水稻25S核糖体RNA2′—O—核糖甲基化位点的测定

核糖 2′ O 甲基化修饰是真核生物核糖体RNA上的一种极为普遍的修饰方式。为了测定水稻 2 5S核糖体RNA上发生甲基化修饰的具体位点 ,设计并纯化了一系列与水稻 2 5S和酵母 2 8S核糖体RNA均配对的引物 ,在测定水稻核糖体RNA甲基化位点的同时 ,将酵母核糖体RNA甲基化位点的测定作为对照 ,在同一条件下 ,分别以水稻及酵母总RNA为模板进行dNTP浓度依赖的引物延伸反应。在测得的水稻甲基化位点中 ,有 3 1个位点是与酵母共有的 ,占酵母 2 8S核糖体RNA的甲基化位点总数的 80 %以上。另外 ,通过与已经测定的拟南芥 2 5S核糖体RNA上的甲基化位点进行比较 ,在水稻中又确定了与拟南芥相同的 5 4个甲基化位点。最终在水稻 2 5S核糖体RNA中 ,初步确定了 85个甲基化位点 ,并绘制了水稻 2 5S核糖体RNA的甲基化位点分布图。这些结果表明在不同的真核生物中 ,核糖体RNA上大部分位点核糖的甲基化修饰是保守的 ,而且亲缘关系越近 ,其保守性越强。结果还表明 ,高等植物核糖体RNA上有大量的核糖甲基化修饰位点 ,并且其中相邻的位点均被甲基化修饰的数量明显高于其他生物。所测得的甲基化位点将为进一步寻找植物中新的C/D框小分子核仁RNA(sonRNA)提供重要的依据     

Larval habitat segregation between the molecular forms of the mosquito Anopheles gambiae in a rice field area of Burkina Faso, West Africa

In West Africa, lineage splitting between the M and S molecular forms of the major Afro-tropical malaria mosquito, Anopheles gambiae (Diptera: Culicidae), is thought to be driven by ecological divergence, occurring mainly at the larval stage. Here, we present evidence for habitat segregation between the two molecular forms in and around irrigated rice fields located within the humid savannahs of western Burkina Faso. Longitudinal sampling of adult mosquitoes emerging from a range of breeding sites distributed along a transect extending from the heart of the rice field area into the surrounding savannah was conducted from June to November 2009. Analysis revealed that the two molecular forms and their sibling species Anopheles arabiensis are not randomly distributed in the area. A major ecological gradient was extracted in relation to the perimeter of the rice fields. The M form was associated with larger breeding sites mostly consisting of rice paddies, whereas the S form and An. arabiensis were found to depend upon temporary, rain-filled breeding sites. These results support hypotheses about larval habitat segregation and confirm the suggestion that the forms have different larval habitat requirements. Segregation appears to be clearly linked to anthropogenic permanent habitats and the community structure they support.     

Molecular and cytological characterization of 5S rDNA in Oryza species: genomic organization and phylogenetic implications.

We investigated the molecular characteristics and chromosomal organization of 5S rDNA in the genus Oryza, including diploid and tetraploid species. A phylogenetic tree of Oryza species was constructed based on the non-transcribed spacer sequences of 5S rDNA, and some novel relationships were discovered. Specifically, comparative sequence analysis of 5S rDNA in several wild rice species showed unique characteristics inconsistent with the model of concerted evolution: (1) multiple distinct 5S rDNA types were detected within a species, leading to intraspecific divergence of 5S rDNA; (2) multiple identical 5S rDNA types were shared among species, resulting in interspecies clustering of 5S rDNA types; and (3) intraspecific nucleotide diversity was detected within a 5S rDNA class. Our results obtained by fluorescence in situ hybridization revealed that each rice species studied contained only one 5S rDNA locus with two hybridization sites, which were located on either chromosome 7 or chromosome 11. These results suggest that different 5S rDNA classes within the rice genome were arranged together and that one pair of 5S rDNA loci from a diploid progenitor of the tetraploid species might have been lost during evolution. Taken together, our data show that 5S rDNA in rice species is more informative at the gene level than at the chromosome level.     

