Characterization of tomato Cycling Dof Factors reveals conserved and new functions in the control of flowering time and abiotic stress responses.

DNA binding with One Finger (DOF) transcription factors are involved in multiple aspects of plant growth and development but their precise roles in abiotic stress tolerance are largely unknown. Here we report a group of five tomato DOF genes, homologous to Arabidopsis Cycling DOF Factors (CDFs), that function as transcriptional regulators involved in responses to drought and salt stress and flowering-time control in a gene-specific manner. SlCDF1?5 are nuclear proteins that display specific binding with different affinities to canonical DNA target sequences and present diverse transcriptional activation capacities in vivo. SlCDF1?5 genes exhibited distinct diurnal expression patterns and were differentially induced in response to osmotic, salt, heat, and low-temperature stresses. Arabidopsis plants overexpressing SlCDF1 or SlCDF3 showed increased drought and salt tolerance. In addition, the expression of various stress-responsive genes, such as COR15, RD29A, and RD10, were differentially activated in the overexpressing lines. Interestingly, overexpression in Arabidopsis of SlCDF3 but not SlCDF1 promotes late flowering through modulation of the expression of flowering control genes such as CO and FT. Overall, our data connect SlCDFs to undescribed functions related to abiotic stress tolerance and flowering time through the regulation of specific target genes and an increase in particular metabolites

Delayed flowering in pineapples (Ananas comosus (L.) Merr.) caused by co-suppression of the ACACS2 gene

Natural flower induction is a major pineapple industry problem. It usually occurs when shortening days and low temperatures give raise to increased ethylene production in the leaf tissue and plant stem apex which in turn stimulates flowering. Natural flowering fruit matures 4 to 6 weeks ahead of the normal summer harvest resulting in the need for extra harvest passes and considerable yield losses. Ethylene is produced through the sequential action of ACC synthase and ACC oxidase. Our team has cloned an ACC synthase gene from pineapple (ACACS2), which is expressed in meristems and activated under the environmental conditions that induce flowering in nature. Genetic constructs have been produced containing ACACS2 in sense orienta¬tion to induce silencing of the host gene in the plant by co-suppression mechanisms. Two independent lines of transgenic plants have been produced and field trials have been conducted in Queensland for four years in order to study the characteristics of the transgenic lines. We have identified a group of transgenic plants demonstrating inherited flowering delay and confirmed co-suppression of the ACACS2 gene due to methylation.

Gibberellic acid in vegetative and reproductive development of Phalaenopsis orchid hybrid genus

O setor de floricultura movimenta cerca de um bilhão de dólares no Brasil e o desenvolvimento de técnicas direcionadas ao controle do florescimento é necessário. O presente trabalho avaliou a influência do ácido giberélico (GA3) no desenvolvimento vegetativo e reprodutivo de plantas jovens de Phalaenopsis FSNT 'Dai-Itigo', híbrido de coloração rósea. A aplicação do GA3 foi feita via pulverização foliar nas concentrações de 0, 125, 250, 500 e 1.000 mg L -1. O comprimento das folhas aumentou consideravelmente com o uso do GA3 em baixas concentrações, porém houve diminuição da largura foliar das plantas tratadas com esse fitorregulador. A aplicação do GA3 na concentração de 125 mg L -1 apresentou os melhores resultados para a promoção do florescimento e qualidade da floração deste híbrido de orquídea. Nesse tratamento, aproximadamente 50% das plantas pulverizadas apresentaram floração cerca de 6-12 meses antes da floração das plantas sem aplicação do fitorregulador. em conclusão, a qualidade da floração e das flores foi melhor no tratamento com 125 mg L -1 de GA3.

