Effect of temperature on rice growth in nutrient solution and in acid sulphate soils from Vietnam

Climatic and soil factors are limiting rice growth in many countries. In Vietnam, a steep gradient of temperature is observed from the North to the South, and acid sulphate soils are frequently devoted to rice production. We have therefore attempted to understand how temperature affects rice growth in these problem soils, by comparison with rice grown in nutrient solution. Two varieties of rice, IR64 and X2, were cultivated in phytotrons at 19/21°C and 28/32°C (day/night) for 56 days, after 3 weeks preculture in optimal conditions. Two soils from the Mekong Delta were tested. Parallel with the growing experiments, these two soils were incubated in order to monitor redox potential (E _h ), pH, soluble Al and Fe, soluble, and available P. Tillering retardation at 20°C compared to 30°C was similar in nutrient solutions and in soils. The effect of temperature on increasing plant biomass was more marked in solutions than in soils. The P concentrations in roots and shoots were higher at 20°C than at 30°C, to such an extent that detrimental effect was suspected in plants grown in solution at the lowest temperature. The translocation of Fe from roots to shoots was stimulated upon rising temperature, both in solutions and in soils. This led to plant death on the most acid soil at 30°C. Indeed, the accumulation of Fe in plants grown on soils was enhanced by the release of Fe²⁺ due to reduction of Fe(III)-oxihydroxides. Severe reducing conditions were created at 30°C: redox potential (E _h ) dropped rapidly down to about 0 V. At 20°C, E _h did not drop below about 0.2 V, which is a value well in the range of Fe(III)/Fe(II) buffering. Parallel to E _h drop, pH increased up to about 6–6.5 at 30°C, which prevented plants from Al toxicity, even in the most acid soil. Phosphate behavior was obviously related to Fe-dynamics: more reducing conditions at 30°C have resulted in enhancement of available P, especially in the most acid soil.     

Comparative effects of organic and inorganic nitrogen sources applied to a flooded soil on rice yield and availability of N

A pot experiment was conducted to study the effect of organic and inorganic nitrogen (N) sources on the yield and N uptake of rice from applied and native soil-N. The residual effect of these N sources on a succeeding wheat crop was also studied. Organic N was applied in the form of ¹⁵N-labelled Sesbania aculeata L., a legume, and inorganic N in the form of ¹⁵N-labelled ammonium sulphate. The two sources were applied to the soil separately or together at the time of transplanting rice. Recovery of N by rice from both the applied sources was quite low but both sources caused significant increases in biomass and N yield of rice. Maximum increase was recorded in soil treated with organic N. The residual value of the two materials as source of N for wheat was not significant; the wheat took up only a small fraction of the N initially applied. Loss of N occurred from both applied N sources, the losses being more from inorganic N. Both applied N sources caused a substantial increase in the availability of soil-N to rice and wheat; most of this increase was due to organic N and was attributed to the so-called ‘priming’ effect or ANI (added nitrogen interaction) of the applied material.     

Effects of foliar applied gibberellic acid and benzyladenine upon yield components in sunflower (Helianthus annuus L.)

Sunflower yield is determined by seed number/m^–2 and by achene weight. Frequently, a high percentage of empty achenes in the inner portion of the capitulum, probably due to a reduced vascularization of that section of the flower head, decreases final yield. The objective of the present research is to determine if foliarly applied gibberellic acid (GA) and benzyladenine (BA) can enhance the vascularization in the inner portion of the capitulum, improving photoassimilate translocation. Field experiments were conducted during 1989/90 with hybrid SPS 894 and during 1990/91 with hybrid ACA 882. GA (150 mg/l^–1), BA (150 and 250 mg/l^–1) and GA 150+BA 150 mg/l^–1 each were foliarly applied 20, 40, or 60 days after emergence. For both seasons and hybrids plant growth regulator (PGRs) applications significantly reduced the percentage of empty achenes, increased achene weight, achene weight (× 1000) and achene number in the inner portion of the capitulum and in the middle and outer portion during 1990/91. A 25% increase in seed yield was achieved due to PGR application and the capitulum partition index (achene weight/receptacle weight^–1, CPI) was significantly increased due to an improvement in photoassimilate distribution. A distribution model was derived showing that preferential allocation of photoassimilates in the outer portion of the capitulum can be modified by PGR application, demonstrating that photoassimilate distribution is under hormonal control.Consejo Nacional de Investigaciones Científicas y Tecnológicas (CONICET)     

