Relationship between potassium fertilization and panicle blast severity in upland rice

The influence of K2O (0, 40, 80, 120 kg ha-1) at varying rates of N application (0, 30, 60 kg ha-1) at planting, on panicle blast (Pyricularia grisea (Cooke) Sacc.) was studied in a field experiment conducted during three consecutive years with the upland rice cultivar Douradão. Panicle blast severity decreased with increasing rates of potassium in the absence of nitrogen (N0). The relationship between panicle blast and K rates was quadratic at 30 kg ha-1 of nitrogen. Significant response to K fertilization was not obtained at 60 kg ha-1 of nitrogen in relation to panicle blast severity.

A fully validated method for the determination of arsenic species in rice and infant cereal products

A full validation of inorganic arsenic (iAs), methylarsonic acid (MA), and dimethyl arsinic acid (DMA) in several types of rice and rice-based infant cereals is reported. The analytical method was developed and validated in two laboratories. The extraction of the As species was performed using nitric acid 0.2 % and hydrogen peroxide 1 %, and the coupled system liquid chromatography-inductively coupled plasma-mass spectrometry (LCICP-MS) was used for speciation measurements. Detection limit (DL), quantification limit, linearity, precision, trueness, accuracy, selectivity, as well as expanded uncertainty for iAs, MA, and DMA were established. The certified reference materials (CRMs) (NMIJ 7503a, NCS ZC73008, NIST SRM 1568a) were used to check the accuracy. The method was shown to be satisfactory in two proficiency tests (PTs). The broad applicability of the method is shown from the results of analysis of 29 samples including several types of rice, rice products, and infant cereal products. Total As ranged from 40.1 to 323.7 μg As kg1. From the speciation results, iAs was predominant, and DMA was detected in some samples while MA was not detected in any sample.

Development of blast resistant somaclones of the upland rice cultivar araguaia

The degree of blast resistance of upland rice (Oryza sativa L.) cultivar Araguaia has decreased over time causing significant yield losses. The major objective of this study was to obtain blast (Pyricularia grisea) resistant somaclones, adapting greenhouse and field selection procedures. Rice blast resistance and agronomic traits were assessed in R2 to R6 generations derived from regenerant plants (R1) from immature panicles of Araguaia. The evaluation and selection procedures include testing of early segregating populations and fixed lines in the advanced generations, under natural field conditions, and artificial inoculations in the greenhouse, with prevalent races IB-1 and IB-9 of P. grisea. Somaclones with both vertical resistance and slow blasting resistance were obtained. Twenty of 31 somaclones developed with a high degree of vertical resistance and fan shaped plant type maintained resistance in field and blast nursery tests in the R6 generation. Greenhouse selection with two specific physiologic races yielded 44 somaclones with slow blasting resistance, similar plant type and yield potential as that of Araguaia.

Carbon isotope discrimination and yield of upland rice as affected by drought at flowering

Field experiments involving upland rice genotypes, sown in various dates in late season, were carried out to assess the relationship of carbon isotope discrimination with grain yield and drought resistance. In each one of the three years, one trial was kept under good water availability, while other suffered water shortage for a period of 18-23 days, encompassing panicle emergence and flowering. Drought stress reduced carbon isotope discrimination measured on soluble sugars (deltas) extracted from stem uppermost internode at the end of the imposition period, but had relatively less effect on bulk dry matter of leaves, sampled at the same period, or that of uppermost internodes and grains, sampled at harvest. The drought-induced reduction in deltas was accompanied of reduced spikelet fertility and grain yield. In the three trials subjected to drought, genotypes with the highest yield and spikelet fertility had the lowest deltas. However, this relationship was weak and it was concluded that deltas is not a sufficiently reliable indicator of rice drought resistance to be useful as a screening test in breeding programs. On the other hand, grain yield and spikelet fertility of genotypes which were the soonest to reach 50% flowering within the drought imposition period, were the least adversely affected by drought. Then, timing of drought in relation to panicle emergence and to flowering appeared to be a more important cause of yield variation among genotypes than variation in deltas.

