Selection of efficient vesicular-arbuscular mycorrhizal fungi for wetland rice (Oryza sativa L.)

Summary We tested the response of the wetland rice cultivar Prakash to inoculation with ten vescular-arbucular mycorrhizal (VAM) fungi (three selected from the first screening and seven isolated from local paddy fields) in a pot experiment under flooded conditions in order to select the most efficient mycorrhizal fungi to inoculate the rice nursery. A sandy clay loam soil was used as the substrate, fertilized with the recommended N and K levels (100 kg N ha^–1 as ammonium sulphate and 50 kg K ha^–1 as muriate of potash) and half the recommended level of P (25 kg ha^–1 as super phosphate). The inoculation was made into dry nursery beds and the beds were flooded when the seedlings were about 25 cm high, in 15 days. Twenty-eight-day old seedlings were transferred to pots filled with well puddled soil flooded with 5 cm of standing water. Based on the increase in grain yield and total biomass, Glomus intraradices and Acaulospora sp. were considered efficient and suitable for inoculation into rice nurseries.     

Physiological enhancement of early growth of rice seedlings (Oryza sativa L.) by production of phytohormone of N2-fixing methylotrophic isolates

Three plant-growth promoting, N₂-fixing methylotrophic strains isolated from rice cultivars (Oryza sativa L.), viz, Methylobacterium sp. CBMB20, Enterobacter sp. CBMB30, Burkholderia sp. CBMB40, were selected, and their activities in promoting the early growth of rice were studied. Seeds treated with the methylotrophic strains improved seed germination, seedling vigor index (SVI), and biomass of rice seedlings. The methylotrophic population in the treated seedlings increased in the vegetative stages when compared to seeding stages. Treated seedlings showed a higher accumulation of plant hormones viz trans-zeatin riboside, isopentenyladenosine, and indole-3-acetic acid than untreated seedlings. Plant hormones were detected immunologically using the phytodetek kit. Conformational evidence suggested that cytokinins were produced by the epiphytic bacteria colonizing the plants rather than by the plants themselves. In addition, the inoculated early stage rice seedlings also exhibited a wide range of acetylene reduction activity. The results suggest the potential use of these bacteria to stimulate germination, SVI, and biomass production, which is mediated by production of plant hormone accumulation and nitrogen fixation.     

Differential colonization of Azospirillum lipoferum on roots of two varieties of rice (Oryza sativa L.)

Summary Seedlings of rice (IR42 and IR50) were aseptically dipped into Azospirillum lipoferum strain 34H suspension under dark, and the presence of bacteria on the differentiating regions of rice roots was observed by scanning electron microscopy. The bacterium did not colonize the root tips of IR42, while it colonized this region in the case of IR50, within 24 h after inoculation. In the early stages, most of the bacteria were embedded in the ruptured mucigel below the root cap cells of IR42. Mucigel was hardly detectable in IR50. While the root hair primordia of IR50 were colonized heavily with the bacterium within 24 h, the root hairs of IR42 were colonized 48 and 72 h after inoculation. This phenomenon in relation to plant varietal differences was discussed.     

Sources of nitrogen and yield advantages for monocropping and mixed cropping with cowpeas (Vigna unguiculata L.) and upland rice (Oryza sativa L.)

Summary Sources of N used by cropped and intercropped cowpeas and rice were determined by the ¹⁵N isotope-dilution technique. The biological efficiency of intercropping cowpeas and rice was assessed by calculating the land equivalent ratio of dry matter yield, total N, and uptake of N. A reduced N uptake by both cowpeas and rice during mixed cropping was attributed to mutual competition, with both crops competing effectively for the scarce environmental resources. The lack of a significant difference in the uptake of fertilizer and soil N by mixed-crop rice and cowpeas is an indication that the soil N was sufficient and that the mixed cropping did not create any imbalance in soil and fertilizer N uptake. The land equivalent ratio ranged between 120% and 180% for shoot dry matter and total N, showing that biological efficiency was increased by intercropping cowpeas with rice. The proportion of N derived from the atmosphere by mono- and intercropped cowpeas was not significantly different, showing that the potential of cowpeas to fix N₂ was independent of the cropping system, since the rice did not stimulate the cowpeas to fix more N₂.     