水稻品种和施肥时间对单季稻田中白背飞虱卵被寄生率的影响

为明确水稻品种和施肥时间对白背飞虱Sogatella furcifera (Horvath)卵被稻飞虱缨小蜂Anagrus spp.寄生率的影响,采用产卵苗诱集法和田间双因子设计进行实验研究。与秀水11水稻品种相比,协优413品种上的白背飞虱卵块位置较高且卵块较大。诱集实验表明白背飞虱卵的被寄生率显著受用于诱集的产卵水稻品种的影响, 在协优413上所产卵的被寄生率（24%）显著高于秀水11上的相应值（12%）,而不受由田间品种和氮肥施用时间组合而形成的诱集苗所置放的栖境条件的影响。田间种植的水稻品种对其稻株上的白背飞虱卵被寄生率的影响大于施肥时间, 在卵高峰期,这种影响更为显著, 协优413上的被寄生率（17%）显著高于秀水11上的相应值（9%）。田间寄生率和诱集寄生率二者的反正弦平方根转换值间存在着极显著的直线相关。这些结果表明,水稻品种是加强白背飞虱自然生物防治的重要因子。     

Detection ofBurkholderia pseudomallei in rice fields with PCR-based technique

Burkholderia pseudomallei Ara- in rice fields was detected using PCR-based techniques with 16S RNA and flagella gene primer sets. The sensitivity of these PCRs was at least 1 CFU/mL of B. pseudomallei Ara- preincubated into Ashdown's medium for 6 h. B. pseudomallei Ara- DNA from watery soil were more detectable than from dry soil. The distribution of this DNA was mainly found at a depth of 300-600 mm under crop-covered fields, but not detected in the location of soil close to the land surface. The results suggest that PCR based on 16S RNA and flagella gene primer sets can be applied to investigate the presence of B. pseudomallei Ara- in contaminated soil of rice fields.     

Reduced Dependence of Crested Ibis on Winter-Flooded Rice Fields: Implications for Their Conservation

The Crested Ibis Nipponia nippon was once thought to be extinct in the wild until seven birds were discovered in a remote mountain village in China in 1981. Studies suggested that winter-flooded rice fields play an essential role in nest site selection by the Crested Ibis and hence in their survival. Considerable efforts were therefore made to conserve the winter-flooded rice fields, but these have caused conflicts between the agricultural and conservation communities. The population and geographical range of the wild Crested Ibis has expanded greatly since 1981, but there is no spatial information on the winter-flooded rice fields, nor on the current association of nest sites and winter-flooded rice fields. We mapped winter-flooded rice fields across the entire current range of Crested Ibis using innovative remote sensing and geographical information systems (GIS) techniques. The spatial relationships between the nest site clusters and winter-flooded rice fields were quantified using Ward''s hierarchical clustering method and Ripley''s K-function. We show that both have significantly clumped distribution patterns and that they are positively associated. However, the dependence of Crested Ibis on the winter-flooded rice fields varied significantly among the nest site clusters and has decreased over the years, indicating the absence of winter-flooded rice fields is not constraining their recovery and population expansion. We therefore recommend that efforts should be made to protect the existing winter-flooded rice fields and to restore the functionality of natural and semi-natural wetlands, to encourage both in-situ conservation and the re-introduction of the Crested Ibis. In addition, we recommend that caution should be exercised when interpreting the habitat requirements of species with a narrow distribution, particularly when that interpretation is based only on their current habitat.     

Compensation of rodent pests after removal: control of two rat species in an irrigated farming system in the Red River Delta, Vietnam

Rodent pests have a strong capacity to recover rapidly from imposed reductions in abundance, but it is unclear how populations compensate to removal when farmers apply rodent control. The response of two rat species to rodent control was monitored using regular live-trapping in an irrigated lowland mixed rice agroecosystem. Rice field rats, Rattus argentiventer (52% of rodent captures), and lesser rice field rats, Rattus losea (29%), were removed using trap-barrier systems (TBS) at two sites while rice crops were present. The TBS was a plastic fence that enclosed a small field planted 3 weeks prior to the surrounding fields and set with multiple-capture cage-traps to capture rats that were attracted to the early-planted crop. Demographic responses of rats were compared to two untreated sites. There was a reduction in abundance of rodents on treated sites relative to untreated sites during the summer rice crop (by 45% and 28% for R. argentiventer and R. losea, respectively) an increase in abundance during the winter season (31% and 69%), and a mixed response during the spring rice crop (39% decrease and 41% increase). There was an increase in the proportion of juveniles captured on treated sites relative to untreated sites post-treatment (148% and 158%) and the body mass was lower on treated sites post-treatment (males: 13% and 41%; females: 22% and 22%). Older, larger animals were removed by the TBS and the rodent populations were compensating through high recruitment of young and high immigration into treated sites. No clear patterns were observed in the occurrence of adult breeding females for either R. argentiventer or R. losea. Rodent management, should therefore, occur over large areas (> 100 ha) to reduce the chance of reinvasion.     

