seasonal changes in eggshell conductance and hatchability of goose eggs by dynamic control of egg water loss

1. The water loss for optimal hatchability of goose eggs was 12% of initial egg mass.

2. Mass specific eggshell water vapour conductance, G_sp, increased by approximately 50% over about 13 weeks during the winter breeding cycle. A similar increase occurred over 8 weeks during summer.

3. Constant setting of humidity in goose incubators and changes in mean G_sp may cause sub‐optimal egg water loss which increases late mortality.

4. Mean G_sp was measured for every batch from each flock. Humidity was changed accordingly in order to reach optimal water loss. A significant improvement of 6% in hatchability and of 8% in Class‐A goslings was noted during 14 successive weeks compared with the previous 11 weeks.

5. Dynamic humidity control for optimising water loss according to mean batch G_sp thus increases hatchability.     

Local and Systemic Activity of BABA (DL-3-aminobutyric Acid) Against Plasmopara viticola in Grapevines

The non-protein amino acid BABA (DL-3-amino-n-butanoic acid, -aminobutyric acid) is reported here to induce local and systemic resistance against downy mildew in grape leaves. Leaf discs of susceptible cultivars placed on BABA solutions and inoculated with Plasmopara viticola on the counter surface produced brownish restricted lesions below the inoculation site (Hypersensitive-like response, HR) which failed to support fungal sporulation. Histochemical analyses of such HR lesions revealed the accumulation of lignin-like deposits in the host cells. In contrast, water-treated inoculated discs produced expanded chlorotic lesions with profuse sporulation in which no lignin accumulation was observed. Mock-inoculated BABA-treated leaf discs showed no HR or lignin accumulation. Concentrations as low as 25µg/ml (0.25mM) of BABA sufficed to prevent tissue colonization with the fungus. Five other isomers of aminobutyric acid, namely L-2 aminobutyric acid, 2-amino isobutyric acid, DL-2-aminobutyric acid (AABA), DL-3-amino isobutyric acid, and 4-aminobutyric acid (GABA) gave no protection against the downy mildew fungus. Of the two (R and S) enantiomers of BABA only the R form was active in producing HR, suggesting a specific stereostructure requirement for activity. BABA could stop fungal colonization even when applied post-infectionally to leaf discs. Resistance of BABA-pulse-loaded leaf discs persisted for more than 14 days. BABA provided systemic protection against the disease when applied via the root system or via the lower leaves of grape plants. Application of ¹⁴C-BABA to a single leaf of intact plants showed the accumulation of the ¹⁴C label in upper leaves (and root tips), suggesting sink-oriented transport.     

Removal of duvatrienediols from the surface of tobacco leaves increases their susceptibility to blue mold

Dipping leaf strips of greenhouse or field-grown tobacco (Nicotiana tabacum L., cv. Ky-14) plants into acetone for 1 s, prior to inoculation with sporangia of Peronospora tabacina Adam, increased their susceptibility to blue mold. Disease severity and sporangial production on leaf discs from acetone-treated leaves were markedly increased compared to those on discs from untreated leaves. Treatment with acetone also decreased variation in susceptibility of leaves from plants of various ages. Disease severity on discs obtained from attached leaves which were dipped in acetone for 1 s was three times greater up to 15 days after dipping than on discs from leaves that were not dipped. TLC and GLC analyses of the acetone extracts indicated that 95% or more of the major cuticular diterpenoids, α- and β-4,8,13-duvatriene-1,3-diols (DVT), were removed from the surface by dipping for 1 s. These compounds had not reappeared on the leaf surface 15 days after leaves were dipped in acetone for 1 s. Aqueous suspensions of the acetone-soluble constituents as well as authentic DVT, inhibited sporangial germination of P. tabacina (ED₁₀₀ = 25 ppm) and the antifungal activity was accounted for by DVT. When DVT was removed from a leaf surface and added back to the same leaf strip, the resistance of the leaf tissue was restored. As tobacco plants aged, their susceptibility to blue mold decreased and the quantity of DVT on the leaf surface increased. The data support a role for DVT in the resistance of tobacco against blue mold.