Phenotypic virulence analysis of Pyricularia grisea isolates from Brazilian upland rice cultivars

Phenotypic virulence analysis was made on population of Pyricularia grisea isolates collected from 10 upland cultivars in three distinct rice breeding sites, with the objective of studying the degree of similarity in the phenotypic virulence among the isolates, the composition of races, and their virulence pattern. Sixteen races were identified based on the reaction type on eight standard international differentials, the predominant ones being IB9 and IB41. The virulence frequency was high on IAC47 and IAC165 among medium and early maturing cultivars, respectively. The frequency of isolates virulent was greater on upland rice cultivars (51.1%) than on irrigated rice cultivars (21.8%). Both virulent and avirulent isolates were present in the population of P. grisea to the known genes in the near isogenic lines. Of72test isolates, 94.4% were virulent for genes Pi3 and Pi4a. Thevirulence frequencies were relatively lower in decreasing order on Pi1, Pi4b and Pi2. Thecoefficient of similarity ranged from 0.28 to1.0 among the isolates pertaining to different races, while within the race IB9, it varied from 0.56 to1.0. Considering the coefficient of similarity of 0.81, 72% of isolates of race IB9 exhibited similar pattern of virulence.

Defining molecular components of symbiotic phosphate uptake in rice

Phosphate (Pi) acquisition of crops via arbuscular mycorrhizal (AM) symbiosis acquires increasing importance due to the limited rock Pi reserves and the demand for environmentally sustainable agriculture. However, the symbiotic Pi uptake machinery has not been characterized in any monocotyledonous plant species. Among these, rice is the primary staple food for more than half of the human population and thus central for future food security. However, the relevance of the AM symbiosis for rice Pi nutrition is presently unclear. Here, we show that 70% of the overall Pi acquired by rice is delivered via the symbiotic route. To better understand this pathway we combined genetic, molecular and physiological approaches to determine the specific functions of the two rice Pi transporters, PT11 and PT13, which are expressed only during AM symbiosis. The PT11 lineage of proteins is present in mono- and dicotyledons whereas PT13, while found across the Poaceae, is absent from dicotyledons. Surprisingly, mutations in either PT11 or PT13 affected fungal colonization and arbuscule formation demonstrating that both genes are essential for AM symbiosis between rice and Glomus intra.rad.ices. Importantly, for symbiotic Pi uptake, only PT11 is necessary and sufficient. We found that mycorrhizal rice, remarkably, received almost all Pi via the symbiotic route. Such dominating mycorrhizal Pi uptake was found in plants grown under controlled conditions as well as in field soils, suggesting that the AM symbiosis is relevant for the Pi nutrition of field grown rice. Development of smaller arbuscules in PT11 mutants suggested that symbiotic Pi signaling is required for fungal nourishment by the plant. However, co-culture of mutant with wild type nurse plants did not restore normal arbuscule size in mutant roots, indicating that other factors than malnutrition accounted for the altered arbuscule phenotype. Surprisingly, the loss of PT13 did not affect symbiotic Pi uptake although it impacted arbuscule morphology, suggesting that PT13 is involved in signaling during arbuscule development. However, induction of PT13 was not only monitored in arbusculated cells but also in inner cortex cells of non-inoculated roots of plants grown under high Pi fertilization conditions. According to preliminary observations, PT13 localized at the tonoplast in arbusculated and non-arbusculated cells, suggesting that it might be involved in transporting Pi into the vacuole, possibly for maintaining cellular Pi homeostasis. The further investigation showed that fungal colonization level was significantly affected in the crown roots of two ptlS mutant alleles, but not in large lateral roots, implying the possible role of PT13 for maintaining Pi homeostasis in the crown roots. - L'acquisition de phosphate (Pi) par les plantes cultivées s'effectue grâce à une symbiose mycorhizienne arbasculaire (AM). L'étude de cette symbiose devient fondamentale puisque d'une part, les réserves en phosphate minéral sont limitées, et, d'autre part, la demande pour une agriculture écologiquement soutenable se renforce. La machinerie d'absorption symbiotique du phosphate n'est cependant pas encore élucidée chez les plantes monocotylédones. Parmi celles-ci, le riz occupe une place primordiale. Aliment de base pour plus de la moitié de la population mondiale, il revêt de ce fait une dimension essentielle en termes de sécurité alimentaire. Pourtant, l'importance de la symbiose AM chez le riz dans le processus d'acquisition du phosphate n'est, encore de nos jours, que peu comprise. Dans cette étude, nous montrons que 70% du phosphate acquis par le riz est mis à disposition de la plante grâce à la symbiose AM. Afin de mieux comprendre ce mécanisme, nous avons employé des approches physiologiques et génétiques nous permettant de déterminer les fonctions spécifiques de deux transporteurs de Pi, PT11 et PT13, présents chez le riz et exprimés uniquement durant la symbiose AM. La famille de gènes à laquelle appartient PT11 est