Relative significance of soil and nitrogenous fertilizer in nitrogen nutrition and growth of wetland rice (Oryza sativa L.)

Summary A pot experiment was conducted to compare the yields from five commercially cultivated varieties (Bas-198, Bas-370, Bas-Pak, Bas-385, and IR-6) of rice (Oryza sativa L.) and to establish the relative significance of soil N and fertilizer N (¹⁵N-labelled ammonium sulphate) in affecting crop performance. Another aim was to study the interaction of fertilizer N and soil N as influenced by different rice varieties. Among the five varieties tested, Bas-Pak gave the maximum dry matter and N yield. The N-use efficiency (percentage of applied N taken up by the plants) of different varieties ranged between 33.7 and 43.7%, Bas-Pak being the most efficient. Significant losses of fertilizer N occurred from the soil-plant system. The maximum N loss (52.1% of applied N) was observed with IR-6 and the minimum loss (39.2%) with Bas-Pak. A substantial increase in the uptake of soil N following the application of fertilizer and an interaction between the two N sources were observed with all varieties except Bas-385. The interaction was attributed to greater root proliferation following the application of fertilizer. It was concluded that a varietey with greater potential to use soil N is likely to give a better yield and that, of the two N sources, the availability of soil N was more important in determining the yield performance of different varieties of rice.     

The influence of inorganic nutrient fertilization on the growth,nutrient composition and vesicular-arbuscular mycorrhizal colonization of pretransplant rice (Oryza sativa L.) plants

Summary The effects of P, N and Ca+Mg fertilization on biomass production, leaf area, root length, vesiculararbuscular mycorrhizal (VAM) colonization, and shoot and root nutrient concentrations of pretransplant rice (Oryza sativa L.) plants were investigated. Mycorrhizal plants generally had a higher biomass and P, N, K, Ca, Mn, Fe, Cu, Na, B, Zn, Al, Mg, and S shoot-tissue nutrient concentrations than non-mycorrhizal plants. Although mycorrhizal plants always had higher root-tissue nutrient concentrations than non-mycorrhizal plants, they were not significantly different, except for Mn. N fertilization stimulated colonization of the root system (colonized root length), and increased biomass production and nutrient concentrations of mycorrhizal plants. Biomass increases due to N were larger when the plants were not fertilized with additional P. P fertilization reduced the colonized root length and biomass production of mycorrhizal plants. The base treatment (Ca+Mg) did not significantly affect biomass production but increased the colonized root length. These results stress the importance of evaluating the VAM rice symbiosis under various fertilization regimes. The results of this study suggest that pretransplant mycorrhizal rice plants may have a potential for better field establishment than non-mycorrhizal plants.     

Effect of methods of application on the efficiency of urea broadcast onto lowland rice (Oryza sativa L.)

Summary We evaluated the effect of different methods of application on the efficiency of urea broadcast at a rate of 100 kg N ha^-1 onto lowland rice (Oryza sativa L. var. SPR 60) in a field experiment conducted on a Phimai soil (Fluvic Tropaquepts) during the dry season of 1989. Analysis of the floodwater on the first day after the fertilizer application showed a high initial concentration of urea-N. Addition of the urease inhibitor N-(n-butyl) thiophosphoric triamide (nBTPT), broadcast with the urea into the floodwater, caused an apparent reduction in the rate of urea disappearance and a subsequent accumulation of NH₃–N in the floodwater; this ureas inhibitor also suppressed the rise in floodwater pH, with a resultant reduction in the partial pressure of ammonia (pNH₃) compared with the unamended urea application. The use of nBTPT did not decrease the N loss from broadcast urea not did it increase the grain yield. Among the different methods of applying broadcast urea that we tested, the broadcast application of granular urea onto drained soil shortly after removing floodwater followed by flooding 2 days later appeared to be a good N management practice, offering considerable potential for improving the efficiency of urea applied to lowland rice crops.     

Effects of inoculation with N2-fixing cyanobacteria on the nitrogenase activity in soil and rhizosphere of wetland rice (Oryza sativa L.)

