The effect of cyanobacteria and azolla on the performance of rice under different levels of fertilizer nitrogen

Cyanobacteria and/or azolla were inoculated, with urea at 0, 72 or 144 kg N/ha, in plots in which azolla-free Indica rice var. IR 28 was grown. Productive tillers, yield and nitrogen contents of grain and straw positively responded to inoculation with cyanobacteria or azolla, even with fertilizer-N up to 144 kg N/ha. Inoculation improved colonization by cyanobacteria. Azolla were superior to the asymbiotic cyanobacteria in enhancing rice performance. Urea at a rate of 72 kg N/ha was found to support the best colonizations when applied with cyanobacteria or azolla or, to give maximum rice yields, both inoculants.     

Studies on the availability of Azolla N and urea N for rice growth using 15N

Field experiments (20 m² plots) were conducted to compare Azolla and urea as N sources for rice (Oryza sativa L.) in both the wet and dry seasons. Parallel microplot (1 m²) experiments were conducted using ¹⁵N. A total of approximately 60 kg N ha^-1 was applied as urea, Azolla, or urea plus Azolla. Urea or Azolla applied with equal applications of 30 kg N ha^-1 at transplanting (T) and at maximum tillering (MT) were equally effective for increasing rice grain yields in both seasons. Urea at 30 kg N ha^-1 at T and Azolla 30 kg N ha^-1 at MT was also equally effective. Urea applied by the locally recommended best split (40 kg at T and 20 kg at MT) gave a higher yield in the wet season, but an equal yield in the dry season. The average yield increase was 23% in the wet season, and 95% in the dry season. The proportion of the N taken up by the rice plants which was derived from urea (%NdfU) or Azolla (%NdfAz) was essentially identical for the treatments receiving the same N split. Recovery of ¹⁵N in the grain plus straw was also very similar. Positive yield responses to residual N were observed in the succeeding rice crop following both the wet and dry seasons, but the increases were not always statistically significant. Recovery of residual ¹⁵N ranged from 5.5 to 8.9% for both crops in succeeding seasons. Residual recovery from the urea applications was significantly higher than from Azolla in the crop succeeding the dry season crop. Azolla was equally effective as urea as an N source for rice production on a per kg N basis.     

Epiphytic cyanobacteria on Chara vulgaris are the main contributors to N(2) fixation in rice fields

The distribution of nitrogenase activity in the rice-soil system and the possible contribution of epiphytic cyanobacteria on rice plants and other macrophytes to this activity were studied in two locations in the rice fields of Valencia, Spain, in two consecutive crop seasons. The largest proportion of photodependent N(2) fixation was associated with the macrophyte Chara vulgaris in both years and at both locations. The nitrogen fixation rate associated with Chara always represented more than 45% of the global nitrogenase activity measured in the rice field. The estimated average N(2) fixation rate associated with Chara was 27.53 kg of N ha(-1) crop(-1). The mean estimated N(2) fixation rates for the other parts of the system for all sampling periods were as follows: soil, 4.07 kg of N ha(-1) crop(-1); submerged parts of rice plants, 3.93 kg of N ha(-1) crop(-1); and roots, 0.28 kg of N ha(-1) crop(-1). Micrographic studies revealed the presence of epiphytic cyanobacteria on the surface of Chara. Three-dimensional reconstructions by confocal scanning laser microscopy revealed no cyanobacterial cells inside the Chara structures. Quantification of epiphytic cyanobacteria by image analysis revealed that cyanobacteria were more abundant in nodes than in internodes (on average, cyanobacteria covered 8.4% +/- 4.4% and 6.2% +/- 5.0% of the surface area in the nodes and internodes, respectively). Epiphytic cyanobacteria were also quantified by using a fluorometer. This made it possible to discriminate which algal groups were the source of chlorophyll a. Chlorophyll a measurements confirmed that cyanobacteria were more abundant in nodes than in internodes (on average, the chlorophyll a concentrations were 17.2 +/- 28.0 and 4.0 +/- 3.8 microg mg [dry weight] of Chara(-1) in the nodes and internodes, respectively). These results indicate that this macrophyte, which is usually considered a weed in the context of rice cultivation, may help maintain soil N fertility in the rice field ecosystem.     