Dynamics of nitrogen uptake and partitioning in early and late fruit ripening peach (Prunus persica) tree genotypes under a mediterranean climate

The dynamics of N uptake and N partitioning in peach (Prunus persica, Batsch) trees of a very early (cv. Flordastar) and a very late (cv. Tudia) fruit ripening varieties grown under a mediterranean climate was assessed during one season. Labelled N was applied to two-year old potted trees which were destructively harvested at regular intervals during the vegetative and reproductive cycle. Tree phenology as well as vegetative and reproductive growth of the two genotypes strongly differed: bud burst started in late January in Flordastar and late March in Tudia. Leaf senescence in Flordastar was almost complete by mid October, while Tudia still retained a significant fraction of leaves at the December harvest. Fruit yield averaged 4.0 and 6.9 kg tree^–1 (fresh weight) in cv. Flordastar and Tudia, respectively, and fruit size was within commercial standards for the two genotypes. After growth resumption, shoot and fruit growth mainly relied on N remobilised from reserves, which accounted for 72–80% of total N in new growth. Nitrogen uptake by both genotypes was relatively low in the first month after bud burst, then was more rapid until the end of the season. Total labelled N uptake did not differ between the two genotypes and accounted on average for 65–70% of total N supplied. The kinetics of labelled N uptake were similar in the two varieties despite the great difference in the timing of their fruit ripening. Leaves were the main sink for N during much of the experimental period. The fruits, when present, also used a significant fraction of the absorbed N, which was almost constant until fruit ripening in Flordastar. Nitrogen partitioning to leaves declined progressively after summer, when a greater fraction of the absorbed N was recovered in the twigs, the trunk, the fine roots and especially in the coarse roots. The data provide evidence for guiding the kinetics of N supply to peach orchards under a Mediterranean climate.     

Differential effects of coinoculations with Pseudomonas jessenii PS06 (a phosphate-solubilizing bacterium) and Mesorhizobium ciceri C-2/2 strains on the growth and seed yield of chickpea under greenhouse and field conditions

In the course of a project carried out in two regions of Spain, Castilla y León and Andalucía, aiming to find useful biofertilizers for staple grain-legumes, an efficient rhizobia nodulating chickpea (termed as C-2/2) and a powerful in vitro phosphate-solubilizing bacterial strain (termed as PS06) were isolated. Analyses of their 16S rDNA sequence indicated that they belong to the bacterial species Mesorhizobium ciceri and Pseudomonas jessenii, respectively. Greenhouse and field experiments were carried out in order to test the effect of single and dual inoculations on chickpea (ecotype ILC-482) growth. Under greenhouse conditions, plants inoculated with Mesorhizobium ciceri C-2/2 alone had the highest shoot dry weight. The inoculation treatment with P. jessenii PS06 yielded a shoot dry weight 14% greater than the uninoculated control treatment, but it was not correlated with shoot P contents. However, the co-inoculation of C-2/2 with PS06 resulted in a decrease in shoot dry weight with respect to the inoculation with C-2/2 alone. Under field conditions, plants inoculated with M. ciceri C-2/2, in single or dual inoculation, produced higher nodule fresh weight, nodule number and shoot N content than the other treatments. Inoculation with P. jessenii PS06 had no significant effect on plant growth. However, the co-inoculation treatment ranked the highest in seed yield (52% greater than the uninoculated control treatment) and nodule fresh weight. These data suggest that P. jessenii PS06 can act synergistically with M. ciceri C-2/2 in promoting chickpea growth. The contrasting results obtained between greenhouse and field experiments are discussed.     