Compensation of rodent pests after removal: control of two rat species in an irrigated farming system in the Red River Delta,Vietnam

Rodent pests have a strong capacity to recover rapidly from imposed reductions in abundance, but it is unclear how populations compensate to removal when farmers apply rodent control. The response of two rat species to rodent control was monitored using regular live-trapping in an irrigated lowland mixed rice agroecosystem. Rice field rats, Rattus argentiventer (52% of rodent captures), and lesser rice field rats, Rattus losea (29%), were removed using trap-barrier systems (TBS) at two sites while rice crops were present. The TBS was a plastic fence that enclosed a small field planted 3 weeks prior to the surrounding fields and set with multiple-capture cage-traps to capture rats that were attracted to the early-planted crop. Demographic responses of rats were compared to two untreated sites. There was a reduction in abundance of rodents on treated sites relative to untreated sites during the summer rice crop (by 45% and 28% for R. argentiventer and R. losea, respectively) an increase in abundance during the winter season (31% and 69%), and a mixed response during the spring rice crop (39% decrease and 41% increase). There was an increase in the proportion of juveniles captured on treated sites relative to untreated sites post-treatment (148% and 158%) and the body mass was lower on treated sites post-treatment (males: 13% and 41%; females: 22% and 22%). Older, larger animals were removed by the TBS and the rodent populations were compensating through high recruitment of young and high immigration into treated sites. No clear patterns were observed in the occurrence of adult breeding females for either R. argentiventer or R. losea. Rodent management, should therefore, occur over large areas (> 100 ha) to reduce the chance of reinvasion.     

DNA uptake by imbibition and expression of a foreign gene in rice

Uptake of DNA by imbibition of dry and viable rice ( Oryza sativa L.) embryos from a DNA solution and expression of a foreign gene were detected using two different vectors contaíning gusA (β-glucuronidase) and hpt (hygromycin phosphotransferase) as reporter genes. The frequency of transient expression of gusA and hpt genes using the CaMV35S promoter was about 30 to 50%. The main sites of gusA gene expression were meristems of roots and vascular bundles of leaves. Also, DNA uptake, integration and expression of the hpt gene in selected rice were investigated by various PCR methods and Southern blot analysis of genomic DNA. It was shown that the hygromycin phosphotransferase (HPT) DNA was present in the rice genome in an integrated form and not as a plasmid form.     

Vertical distribution of the methanotrophic community after drainage of rice field soil

Anoxic soils, such as flooded rice fields, are major sources of the greenhouse gas CH(4) while oxic upland soils are major sinks of atmospheric CH(4). Nevertheless, CH(4) is also consumed in rice fields where up to 90% of the produced CH(4) is oxidized in a narrow oxic zone around the rice roots and in the soil surface layer before it escapes into the atmosphere. After 1 day drainage of rice field soil, CH(4) oxidation was detected in the top 2-mm soil layers, but after 8 days drainage the zone of CH(4) oxidation extended to 8 mm depth. Simultaneously, the potential for CH(4) production decreased, but some production was still detectable after 8 days drainage throughout the soil profile. The vertical distribution of the methanotrophic community was also monitored after 1 and 8 days drainage using denaturing gradient gel electrophoresis after PCR amplification with primer sets targeting two regions on the 16S rRNA gene that are relatively specific for methylotrophic alpha- and gamma-Proteobacteria, and targeting two functional genes encoding subunits of key enzymes in all methanotrophs, i.e. the genes for the particulate methane monooxygenase (pmoA) and the methanol dehydrogenase (mxaF). Drainage stimulated the methanotrophic community. Eight days after drainage, new methanotrophic populations appeared and a distinct methanotrophic community developed. The population structure of type I and II methanotrophs was differently affected by drainage. Type II methanotrophs (alpha-Proteobacteria) were present throughout the soil core directly after drainage (1 day), and the community composition remained largely unchanged with depth. Only two new type II populations appeared after 8 days of drainage. Drainage had a more pronounced impact on the type I methanotrophic community (gamma-Proteobacteria). Type I populations were not or only weakly detected 1 day after drainage. However, after 8 days of drainage, a large diversity of type I methanotrophs were detected, altough they were not evenly distributed throughout the soil core but dominated at different depths. A distinct type I community structure had developed within each soil section between 0 and 20 mm soil depth, indicating the widening of suitable habitats for methanotrophs in the rice field soil within 1 week of drainage.     

