拟南芥AtCIPK23基因对烟草抗旱能力的影响

为了探索拟南芥AtCIPK23基因对烟草耐旱能力的影响,对3个转AtCIPK23基因阳性纯合株系KA13、KA14和KA44与野生型烟草K326（对照）进行了自然干旱处理,测定离体叶片的失水速率、叶绿素含量、相对电导率、脯氨酸和可溶性糖含量,并分析了转基因及野生型材料对活性氧的清除能力,对活性氧清除基因NtSOD、NtCAT和NtAPX及干旱胁迫相关基因NtDREB、NtLEA5和NtCDPK2的表达量进行检测。结果表明：（1）转基因烟草离体叶片的失水速率明显低于K326;自然干旱7 d后,野生型K326出现了明显的干旱胁迫症状;干旱7 d进行复水后,转基因株系的复水存活率明显高于K326。（2）转基因株系中的叶绿素、脯氨酸及可溶性糖含量比K326显著提高,电导率则明显降低。（3）野生型烟草K326中H₂O₂的积累量明显高于3个转基因株系,转基因株系中ROS清除机制的3个关键基因NtSOD、NtCAT和NtAPX被诱导上调表达。（4）抗旱相关基因NtDREB、NtLEA5和NtCDPK2仅在转基因烟草中受干旱诱导。研究认为,AtCIPK23基因可能具有提高植物抗旱能力的功能。     

Fluxes of greenhouse gases from Andosols under coffee in monoculture or shaded by <Emphasis Type="Italic">Inga densiflora</Emphasis> in Costa Rica

The objective of this study was to evaluate the effect of N fertilization and the presence of N₂ fixing leguminous trees on soil fluxes of greenhouse gases. For a one year period, we measured soil fluxes of nitrous oxide (N₂O), carbon dioxide (CO₂) and methane (CH₄), related soil parameters (temperature, water-filled pore space, mineral nitrogen content, N mineralization potential) and litterfall in two highly fertilized (250 kg N ha⁻¹ year⁻¹) coffee cultivation: a monoculture (CM) and a culture shaded by the N₂ fixing legume species Inga densiflora (CIn). Nitrogen fertilizer addition significantly influenced N₂O emissions with 84% of the annual N₂O emitted during the post fertilization periods, and temporarily increased soil respiration and decreased CH₄ uptakes. The higher annual N₂O emissions from the shaded plantation (5.8 ± 0.3 kg N ha⁻¹ year⁻¹) when compared to that from the monoculture (4.3 ± 0.1 kg N ha⁻¹ year⁻¹) was related to the higher N input through litterfall (246 ± 16 kg N ha⁻¹ year⁻¹) and higher potential soil N mineralization rate (3.7 ± 0.2 mg N kg⁻¹ d.w. d⁻¹) in the shaded cultivation when compared to the monoculture (153 ± 6.8 kg N ha⁻¹ year⁻¹ and 2.2 ± 0.2 mg N kg⁻¹ d.w. d⁻¹). This confirms that the presence of N₂ fixing shade trees can increase N₂O emissions. Annual CO₂ and CH₄ fluxes of both systems were similar (8.4 ± 2.6 and 7.5 ± 2.3 t C-CO₂ ha⁻¹ year⁻¹, −1.1 ± 1.5 and 3.3 ± 1.1 kg C-CH₄ ha⁻¹ year⁻¹, respectively in the CIn and CM plantations) but, unexpectedly increased during the dry season.     

Stability of transgene expression, field performance and recombination breeding of transformed barley lines