Evaluation of the water fern Azolla microphylla for mosquito population management in the rice-land agro-ecosystem of south India

The floating water fern Azolla microphylla Kaulfess was evaluated as a biocontrol agent against mosquitoes breeding in rice fields in Tamil Nadu, South India. Anopheles subpictus Grassi, Culex pseudovishnui Colless and Culex tritaeniorhynchus Giles were the predominant species of mosquitoes, with peak densities of late instar larvae and pupae occurring during the second week after transplantation of rice seedlings of short-term (c. 80 days from transplantation to harvest) or medium-term (c. 95 days) varieties. Immature mosquito populations were reduced by mats of Azolla microphylla covering more than 80% of the water surface. However, since 80% coverage by Azolla was achieved only 13-14 days after rice transplantation, its usefulness for mosquito control was limited. Azolla may have a greater potential in an integrated control programme, or in areas where long-term varieties of rice are predominantly grown.     

Efficiency of rice fertilization schedules including cyanobacteria under soil application of phosphate and molybdate

Productive tillering, grain and straw yields and N-contents of indica rice IR-28 were increased more by urea fertilizer at 48 or 96 kg N/ha along with Inoculation by cyanobacteria than by using 144 kg N/ha without inoculation. Fertilizer N-use efficiency was increased by the cyanobacteria and decreased with increasing amount of urea fertilizer. Further enhancement was obtained by soil application of calclum superphosphate at 36 kg P₂O₅/ha or sodium molybdate at 1 ppm Mo along with cyanobacteria and/or urea. Application of phosphate, however, slightly diminished the enhancement by molybdate.The author is with the Sakha Agricultural Research Station, Kafr El-Sheikh, Egypt.     

Genetic diversity among and within cultured cyanobionts of diverse species of <Emphasis Type="Italic">Azolla</Emphasis>

The cyanobionts isolated from 10 Azolla accessions belonging to 6 species (Azolla mexicana, A. microphylla, A. rubra, A. caroliniana, A. filiculoides, A. pinnata) were cultured under laboratory conditions and analyzed on the basis of whole cell protein profiles and molecular marker dataset generated using repeat sequence primers (STRR(mod) and HipTG). The biochemical and molecular marker profiles of the cyanobionts were compared with those of the free-living cyanobacteria and symbiotic Nostoc strains from Anthoceros sp., Cycas sp. and Gunnera monoika. Cluster analysis revealed the genetic diversity among the selected strains, and identified 3 distinct clusters. Group 1 included cyanobionts from all the 10 accessions of Azolla, group 2 comprised all the symbiotic Nostoc strains, while group 3 included the free-living cyanobacteria belonging to the genera Nostoc and Anabaena. The interrelationships among the Azolla cyanobionts were further revealed by principal component analysis. Cyanobionts from A. caroliniana-A. microphylla grouped together while cyanobionts associated with A. mexicana-A. filiculoides along with A. pinnata formed another group. A. rubra cyanobionts had intermediate relationship with both the subgroups. This is the first study analyzing the diversity existing among the cultured cyanobionts of diverse Azolla species through the use of biochemical and molecular profiles and also the genetic distinctness of these free-living cyanobionts as compared to cyanobacterial strains of the genera Anabaena and Nostoc.     