Binding Site Concentration Explains the Differential Susceptibility of Chilo suppressalis and Sesamia inferens to Cry1A-Producing Rice

Chilo suppressalis and Sesamia inferens are two important lepidopteran rice pests that occur concurrently during outbreaks in paddy fields in the main rice-growing areas of China. Previous and current field tests demonstrate that the transgenic rice line Huahui 1 (HH1) producing a Cry1Ab-Cry1Ac hybrid toxin from the bacterium Bacillus thuringiensis reduces egg and larval densities of C. suppressalis but not of S. inferens. This differential susceptibility to HH1 rice correlates with the reduced susceptibility to Cry1Ab and Cry1Ac toxins in S. inferens larvae compared to C. suppressalis larvae. The goal of this study was to identify the mechanism responsible for this differential susceptibility. In saturation binding assays, both Cry1Ab and Cry1Ac toxins bound with high affinity and in a saturable manner to midgut brush border membrane vesicles (BBMV) from C. suppressalis and S. inferens larvae. While binding affinities were similar, a dramatically lower concentration of Cry1A toxin binding sites was detected for S. inferens BBMV than for C. suppressalis BBMV. In contrast, no significant differences between species were detected for Cry1Ca toxin binding to BBMV. Ligand blotting detected BBMV proteins binding Cry1Ac or Cry1Ca toxins, some of them unique to C. suppressalis or S. inferens. These data support that reduced Cry1A binding site concentration is associated with a lower susceptibility to Cry1A toxins and HH1 rice in S. inferens larvae than in C. suppressalis larvae. Moreover, our data support Cry1Ca as a candidate for pyramiding efforts with Cry1A-producing rice to extend the activity range and durability of this technology against rice stem borers.     

Trans-activation and stable integration of the maize transposable element Ds cotransfected with the Ac transposase gene in transgenic rice plants

To develop an efficient gene tagging system in rice, a plasmid was constructed carrying a non-autonomous maize Ds element in the untranslated leader sequence of a hygromycin B resistance gene fused with the 35S promoter of cauliflower mosaic virus. This plasmid was cotransfected by electroporation into rice protoplasts together with a plasmid containing the maize Ac transposase gene transcribed from the 35S promoter. Five lines of evidence obtained from the analyses of hygromycin B-resistant calli, regenerated plants and their progeny showed that the introduced Ds was trans-activated by the Ac transposase gene in rice. (1) Cotransfection of the two plasmids is necessary for generation of hygromycin B resistant transformants. (2) Ds excision sites are detected by Southern blot hybridization. (3) Characteristic sequence alterations are found at Ds excision sites. (4) Newly integrated Ds is detected in the rice genome. (5) Generation of 8 by target duplications is observed at the Ds integration sites on the rice chromosomes. Our results also show that Ds can be trans-activated by the transiently expressed Ac transposase at early stages of protoplast culture and integrated stably into the rice genome, while the cotransfected Ac transposase gene is not integrated. Segregation data from such a transgenic rice plant carrying no Ac transposase gene showed that four Ds copies were stably integrated into three different chromosomes, one of which also contained the functional hph gene restored by Ds excision. The results indicate that a dispersed distribution of Ds throughout genomes not bearing the active Ac transposase gene can be achieved by simultaneous transfection with Ds and the Ac transposase gene.     

Trans-activation and stable integration of the maize transposable element Ds cotransfected with the Ac transposase gene in transgenic rice plants

To develop an efficient gene tagging system in rice, a plasmid was constructed carrying a non-autonomous maize Ds element in the untranslated leader sequence of a hygromycin B resistance gene fused with the 35S promoter of cauliflower mosaic virus. This plasmid was cotransfected by electroporation into rice protoplasts together with a plasmid containing the maize Ac transposase gene transcribed from the 35S promoter. Five lines of evidence obtained from the analyses of hygromycin B-resistant calli, regenerated plants and their progeny showed that the introduced Ds was trans-activated by the Ac transposase gene in rice. (1) Cotransfection of the two plasmids is necessary for generation of hygromycin B resistant transformants. (2) Ds excision sites are detected by Southern blot hybridization. (3) Characteristic sequence alterations are found at Ds excision sites. (4) Newly integrated Ds is detected in the rice genome. (5) Generation of 8 by target duplications is observed at the Ds integration sites on the rice chromosomes. Our results also show that Ds can be trans-activated by the transiently expressed Ac transposase at early stages of protoplast culture and integrated stably into the rice genome, while the cotransfected Ac transposase gene is not integrated. Segregation data from such a transgenic rice plant carrying no Ac transposase gene showed that four Ds copies were stably integrated into three different chromosomes, one of which also contained the functional hph gene restored by Ds excision. The results indicate that a dispersed distribution of Ds throughout genomes not bearing the active Ac transposase gene can be achieved by simultaneous transfection with Ds and the Ac transposase gene.     