Stable inheritance of the transgene, consistent expression and competitive agronomic properties of transgenic crops are important parameters for successful use of the latter. These properties have been analyzed with 18 homozygous transgenic barley lines of the cultivar Golden Promise. The lines originated from three independent primary transformants obtained by the biolistic method with three plasmids containing respectively, the bar gene, the uidA gene and the gene for a protein-engineered heat-stable (1,3–1,4)-β-glucanase. Three production levels of recombinant β-glucanase were identified in homozygous transgenic T₃ plants, and these remained constant over a 3-year period. In micro-malting experiments, the heat-stable enzyme reached levels of up to 1.4 μg·mg⁻¹ protein and survived kiln drying at levels of 70–100%. In the field trials of 1997 and 1998 the transgenic lines had a reduced 1000-grain weight as well as variable yield depressions compared to the Golden Promise progenitor. In 1999 large-scale propagations of the lines with the highest recombinant enzyme synthesis during germination and of Golden Promise were studied at three different locations. In an irrigated field transgenic lines yielded approximately 6 t·ha⁻¹ and Golden Promise 7.7 t·ha⁻¹. Cross-breeding was carried out to transfer the transgene into a more suitable genetic background. Crosses of the semi-dwarf ari-e mutant Golden Promise gave rise to the four morphological phenotypes nutans, high erect, erect, and ari-e. Two improvements were achieved: (1) F₃ lines homozygous for the expression of heat-stable (1,3−1,4)-β-glucanase were found among lines that were homozygous for each of the four morphological phenotypes; (2) improved 1000-grain weights and yields with respect to those of the original transformants were observed in some F₄ lines homozygous for the morphological phenotypes and for the transgene. In the case of a homozygous nutans line, the transgenic plants had a higher 1000-grain weight than those lacking the transgene. Like mutants providing useful output traits, transgenic plants will often have to be improved by relocating the gene into more suitable genotypes. Received: 6 March 2000 / Accepted: 14 April 2000     

Overexpression of farnesyl pyrophosphate synthase (<Emphasis Type="Italic">FPS</Emphasis>) gene affected artemisinin content and growth of <Emphasis Type="Italic">Artemisia annua</Emphasis> L.

Transgenic plants of Artemisia annua L., a medicinal plant that produces the compound artemisinin which has an anti-malarial activity, were developed following Agrobacterium tumefaciens-mediated transformation of leaf explants. A. tumefaciens strain EHA105 carrying either pCAMBIA1301 or pCAMBIAFPS was used. Both plasmids harbored the hygromycin phosphotransferase II (hptII) gene as a selectable gene, but the latter plasmid also harbored the gene encoding for farnesyl pyrophosphate synthase (FPS), a key enzyme for artemisinin biosynthesis. Shoot regeneration was observed either directly from leaf sections or via intervening callus when explants were incubated on solidified Murashige and Skoog (MS) (1962) medium containing 0.1 mg l⁻¹ α-naphthaleneacetic acid (NAA), 1 mg l⁻¹ N⁶-benzyladenine (BA), 30 mg l⁻¹ meropenem and 10 mg l⁻¹ hygromycin. Applying vacuum infiltration dramatically increased transformation efficiency up to 7.3 and 19.7% when plasmids with and without FPS gene were used, respectively. All putative transgenic regenerants showed positive bands of hptII gene following Southern blot analysis. Expression of FPS was observed in all transgenic lines, and FPS over-expressed lines exhibited higher artemisinin content and yield, of 2.5- and 3.6-fold, respectively, than that detected in wild-type plants. A relatively high correlation (R ² = 0.78) was observed between level of expression of FPS and artemisinin content. However, gene silencing was detected in some transgenic lines, especially for those lines containing two copies of the FPS transgene, and with some lines exhibiting reduced growth.     

Seasonal accumulation and partitioning of nitrogen by lentil

Although wheat (Triticum aestivum L.) is the dominant crop of the semi-arid plains of Canada and the western United States, lentil (Lens culinaris Medik.) has become an important alternative crop. Sources and seasonal accumulation of N must be understood in order to identify parameters that can lead to increased N₂-fixing activity and yield. Inoculated lentil was grown in a sandy-loam soil at an irrigated site in Saskatchewan, Canada. Wheat was used as the reference crop to estimate N₂ fixation by the A-value approach. Lentil and wheat received 10 and 100 kg N ha⁻¹ of ammonium nitrate, respectively. Crops were harvested six times during the growing season and plant components analyzed. During the first 71 days after planting the wheat had a higher daily dry matter and N accumulation compared to lentil. However, during the latter part of the growing season, daily dry matter and N accumulation were greater for lentil. The maximum total N accumulation for lentil at maturity was 149 kg ha⁻¹. In contrast, wheat had a maximum N accumulation of 98 kg ha⁻¹ in the Feekes 11.1 stage, or 86 days after planting. The maximum daily rates of N accumulation were 3.82 kg N ha⁻¹ day⁻¹ for lentil and 2.21 kg N ha⁻¹ day⁻¹ for wheat. The percentage of N derived from N₂ fixation (% Ndfa) ranged from 0 at the first harvest to 92 % at final harvest. Generative plant components had higher values for % Ndfa than the vegetative components which indicates that N in the reproductive plant parts was derived largely from current N₂ fixation and lentil continued to fix N until the end of the pod fill stage. At final harvest, lentil had derived 129 kg N ha⁻¹ from N₂ fixation with maximum N₂-fixing activity (4.4 kg N ha⁻¹ day⁻¹) occurring during the early stages of pod fill. Higher maximum rates of N₂-fixing activity than net N accumulation (3.82 kg N ha⁻¹ day⁻¹) may have been caused by N losses like volatilization. In addition, lentil provided a net N contribution to the soil of 59 kg ha⁻¹ following the removal of the grain.     