Summary A greenhouse experiment was conducted with wetland rice (Oryza sativa cv. IR-50) in a clay-loam soil (Fluventic Eutrochrept) to study the effect of cyanobacterial inoculation a mixed culture of Aulosira fertilissima, Nostoc muscorum, Nostoc spp., and Anabaena spp., applied at the rate of 0.15 g (dry weight pot^-1 or 43 kg ha^-1) on acetylene reduction activity in soil and the root system (excised root), and the grain and straw yield. The effects of applying P (40 kg ha^-1), N (60 kg ha^-1), and P+N to the soil were also evaluated. Cyanbacterial inoculation significantly increased (more than 200% on average) photo-dependent acetylene reduction activity in soils, particularly where the indigenous activity was considerably low, i.e. under unfertilized and N-fertilized conditions. The effect of inoculation was prominent at the maximum tillering and grain formation stages of the crop. This inoculation benefit was, however, marginal in P-applied soils (P and P+N), where the indigenous activity was stimulated more than threefold. The inoculation led to a remarkable increase in root-associative acetylene reduction activity after the maximum tillering stage of the crop, particularly with applied N but for other treatments this inoculation effect was not significant. Cyanobacterial inoculation also increased the grain and straw yield of the crop when N was not applied. The grain and straw yield was significantly correlated with the acetylene reduction activity in flooded soils and in the root system during the tillering and maximum tillering stages of rice growth, respectively.     

Genetic Erosion from Modern Varieties into Traditional Upland Rice Cultivars (Oryza sativa L.) in Northern Thailand

The purpose of this study was to assess the extent of genetic erosion of traditional upland germplasm in northern Thailand as a result of gene-flow from distinct strains carrying different genotypes. Even modern variety specific markers have not been developed, there is a comparative population in Laos. Thus, both populations were compared with various characters to evaluate gene-flow from modern variety to landraces. Glutinous and glabrous strains are predominated in Laos. However, such strains were drastically decreased in north–east Thailand. Gene diversity is higher in Thailand, compared to Laos at seven isozyme loci. This was a result of the higher frequencies of Indica strains and heterozygotes in Thailand. Plastid type was also determined by using an INDEL marker. Nearly half of Indica strains carried the Japonica plastid. Heterozygotes also tended to carry Japonica cytoplasm. Such nuclear–cytoplasm substituted strains and heterozygotes were probably generated by natural hybridization. Japonica strains tended to be a maternal donor rather than Indica ones. Or Indica strains would easily release pollens, which grow outside of upland fields.     

Endophytic establishment of Azorhizobium caulinodans through auxin-induced root tumors of rice (Oryza sativa L.)

Rice seedlings developed nodule-like tumors (para-nodules) along primary and secondary roots when treated with the auxin 2,4-dichlor-phenoxy-acetic acid (2,4-D). Histologically, these tumors appeared as cancerous out-grown lateral-root primordes and were thus comparable with stem nodules of the legume Sesbania rostrata. Azorhizobium caulinodans (a diazotroph known as a specific endophyte of Sesbania rostrata) was introduced and became established inside rice para-nodules and in root tissues around tumor bases. The infection with A. caulinodans followed a typical crack-entry invasion at places where paranodule tumors had emerged through the root cortex and epidermis. The bacteria settled with high cell densities in intercellular spaces of the induced tumors and between root cortical cells. Infection of plant cells took place both in the epidermis and in cortical tissue. Intracellularly established A. caulinodans was found inside the cytoplasm, surrounded by membrane-like structures. N₂ fixation by tumor-inhabiting Azorhizobium sp. was increased at low O₂ tensions (1.5–3 kPa) compared with an untreated control. Only a little activity remained at O₂ tensions of 5 kPa and above. The present results confirm that root-tumor induction offers a suitable method of establishing diazotrophs endophytically in the roots of gramineous crops.     

