Effect of successive cuttings on uptake and partitioning of 15N among plant parts of Leucaena leucocephala

Summary We studied the effect of three successive cuttings on N uptake and fixation and N distribution in Leucaena leucocephala. Two isolines, uninoculated or inoculated with three different Rhizobium strains, were grown for 36 weeks and cut every 12 weeks. The soil was labelled with 50 ppm KNO₃ enriched with 10 atom % ¹⁵N excess soon after the first cutting. Except for the atom % ¹⁵N excess in branches of K28 at the second cutting, both the L. leucocephala isolines showed similar patterns of total N, fixed N₂, and N from fertilizer distribution in different parts of the plant at each cutting. The Rhizobium strain did not influence the partitioning of ¹⁵N among the different plant parts. Significant differences in ¹⁵N enrichment occurred in different parts. Live nodules of both isolines showed the lowest atom % ¹⁵N excess values (0.087), followed by leaves (0.492), branches (0.552), stems (0.591), and roots (0.857). The roots contained about 60% of the total plant N and about 70% of the total N derived from fertilizer over the successive cuttings. The total N₂ fixed in the roots was about 60% of that fixed in the whole plant, while the shoots contained only 20% of the fixed N₂. We conclude that N reserves in roots and nodules constitute another N source that must be taken into account when estimating fixed N₂ or the N balance after pruning or cutting plants. ¹⁵N enrichment declined up to about fivefold in the reference and the N₂-fixing plants over 24 weeks following the ¹⁵N application. The proportion and the amounts of N derived from fertilizer decreased, while the amount derived from N₂ fixation increased with time although its proportion remained constant.     

From on-farm solid-set sprinkler irrigation design to collective irrigation network design in windy areas

In this paper, a contribution to the design of collective pressurised irrigation networks in solid-set sprinkler-irrigated windy areas is presented. The methodology is based on guaranteeing minimum on-farm performance, using a historical hourly wind speed database and a ballistic solid-set irrigation simulation model. The proposed method was applied to the Montesnegros Irrigation District (central Ebro basin, Spain). The district irrigates an area of 3493 ha using an on-demand schedule. The average wind speed in the area is 2.8 m s⁻¹. An analysis of district water records showed that farmers often reduce water demand when the wind speed is high, but their irrigation decision making is limited by the capacity of the irrigation network and by the unpredictable character of local winds. Simulations were performed for 11 irrigation seasons, 2 triangular sprinkler spacings (18 m × 18 m and 18 m × 15 m), and 2 sprinkler models. The percentage of monthly suitable time for irrigation was determined for four management strategies. The first one was based on a wind speed threshold (3 m s⁻¹), while the other three were based on three levels (standard, relaxed and restrictive) of two irrigation performance parameters: the Christiansen Uniformity Coefficient (CU) and the Wind Drift and Evaporation Losses (WDEL). The standard strategy classified the time as suitable for irrigation when CU ≥ 84% and WDEL ≤ 20%. The thresholds limits of the irrigation parameters for the relaxed strategy were CU ≥ 80% and WDEL ≤ 25%. Finally, the restrictive strategy used thresholds of CU ≥ 90% and WDEL ≤ 15%. The suitable time for the first strategy (56%) was always lower than for the standard and the relaxed strategies (with respective average values of 75 and 86%), and higher than for the restrictive strategy (30%). In order to design the collective network, the hydrant operating time was equalled to the suitable time for irrigation. The differences in the cost of the collective network plus the on-farm equipment were particularly relevant between the restrictive strategy and the other three. Differences in suitable operating time were clear between sprinkler spacings, and less evident between sprinkler models. The application of the proposed methodology may be limited by the availability of historical wind speed records and CU estimates for different combinations of sprinkler models, sprinkler spacings and wind speed.     

Improved genetic parameter estimations in zoysiagrass by implementing post hoc blocking

Randomized complete block (RCB) design is the most widely used experimental design in biological sciences. As number of treatments increases, the block size become larger and it looses the capacity to control the variance within block, which is its original purpose. A method known as post hoc blocking could be used in these cases to improve the genetic parameter estimation and thus obtain an unbiased assessment of the performance of a given treatment. In trufgrass breeding, as other breeding program, this is a common challenge. The goal of this study was to test the capacity of different post hoc blocking designs to improve the genetic parameter estimation of zoysiagrass (Zoysia spp.). We evaluated two post hoc blocking designs; row–column (R–C) and incomplete block (IB) designs on five genotype trials located in Florida. The results showed that post hoc R–C design had superior model fitting than both the original RCB and the post hoc IB designs when studied at the single measurement level and at the site level. The narrow-sense heritability (0.24–0.40) and the genotype-by-measurement correlation (0.57–0.99) did not change significantly when R–C was compared to the original RCB design. The ranking of the top performing genotypes changed considerably when comparing RCB to R–C design, but the degree depended on the location analyzed. We conclude that the change in the ranking of the top (potentially select individuals) is coming from the better control of intra-block environmental variation, and this could potentially have a significant impact on the breeding selection process.     