Influence of Azolla management on the growth,yield of rice and soil fertility

A field experiment conducted at Central Rice Research Institute, Cuttack, during three successive seasons showed that with the 120-day-duration variety Ratna two dual crops ofAzolla pinnata R. Brown (Bangkok isolate) could be achieved 25 and 50 days after transplanting (DAT) by inoculating 2.0 t ha⁻¹ of fresh Azolla 10 and 30 DAT respectively. One basal crop of Azolla could also be grown using the same inoculum 20 days before transplanting (DBT) in fallow rice fields. The three crops of Azolla grown—once before transplanting and twice after transplanting—gave an average total biomass of 38–63 and 43–64 t ha⁻¹ fresh Azolla containing 64–90 and 76–94 kg N ha⁻¹ respectively in the square and rectangular spacings. Two crops of Azolla grown only as a dual crop, on the other hand, gave 26–39 and 29–41 t ha⁻¹ fresh Azolla which contained 44–61 and 43–59 kg N ha⁻¹ respectively. Growth and yield of rice were significantly higher in Azolla basal plus Azolla dual twice incorporated treatments than in the Azolla dual twice incorporation, Azolla basal plus 30 kg N ha⁻¹ urea and 60 kg N ha⁻¹ urea treatments. Azolla basal plus 30 kg N ha⁻¹ urea and 60 kg N ha⁻¹ urea showed similar yields but Azolla dual twice incorporation was significantly lower than those. The different spacing with same plant populations did not affect growth and yield significantly, whereas Azolla growth during dual cropping was 8.3 and 64% more in the rectangular spacing than in the square spacing in Azolla basal plus Azolla dual twice incorporation and Azolla dual twice incorporation treatments.     

Biological N2 Fixation in wetland rice fields: Estimation and contribution to nitrogen balance

This paper 1) reviews improvements and new approaches in methodologies for estimating biological N₂ fixation (BNF) in wetland soils, 2) summarizes earlier quantitative estimates and recent data, and 3) discusses the contribution of BNF to N balance in wetland-rice culture.Measuring acetylene reducing activity (ARA) is still the most popular method for assessing BNF in rice fields. Recent studies confirm that ARA measurements present a number of problems that may render quantitative extrapolations questionable. On the other hand, few comparative measures show ARA's potential as a quantitative estimate. Methods for measuring photodependent and associative ARA in field studies have been standardized, and major progress has been made in sampling procedures. Standardized ARA measurements have shown significant differences in associative N₂ fixation among rice varieties.The ¹⁵N dilution method is suitable for measuring the percentage of N derived from the atmosphere (% Ndfa) in legumes and rice. In particular, the ¹⁵N dilution technique, using available soil N as control, appears to be a promising method for screening rice varieties for ability to utilize biologically fixed N. Attempts to adapt the ¹⁵N dilution method to aquatic N₂ fixers (Azolla and blue-green algae [BGA]) encountered difficulties due to the rapid change in ¹⁵N enrichment of the water.Differences in natural ¹⁵N abundance have been used to show differences among plant organs and species or varieties in rice and Azolla, and to estimate Ndfa by Azolla, but the method appears to be semi-quantitative.Recent pot experiments using stabilized ¹⁵N-labelled soil or balances in pots covered with black cloth indicate a contribution of 10–30 kg N ha^-1 crop^-1 by heterotrophic BNF in flooded planted soil with no or little N fertilizer used.Associative BNF extrapolated from ARA and ¹⁵N incorporation range from 1 to 7 kg N ha^-1 crop^-1. Straw application increases heterotrophic and photodependent BNF. Pot experiments show N gains of 2–4 mg N g^-1 straw added at 10 tons ha^-1.N₂ fixation by BGA has been almost exclusively estimated by ARA and biomass measurements. Estimates by ARA range from a few to 80 kg N ha^-1 crop^-1 (average 27 kg). Recent extensive measurements show extrapolated values of about 20 kg N ha^-1 crop^-1 in no-N plots, 8 kg in plots with broadcast urea, and 12 kg in plots with deep-placed urea.Most information on N₂ fixed by Azolla and legume green manure comes from N accumulation measurements and determination of % Ndfa. Recent trials in an international network show standing crops of Azolla averaging 30–40 kg N ha^-1 and the accumulation of 50–90 kg N ha^-1 for two crops of Azolla grown before and after transplanting rice. Estimates of % Ndfa in Azolla by ¹⁵N dilution and delta ¹⁵N methods range from 51 to 99%. Assuming 50–80% Ndfa in legume green manures, one crop can provide 50–100 kg N ha^-1 in 50 days. Few balance studies in microplots or pots report extrapolated N gains of 150–250 kg N ha^-1 crop^-1.N balances in long-term fertility experiments range from 19 to 98 kg N ha^-1 crop^-1 (average 50 kg N) in fields with no N fertilizer applied. The problems encountered with ARA and ¹⁵N methods have revived interest in N balance studies in pots. Balances are usually highest in flooded planted pots exposed to light and receiving no N fertilizer; extrapolated values range from 16 to 70 kg N ha^-1 crop^-1 (average 38 kg N). A compilation of balance experiments with rice soil shows an average balance of about 30 kg N ha^-1 crop^-1 in soils where no inorganic fertilizer N was applied.Biological N₂ fixation by individual systems can be estimated more or less accurately, but total BNF in a rice field has not yet been estimated by measuring simultaneously the activities of the various components in situ. As a result, it is not clear if the activities of the different N₂-fixing systems are independent or related. A method to estimate in situ the contribution of N₂ fixed to rice nutrition is still not available. Dynamics of BNF during the crop cycle is known for indigenous agents but the pattern of fixed N availability to rice is known only for a few green manure crops.     