Soil-Atmosphere Exchange of N<Subscript>2</Subscript>O and NO in Near-Natural Savanna and Agricultural Land in Burkina Faso (W. Africa)

In a combined field and laboratory study in the southwest of Burkina Faso, we quantified soil-atmosphere N₂O and NO exchange. N₂O emissions were measured during two field campaigns throughout the growing seasons 2005 and 2006 at five different experimental sites, that is, a natural savanna site and four agricultural sites planted with sorghum (n = 2), cotton and peanut. The agricultural fields were not irrigated and not fertilized. Although N₂O exchange mostly fluctuated between −2 and 8 μg N₂O–N m⁻² h⁻¹, peak N₂O emissions of 10–35 μg N₂O–N m⁻² h⁻¹ during the second half of June 2005, and up to 150 μg N₂O–N m⁻² h⁻¹ at the onset of the rainy season 2006, were observed at the native savanna site, whereas the effect of the first rain event on N₂O emissions at the crop sites was low or even not detectable. Additionally, a fertilizer experiment was conducted at a sorghum field that was divided into three plots receiving different amounts of N fertilizer (plot A: 140 kg N ha⁻¹; plot B: 52.5 kg N ha⁻¹; plot C: control). During the first 3 weeks after fertilization, only a minor increase in N₂O emissions at the two fertilized plots was detected. After 24 days, however, N₂O emission rates increased exponentially at plot A up to a mean of 80 μg N₂O–N m⁻² h⁻¹, whereas daily mean values at plot B reached only 19 μg N₂O–N m⁻² h⁻¹, whereas N₂O flux rates at plot C remained unchanged. The calculated annual N₂O emission of the nature reserve site amounted to 0.52 kg N₂O–N ha⁻¹ a⁻¹ in 2005 and to 0.67 kg N₂O–N ha⁻¹ a⁻¹ in 2006, whereas the calculated average annual N₂O release of the crop sites was only 0.19 kg N₂O–N ha⁻¹ a⁻¹ and 0.20 kg N₂O–N ha⁻¹ a⁻¹ in 2005 and 2006, respectively. In a laboratory study, potential N₂O and NO formation under different soil moisture regimes were determined. Single wetting of dry soil to medium soil water content with subsequent drying caused the highest increase in N₂O and NO emissions with maximum fluxes occurring 1 day after wetting. The stimulating effect lasted for 3–4 days. A weaker stimulation of N₂O and NO fluxes was detected during daily wetting of soil to medium water content, whereas no significant stimulating effect of single or daily wetting to high soil water content (>67% WHC_max) was observed. This study demonstrates that the impact of land-use change in West African savanna on N trace gas emissions is smaller—with the caveat that there could have been potentially higher N₂O and NO emissions during the initial conversion—than the effect of timing and distribution of rainfall and of the likely increase in nitrogen fertilization in the future.     

A laboratory study on feeding plasticity of the shredder <Emphasis Type="Italic">Sericostoma vittatum</Emphasis> Rambur (Sericostomatidae)

Since litter input and availability of leaves in many streams is highly seasonal in Portugal, we investigated whether Sericostoma vittatum, a typical shredder, was able to grow using alternative food sources. To test this hypothesis we fed S. vittatum with Alnus glutinosa (alder, CPOM, coarse particulate organic matter), leaf powder from A. glutinosa and Acacia dealbata and FPOM (fine particulate organic matter) from a 5th and a >6th order river, the macrophyte Myriophyllum aquaticum and biofilm. Growth in S. vittatum was significantly influenced by the food item given (ANOVA, P = 0.0082). The food item promoting the highest growth was A. glutinosa, in the form of FPOM (6.48% day⁻¹) and CPOM (4.24% day⁻¹); all other forms of FPOM and biofilm provided relatively low growth rates (0.77–1.77% day⁻¹). The macrophyte M. aquaticum was also used as food source by S. vittatum and promoted intermediate growth (1.96% day⁻¹). Neither nitrogen, phosphorus nor caloric content was correlated with growth. However, since higher growth was achieved with alder, in the form of CPOM and FPOM, we concluded that the chemical content of food was more important for S. vittatum than the physical form of such food. This may partially explain why shredders are able to survive when leaves are scarce in streams. Handling editor: K. Martens     