利用rBE3和rBE4系统进行水稻基因单碱基编辑的特异性和遗传稳定性分析

对基于rBE3(Rice base editor)和rBE4碱基编辑系统创制获得Os SERK1(D428N)和pi-ta(S918F)等基因的单碱基编辑突变体材料进行编辑特异性和遗传稳定性分析,旨在全面了解和更好地利用该碱基编辑系统。首先对Os SERK1(D428N)和pi-ta(S918F)sg RNA的潜在脱靶位点进行预测,并对T0代材料中的各脱靶位点进行PCR扩增和Sanger测序检测;同时对该两个基因的突变体材料自交获得的T1代植株的靶位点序列和外源T-DNA分离进行检测。结果显示各T0代材料均未检测到潜在脱靶位点发生单碱基编辑;此外,Os SERK1(D428N)和Os08g07774含有相同的sg RNA位点,且两个位点均能发生单碱基编辑;rBE3或rBE4系统介导产生的碱基编辑可稳定遗传至T1代,并在T1代可获得无外源T-DNA的纯合突变体。上述结果表明由rBE3或rBE4介导的碱基编辑具有较高的特异性,可进行多位点编辑,引入的碱基替换可稳定遗传至后代。     

Feeding frequency and survival of Anopheles gambiae in a rice-growing area in Ghana

Mortality rates, determined by dissection, of predominantly M form female Anopheles gambiae (Diptera: Culicidae) were estimated. Mosquitoes were collected in tent traps and light traps in an irrigation project village in Ghana in June and July 2010, when much of the area was flooded. Both M and S form larvae were collected from rice fields (74 of 80 specimens were M form). Adults were collected in equal proportions from the two traps (90 of 107 specimens from the light trap and 106 of 116 specimens from the tent trap were M form). During the study, collection numbers rose from 105 to 972 per night. A total of 1787 of the 15 431 An. gambiae collected were dissected. Of these, 953 (53%) were found to have taken their first bloodmeal, either as virgins or following mating. The age profiles of mosquitoes collected alive and dead, respectively, were similar. Eighteen of 2933 (0.61 ± 0.49%) specimens were found to be positive for sporozoites in an enzyme-linked immunosorbent assay (ELISA). Lagged cross correlations among the different age groups implied that the mosquitoes fed on days 2 and 4 following emergence prior to oviposition and every 2.65 ± 0.17 days thereafter. The best model to describe the observed population patterns implied a daily mortality of 84%. The results are discussed in relation to possible mosquito control measures for the village.     

Evidence and population diversity of the rice yellow mottle virus (RYMV) insect vectors and some of their natural antagonists in paddy fields in the north of Cameroon

From 2002 to 2004 two sampling methods were combined to assess the population of insects known as vectors of rice yellow mottle virus (RYMV) in the three major irrigated rice ecosystems in the north of Cameroon (Lagdo, Maga, Yagoua) and in lowland rice fields. Samplings were conducted by means of sweep net and D-Vac (the sucking trap) fortnightly in 2002 and 2003 until mid-October in lowland rice fields while in the irrigated fields, samplings occurred between December and April. Simultaneously, rearing on death concerned insects was conducted in the laboratory to identify the parasitoid insect species. From samples obtained at the different sites: (1) the dominant structure of the RYMV insect vectors was analysed according to the rice phenology; (2) the diversity and the occurrence of potential major groups of predators and parasitoids were assessed. Among the RYMV insect vectors sampled: Chaetocnema pulla Chapuis (Coleoptera: Chrysomelidae), Chnootriba similis Mulsant (Coleoptera: Coccinellidae), Trichispa sericea Guerin-Meneville (Coleoptera: Chrysomelidae), Locris rubra Fabricius (Hemiptera: Cicadellidae), Oxya hyla Stål (Orthoptera: Acrididae) and Conocephalus longipennis (de Haan) (Orthoptera: Tettigoniidae) were the most encountered insect species during the rice growing seasons. When investigating the arthropods acting in the rice fields as predators, spiders (Araneae) came at the top position with a high concurrency of Pardosa spp. (>42%) at all sites. At the sites Maga and Yagoua, the carabid beetles species Abacetus crenulatus Dejean and Abacetus foveolatus Chaudoir were the most numerous whereas the Lagdo site was highly colonised by Clivina erythropyga Putzeys, Paederus sabaeus Erichson and Stenus ravus Puthz were the most abundant staphylinid beetles. The rearing of dead RYMV insect vectors gave results on the emergence of the following parasitoids: Eurytoma spp., Pediobius spp., Tetrastichus spp., Telenomus spp.     