Rice yields as influenced by Azolla N2 fixation and urea N-fertilization

Application of 0, 30, 60, 90 and 120 kg N ha^–1 of urea (U) in split doses with (and without)Azolla pinnata, R. Brown was studied for three consecutive seasons under planted field condition. Fresh weight (FW), acetylene reduction activity (ARA) and N yield of Azolla were found to be maximum 14 days after inoculation (DAI). Among the different treatments, maximum Azolla growth was recorded in no N control. The FW, ARA and N yield of Azolla were inhibited increasingly with the increase in N levels. Irrespective of season, FW and N yield of Azolla were inhibited only a small extent with 90 kg N ha^–1 U, beyond which the inhibition was pronounced. ARA was inhibited only slightly up to 60 kg N ha^–1 of U. Grain yield and crop N uptake of rice increased significantly up to 90 kg N ha^–1 of U (alone or in combination with Azolla) in the dry seasons (variety IR 36) and up to 60 kg N ha^–1 U in the wet season (variety CR 1018).     

Evaluation of the availability ofAzolla-N and urea-N to rice using15N

Summary The symbiotic association of the water fernAzolla with the blue-green algaAnabaena azollae can fix 30–60 kg N ha^–1 per rice cropping season. The value of this fixed N for rice production, however, is only realized once the N is released from theAzolla biomass and taken up by the rice plants. The availability of N applied asAzolla or as urea was measured in field experiments by two¹⁵N methods. In the first,Azolla caroliniana (Willd.) was labelled with¹⁵N in nutrient solution and incorporated into the soil at a rate of 144 kg N ha^–1. The recovery ofAzolla-N in the above ground parts of rice [Oryza sativa (L) cv. Nucleoryza] was found to be 32% vs. 26% for urea applied at a rate of 100 kg N/ha; there was no significant difference in recovery. In the second, 100 kg N/ha of¹⁵N-urea was applied separately or in combination with either 250 or 330 kg N ha^–1 of unlabelledAzolla. At the higher rate, the recovery ofAzolla-N was significantly greater than that of urea. There was a significant interaction when both N sources were applied together, which resulted in a greater recovery of N from each source in comparison to that source applied separately. Increasing the combined urea andAzolla application rate from 350 kg N ha^–1 to 430 kg N ha^–1 increased the N yield but had no effect on the dry matter yield of rice plants. The additional N taken up at the higher level of N application accumulated to a greater extent in the straw compared to the panicles. Since no assumptions need to be made about the contribution of soil N in the method using¹⁵N-labelledAzolla, this method is preferable to the¹⁵N dilution technique for assessing the availability ofAzolla-N to rice. Pot trials usingAzolla stored at –20°C or following oven-drying showed that both treatments decreased the recovery of N by one third in comparison to freshAzolla.     