Microbial N Turnover and N-Oxide (N<Subscript>2</Subscript>O/NO/NO<Subscript>2</Subscript>) Fluxes in Semi-arid Grassland of Inner Mongolia

Gross rates of N mineralization and nitrification, and soil–atmosphere fluxes of N₂O, NO and NO₂ were measured at differently grazed and ungrazed steppe grassland sites in the Xilin river catchment, Inner Mongolia, P. R. China, during the 2004 and 2005 growing season. The experimental sites were a plot ungrazed since 1979 (UG79), a plot ungrazed since 1999 (UG99), a plot moderately grazed in winter (WG), and an overgrazed plot (OG), all in close vicinity to each other. Gross rates of N mineralization and nitrification determined at in situ soil moisture and soil temperature conditions were in a range of 0.5–4.1 mg N kg⁻¹ soil dry weight day⁻¹. In 2005, gross N turnover rates were significantly higher at the UG79 plot than at the UG99 plot, which in turn had significantly higher gross N turnover rates than the WG and OG plots. The WG and the OG plot were not significantly different in gross ammonification and in gross nitrification rates. Site differences in SOC content, bulk density and texture could explain only less than 15% of the observed site differences in gross N turnover rates. N₂O and NO _x flux rates were very low during both growing seasons. No significant differences in N trace gas fluxes were found between plots. Mean values of N₂O fluxes varied between 0.39 and 1.60 μg N₂O-N m⁻² h⁻¹, equivalent to 0.03–0.14 kg N₂O-N ha⁻¹ y⁻¹, and were considerably lower than previously reported for the same region. NO _x flux rates ranged between 0.16 and 0.48 μg NO _x -N m⁻² h⁻¹, equivalent to 0.01–0.04 kg NO _x -N ha⁻¹ y⁻¹, respectively. N₂O fluxes were significantly correlated with soil temperature and soil moisture. The correlations, however, explained only less than 20% of the flux variance.     

Nutrient losses during a winter low-intensity prescribed fire in a Mediterranean forest

Losses of N, P, K and Ca were measured during a prescribed burning in a French MediterraneanPinus halepensis forest, with understorey ofQuercus coccifera. Nutrient loss was measured by difference between the quantity of a nutrient in the fuel before burning and that found in the postfire remains which were harvested or recovered in small trays. Reduction in fuel weight amounted to 77%, losses from initial fuel elements amounted to 77% for N, 54% for K and 35% for P. No significative loss of Ca was measured. Burning resulted in 7 t ha⁻¹ fuel reduction and in loss of 55 kg ha⁻¹N, 8.5 kg ha⁻¹K and 1.0 kg ha⁻¹P.     

Seasonal variation in the nitrogenase activity of aPanicum maximum var.trichoglume pasture and identification of associated bacteria

Summary The influence of seasonal variation on nitrogenase (N₂-ase) activity of undisturbed soil-plant cores ofPanicum maximum var.trichoglume was measured using the C₂H₂ reduction assay. The largest N₂-ase activity in the field, 14.7 g N ha⁻¹ day⁻¹, occurred in spring when soil moisture was high, soil temperature was low and nitrogenous fertiliser influence was at a minimum. The potential N₂-ase activity of the cores, measured under controlled conditions, reached a maximum of 27.2 g N ha⁻¹ day⁻¹ and averaged 26.3 g N ha⁻¹ day⁻¹ over the 14 month sampling period. N₂-ase activity was positively correlated (P=0.05) with field soil moisture and negatively correlated with field soil temperature (r=0.59 and −0.78 respectively). Multiple regression showed that 69% of the variation of N₂-ase activity in the field was associated with the combined effects of soil moisture and soil temperature. Nitrogen fixing bacteria were isolated from the roots ofP. maximum and based upon morphology, biochemical tests and fluorescent antibody reaction, were found to be closely related toAzospirillum lipoferum.     