présente chez les monocotylédones ainsi que chez les dicotylédones tandis que PT13, bien que retrouvé au sein des Poaceae, est absent chez les dicotylédones. Etonnamment, des versions mutées de PT11 ou de PT13 affectent la colonisation par le champignon endo-mycorhizien ainsi que la formation d'arbuscules, démontrant l'importance de ces deux gènes dans la symbiose AM entre le riz et Glomus intraradices. Il est à noter que seul PT11 se révèle nécessaire et suffisant pour l'apport de Pi grâce à la symbiose. Nous avons observé que la presque totalité du phosphate dont dispose le riz lors d'une symbiose AM provient du champignon. De telles proportions ont été observées tant chez des plantes cultivées en conditions contrôlées que chez des plantes cultivées dans les champs. Cela suggère l'importance de la symbiose AM dans le processus d'acquisition du Pi chez le riz cultivé à l'extérieur. Le développement d'arbuscules plus petits chez le mutant PT11 tend à montrer qu'une voie signalétique impliquant le Pi symbiotique est nécessaire pour l'entretien du champignon par la plante. Toutefois, une co-culture du mutant avec des plantes sauvages ne permet pas de restaurer des arbuscules de taille normale dans les racines du mutant. Ce résultat indique le rôle de facteurs autres que la malnutrition aboutissant à la formation d'arbuscules altérés. Si la perte de PT13 n'affecte pas l'acquisition de phosphate symbiotique, la morphologie de l'arbuscule est, quant à elle, modifiée. Ceci suggère un rôle de PT13 durant le développement de l'arbuscule. Or, l'induction de PT13 est non seulement détectée dans des cellules contenant des arbuscules mais également dans des cellules du cortex, ceci chez des plantes cultivées sans champignon mais dans des conditions de fortes concentrations en engrais phosphaté. En accord avec des observations précédentes, PT13 est localisé au niveau du tonoplaste des cellules contenant ou non des arbuscules. Ceci suggère que PT13 pourrait être impliqué dans le transport du Pi vers la vacuole, éventuellement pour maintenir une certaine homéostasie du phosphate. Dans cette étude, nous démontrons également que le niveau de colonisation par le champignon est affecté de manière significative dans les racines principales des deux allèles du mutants ptl3, mais pas dans les grosses racines latérales. Cela impliquerait un rôle possible de PT13 dans le maintien de l'homéostasie du phosphate dans les racines principales. RESUME POUR UN LARGE PUBLIC Le phosphate (Pi), l'un des éléments minéraux essentiel au développement des plantes, se trouve généralement en faible quantité dans le sol, limitant ainsi la croissance des plantes. Le rendement de la production agricole dépend dès lors de l'addition d'engrais contenant du phosphate inorganique (Pi), obtenu à partir de ressources minières riches en phosphate. Or, ces ressources devraient être épuisées d'ici la fin du siècle. Les racines des plantes possèdent des transporteurs de phosphate efficaces leur permettant d'acquérir rapidement le Pi présent dans le sol. Comme le Pi s'avère immobile dans le sol, l'absorption rapide par les racines crée des zones pauvres en Pi autour des systèmes racinaires. Pour surmonter cet obstacle, les plantes ont développé une symbiose avec des champignons endomycorhiziens, la symbiose mycorhizienne arbusculaire (AM). Cette association leur donne accès à d'autres ressources en phosphate puisque le mycélium de ces champignons se développe sur une surface 100 fois supérieure à celle des racines. Cela augmente considérablement la surface de nutrition, dépassant ainsi la zone appauvrie en Pi. Le phosphate, transporté grâce au champignon jusqu'à l'intérieur des racines, est fourni à la plante par le biais de structures établies à l'intérieur des cellules végétales, appelées arbuscules. De leur côté, les plantes possèdent des transporteurs spécifiques afin de recevoir le Pi fourni par les champignons. A l'heure actuelle, la machinerie nécessaire à cette absorption a été uniquement décrite chez des plantes dicotylédones. Or, comprendre l'apport de phosphate par les champignons mycorhiziens s'avère particulièrement pertinent dans le cas des espèces monocotylédones cultivées telles que les céréales. Ces dernières constituent en effet la majeure partie de l'alimentation humaine. Parmi les céréales, le riz demeure l'aliment de base de la population mondiale, d'où son importance en terme de sécurité alimentaire. Durant mon travail de thèse, j'ai identifié et caractérisé le transporteur du riz impliqué dans l'apport de phosphate par ce type de symbiose AM. J'ai également démontré que le riz, lorsqu'il vit en symbiose, bénéficie de la presque totalité du Pi transporté par le champignon. Environ 40% de la production globale de riz est cultivée dans des conditions permettant la symbiose avec des mycorhizes arbusculaires. Les variétés de riz adaptées à ces conditions aérobiques deviennent des alternatives favorables aux cultivars actuels nécessitant une forte irrigation. Elles se révèlent en effet plus tolérantes aux pénuries d'eau et permettent l'utilisation de pratiques agricoles moins intensives. Les données présentées dans cette étude enrichissent nos connaissances concernant l'absorption du phosphate chez le riz grâce à la symbiose AM. Ces connaissances peuvent s'avérer décisives pour le développement de cultivars du riz plus adaptés à une agriculture écologiquement soutenable.