Frequency and pattern of transposition of the maize transposable element Ds in transgenic rice plants

Two kinds of T-DNA constructs, I-RS/dAc-I-RS and Hm(R)Ds, carrying a non-autonomous transposable element of Ac of maize were introduced into rice plants by Agrobacterium-mediated gene transfer. Six transgenic rice plants identified as containing a single copy of the element were crossed with two transgenic rice plants carrying a gene for Ac transposase under the control of the cauliflower mosaic virus 35S promoter. In F2 progenies, excision of the element was detected by PCR analysis and re-integration of the element was investigated by Southern blot analysis. The frequency of the excision of the element was found to vary from 0 to 70% depending on the crossing combination. The frequency of the number of individual transposition events out of the total number of F2 plants with germinal excision was 44% in one crossing combination and 38% in the other. In the most efficient case, 10 plants with independent transposition were obtained out of the 49 F2 plants tested. Linkage analysis of the empty donor site and the transposed Ds-insertion site in F3 plants demonstrated that one of five Ds-insertion sites was not linked to the empty donor site. The transgenic rice obtained in this study can be used for functional genomics of rice.     

水稻生育期对褐飞虱和白背飞虱卵巢发育及起飞行为的影响

研究了取食分蘖初期、拔节期和孕穗抽穗期稻株上的褐飞虱和白背飞虱成虫的卵巢发育和起飞情况。其中,水稻生育期对褐飞虱的影响相对较小,不同生育期稻株上试虫的卵巢发育级别羽化后的4d内均无显著差异,羽化后第5天的褐飞虱在孕穗抽穗期卵巢发育最快,分蘖初期最慢,拔节期居中。褐飞虱的起飞率在各个生育期均无显著差异。白背飞虱卵巢发育、起飞率均受水稻生育期的显著影响,在水稻分蘖初期,卵巢发育最快,孕穗抽穗期则卵巢发育最慢,拔节期居中,水稻分蘖初期,起飞率最低,孕穗抽穗期起飞率最高,拔节期居中。结果表明,水稻生育期对两种飞虱卵巢发育和起飞行为的影响明显不同。认为水稻生育期对两种飞虱卵巢发育与起飞行为的不同影响,应是水稻田间白背飞虱发生较早、褐飞虱发生较晚的一个重要原因。     

Field assessments of gene flow from transgenic to cultivated rice (Oryza sativa L.) using a herbicide resistance gene as tracer marker

Development of plant genetic engineering has led to the deployment of transgenic crops and, simultaneously, to the need for a thorough assessment of the risks associated with their environmental release. This study investigated the occurrence of gene flow from transgenic rice to non-transgenic rice plants under agronomic conditions using a herbicide resistance gene as a tracer marker. Two field experiments were established in the paddy fields of two main Mediterranean rice-growing areas of Spain and Italy. In both locations analyses of phenotypic, molecular and segregation data showed that pollination of recipient plants with pollen of the transgenic source occurred at a significant frequency. A gene flow slightly lower than 0.1% was detected in a normal side-by-side plot design. Similar results were found in a circular plot when the plants were placed at 1-m distance from the transgenic central nucleus. A strong asymmetric distribution of the gene flow was detected among this circle and highest values (0.53%) were recorded following the direction of the dominant wind. A significant lowest value (0.01%) was found in the other circle (5 m from the transgenic plants) as was expected according to the characteristics of rice pollen. Such circular-field trial designs could also prove to be very useful in studying the gene flow to other commercial cultivars of rice with the aim of establishing strategies to prevent pollen dispersal from commercial transgenic fields to the neighbouring conventional fields. Received: 23 February 2001 / Accepted: 31 March 2001