A disponibilidade do fósforo de oiversos fosfatos estudada por meio do método de Neubauer"

In order to study the phosphorus availability from various phosphates fertilizers an experiment was performed according to the biological seedling method of Neubauer. The physico-chemical properties of the soil "terra roxa-misturada", a red soil derived from basaltic rocks are given in the Portuguese text. Rice (Oryza sativa, L.) instead of rye (Secale cereale, L.) was used. Five replications of each of the following treatments were made: 1 - check, with 350 g of sand 2 - 350 g of sand plus 100 g of soil 3 - 350 g of sand and plus 100 g of soil plus 40 mg of P2O5, from superphosphate. 4 - 350 g of sand plus 100 g of soil plus 40 mg of P2O5. from Olinda (Brazil) phosphorite. 5 - 350 g of sand plus 100 g of soil plus 40 mg of P2O5 from Florida (U. S. A.) phosphorite. 6 - 350 g os sand plus 100 g of soil plus 40 mg of P2O5 from Hyperphosphate, a commertial name of a North African (Gafsa) phosphorite. 7 - 350 g of sand plus 100 g of soil plus 40 mg of P2O5 from Araxá (Brazil) apatite. After 18 days of growth, the roots and tops of rice seedlings were harvested and analysed for phosphorus, and the results are summarized in table 1. Table 1 - Milligrams of P2O5 determined in rice seedlings. Treatments Mean of 5 replications mg of P2O5 1 ..................... 24.196 2 ..................... 23.850 3 ..................... 30.724 4 ..................... 27.620 5 ..................... 27.480 6..................... 30.210 7 ..................... 26.032 The least significant difference at the 5% level by Tukey's procedure for comparisons among the treatments means is 1.365 mg of P(2)0. It is interesting to observe that rice plants did not take any phosphorus from the soil according to he data of the treatments n.° 1 and n.° 2. This can be explained by the high phosphorus fixing capacity of the soil "terra roxa misturada".

Matointerferência em arroz de sequeiro. I: efeitos do espaçamento e da adubação nitrogenada nas concentrações de nutrientes no arroz e nas plantas daninhas

O trabalho insere-se num estudo global sobre os efeitos do espaçamento entre sulcos de semeadura e da dose de adubação nitrogenada sobre as relações competitivas entre a cultura do arroz de sequeiro (Oryza sativa L.) e uma comunidade de plantas daninhas. Para tanto, os tratamentos foram dispostos num esquema fatorial 2x2x3, onde constituiram variáveis: duas condições de manejo da comunidade infestante - sem controle e com controle do mato durante todo o ciclo do arroz - dois espaçamentos entre sulcos de semeadura - 0,40 e 0,60 m - e três doses de nitrogênio - 2,4, e 7,2 de N/m de sulco. O experimento foi montado sobre solo Latossolo Vermelho Escuro fase arenosa e obedeceu o delineamento experimental de blocos ao acaso com 4repetições. Foram realizadas duas avaliações: por ocasião do perfilhamento e por ocasião do florescimento da cultura. Em comparação com as plantas daninhas, os teores de N, P, Fe, Mn e Zn foram mais elevados no arroz; os de Ca e Mg foram menos elevados e os teores de K e Cu foram inferiores aos de D. horizontalis e superiores aos de 7. hirsuta. Por ocasião do florescimento, o arroz apresentou teores mais elevados de P, Cu, Mn e Zn, enquanto que a comunidade infestante apresentou maiores teores de N, K, Ca, Mg e Fe. Os efeitos do espaçamento e da fertilização nitrogenada apresentaram aspectos distintos de acordo com o nutriente, a espécie envolvida e a época da avaliação. De um modo geral, os efeitos das plantas daninhas foram mais acentuados no espaçamento de 0,60 m.

An ATP-binding cassette subfamily G full transporter is essential for the retention of leaf water in both wild barley and rice.

Land plants have developed a cuticle preventing uncontrolled water loss. Here we report that an ATP-binding cassette (ABC) subfamily G (ABCG) full transporter is required for leaf water conservation in both wild barley and rice. A spontaneous mutation, eibi1.b, in wild barley has a low capacity to retain leaf water, a phenotype associated with reduced cutin deposition and a thin cuticle. Map-based cloning revealed that Eibi1 encodes an HvABCG31 full transporter. The gene was highly expressed in the elongation zone of a growing leaf (the site of cutin synthesis), and its gene product also was localized in developing, but not in mature tissue. A de novo wild barley mutant named "eibi1.c," along with two transposon insertion lines of rice mutated in the ortholog of HvABCG31 also were unable to restrict water loss from detached leaves. HvABCG31 is hypothesized to function as a transporter involved in cutin formation. Homologs of HvABCG31 were found in green algae, moss, and lycopods, indicating that this full transporter is highly conserved in the evolution of land plants.