Spatial distribution of methane-oxidizing activity in a flooded rice soil

In a study on spatial distribution of methane oxidation in an unplanted flooded field, methane-oxidizing activity, analysed in soil samples under laboratory conditions, decreased with increasing depth (25 cm and beyond). In a flooded field planted to rice, rates of methane oxidation followed the order : rhizosphere (collected from roots at 10-20 cm depth) > surface soil at (0-1 cm) > subsurface soil at 10-20 cm depth, diagonally 10-15 cm away from the centre of hill. Application of ammonium sulfate and, to a lesser extent, urea to surface, rhizosphere and subsurface soil samples from flooded field planted to rice effected a distinct inhibition of methane oxidation. Nitrification inhibitors (thiourea, sodium thiosulfate and dicyandiamide) were also effective in inhibiting methane oxidation. Both surface and rhizosphere soil samples harbored higher populations of methane-oxidizing bacteria than the subsurface soil. Inhibition of methane oxidation in surface and rhizosphere soil samples concomitant with the suppression of autotrophic ammonium oxidizers by nitrification inhibitors implicates an active involvement of autotrophic ammonium oxidizers in methane oxidation.     

无藻萍的RAPD分析及其在三膘组满江红种间关系研究中的应用

从满江红Azolla Lam.萍-藻共生体中提取DNA进行的RAPD系统分析通常忽视了满江红样品的异质性。本研究通过获得无藻的满江红,比较有藻萍、无藻萍和离体藻之间的RAPD指纹图谱。发现从有藻萍中提取DNA的扩增反应来源于萍藻双方DNA的共同影响。依引物和植物样本的不同,共生双方对扩增产物的贡献结果不同,说明了用无藻萍进行RAPD检测的重要性。对满江红三膘组5个种的11个无藻萍样本进行了RAPD分析,由9个引物产生的127个DNA多态片段用于计算样本间的Jaccard相似系数和UPGMA树状聚类图。结果     

Availability of nitrogen from 15N-labelled Azolla pinnata and urea to flooded rice

In view of the recently generated interest in Azolla and the high cost of N fertilizers, this field study was aimed at measuring the availability of Azolla-N applied in two split application in comparison to urea-N. Azolla was cultivated and labelled with ¹⁵N isotope in the field. A total of about 60 kg N ha^-1 was applied as Azolla, urea or Azolla and urea in combination, in two equal splits at transplanting and at maximum tillering, i.e. 30 days after transplanting (30 DAT).The recovery by the crop of Azolla-N applied at 30 DAT was significantly higher than that applied at transplanting, viz. 30.2% and 20.2%, respectively. The recoveries of urea-N applied at the same stages were similarly low, viz. 22.5% at transplanting and 38.6% at 30 DAT. Total recoveries of fertilizer N at the time of harvest were 26.8% from Azolla, 30.7% from urea applied in the same two splits and 49.1% from urea applied in locally recommended three splits. Recoveries of labelled Azolla-N in succeeding rice crop were twice higher than those of labelled urea-N. The recoveries ranged from 1.9 to 2.1% from urea-N and 4.0 to 4.9% from Azolla-N. There were no differences in residual ¹⁵N recovery in the succeeding crop between Azolla and urea either applied at transplanting or at 30 DAT.     

Effect of different nitrogen sources on growth,acetylene reduction activity ofAzolla pinnata and yield of rice

The effect of nitrogenous sources like ammonium sulphate (AS), prilled urea (U), urea super granule (USG) and farm yard manure (FYM) was studied on the fresh biomass (FB) and acetylene reduction activity (ARA) ofAzolla pinnata, R. Brown (Bangkok isolate), grown as a dual crop with rice, and rice yield in three successive seasons. Irrespective of the N-sources and seasons, the FB and ARA of Azolla were observed to be maximum on 14th day after Azolla inoculation (DAI). The different N-sources had significant effect on the ARA and to a lesser extent on the FB of Azolla. The treatment without fertilizer-N (control) exhibited highest ARA, FB and total N-content of Azolla. These were inhibited to a lesser extent by USG and FYM, though used at higher rates of 75 kg N ha⁻¹ and 90 kgN ha⁻¹ respectively, compared to that by AS and U, used at lower rates of 45kg N ha⁻¹ each.     