Changes in the amino acid composition and protein modification in phloem sap of rice

Pure phloem sap was collected from insects feeding on rice (Oryza sativa L.) leaves by a laser technique similar to the aphid stylet technique. Rapid circulation of nitrogen in the sieve tubes was demonstrated directly using ¹⁵N as a tracer. Application to the roots of the metabolic inhibitors of amino acids, aminooxyacetate and methioninesulfoximine, changed the amino acid composition in the sieve tubes. Feeding methionine to leaf tips resulted in its bulk transfer into the sieve tubes. In vitro experiments confirmed the existence of protein kinases in the pure rice phloem sap. The phosphorylation status of the sieve tube sap proteins was affected by the light regime. The possibility that changes in chemical composition or protein modification such as phosphorylation in the sieve tubes might affect plant growth are discussed.Analysis of pure phloem sap collected from rice plants by insect laser technique has shown dynamic changes in the chemical composition and the quality of proteins in the sap.     

Nodulation of lucerne (Medicago sativa L.) in an acid soil: Effects of inoculum size and lime-pelleting

The effects of inoculum level and lime-pelleting were studied in an acid soil with respect to the nodulation and growth of lucerne (Medicago sativa cv Resis) and the population dynamics of Rhizobium meliloti. In small root-boxes (rhizotrons), the in-situ survival of inoculated rhizobia was studied in the micro-environment around the seed for a period of 12 days after sowing. During the initial 24 hours, a strong increase in rhizobial numbers was measured, concomitantly with the development of roots. As a result of lime-pelleting, rhizobial numbers were higher only at 3 days after sowing (P<0.05). Later, this difference diminished steadily. Addition of lime did not increase the adhesion of the rhizobia to the seedling tap root. Plant responses to inoculation were studied in pots. To obtain optimal nodulation, the soil had to be neutralized around the seed with lime and at least 10⁵ cells of R. meliloti were required. With more than 10⁵ rhizobia per seed, lime-pelleting increased the number of crown-nodulated seedlings from 24% to 77%. Higher numbers of rhizobia could not compensate the effect of lime. A strong correlation was found between crown nodulation, nitrogen content and dry weight of the shoots.     

Soil organic matter and the indigenous nitrogen supply of intensive irrigated rice systems in the tropics

Soil organic matter (SOM) has been proposed as an index of N supply in paddy soils although field validations are few. We evaluated the relationship between the indigenous N supply (N _i ) of the soil-floodwater system and soil organic carbon (SOC) or total N (N _t ) in surface soil of long-term fertility experiments (LTFEs) at 11 sites, in 42 farmer's fiels with similar soil type, and in the same field in ten consecutive rice (Oryza sativa L.) crops. The N _i was estimated by crop N uptake from plots without applied N (N _o plots) under otherwise favorable growth conditions. There was a tight linear correlation between yields and N uptake in N _o plots and tremendous variation in both parameters among LTFE sites, farmer's fields, and in the same field over time. Correlation between N _i and SOC or N _t explained little of this variation. Factors likely to contribute to the poor correlation were: (1) inputs of N from sources other than N mineralization of SOM in surface soil, (2) degree of congruence between soil N supply and crop demand, which is sensitive to soil drying, length of fallow, crop rotation, and residue management, and (3) differences in SOM quality related to intensive cropping in submerged soil. Better understanding of the processes governing the N _i of tropical lowland rice systems would contribute to the development of crop management practices that optimize utilization of indigenous N resources.     