Changes in hydrophilic and lipophilic antioxidant activity and related bioactive compounds during postharvest storage of yellow and purple plum cultivars

Eight plum cultivars (four dark-purple and four yellow) were harvested at the commercial ripening stage, and changes of fruit quality properties were evaluated during cold storage and subsequent shelf-life, with special emphasis on bioactive compounds (phenolics, anthocyanins and carotenoids) and antioxidant activity (TAA). From the eight plum cultivars, four showed the typical climacteric ripening pattern (‘Blackamber’, ‘Larry Ann’, ‘Golden Globe’ and ‘Songold’) while four behaved as suppressed-climacteric types (‘Golden Japan’ ‘Angeleno’, Black Diamond’ and ‘TC Sun’), the latter being described for the first time. At harvest, large variations in phytochemicals and antioxidant activity were found among cultivars in peel and pulp tissues, although phytochemical concentration and antioxidant activity were higher in the peel than in the flesh (2–40-fold depending on the bioactive compound). During storage, increases in total phenolics for all cultivars (peel and pulp), in total anthocyanin content in the peel of the dark-purple plums, and total carotenoids in the peel and pulp of the yellow cultivars were observed. This behaviour of the bioactive compounds was reflected in TAA changes, since hydrophilic-TAA (H-TAA) was correlated with both phenolics and anthocyanins, while lipophilic-TAA (L-TAA) was correlated with carotenoids. L-TAA comprised about 30–50% of the TAA in plum tissues. Carotenoids and phenolics (and among them the anthocyanins) could be the main lipophilic and hydrophilic compounds contributing to L-TAA and H-TAA, respectively. No significant loss of bioactive compounds and TAA occurred during prolonged plum storage. Moreover, for a better evaluation of the antioxidant potential of plums, the contribution to carotenoids should not be overlooked.     

Enhanced anther culture efficiency of indica rice (Oryza sativa L.) through modification of the culture media

The production of microspore-derived green plants from anther culture of indica rice is generally very low compared with japonica cultivars. A modified anther culture medium, consisting of a higher KNO₃ content (31 mm ) and casein hydrolysate (CH, 500 mg/1) but without ammonium salts, was tested in comparison with a medium consisting of the widely-used N6 medium nitrogen background, using four indica × indica F₁ hybrids as test materials. Green plant regeneration frequency was at least three-fold higher in the microspore-calli derived from the former medium than in those derived from the modified N6 medium. More than 700 microspore-derived plants were raised in the field. Another study was carried out using indica × japonica and indica × javanica F₁ hybrids. The results indicated that a medium with higher (3.5 mm ) ammonium sulphate may induce a higher frequency of anthers with microspore-calli but not necessarily lead to a larger number of green-plant regenerating calli. Subsequently, using the indica cv. ‘IR-43’ as the test material, use of a lower level (1.75 mm ) of (NH₄)₂SO₄, in addition to KNO₃ (31 mm ), was found to be better than CH (500 mg/l) for anther-response as well as green plant regenerability of the derived microspore-calli. Nitrate-nitrogen or ammonium-nitrogen alone elicited poor response. Twenty-five media involving combinations of KNO₃ (20–34 mm ) and (NH₄)₂SO₄ (1–3 mm ) were tested for their effects on anther response. Combinations involving KNO₃ (31–34 mM) and (NH₄)₂SO₄ (2.0–2.5 mm ) were found superior not only for achieving greater anther response but also, for subsequent green-plant regeneration. This contrasts with the 28 mm of KNO₃ and 3.5 mm of (NH₄)₂SO₄ in the widely-used N6 medium developed for japonica rice. Other than potassium nitrate and ammonium sulphate, and potassium phosphate to some extent, the levels of other inorganic salts tested did not make any significant difference to the process of anther response. Based on these results, modified media with three levels of ammonium sulphate were tested for anther culture efficiency of indica × japonica and indica × javanica derivatives (F₃s). Microspore-calli derived from a medium of a lower (1.75 mm ) level of (NH₄)₂SO₄ showed a higher regeneration potential overall than those derived from a higher (3.5 mm ) level. A revised medium has been suggested, on the basis of these results, for the realization of improved anther culture efficiency and, consequently, improved feasibility of using doubled haploids in genetic and breeding research with indica rice.     

Callus Formation and Plant Regeneration from Protoplasts Derived from Suspension Culture of Rice (Oryza sativa L. cv. 'Taipei 177')

Protoplasts were isolated from young inflorescence-derived suspension cultures of a japonica rice cultivar ‘Taipei 177’. The isolated protoplasts which were cultured either in liquid, agar on Sea plaque agarose underwent sustained division. Maximum plating efficiency of 1.06% occurred in a medium containing macroelements of KM, microelements and vitamins of B5, 0.5 % Sea plaque agarose, 1.0 mg/l of 2,4-D, and glucose as an osmotic stabilizer. Green and albino plants were regenerated from the protocalli in MS semisolid medium containing 4 mg/l BAP, 0.5 mg/l NAA and 500 mg/l casein hydrolysate (MS_18–2).     