Effect of rhizobia and rock biofertilizers with <Emphasis Type="Italic">Acidithiobacillus</Emphasis> on cowpea nodulation and nutrients uptake in a tableland soil

Chemical fertilizers have been used in the cultivation of plants due to their high solubility and effect on crops yield. Biofertilizers with phosphate rock (PR) and potash rock (KR) plus sulfur inoculated with Acidithiobacillus may improve plant growth and contribute to addition of available P and K in soil. The effectiveness of biofertilizers from phosphate and potash rocks mixed with sulfur and Acidithiobacillus was studied in a Typic Fragiuldult soil of the Brazilian Northeast Tableland. Cowpea (cv. “IPA 206”) was grown with and without rhizobia inoculation. Treatments were: (a) phosphate rock (1000 kg ha⁻¹); (b) Biofertilizers-B_P (250 and 500 kg ha⁻¹); (c) triple superphosphate-TSP (250 kg ha⁻¹); (d) potash rock (1000 kg ha⁻¹); (e) biofertilizer-B_K (250; 500 and 750 kg ha⁻¹); (f) potassium chloride-KCl (250 kg K₂0 ha⁻¹); (g) control without P or K fertilization (P₀K₀). The soil was maintained under water submersion covered with black plastic (solarization process) for a period of 30 days. Biofertilizers (B_p and B_K) and soluble fertilizers increased plant growth and NPK uptake. Biofertilizers reduced soil pH, especially when applied in highest rates. Biofertilizers and TSP+KCl showed the best values of available P and K in soil. Rhizobial inoculation was effective on cowpea, but no nodules were formed by bacteria native from the soil, probably due to the effect of the solarization process. From obtained PK biofertilizers could be used as alternative for cowpea fertilization in Tableland soils.     

Growth and NPK uptake of high-yielding cotton grown at different nitrogen levels in a permanent-plot experiment

Growth and N, P, K uptake of Acala SJ-2 cotton (Gossypium hirsutum) were investigated in an irrigated permanent-plot field (Typic chromoxerert) at Bet Dagan, Israel, under semi-arid conditions using different nitrogen levels: 0, 60, 120, 180 and 240 kg N ha⁻¹. The total dry matter accumulation at these levels was 9.0, 10.7, 15.1, 17.1 and 15.6 ton ha⁻¹, respectively. The uptake of N, P and K was 110, 144, 267, 322 and 301 kg N ha⁻¹∶31, 34, 46, 44 and 38 kg P ha⁻¹; and 120, 151, 208, 251 and 230 kg K ha⁻¹, respectively. Dry matter production, as well as N, P, K uptake by the cotton plants were greatly increased by raising the N application levels to 120 or 180 kg N ha⁻¹, but the pattern of accumulation and relative distribution of dry matter and NPK among plant organs were not considerably affected. Joint contribution from the Dept. of Soil Chemistry and Plant Nutrition, ARO, the Volcani Center, Bet Dagan, Israel (No. 1413-E, 1985 series)     

Isolation of the maize <Emphasis Type="Italic">Zpu1</Emphasis> gene promoter and its functional analysis in transgenic tobacco plants

By screening a genomic library of maize, a 2.2 kb 5′ flanking fragment of Zpu1 gene, encoding the pullulanase-type starch debranching enzyme, was isolated. Promoter fragments of various lengths, including the full 5′ flanking sequence (−2267 to −1) (Z1), a 3′ deletion (−2267 to −513) (Z5) and three 5′ deletions extending to −1943 (Z2), −1143 (Z3) and −516 (Z4) upstream of the translational initiation codon (ATG), were fused to the GUS reporter gene and introduced into tobacco. When these constructs were tested in transgenic tobacco plants, seed-preferred GUS activity was observed in pZ1-transgenic lines. In pZ2-transgenic lines, the GUS activity was not only restricted to seeds, but was also detected in calyxes, petals, stamens and mature leaves. At the same time, negligible GUS activity was detected in roots, stems, young leaves, stigmas and ovaries from the transgenic tobacco plants, which had integrated the full isolated sequence of Zpu1 promoter or its deletions. Deletion analysis indicated that the promoter contained a putative positive cis-regulatory element and the proximal region (−516 to −1) was essential for directing the expression of gus reporter gene. Analysis of GUS activity during the fruit development and seed germination suggested that Zpu1 promoter is active both in starch anabolism and in starch catabolism, which is consistent with the function of the endogenous gene in maize. GUS activity in leaves under light and darkness confirmed that Zpu1 promoter functions in the starch degradation of photosynthetic tissues in the dark phase of the diurnal cycle.     