Netuptake of sulphate and its transport to the shoot in tobacco plants fumigated with H2S or SO2

In tobacco plants the net uptake of sulphate and its transport to the shoot were determined after cultivation with low, normal, and high sulphate supply. The relative amount of the sulphate taken up that was transported to the shoot was used as a measure of xylem loading. Net uptake of sulphate and its transport to the shoot were low in tobacco plants grown with low sulphate, and high in plants cultivated with high sulphate. Xylem loading, however, was relatively low in tobacco plants grown with high sulphate and relatively high in tobacco plants grown with low sulphate supply. Pre-culture in low sulphate containing nutrient solution also resulted in a high proportion of the absorbed sulphate being transported into the xylem if normal sulphateconcentration was supplied afterwards. Fumigation with H₂S or SO₂ reduced net uptake of sulphate in tobacco plants grown with normal, but not with high sulphate supply. Sulphate transport to the shoots was diminished by H₂S or SO₂ fumigation in tobacco plants grown with normal and high sulphate supply. Also the relative amount of the sulphate taken up that was transported to the shoot was lowered by fumigation with H₂S or SO₂ in tobacco plants grown with normal sulphate supply. Apparently, the diminished sulphate transport to the shoot upon H₂S or SO₂ fumigation can only partially be explained by a smaller sulphate uptake. Sulphur nutrition of tobacco plants also seems to be controlled by xylem loading of sulphate. The possible role of glutathione as a signal regulating sulphur nutrition of tobacco plants upon fumigation with H₂S and SO₂ is discussed.     

Effect of Sesbania,Azolla and rice straw incorporation on the kinetics of NH4, K,Fe, Mn,Zn and P in some flooded rice soils

In a greenhouse study, with and without rice plants, of five flooded Philippine rice soils whose organic C (OC) content varied from 0.5 to 3.6%, incorporation ofSesbania rostrata, Azolla microphylla and rice straw affected the kinetics of soil solution NH ₄ ⁺ −N, K⁺, Fe²⁺, Mn²⁺, Zn²⁺, and P. Sesbania and Azolla increased NH ₄ ⁺ −N concentration above the control treatment, whereas rice straw depressed it. In all soils Azolla released less NH ₄ ⁺ −N than Sesbania. The apparent net N release depended on the soil and ranged from 44–81% for Sesbania and 27–52% for Azolla. These effects persisted throughout the growth of IR36. Soil solution and exchangeable NH ₄ ⁺ −N increased initially but levelled off between 30 to 80 days and between 20 to 40 days after flooding (DF), respectively. With rice, soil solution NH ₄ ⁺ −N concentration, reached a peak at 15–40 DF and declined to very low levels (<4mg L⁻¹). In the 3 soils of low OC content nitrogen derived from green manure ranged from 34–53% and the apparent revovery of added green manure N varied from 29–67%. Almost all N released from both Azolla and Sesbania were recovered in the rice plant in all soils except Concepcion with only 77%. The concentration of K⁺, Fe²⁺, Mn²⁺ and P in the soil solution were higher with rice straw than Sesbania and Azolla in all soils except Hanggan which showed no change in Fe²⁺ and Mn²⁺ but increased K⁺ and P. In general, rice straw, Sesbania and Azolla decreased Zn²⁺ concentration in all soils.     

Characterization and use in ELISA of a new monoclonal antibody for quantitation of abscisic acid in senescing rice leaves

A new monoclonal antibody (mAb) was generated against abscisic acid (ABA), and an indirect enzyme-linked immunosorbent assay (ELISA) using this mAb was developed for convenient quantitative analysis of ABA levels in rice leaf extracts. The mAb, raised against (+-)-ABA conjugated to bovine serum albumin (BSA) through its carboxyl group (C₁), reacted preferentially with the (+)-ABA enantiomer, and equally well with both free and methyl-ester (+-)-ABA. Cross-reactivity with several ABA-related compounds was negligible. Linearity was obtained between 3 and 1000 pmo1 of (+)-ABA. The ABA-mAb was further used to quantitate pmol quantities of (+)-ABA in attached and detached rice leaves. Results obtained with such ELISA quantitation showed an increase in the free ABA content of detached rice leaves at progressive stages of senescence, which was regarded as a senescence-related response. This quantitation compared favorably with other presently used techniques for ABA determination, with regard to their detection limits, cost and assay time. The results suggest that the combination of a specific mAb with a sensitive ELISA technique is quite promising for quantitation of ABA.     