Fate and interaction with native soil N of ammonium N applied to wetland rice (Oryza sativa L.) grown under saline and non-saline conditions

Summary The effect of salts on the balance of fertilizer N applied as ¹⁵N-labelled ammonium sulphate and its interaction with native soil N was studied in a pot experiment using rice (Oryza sativa L.) as a test crop. The rice crop used 26%–40% of the applied N, the level of applied N and salts showing no significant bearing on the uptake of fertilizer N. Losses of fertilizer N ranged between 54% and 68% and only 5%–8% of the N was immobilized in soil organic matter. Neither the salts nor the rate of N application had any significant effect on fertilizer N immobilization. The effective use of fertilizer N (fertilizer N in grain/fertilizer N in whole plant) was, however, better in the non-saline soil. The uptake of unlabelled N (N mineralized from soil organic matter and that originating from biological N₂ fixation in thes rhizosphere) was inhibited in the presence of the salts. However, in fertilized soil, the uptake of unlabelled N was significantly enhanced, leading to increased A values [(1-% Ndff/% Ndff)x N fertilizer applied, where Ndff is N derived from fertilizer], an index of interaction with the added N. This added N interaction increased with increasing levels of added N. Since the extra unlabelled N taken up by fertilized plants was greater than the fertilizer N immobilized, and the root biomass increased with increasing levels of added N, a greater part of the added N interaction was considered to be real, any contribution by an apparent N interaction (pool substitution or isotopic displacement) to the total calculated N interaction being fairly small. Under saline conditions, for the same level of fertilizer N addition, the added N interaction was lower, and this was attributed to a lower level of microbial activity, including mineralization of native soil N, rootdriven immobilization of applied N, and N₂ fixation.     

Fate of Azolla spp. and urea nitrogen applied to wetland rice (Oryza sativa L.)

Summary Using ¹⁵N, the fate of N applied to wetland rice either as Azolla or urea was studied in a field at the International Rice Research Institute (IRRI). In bigger plots nearby, yield response and N uptake were also determined with unlabelled N sources. Azolla microphylla was labelled by repeated application of labelled ammonium sulfate. Labelled and unlabelled N were used alternately in applications of Azolla or urea 0 and 42 days after transplanting, in order to determine the effect of the time of application on the availability of Azolla N. The quantities of Azolla N incorporated were 23% more than those of urea N (30 kg N ha^–1) in the isotope plots or 7% less in the yield response plots. Grain yield and total N uptake by the rice plants in the yield-response plots were higher in the urea-treated plots than in the Azolla-treated plots, but the physiological effect of Azolla N (grain yield response/increase in N uptake) was higher than that of rea. The labelled N balance was studied after the first and second crops of rice. Losses of labelled N after the first crop were higher from urea (30%–32%) than from Azolla (0%–11 %). Losses in N applied as a side dressing 42 days after transplanting were less than those of N applied basally. No further losses of ¹⁵N occurred after the first crop. The recovery of Azolla ¹⁵N in the first crop of rice was 39% from the basal application and 63% from the side dressing. The recovery of urea ¹⁵N was 27% from the basal application and 48% from the side dressing. Recoveries of residual N from both Azolla and urea during the second rice crop were similar. Laboratory incubation of the Azolla used and the changes in labelled exchangeable N in the soil showed that at least 65% of Azolla N (4.7% N content) was mineralized within 10 days.     

The fragrance (fgr) gene in natural populations of wild rice (Oryza rufipogon Griff.)

The Asian cultivated rice, Oryza sativa L. (spp. indica or japonica), is assumed to have originated from one or both of the two wild Asian species, O. rufipogon Griff. and O. nivara Sharma and Shastry. They occur throughout the monsoon Asia and west Oceania. Fragrance is the most important trait among the domesticated characters of basmati and jasmine rice of Asia. The gene for fragrance in a scented rice shows the presence of a mutated portion (i.e., an eight base pair deletion in exon 7) that result in its loss of fragrance. In the present study, 229 wild rice O. rufipogon accessions were genotyped for this locus using a PCR assay. The wild rice species contained the mutated allele of the fgr gene at a low frequencies of 0.23. The surveyed populations were in Hardy–Weinberg equilibrium. This observation supports the hypothesis that the allele for fragrance was already present in the wild rice, and that this trait appeared in scented rice cultivars because of selection by the farmers of genotypes possessing this character during the process of domestication.     