Molecular characterization and the effect of salinity on cyanobacterial diversity in the rice fields of Eastern Uttar Pradesh, India

Background

Salinity is known to affect almost half of the world's irrigated lands, especially rice fields. Furthermore, cyanobacteria, one of the critical inhabitants of rice fields have been characterized at molecular level from many different geographical locations. This study, for the first time, has examined the molecular diversity of cyanobacteria inhabiting Indian rice fields which experience various levels of salinity.

Results

Ten physicochemical parameters were analyzed for samples collected from twenty experimental sites. Electrical conductivity data were used to classify the soils and to investigate relationship between soil salinity and cyanobacterial diversity. The cyanobacterial communities were analyzed using semi-nested 16S rRNA gene PCR and denaturing gradient gel electrophoresis. Out of 51 DGGE bands selected for sequencing only 31 which showed difference in sequences were subjected to further analysis. BLAST analysis revealed highest similarity for twenty nine of the sequences with cyanobacteria, and the other two to plant plastids. Clusters obtained based on morphological and molecular attributes of cyanobacteria were correlated to soil salinity. Among six different clades, clades 1, 2, 4 and 6 contained cyanobacteria inhabiting normal or low saline (having EC < 4.0 ds m^-1) to (high) saline soils (having EC > 4.0 ds m^-1), however, clade 5 represented the cyanobacteria inhabiting only saline soils. Whilst, clade 3 contained cyanobacteria from normal soils. The presence of DGGE band corresponding to Aulosira strains were present in large number of soil indicating its wide distribution over a range of salinities, as were Nostoc, Anabaena, and Hapalosiphon although to a lesser extent in the sites studied.

Conclusion

Low salinity favored the presence of heterocystous cyanobacteria, while very high salinity mainly supported the growth of non-heterocystous genera. High nitrogen content in the low salt soils is proposed to be a result of reduced ammonia volatilization compared to the high salt soils. Although many environmental factors could potentially determine the microbial community present in these multidimensional ecosystems, changes in the diversity of cyanobacteria in rice fields was correlated to salinity.     

Identification of a common cyanobacterial symbiont associated with Azolla spp. through molecular and morphological characterization of free-living and symbiotic cyanobacteria.

Symbiotically associated cyanobacteria from Azolla mexicana and Azolla pinnata were isolated and cultured in a free-living state. Morphological analyses revealed differences between the free-living isolates and their symbiotic counterparts, as did restriction fragment length polymorphism (RFLP) analyses with both single-copy glnA and rbcS gene probes and a multicopy psbA gene probe. RFLP analyses with Anabaena sp. strain PCC 7120 nifD excision element probes, including an xisA gene probe, detected homologous sequences in DNA extracted from the free-living isolates. Sequences homologous to these probes were not detected in DNA from the symbiotically associated cyanobacteria. These analyses indicated that the isolates were not identical to the major cyanobacterial symbiont species residing in leaf cavities of Azolla spp. Nevertheless, striking similarities between several free-living isolates were observed. In every instance, the isolate from A. pinnata displayed banding patterns virtually identical to those of free-living cultures previously isolated from Azolla caroliniana and Azolla filiculoides. These results suggest the ubiquitous presence of a culturable minor cyanobacterial symbiont in at least three species of Azolla.     

Identification of a common cyanobacterial symbiont associated with Azolla spp. through molecular and morphological characterization of free-living and symbiotic cyanobacteria.

Symbiotically associated cyanobacteria from Azolla mexicana and Azolla pinnata were isolated and cultured in a free-living state. Morphological analyses revealed differences between the free-living isolates and their symbiotic counterparts, as did restriction fragment length polymorphism (RFLP) analyses with both single-copy glnA and rbcS gene probes and a multicopy psbA gene probe. RFLP analyses with Anabaena sp. strain PCC 7120 nifD excision element probes, including an xisA gene probe, detected homologous sequences in DNA extracted from the free-living isolates. Sequences homologous to these probes were not detected in DNA from the symbiotically associated cyanobacteria. These analyses indicated that the isolates were not identical to the major cyanobacterial symbiont species residing in leaf cavities of Azolla spp. Nevertheless, striking similarities between several free-living isolates were observed. In every instance, the isolate from A. pinnata displayed banding patterns virtually identical to those of free-living cultures previously isolated from Azolla caroliniana and Azolla filiculoides. These results suggest the ubiquitous presence of a culturable minor cyanobacterial symbiont in at least three species of Azolla.     