Enhanced glucotropaeolin production in hairy root cultures of Tropaeolum majus L. by combining elicitation and precursor feeding

We have studied the effects of precursor amino acids (phenylalanine and cystein), elicitors (salicylic and acetylsalicylic acids, methyl jasmonate, β-aminobutyric acid, yeast extract), PAL-inhibitor (1-amino-2-phenylethylphosphonic acid) applied alone or in combination on glucotropaeolin production and myrosinase activity in hairy root cultures of Tropaeolum majus. Short, 24-h treatment, and subsequent transfer of hairy roots to the fresh medium enabled avoiding detrimental effect of studied stimulators on biomass growth. In control cultures the highest glucotropaeolin content, 58.1 ± 6.7 mg g⁻¹ DW, was detected on the 3rd day after transfer of the roots to the fresh medium but glucotropaeolin yield (mg 100 ml^–1 culture volume) had been increasing until the 9th day after transfer as a result of continuous biomass growth. Glucotropaeolin content and yield were 2-fold enhanced after treatment with precursor amino acids or PAL-inhibitor alone, but their combination additively led to 4-fold increase in glucotropaeolin production. Among the studied elicitors acetylsalicylic acid induced the highest, 3-fold increase in glucotropaeolin production, it also enhanced myrosinase activity, but to a smaller extend (by about 50%). Acetylsalicylic acid also potentiated induced by precursors, PAL-inhibitor, methyl jasmonate and yeast extract production of glucotropaeolin. The highest, 4.8-fold increase in glucotropaeolin production was found after combined acetylsalicylic acid and precursors treatment. Additive effect of acetylsalicylic acid-combined treatment on myrosinase activity was not detected. The obtained results indicate that amino acid precursors, phenylalanine and cystein, availability may be a limiting factor in the process of stimulation of glucotropaeolin production in T. majus hairy root cultures.     

Controlled Expression of Recombinant Proteins in <Emphasis Type="Italic">Physcomitrella patens</Emphasis> by a Conditional Heat-shock Promoter: a Tool for Plant Research and Biotechnology

The ability to express tightly controlled amounts of endogenous and recombinant proteins in plant cells is an essential tool for research and biotechnology. Here, the inducibility of the soybean heat-shock Gmhsp17.3B promoter was addressed in the moss Physcomitrella patens, using β-glucuronidase (GUS) and an F-actin marker (GFP-talin) as reporter proteins. In stably transformed moss lines, Gmhsp17.3B-driven GUS expression was extremely low at 25 °C. In contrast, a short non-damaging heat-treatment at 38 °C rapidly induced reporter expression over three orders of magnitude, enabling GUS accumulation and the labelling of F-actin cytoskeleton in all cell types and tissues. Induction levels were tightly proportional to the temperature and duration of the heat treatment, allowing fine-tuning of protein expression. Repeated heating/cooling cycles led to the massive GUS accumulation, up to 2.3% of the total soluble proteins. The anti-inflammatory drug acetyl salicylic acid (ASA) and the membrane-fluidiser benzyl alcohol (BA) also induced GUS expression at 25 °C, allowing the production of recombinant proteins without heat-treatment. The Gmhsp17.3B promoter thus provides a reliable versatile conditional promoter for the controlled expression of recombinant proteins in the moss P. patens.     