稻草覆盖还田对水稻氮素吸收和氮肥利用率的影响

以杂交稻“天优998”为材料, 设置覆盖还田和不还田2种稻草还田处理, 4种氮肥处理, 进行了连续2年的田间试验, 研究稻草还田对水稻氮素吸收和氮肥利用率的影响。结果表明: 稻草还田显著促进了水稻对氮素的吸收, 其总吸氮量比稻草不还田处理增加13.7%~20.3%, 但对不同生育阶段吸氮量占总吸氮量的比例影响不大。与稻草不还田处理相比, 稻草还田处理的水稻在分蘖中期(MT)、穗分化始期(PI)和抽穗期(HD)叶片叶绿素含量显著提高[实地养分管理(N1)的MT时期除外]。稻草还田可提高氮肥吸收利用率(RE)、农学利用率(AE)和氮肥偏生产力(PFP)。两年平均, 稻草还田处理的RE比稻草不还田处理提高8.23个百分点, 相对提高23.4%, 达极显著水平。AE、RE、PFP与MT~PI期间的吸氮量呈正相关, 其中AE和PFP达极显著水平。RE与HD~MA期间的吸氮量呈极显著正相关。在MT施氮、促进MT~PI期间的氮素吸收对提高AE、RE和PFP有重要意义。创造条件增加抽穗后的氮素吸收, 对于提高RE、减少氮肥损失和面源污染至关重要。     

水稻雄蕊导管发育研究

本文以水稻粤泰保持系为材料，采用ATPase定位方法研究了单核至三核花粉期雄蕊导管的发育。单核边位花粉期，分化的导管胞质变浓厚、细胞核染色质浓缩、核转变为新月形并出现凋亡小体；此外，导管内物质也可以大分子形式直接撤离。单核边位晚期，雄蕊导管发育成熟。在二核和三核花粉期，导管内重新出现具有ATPase活性的物质，同样的现象也存在于农垦58s可育花药成熟导管中。以上结果暗示水稻雄蕊导管分化存在程序性死亡及非程序性死亡多种途径，水稻成熟导管内具有ATPase活性物质可能是一种普遍现象。     

Yield and nitrogen response of lowland rice (Oryza sativa L.) to Sesbania rostrata and Aeschynomene afraspera green manure in different marginally productive soils in the Philippines

Organic-N fertilizers in the form of flood-tolerant, leguminous, stem-nodulating Sesbania rostrata and Aeschynomene afraspera may be useful alternatives to resource-poor rice farmers if applied as green manure. Therefore, the accumulation of N by these green manure species and their effect on the performance and yield of wetland rice (IR 64) was examined at four different sites in Luzon, Philippines. Soils deficient in N, P, and K were selected and compared with the fertile Maahas clay of the International Rice Research Institute (IRRI) at Los Baños. The green manure plants were grown under flooded conditions for 49 days in the wet season of 1987, chopped, and then ploughed in before transplanting rice seedlings. In a second experiment, the effect of S. rostrata green manure was studied under rainfed conditions. All green manure treatments were compared to an urea treatment (60 kg N ha^–1) and an untreated control. Both legumes developed well, even on the marginally productive soils. S. rostrata accumulated up to 190 kg N ha^–1 and A. afraspera even accumulated 196 kg N ha^–1 in the shoots. In all treatments, green manure increased grain yield significantly (P=0.05) over the untreated control, by 1.3–1.7 Mg ha^–1. The yields were comparable to those obtained with 60 kg N ha^–1 of urea fertilizer. S. rostrata caused the highest grain yield, of 6.5 Mg ha^–1 on the Maahas clay soil of IRRI. The apparent release of exchangeable NH ₄ ⁺ -N in the soils after green manuring and the rice grain yield response showed that both green manure species may provide sufficient available N throughout the development of IR 64 in the wet season. In the rainfed marginal soil site, green manure with S. rostrata produced even higher rice grain yields than urea. Green manure therefore seems particularly attractive for poor farmers on marginally productive soils, at least as a temporary strategy to improve yield and yield sustainability.     