Glomus intraradices induces changes in root system architecture of rice independently of common symbiosis signaling

? Arbuscular mycorrhizal fungi colonize the roots of most monocotyledons and dicotyledons despite their different root architecture and cell patterning. Among the cereal hosts of arbuscular mycorrhizal fungi, Oryza sativa (rice) possesses a peculiar root system composed of three different types of roots: crown roots; large lateral roots; and fine lateral roots. Characteristic is the constitutive formation of aerenchyma in crown roots and large lateral roots and the absence of cortex from fine lateral roots. Here, we assessed the distribution of colonization by Glomus intraradices within this root system and determined its effect on root system architecture. ? Large lateral roots are preferentially colonized, and fine lateral roots are immune to arbuscular mycorrhizal colonization. Fungal preference for large lateral roots also occurred in sym mutants that block colonization of the root beyond rhizodermal penetration. ? Initiation of large lateral roots is significantly induced by G. intraradices colonization and does not require a functional common symbiosis signaling pathway from which some components are known to be needed for symbiosis-mediated lateral root induction in Medicago truncatula. ? Our results suggest variation of symbiotic properties among the different rice root-types and induction of the preferred tissue by arbuscular mycorrhizal fungi. Furthermore, signaling for arbuscular mycorrhizal-elicited alterations of the root system differs between rice and M. truncatula.

Influencia de las técnicas de control del arroz salvaje sobre la población de quironómidos

Las larvas de quironómidos (Cricotupus sylvestris) constituyen la plaga invertebrada más importante que afecta al establecimiento del cultivo en aquellos arrozales donde se realiza siembra directa sobre una lámina de agua. En el delta del Ebro, Catalá MM et al., 2008a y 2008 b, demostraron como largos periodos de inundación favorecen el incremento de la población de larvas de quironómidos, hasta tal punto que se llega a destruir prácticamente la totalidad de semillas y plántulas si coincide el periodo de máxima población de la plaga con el de mayor sensibilidad del cultivo. Los campos de arroz infestados de arroz salvaje (Oryza sativa L.) requieren largos periodos de inundación para favorecer la nascencia del mismo y proceder a su destrucción, que puede ser mecánica o química. Este método de eliminación de arroz salvaje se denomina “falsa siembra”. Por otra parte se sabe que no hay insecticidas autorizados en el cultivo del arroz con suficiente eficacia para controlar las elevadas poblaciones de quironómidos (trabajos no publicados del autor), lo que nos lleva a estudiar la plaga dentro del ámbito agronómico del cultivo para disminuir la misma a niveles poblacionales suficientemente bajos que permitan conseguir una buena densidad de planta. Así pues, este trabajo pretende estudiar el efecto que tienen las distintas técnicas de control del arroz salvaje sobre la población de quironómidos.

Segregation in a mycorrhizal fungus alters rice growth and symbiosis-specific gene transcription

Arbuscular mycorrhizal fungi (AMF) form symbioses with the majority of plants, improving plant nutrition and diversity. Evidence exists suggesting that AMF contain populations of genetically different nucleotypes coexisting in a common cytoplasm. This potentially has two important consequences for their genetics. First, by random distribution of nuclei at spore formation, new offspring of an AMF could receive different complements of nucleotypes compared to the parent or siblings-we consider this as segregation. Second, genetic exchange between AMF would allow the mixing of nuclei, altering nucleotype diversity in new spores. Because segregation was assumed not to occur and genetic exchange has only recently been demonstrated, no attempts have been made to test whether this affects the symbiosis with plants. Here, we show that segregation occurs in the AMF Glomus intraradices and can enhance the growth of rice up to five times, even though neither parental nor crossed AMF lines induced a positive growth response. This process also resulted in an alteration of symbiosis-specific gene transcription in rice. Our results demonstrate that manipulation of AMF genetics has important consequences for the symbiotic effects on plants and could be used to enhance the growth of globally important crops.