Improvement of growth,under field conditions,of wheat inoculated with <Emphasis Type="Italic">Pseudomonas chlororaphis</Emphasis> subsp.<Emphasis Type="Italic"> aurantiaca</Emphasis> SR1

Effects of inoculation of wheat (Triticum aestivum L.) with the rhizobacterium Pseudomonas chlororaphis subsp. aurantiaca strain SR1 (termed SR1) were studied at an experimental field site in Río Cuarto, Argentina. Treatments involved SR1 inoculation with or without nitrogen/phosphorus fertilization. Inoculation produced a significant increase in plant height and root length in early growth stages. Inoculation plus fertilization with 40 kg ha⁻¹ urea/30 kg ha⁻¹ diamonic phosphate (“50% dose”) gave a yield increase of 636 kg ha⁻¹ relative to control, and an increase of 472 kg ha⁻¹ relative to fertilization with 80 kg ha⁻¹ urea/60 kg ha⁻¹ phosphate without inoculation. SR1 inoculation without fertilization, compared to control, produced increases of 6% in weight of 1,000 grains, 13% in number of spikes per plant, and 30% in number of grains per spike. Inoculation plus 50% dose fertilization also improved these parameters. Results of the study indicate that inoculation of wheat with SR1 improves various growth and yield parameters, and allows reduced dosage of nitrogen/phosphorus fertilizers in the field.     

Tillage and Nitrogen Application Effects on Nitrous and Nitric Oxide Emissions from Irrigated Corn Fields

A 2-year study was conducted to investigate the potential of no-till cropping systems to reduce N₂O and NO emissions under different N application rates in an irrigated corn field in northeastern Colorado. Flux measurements were begun in the spring of 2003, using vented (N₂O) and dynamic (NO) chambers, one to three times per week, year round, within plots that were cropped continuously to corn (Zea mays L.) under conventional-till (CT) and no-till (NT). Plots were fertilized at planting in late April with rates of 0, 134 and 224 kg N ha⁻¹ and corn was harvested in late October or early November each year. N₂O and NO fluxes increased linearly with N application rate in both years. Compared with CT, NT did not significantly affect the emission of N₂O but resulted in much lower emission of NO. In 2003 and 2004 corn growing seasons, the increase in N₂O-N emitted per kg ha⁻¹ of fertilizer N added was 14.5 and 4.1 g ha⁻¹ for CT, and 11.2 and 5.5 g ha⁻¹ for NT, respectively. However, the increase in NO-N emitted per kg ha⁻¹ of fertilizer N added was only 3.6 and 7.4 g ha⁻¹ for CT and 1.6 and 2.0 g ha⁻¹ for NT in 2003 and 2004, respectively. In the fallow season (November 2003 to April 2004), much greater N₂O (2.0–3.1 times) and NO (13.1–16.8 times) were emitted from CT than from NT although previous N application did not show obvious carry-over effect on both gas emissions. Results from this study reveal that NT has potential to reduce NO emission without an obvious change in N₂O emission under continuous irrigated corn cropping compared to CT.     

Total nitrogen and phosphorus balance sheet in a typic ustochrept soil under intensive cropping and fertilizer use

Summary Balance sheets were computed for total nitrogen and phosphorus in plough layer (0–15 cm) of a Typic Ustochrept soil under continuous multiple cropping for seven years (1971–72 to 1977–78) with a fixed rotation of pearl millet (Pennisetum typhoideum L.) wheat (Triticum aestivum L.) (Vigna sinensis Savi.) The treatments considered of soil test-based rates of N, P and K, applied both singly and in combinations together with farm yard manure, sulphur and zinc superimposed over optimum rates (100%) of NPK. Heavy, losses of N (762–899 kg ha⁻¹) occurred in the plots which received high rates of Nviz. 150% of recommended NPK and 100% NPK plus FYM. Application of N alone accelerated N losses whereas addition of P, PK, PKS to N minimised such losses. Enrichment of P (66 to 198 kg ha⁻¹) occurred in all phosphate-treated plots. A marginal net decrease (29–54 kg ha⁻¹) in P levels was observed in control and N alone treatments.     