Carbon Isotope Discrimination, Leaf Ash Content and Grain Yield in Bread and Durum Wheat Grown under Full-Irrigated Conditions

Integrative physiological criteria, such as carbon isotope discrimination (Δ) and (mineral) ash content (m_a) have been found to be very useful, under drought conditions, to elucidate the association between yield gains and variation of photosynthesis‐related traits and orientate future breeding efforts. Information on this association is scarce under irrigated conditions. The relationships between Δ, m_a and yield were studied in bread wheat (Triticum aestivum L.) and durum wheat (Triticum turgidum L. var. durum) under optimal (drip) irrigation in the arid conditions of north‐west Mexico. Carbon isotope discrimination was analysed on leaves at booting stage and anthesis and on grain at maturity, whereas ash content was measured on the flag leaf at anthesis and maturity. At anthesis, there were differences between bread and durum wheat during grain filling for Δ, but not for m_a. No relationship was found between grain yield and Δ. Leaf ash content at anthesis and maturity showed a broad variability within each species and were associated with grain yield. These results suggest that ash content in leaves could be also used as predictive criteria for yield not only under drought, but also under irrigated conditions, particularly when evaporative demand is high.     

Time-course of nitrogen fixation in two bambara groundnut (Vigna subterranea L. Verdc.) cultivars

TheA-value method, involving the application of a higher¹⁵N rate to a reference non-N₂-fixing plant, was used to assess the magnitude of N₂ fixation in two bambara groundnut cultivars at four growth stages [vegetative, 0–47 days after planting (DAP); early pod-filling, 47–99 DAP; mid-pod-filling, 99–120 DAP; physiological maturity, 120–148 DAP). The cultivars were Ex-Ada, a bunchy type, and CS-88-11, a slightly spreading type. They were grown on a loamy sand. Uninoculated Ex-Ada and CS-88-11 were used as reference plants to measure the N₂ fixed in the inoculated bambara groundnuts. In this greenhouse study, soil was the major source of N in bambara groundnuts during vegetative growth, and during this period it accounted for over 80% of the N accumulaed in the plants. However, N₂ fixation became the major source of plant N during reproductive growth. There were significant differences between the two cultivars in the ability to fix N₂, and at physiological maturity, almost 75% of the N in CS-88-11 was derived from the atmosphere compared to 55% in Ex-Ada. Also, the total N fixed in CS-88-11 at physiological maturity was almost double that in Ex-Ada. Our data indicate that the higher N₂ fixation in CS-88-11 was due to two factors, a higher intensity of N₂ fixation and a longer active period of N₂ fixation. The results also suggest that bambara groundnut genotypes could be selected for higher N₂ fixation in farining systems.     

Field evaluation of N2 fixation and N partitioning in climbing bean (Phaseolus vulgaris L.) using 15N

Summary The common bean (Phaseolus vulgaris L.) is generally regarded as a poor N₂ fixer. This study assessed the sources of N (fertilizer, soil, and fixed N), N partitioning and mobilization, and soil N balance under field conditions in an indeterminate-type climbing bean (P. vulgaris L. cv. Cipro) at the vegetative, early pod-filling, and physiological maturity stages, using the A-value approach. This involved the application of 10 and 100 kg N ha^-1 of ¹⁵N-labelled ammonium sulphate to the climbing bean and a reference crop, maize (Zea mays L.). At the late pod-filling stage (75 days after planting) the climbing bean had accumulated 119 kg N ha^-1, 84% being derived from fixation, 16% from soil, and only 0.2% from the ¹⁵N fertilizer. N₂ fixation was generally high at all stages of plant growth, but the maximum fixation (74% of the total N₂ fixed) occurred during the interval between early (55 days after planting) and late podfilling. The N₂ fixed between 55 and 75 days after planting bas a major source (88%) of the N demand of the developing pod, and only about 11% was contributed from the soil. There was essentially no mobilization of N from the shoots or roots for pod development. The cultivation of common bean cultivars that maintain a high N₂-fixing capacity especially during pod filling, satisfying almost all the N needs of the developing pod and thus requiring little or no mobilization of N from the shoots for pod development, may lead to a net positive soil N balance.     

Mycorrhiza induced resistance in potato plantlets challenged by Phytophthora infestans

Biological control of soil-borne pathogens by arbuscular mycorrhizal (AM) fungi has been repeatedly demonstrated. However, their role in the control of above-ground hemibiotrophic pathogens is less conclusive. Here, we investigated in vitro the impact of an AM fungus on Phytophthora infestans in potato plants. The leaf infection index was decreased in mycorrhizal potato plants. Real-Time Quantitative PCR revealed the induction of two pathogenesis related genes (PR1 and PR2) in the leaves of mycorrhizal plants shortly after infection with P. infestans. These results suggested a systemic resistance in mycorrhizal plants, related to the priming of the two PR genes in potato.