Induction of early blight resistance in tomato by <Emphasis Type="Italic">Quercus infectoria</Emphasis> gall extract in association with accumulation of phenolics and defense-related enzymes

Treatment of tomato leaves with aqueous extract (0.5%) of the galls of Quercus infectoria significantly reduced infection from subsequent inoculation with Alternaria solani, the tomato early blight pathogen. When the leaves were challenge-inoculated with A. solani 3 d after application of Q. infectoria gall extract (QIGE), the percent defoliation decreased from 33.6 to 7.3. Two to three day pre-treatment with QIGE reduced the percent defoliation by 77 percent. The biochemical responses of tomato plants to QIGE were also studied. In tomato plants treated with QIGE, phenolic content increased rapidly, reached the maximum at 2 d after treatment. Phenylalanine ammonia-lyase (PAL) activity increased significantly from 1 d after treatment and the maximum enzyme activity was recorded 2 d after treatment at which period a 3-fold increase in PAL activity was observed when compared to the control. Peroxidase (PO) activity was also significantly increased 1 d after treatment and the maximum activity was reached 2 d after treatment. Peroxidase isozyme analysis indicated that PO-1 was increased dramatically in tomato leaves 1 d after treatment and maintained at the same level throughout the experimental period of 6 d. When tomato leaves were treated with QIGE, a two-fold increase in chitinase and β-1,3-glucanase activities was recorded 2 and 3 d respectively, after treatment. The enhanced activities of defense-related enzymes and elevated levels of phenolics in QIGE-treated tomato plants between 1 and 3 d after treatment suggest that these induced biochemical defenses may be involved in the suppression of early blight by QIGE.     

Lysine is catabolized to 2-aminoadipic acid in <Emphasis Type="Italic">Penicillium chrysogenum</Emphasis> by an omega-aminotransferase and to saccharopine by a lysine 2-ketoglutarate reductase. Characterization of the omega-aminotransferase

The biosynthesis and catabolism of lysine in Penicillium chrysogenum is of great interest because these pathways provide 2-aminoadipic acid, a precursor of the tripeptide δ-L-2-aminoadipyl-L-cysteinyl-D-valine that is an intermediate in penicillin biosynthesis. In vivo conversion of labelled L-lysine into two different intermediates was demonstrated by HPLC analysis of the intracellular amino acid pool. L-lysine is catabolized to 2-aminoadipic acid by an ω-aminotransferase and to saccharopine by a lysine-2-ketoglutarate reductase. In lysine-containing medium both activities were expressed at high levels, but the ω-aminotransferase activity, in particular, decreased sharply when ammonium was used as the nitrogen source. The ω-aminotransferase was partially purified, and found to accept L-lysine, L-ornithine and, to a lesser extent, N-acetyl-L-lysine as amino-group donors. 2-Ketoglutarate, 2-ketoadipate and, to a lesser extent, pyruvate served as amino group acceptors. This pattern suggests that this enzyme, previously designated as a lysine-6-aminotransferase, is actually an ω-aminotransferase. When 2-ketoadipate is used as substrate, the reaction product is 2-aminoadipic acid, which contributes to the pool of this intermediate available for penicillin biosynthesis. The N-terminal end of the purified 45-kDa ω-aminotransferase was sequenced and was found to be similar to the corresponding segment of the OAT1 protein of Emericella (Aspergillus) nidulans. This information was used to clone the gene encoding this enzyme.     

Determination of 1-aminocyclopropane-1-carboxylic acid (ACC) in leaf tissue and xylem sap using capillary column gas chromatography and a nitrogen/phosphorus detector

The Lizada and Yang method, commonly used for analyzing 1-aminocyclopropane-1-carboxylic acid (ACC), the immediate precursor of the plant hormone ethylene, is subject to interference and lacks internal standards. The use of combined gas chromatography-mass spectrometry (GC-MS) overcomes these shortcomings but the method is expensive and unavailable to many laboratories. We describe an alternative physico-chemical method using a capillary column gas chromatograph fitted with a standard nitrogen/phosphorus detector. After forming the N-benzoyl n-propyl derivative, measurements of ACC concentrations in extracts of leaves and in xylem sap of tomato plants using the nitrogen/phosphorus detector were within 10% of those obtained by GC-MS. Concentrations in plants grown in well-drained soil were approximately 0.16 nmol g^–1 fresh weight (leaves) and 0.04–0.01 mmol m^–3 (sap). Flooding the soil for 48–72 h increased these values approximately 9-fold.