Variation in virulence in the rice blast fungus Magnaporthe grisea in São Paulo State

Resistant varieties have been the preferred means to control Magnaporthe grisea, the causal organism of the rice blast disease. The objective of this study was to examine the degree of diversity of the pathogen in different rice growing regions of São Paulo State, Brazil. Blast samples collected from rice varieties in three different regions (Tremembé, Mococa and José Bonifácio) were analyzed for race structure employing the Japanese rice differentials. The highest degree of virulence diversity was observed in Tremembé with 22 different races in three different varieties. Furthermore, no resistance gene in the Japanese differentials was effective to all isolates of M. grisea from São Paulo State.

Genetic diversity and virulence pattern in field populations of Pyricularia grisea from rice cultivar Metica-1

Rice blast is a major yield constraint of the irrigated rice in the State of Tocantins, Brazil. The objective of this investigation was to study the phenotypic and genetic diversity within the pathogen population of Pyricularia grisea in samples collected from four individual farms of rice cultivar Metica-1, under epidemic conditions of leaf blast. A set of 87 isolates was tested on 32 rice genotypes including eight international differentials. Considering 80% similarity in virulence, two groups comprising a total of 81 isolates were recognized, independently of the farms from which they were collected. Eighty percent of the isolates pertained to pathotype ID-14, indicating high cultivar specificity and narrow diversity of virulence in the sample population. The virulence in pathogen population on rice cultivars BR-IRGA 409 and Rio Formoso was low. Analysis of P. grisea isolates using rep-PCR with two primer sequences from Pot2 generated fingerprint profiles of one to nine bands. Cluster analysis revealed the occurrence of six fingerprint groups with similarities ranging from 0.09 to 1. There was no straight relationship between virulence of the isolates based on reaction pattern on 32 genotypes and grouping based on Pot2 rep-PCR analysis of P. grisea isolates collected from 'Metica-1'.