Production of low phytic acid rice by hairpin RNA- and artificial microRNA-mediated silencing of OsMIK in seeds

To produce agronomically competitive rice with nutritionally superior, environmentally safe phytic acid (PA) levels, hairpin RNA (hpRNA)- and artificial microRNA (amiRNA)-mediated gene silencing approaches were explored to reduce both myo-inositol kinase gene (OsMIK) expression and PA accumulation in rice seeds. hpRNA and amiRNA sequences targeted to OsMIK (hpMIK and amiMIK), under the control of a rice Ole18 promoter, were transformed into the rice cultivar Nippon-bare. Fourteen and 21 independent transgenic events were identified containing the hpMIK and amiMIK constructs, respectively, from which five stable homozygous transgenic lines of each were developed together with their null siblings. Southern blotting demonstrated transgene integration into the genome and quantitative real-time PCR showed that gene silencing was restricted to seeds. OsMIK transcripts were significantly reduced in both transgenic amiMIK and hpMIK seeds, which had PA levels reduced by 14.9-50.2 and 38.1-50.7 %, respectively, compared with their respective null siblings. There were no systematic significant differences in agronomic traits between the transgenic lines and their non-transgenic siblings, and no correlation between seed PA contents and decreased rates of seed germination and seedling emergence. The results of the present study suggest that Ole 18-driven OsMIK silencing via hpRNA and amiRNA could be an effective way to develop agronomically competitive low phytic acid rice.

Comparative transcriptomics of rice reveals an ancient pattern of response to microbial colonization.

Glomalean fungi induce and colonize symbiotic tissue called arbuscular mycorrhiza on the roots of most land plants. Other fungi also colonize plants but cause disease not symbiosis. Whole-transcriptome analysis using a custom-designed Affymetrix Gene-Chip and confirmation with real-time RT-PCR revealed 224 genes affected during arbuscular mycorrhizal symbiosis. We compared these transcription profiles with those from rice roots that were colonized by pathogens (Magnaporthe grisea and Fusarium moniliforme). Over 40% of genes showed differential regulation caused by both the symbiotic and at least one of the pathogenic interactions. A set of genes was similarly expressed in all three associations, revealing a conserved response to fungal colonization. The responses that were shared between pathogen and symbiont infection may play a role in compatibility. Likewise, the responses that are different may cause disease. Some of the genes that respond to mycorrhizal colonization may be involved in the uptake of phosphate. Indeed, phosphate addition mimicked the effect of mycorrhiza on 8% of the tested genes. We found that 34% of the mycorrhiza-associated rice genes were also associated with mycorrhiza in dicots, revealing a conserved pattern of response between the two angiosperm classes.

Nonredundant Regulation of Rice Arbuscular Mycorrhizal Symbiosis by Two Members of the PHOSPHATE TRANSPORTER1 Gene Family.

Pi acquisition of crops via arbuscular mycorrhizal (AM) symbiosis is becoming increasingly important due to limited high-grade rock Pi reserves and a demand for environmentally sustainable agriculture. Here, we show that 70% of the overall Pi acquired by rice (Oryza sativa) is delivered via the symbiotic route. To better understand this pathway, we combined genetic, molecular, and physiological approaches to determine the specific functions of two symbiosis-specific members of the PHOSPHATE TRANSPORTER1 (PHT1) gene family from rice, ORYsa;PHT1;11 (PT11) and ORYsa;PHT1;13 (PT13). The PT11 lineage of proteins from mono- and dicotyledons is most closely related to homologs from the ancient moss, indicating an early evolutionary origin. By contrast, PT13 arose in the Poaceae, suggesting that grasses acquired a particular strategy for the acquisition of symbiotic Pi. Surprisingly, mutations in either PT11 or PT13 affected the development of the symbiosis, demonstrating that both genes are important for AM symbiosis. For symbiotic Pi uptake, however, only PT11 is necessary and sufficient. Consequently, our results demonstrate that mycorrhizal rice depends on the AM symbiosis to satisfy its Pi demands, which is mediated by a single functional Pi transporter, PT11.