大田条件下模拟UV-B辐射滤减对水稻生长及内源激素含量的影响

对大田栽培条件下人工模拟的UV-B辐射滤减环境对晚稻品种“协优432”的叶绿素含量、株高、内源激素吲哚乙酸(IAA)、脱落酸(ABA)、赤霉素(GA3)、玉米素(Zeatin)含量的影响研究结果表明,在正常日光处理组与UV-A滤光组之间叶绿素a含量变化显著;正常日光处理组与UV-A滤光组、UV-A UV-B滤光组之间株高无明显变化。UV-A UV-B滤光组IAA、GA3、Zeatin含量分别比正常日光处理组降低了40.42%、48.60%、36.21%;分别比UV-A滤光处理组降低了36.52%、70.94%、54.54%。单因素方差分析及多重比较结果显示,UV-A UV-B滤光组与正常日光处理组之间IAA、GA3含量差异显著;UV-A UV-B滤光组与UV-A滤光处理组之间GA3、Zeatin含量差异显著。UV-A UV-B滤光组ABA含量比正常日光处理组降低了29.74%,比UV-A滤光处理组降低了5.19%,经单因素方差分析后发现差异不显著。     

Effect of presubmergence and green manuring (Sesbania aculeata) on nutrition and yield of wetland rice (Oryza sativa L.) on a sodic soil

Summary A field experiment was conducted to evaluate the effect of 0, 1 and 2 weeks' submergence prior to the transplantation of rice (presubmergence), with and without Sesbania aculeata green manure, on the yield and nutrition of rice in a highly sodic soil [pH 10.3, exchangeable sodium percentage (ESP) 86.4]. Green manuring significantly enhanced the yield of rice and contributed 112 kg N/ha. One week's presubmergence with incorporated green manure improved the yield of rice significantly over that obtained with no previous flooding, giving a yield on a par with that obtained with 2 weeks' submergence. The beneficial effect of presubmergence without green manure on rice yield was of lesser magnitude and was significant only when it was continued for 2 weeks. Cultivation of rice under submerged conditions improved the sodic soil, and green manuring enhanced the process of reclamation by further decreasing the pH value and the exchangeable Na of soil. Green manuring considerably improved organic C, available N, P, and K status of the soil and enhanced the uptake of N, P, Ca, Mg, S, Fe, Mn, and Zn by the crop. Effects of submergence and green manuring on the availability of Fe, Mn, Zn, and P in soil and their role in the nutrition of rice are discussed.     

Dual inoculation of pretransplant stage Oryza sativa L. plants with indigenous vesicular-arbuscular mycorrhizal fungi and fluorescent Pseudomonas spp.

Summary This study examined the response of rice (Oryza sativa L.) plants at the pretransplant/nursery stage to inoculation with vesicular-arbuscular mycorrhizal (VAM) fungi and fluorescent Pseudomonas spp., singly or in combination. The VAM fungi and fluorescent Pseudomonas spp. were isolated from the rhizosphere of rice plants. In the plants grown in soil inoculated with fluorescent Pseudomonas spp. alone, I found increases in shoot growth, and in root length and fine roots, and decreases in root growth, and P and N concentrations. In contrast, in the plants colonized by VAM fungi alone, the results were the reverse of those of the pseudomonad treatment. Dual inoculation of soil with VAM fungi and fluorescent Pseudomonas spp. yielded plants with the highest biomass and nutrient acquisition. In contrast, the plants of the control treatment had the lowest biomass and nutrient levels. The dual-inoculated plants had intermediate root and specific root lengths. The precentages of mycorrhizal colonization and colonized root lengths were significantly lower in the dual-inoculated treatment than the VAM fungal treatment. Inoculation of plants with fluorescent Pseudomonas spp. suppressed VAM fungal colonization and apparently reduced photosynthate loss to the mycorrhizal associates, which led to greater biomass and nutrient levels in dual-inoculated plants compared with plants inoculated with VAM fungi alone. Dual inoculation of seedlings with fluorescent Pseudomonas spp. and VAM fungi may be preferable to inoculation with VAM alone and may contribute to the successful establishment of these plants in the field.     

Effect of green manuring,blue-green algae and neem-cake-coated urea on wetland rice (Oryza sativa L.)

Summary A field trial was set up to examine the effect of green manuring, blue-green algae, and neem-cake-coated urea on a rice crop. Summer green manuring using Sesbania aculeata increased the crop yield. Inoculation of blue-green algae increased the rice grain yield when 60 kg N ha^-1 was applied as prilled urea, but the increase in grain yield was greater when 60 kg N ha^-1 was applied as neem-cake-coated urea. The results of the present study show that applications of green manure, neem-cake-coated urea, and blue-green algae are complementary and that the three treatments can be used together in the rice ecosystem. The green manure and the fertilizer treatments had no effect on the algal flora of the soil.