Cultivar response to fungicide application in relation to rice blast control, productivity and sustainability

Four field trials were conducted, from 1995 to 1997, with the objective of studying the response of four upland cultivars to foliar fungicide application in relation to panicle blast control, grain yield and sustainability. Differential disease control and yield response of cultivars to fungicide treatment were obtained. Losses in grain yield of cultivars IAC 202, Caiapó, Rio Paranaíba and Araguaia due to panicle blast were 44.8%, 27.4%, 24.4% and 18.2%, respectively. Two applications of tricyclazole or benomyl controlled panicle blast, as indicated by lower values of disease progress curve and relative panicle blast severity, and increased grain yield of the cultivar IAC 202. The losses in 100 panicle grain weight and grain yield were significantly reduced by 22.3% and 25.1% in IAC 202 and 23.6% and 20.5% in Caiapó, respectively, with two sprays of tricyclazole. Sustainable value index for yield was maximum with two applications of tricyclazole (0.59), followed by one application at booting (0.46) and at heading (0.40) in cultivar IAC 202. Results showed no yield response of the cultivars Rio Paranaíba and Araguaia to fungicide applications for panicle blast control.

Resistance spectra of six elite breeding lines of upland rice to Pyricularia grisea

The objective of this work was to evaluate the resistance spectra of six elite breeding lines of rice, developed for improved yield and grain quality, in inoculation tests in the greenhouse and in the field. Forty-six isolates of Pyricularia grisea collected from the cultivar Primavera, 31 from the cultivar Maravilha and 19 from six elite breeding lines, totaling 96 were utilized for inoculations. Out of 11 international and 15 Brazilian pathotypes, IC-1, IB-9, and BD-16, respectively, were identified as most frequent isolates collected from the cultivar Primavera. The isolates retrieved from Maravilha belong to four international and 11 Brazilian pathotypes, the predominant ones being IB-9 and IB-49 and BB-1 and BB-21, respectively. Lines CNAs 8711 and CNAs 8983 showed resistant reaction to all test isolates from Maravilha, while CNAs 8983 was susceptible to three isolates of Primavera pertaining to the pathotype IC-1. A majority of isolates exhibiting compatible reaction to Primavera were incompatible to Maravilha and vice-versa.Field assessment of rice blast utilizing the area under disease progress curve as a criterion for measuring disease severity showed significant differences among the six breeding lines. The isolates of P. grisea exhibiting differential reaction on breeding lines can be utilized in pyramiding resistance genes in new upland rice cultivars.

Characterization of OsMIK in a rice mutant with reduced phytate content reveals an insertion of a rearranged retrotransposon.

The rice low phytic acid (lpa) mutant Os-lpa-XS110-1(XS-lpa) has ~45 % reduction in seed phytic acid (PA) compared with the wild-type cultivar Xiushui 110. Previously, a single recessive gene mutation was shown to be responsible for the lpa phenotype and was mapped to a region of chromosome 3 near OsMIK (LOC_Os03g52760) and OsIPK1 (LOC_Os03g51610), two genes involved in PA biosynthesis. Here, we report the identification of a large insert in the intron of OsMIK in the XS-lpa mutant. Sequencing of fragments amplified through TAIL-PCRs revealed that the insert was a derivative of the LINE retrotransposon gene LOC_Os03g56910. Further analyses revealed the following characteristics of the insert and its impacts: (1) the inserted sequence of LOC_Os03g56910 was split at its third exon and rejoined inversely, with its 5' and 3' flanking sequences inward and the split third exon segments outward; (2) the LOC_Os03g56910 remained in its original locus in XS-lpa, and the insertion probably resulted from homologous recombination repair of a DNA double strand break; (3) while the OsMIK transcripts of XS-lpa and Xiushui 110 were identical, substantial reductions of the transcript abundance (~87 %) and the protein level (~60 %) were observed in XS-lpa, probably due to increased methylation in its promoter region. The above findings are discussed in the context of plant mutagenesis, epigenetics and lpa breeding.