A rice cis-natural antisense RNA acts as a translational enhancer for its cognate mRNA and contributes to phosphate homeostasis and plant fitness.

cis-natural antisense transcripts (cis-NATs) are widespread in plants and are often associated with downregulation of their associated sense genes. We found that a cis-NAT positively regulates the level of a protein critical for phosphate homeostasis in rice (Oryza sativa). PHOSPHATE1;2 (PHO1;2), a gene involved in phosphate loading into the xylem in rice, and its associated cis-NATPHO1;2 are both controlled by promoters active in the vascular cylinder of roots and leaves. While the PHO1;2 promoter is unresponsive to the plant phosphate status, the cis-NATPHO1;2 promoter is strongly upregulated under phosphate deficiency. Expression of both cis-NATPHO1;2 and the PHO1;2 protein increased in phosphate-deficient plants, while the PHO1;2 mRNA level remained stable. Downregulation of cis-NATPHO1;2 expression by RNA interference resulted in a decrease in PHO1;2 protein, impaired the transfer of phosphate from root to shoot, and decreased seed yield. Constitutive overexpression of NATPHO1;2 in trans led to a strong increase of PHO1;2, even under phosphate-sufficient conditions. Under all conditions, no changes occurred in the level of expression, sequence, or nuclear export of PHO1;2 mRNA. However, expression of cis-NATPHO1;2 was associated with a shift of both PHO1;2 and cis-NATPHO1;2 toward the polysomes. These findings reveal an unexpected role for cis-NATPHO1;2 in promoting PHO1;2 translation and affecting phosphate homeostasis and plant fitness.

Characterization of the rice PHO1 gene family reveals a key role for OsPHO1;2 in phosphate homeostasis and the evolution of a distinct clade in dicotyledons.

Phosphate homeostasis was studied in a monocotyledonous model plant through the characterization of the PHO1 gene family in rice (Oryza sativa). Bioinformatics and phylogenetic analysis showed that the rice genome has three PHO1 homologs, which cluster with the Arabidopsis (Arabidopsis thaliana) AtPHO1 and AtPHO1;H1, the only two genes known to be involved in root-to-shoot transfer of phosphate. In contrast to the Arabidopsis PHO1 gene family, all three rice PHO1 genes have a cis-natural antisense transcript located at the 5 ' end of the genes. Strand-specific quantitative reverse transcription-PCR analyses revealed distinct patterns of expression for sense and antisense transcripts for all three genes, both at the level of tissue expression and in response to nutrient stress. The most abundantly expressed gene was OsPHO1;2 in the roots, for both sense and antisense transcripts. However, while the OsPHO1;2 sense transcript was relatively stable under various nutrient deficiencies, the antisense transcript was highly induced by inorganic phosphate (Pi) deficiency. Characterization of Ospho1;1 and Ospho1;2 insertion mutants revealed that only Ospho1;2 mutants had defects in Pi homeostasis, namely strong reduction in Pi transfer from root to shoot, which was accompanied by low-shoot and high-root Pi. Our data identify OsPHO1;2 as playing a key role in the transfer of Pi from roots to shoots in rice, and indicate that this gene could be regulated by its cis-natural antisense transcripts. Furthermore, phylogenetic analysis of PHO1 homologs in monocotyledons and dicotyledons revealed the emergence of a distinct clade of PHO1 genes in dicotyledons, which include members having roles other than long-distance Pi transport.

Use of unmodified starches and partial removal of serum to improve Granada medium stability

The use of 1% unmodified rice starch and 1% horse serum instead of 2% soluble starch and 5% serum in Granada medium is described. These components result in a medium of increased stability, preventing spoilage after a few days of storage at room temperature

On the cellular site of two-pore channel TPC1 action in the Poaceae.

The slow vacuolar (SV) channel has been characterized in different dicots by patch-clamp recordings. This channel represents the major cation conductance of the largest organelle in most plant cells. Studies with the tpc1-2 mutant of the model dicot plant Arabidopsis thaliana identified the SV channel as the product of the TPC1 gene. By contrast, research on rice and wheat TPC1 suggested that the monocot gene encodes a plasma membrane calcium-permeable channel. To explore the site of action of grass TPC1 channels, we expressed OsTPC1 from rice (Oryza sativa) and TaTPC1 from wheat (Triticum aestivum) in the background of the Arabidopsis tpc1-2 mutant. Cross-species tpc1 complementation and patch-clamping of vacuoles using Arabidopsis and rice tpc1 null mutants documented that both monocot TPC1 genes were capable of rescuing the SV channel deficit. Vacuoles from wild-type rice but not the tpc1 loss-of-function mutant harbor SV channels exhibiting the hallmark properties of dicot TPC1/SV channels. When expressed in human embryonic kidney (HEK293) cells OsTPC1 was targeted to Lysotracker-Red-positive organelles. The finding that the rice TPC1, just like those from the model plant Arabidopsis and even animal cells, is localized and active in lyso-vacuolar membranes associates this cation channel species with endomembrane function.