Improving nitrogen-fixing systems and integrating them into sustainable rice farming

This paper summarizes recent achievements in exploiting new biological nitrogen fixation (BNF) systems in rice fields, improving their management, and integrating them into rice farming systems. The inoculation of cyanobacteria has been long recommended, but its effect is erratic and unpredictable. Azolla has a long history of use as a green manure, but a number of biological constraints limited its use in tropical Asia. To overcome these constraints, the Azolla-Anabaena system as well as the growing methods were improved. Hybrids between A. microphylla and A. filiculoides (male) produced higher annual biomass than either parent. When Anabaena from high temperature-tolerant A. microphylla was transferred to Anabaena-free A. filiculoides, A. filiculoides became tolerant of high temperature. Azolla can have multiple purposes in addition to being a N source. An integrated Azolla-fish-rice system developed in Fujian, China, could increase farmers' income, reduce expenses, and increase ecological stability. A study using Azolla labeled with ¹⁵N showed the reduction of N losses by fish uptake of N. The Azolla mat could also reduce losses of urea N by lowering floodwater-pH and storing a part of applied N in Azolla. Agronomically useful aquatic legumes have been explored within Sesbania and Aeschynomene. S. rostrata can accumulate more than 100kg N ha^-1 in 45 d. Its N₂ fixation by stem nodules is more tolerant of mineral N than that by root nodules, but the flowering of S. rostrata is sensitive to photoperiod. Aquatic legumes can be used in rainfed rice fields as N scavengers and N₂ fixers. The general principle of integrated uses of BNF in rice-farming systems is shown.     

Effect of varying periods of inoculation ofAzolla pinnata R. Brown on its growth,nitrogen fixation and response to rice crop

Summary Inoculation of water fernAzolla pinnata R. Brown (Bangkok isolate) at the rate of 500kg fresh weight ha⁻¹ in rice fields at weekly intervals after planting in addition to 30 kg N ha⁻¹ as urea showed a decrease in its growth and N₂-fixation with delay in application. Use of Azolla up to 3 weeks after planting (WAP) during wet and 4 WAP during dry season produced significantly more grain yield than 30 kg N ha⁻¹, whereas its application upto one WAP produced more grain yield than 60 kg N ha⁻¹. Grain yield with Azolla applied at the time of planting was similar to that of 60 kg N treatment during the wet season. Higher grain yields in zero and one WAP Azolla treatments resulted due to increase in both number of panicles m⁻² and number of grains/panicle while the subsequent Azolla inoculations increased grain yield mainly by producing more number of grains/panicle. Dry matter and total N yields at maturity of rice crop were more with Azolla application upto 3 WAP during wet and 2 WAP during dry season while the reduction in sterility (%) was observed upto one WAP over 30 kg N ha⁻¹ during both seasons. Number of tillers m⁻² and dry matter production at maximum tillering and flowering were more than 30 kg N ha⁻¹ with the use of Azolla upto one WAP. Increased grain N yield was observed with the use of Azolla upto 4 WAP during two seasons whereas straw N yield increased upto one WAP during wet and 2 WAP during dry season.     

Effect of rates of nitrogen and relative efficiency of sulphur-coated urea and nitrapyrin-treated urea in dry matter production and nitrogen uptake by rice

Summary A field experiment conducted for two rainy seasons (1974 and 1975) on a sandy clay loam soil at the Indian Agricultural Research Institute, New Delhi showed that at 100kg N/ha the apparent recovery of urea nitrogen by the rice crop was only 28%, which was raised to 41.7% by treating urea with Nitrapyrin and to 47.4% by coating urea withneem (Azadirachta indica Juss) cake. The recovery with sulphur-coated urea was 37.7%. Dry matter production nitrogen concentration in plant and uptake by rice were increased as the rate of nitrogen was increased from 0 to 150kg N/ha. Advantage of treating urea with Nitrapyrin or coating withneem cake was seen more in grain than straw yield.