Root growth,macro-nutrient uptake dynamics and soil fertility requirements of a high-yielding winter oilseed rape crop

Shoot growth, root growth and macro-nutrient uptake by a high-yielding (5t/ha grain) winter oilseed rape crop have been measured. Maximum rooting density in the top 20cm of soil was 9.4 cm cm⁻³ and roots reached a depth of at least 1.8 m. Maximum nutrient uptakes were 364 kg ha⁻¹ for N, 43 kg ha⁻¹ for P, 308 kg ha⁻¹ for K, 287 kg ha⁻¹ for Ca and 16 kg ha⁻¹ for Mg. A 30-day drought coincided with the flowering period and root and shoot growth, as well as nutrient uptake rates, were reduced. Nutrient concentrations in the soil solution necessary to sustain the nutrient fluxes into the root system by diffusive supply have been calculated. Peak values were in the range 10 μM for P to 87 μM for N, lower than the observed concentrations, and it was concluded that nutrient transport to roots was not a limitation to uptake by this rape crop.     

Application of Arabidopsis AGAMOUS second intron for the engineered ablation of flower development in transgenic tobacco

To explore a new approach to generating reproductive sterility in transgenic plants, the barnase gene from Bacillus amyloliquefaciens was placed under the control of an 1853-bp nucleotide sequence from the 3′end of the second intron of Arabidopsis AGAMOUS and CaMV 35S (−60) minimal promoter [AG-I-35S (−60)::Barnase], and was introduced into tobacco through transformation mediated by Agrobacterium tumefaciens. All AG-I-35S (−60)::Barnase transgenic plants showed normal vegetative growth and 28% of the transgenic lines displayed complete ablation of flowering. Two transgenic lines, Bar-5 and Bar-15, were 98.1 and 98.4% sterile, respectively, as determined by seed production and germination. When controlled by AG-I-35S (−60) chimeric promoter, barnase mRNA was detected in the reproductive tissues of transgenic tobacco plants, but not in vegetative parts. This study presents the first application of an AG intron sequence in the engineered ablation of sexual reproduction in plants. The AG-I-35S (−60)::Barnase construct can be useful in diminishing pollen and seed formation in plants, providing a novel bisexual sterility strategy for interception of transgene escape and has other potentially commercial use for transgenic engineering.     

Biomass and aboveground net primary production in a subtropical mangrove stand of Kandelia obovata (S., L.) Yong at Manko Wetland, Okinawa, Japan

Biomass and aboveground net primary production (ANPP) in a monospecific pioneer stand of a mangrove Kandelia obovata (S., L.) Yong were quantified. The estimated biomasses in leaves, branches, stems, roots, aboveground and total were 5.61 (3.68%), 28.8 (18.9%), 46.1 (30.2%), 71.8 (47.2%), 80.5 (52.8%) and 152 Mg ha⁻¹ (100%), respectively. Stem phytomass increment per tree was estimated using allometric relationships and stem analysis. Stem volume without bark of harvested trees showed a strong allometric relationship with D _0.1² H (D _0.1, diameter at a height of one-tenth of tree height H) (R ² = 0.924). Annual stem volume increment per tree showed a strong allometric relationship with D _0.1² H (R ² = 0.860). Litterfall rate ranges from 3.87 to 56.1 kg ha⁻¹ day⁻¹ for leaves and 0.177 to 46.2 kg ha⁻¹ day⁻¹ for branches. Seasonal changes of litterfall rate were observed, which showed a peak during wet season (August–September). Total annual litterfall was estimated as 10.6 Mg ha⁻¹ year⁻¹, in which 68.2% was contributed by the leaves. The ANPP in the K. obovata stand was 29.9–32.1 Mg ha⁻¹ year⁻¹, which is ca. 2.8–3.0 times of annual litterfall. The growth efficiency (aboveground biomass increment/LAI) was 5.35–5.98 Mg ha⁻¹ year⁻¹. The low leaf longevity (9.3 months) and high growth efficiency of K. obovata makes it a highly productive mangrove species.     

Identification of the double genome donor in spontaneous triploid tomato plants by RFLP analysis

Independent spontaneous triploid tomato plants (Lycopersicon esculentum Mill.) were collected among diploid hybrids growing in commercial greenhouses. Ploidy levels were verified by counting chromosomes, and the donor of the double genome dose was determined by restriction fragment length polymorphism (RFLP) analysis. The TG101 probe, which is tightly linked to the Tm-2 ^a locus, revealed different restriction patterns between TMV-resistant and TMV-susceptible parent lines. The parent donor which provided two genomes to the triploid was identified by comparing the relative intensity of alleles in the triploid with that in the diploid. The results indicate that both parents can serve as a double genome donor.