Mapping of quantitative trait loci for thermosensitive genic male sterility in indica rice

The objective of this work was to select and use microsatellite markers, to map genomic regions associated with the genetic control of thermosensitive genic male sterility (TGMS) in rice. An F2 population, derived from the cross between fertile and TGMS indica lines, was used to construct a microsatellite-based genetic map of rice. The TGMS phenotype showed a continuous variation in the segregant population. A low level of segregation distortion was detected in the F2 (14.65%), whose cause was found to be zygotic selection. There was no evidence suggesting a cause-effect relationship between zygotic selection and the control of TGMS in this cross. A linkage map comprising 1,213.3 cM was constructed based on the segregation data of the F2 population. Ninety-five out of 116 microsatellite polymorphic markers were assembled into 11 linkage groups, with an average of 12.77 cM between two adjacent marker loci. The phenotypic and genotypic data allowed for the identification of three new quantitative trait loci (QTL) for thermosensitive genic male sterility in indica rice. Two of the QTL were mapped on chromosomes that, so far, have not been associated with the genetic control of the TGMS trait (chromosomes 1 and 12). The third QTL was mapped on chromosome 7, where a TGMS locus (tms2) has recently been mapped. Allelic tests will have to be developed, in order to clarify if the two regions are the same or not.

Growth components and zinc recovery efficiency of upland rice genotypes

The objective of this work was to evaluate Zn use efficiency by upland rice genotypes. The experiment was carried out in a greenhouse, with ten upland rice genotypes grown on an Oxisol (Typic Hapludox) with no application, and with application of 10 mg kg-1 Zn, applied as zinc sulfate. Shoot dry weight, grain yield, Zn harvest index, Zn concentration in shoot and in grain were significantly influenced by soil Zn levels and genotypes. However, panicle number and grain harvest index were significantly affected only by genotype. Genotypes CNA8557, CNA8540 and IR42 produced higher grain yield than other genotypes. Genotypes showed significant variability in Zn recovery efficiency. On average, 13% of the applied Zn was recovered by upland rice genotypes. Genotypes with high Zn recovery efficiency could be used in breeding of Zn efficient upland rice cultivars. Higher level of soil Zn (10 mg kg-1) increased significantly the concentrations of plant Cu and Mn. However, Fe concentrations in plant (shoot and grain) were not influenced by soil Zn levels.

Rhizosphere properties of rice genotypes as influenced by anoxia and availability of zinc and iron

The objective of this work was to study possible mechanisms involved in root-induced changes of rhizosphere physicochemical properties of rice genotypes, under anoxia and low supply of Zn and Fe. Two rice genotypes, including an upland and a lowland ones, were grown in hydroponic medium under adequate and low supply of Zn and Fe, with or without aeration. Anoxia increased shoot dry weight, root length and uptake of Zn and Fe in lowland Amol genotype, but reduced these parameters in upland Gasrol-Dashti genotype. The amount of oxygen released by roots was statistically higher in 'Amol'. The highest acidification potential of roots was observed in the lowland genotype under low supply of Zn, and in the upland genotype under Fe starvation. The highest oxalate (only organic acid detected) exudation from roots was observed in Zn and Fe deficient Gasrol-Dashti genotype. Zinc deficiency caused reduction of alcohol dehydrogenase and stimulation of lactate dehydrogenase activity, particularly in shoot. The ability to induce changes in the rhizosphere properties has a great contribution for the adaptation of both lowland and upland rice genotypes to specific soil conditions.

Impact of increasing mean air temperature on the development of rice and red rice

The objective of this study was to assess the development response of cultivated rice and red rice to different increases in minimum and maximum daily air temperatures, in Santa Maria, Rio Grande do Sul State, Brazil. One hundred years climate scenarios of temperatures 0, +1, +2, +3, +4, and +5ºC, with symmetric and asymmetric increases in minimum and maximum daily air temperatures were created, using the LARS-WG Weather Generator, and a 1969-2003 database. Nine cultivated rice genotypes (IRGA 421, IRGA 416, IRGA 417, IRGA 420, BRS 7 TAIM, BR-IRGA 409, EPAGRI 109, EEA 406 and a hybrid), and two red rice biotypes (awned black hull-ABHRR, and awned yellow hull-AYHRR) were used. The dates of panicle differentiation (R1), anthesis (R4), and all grains with brown hulls (R9) were estimated with a nonlinear simulation model. Overall, the duration of the emergence-R1 phase decreased, whereas the duration of the R1-R4 and R4-R9 phases most often increased, as temperature increased in the climate change scenarios. The simulated rice development response to elevated temperature was not the same, when the increase in minimum and maximum temperature was symmetric or asymmetric.