Acúmulo de sacarose, produtividade e florescimento de cana-de-açúcar sob reguladores vegetais

Plant regulators have been used successfully for early ripening and flowering control in sugarcane (Saccharum spp. Hybrid). However, little information is available about the interactions between new genotypes and two plant regulators with regard to those variables. This study determined the effect of the regulators on the sucrose content, stalk production and flowering of seven sugarcane genotypes. The experiment was installed in March 2004 near Ja-, SP, Brazil A randomized complete block design was used with four blocks with a split plot treatment arrangement where the main plots were the genotypes IAC87-3396, IAC87-3410, IAC89-3124, IAC91-2195, 1AC91-5155, P088-62 and SP80-1842 and the subplots were sulfomethuron-methyl (15g i.a.·ha -1), etefon (480g i.a.·ha-1) and unsprayed control. Pol in cane was evaluated at 0, 21, 42, 63, 84, 105 and 126 days after the plant regulators application (DAA). Flowering, pith, stalk production and sucrose content were evaluated at 126 DAA. In most of the genotypes the use of plant regulators anticipated maturation in 21 days when compared with the unsprayed control. Etefon was more efficient for harvesting sugarcane between 42-84 DAA, whereas sulfomethuron-methyl was between 105 and 126 DDA. Both products controlled flowering. For most genotypes, the ripeners did not affect the productivity of stalks, except for sulfomethuron-methyl that reduced it in SP80-1842, and for ethephon that increased it in IAC91-2195. Sugar productivity of IAC89-3124 increased with both regulators, while it was higher in IAC91-2195 with ethephon application and lower in SP80-1842 with sulfomethuron-methyl use.

Gibberellic acid in vegetative and reproductive development of Phalaenopsis orchid hybrid genus

The flower industry represents about one billion dollars in Brazil and the development of techniques aimed at flowering control is required. This study evaluated the influence of gibberellic acid (GA3) on the vegetative and reproductive development of young plants of Phalaenopsis FSNT 'Dai-Itigo' hybrid pink color. The application of GA3 was made by foliar sprays at concentrations of 0, 125, 250, 500 and 1,000 mg L -1. The length of leaves increased significantly when using GA3 at low concentrations, but leaf width decreased. The application of GA3 at 125 mg L -1 showed the best results for the promotion of flowering and flower quality of this orchid hybrid. In this treatment, about 50% of plants treated with GA3 flowered about 6-12 months before the plants that were non-treated with this plant growth regulator. The quality of flowering and flowers was best with 125 mg L -1 GA3.

Cycling Dof Factors: Molecular and functional characterization of Arabidopsis thaliana AtCDF3 and tomato (Solanum lycopersicum L.) SlCDF3 in response to abiotic stress

El tomate (Solanum lycopersicum L.) es considerado uno de los cultivos hortícolas de mayor importancia económica en el territorio Español. Sin embargo, su producción está seriamente afectada por condiciones ambientales adversas como, salinidad, sequía y temperaturas extremas. Para resolver los problemas que se presentan en condiciones de estrés, se han empleado una serie de técnicas culturales que disminuyen sus efectos negativos, siendo de gran interés el desarrollo de variedades tolerantes. En este sentido la obtención y análisis de plantas transgénicas, ha supuesto un avance tecnológico, que ha facilitado el estudio y la evaluación de genes seleccionados en relación con la tolerancia al estrés. Estudios recientes han mostrado que el uso de genes reguladores como factores de transcripción (FTs) es una gran herramienta para obtener nuevas variedades de tomate con mayor tolerancia a estreses abióticos. Las proteínas DOF (DNA binding with One Finger) son una familia de FTs específica de plantas (Yangisawa, 2002), que están involucrados en procesos fisiológicos exclusivos de plantas como: asimilación del nitrógeno y fijación del carbono fotosintético, germinación de semilla, metabolismo secundario y respuesta al fotoperiodo pero su preciso rol en la tolerancia a estrés abiótico se desconoce en gran parte. El trabajo descrito en esta tesis tiene como objetivo estudiar genes reguladores tipo DOF para incrementar la tolerancia a estrés abiotico tanto en especies modelo como en tomate. En el primer capítulo de esta tesis se muestra la caracterización funcional del gen CDF3 de Arabidopsis, así como su papel en la respuesta a estrés abiótico y otros procesos del desarrollo. La expresión del gen AtCDF3 es altamente inducido por sequía, temperaturas extremas, salinidad y tratamientos con ácido abscísico (ABA). La línea de inserción T-DNA cdf3-1 es más sensible al estrés por sequía y bajas temperaturas, mientras que líneas transgénicas de Arabidopsis 35S::AtCDF3 aumentan la tolerancia al estrés por sequía, osmótico y bajas temperaturas en comparación con plantas wild-type (WT). Además, estas plantas presentan un incremento en la tasa fotosintética y apertura estomática. El gen AtCDF3 se localiza en el núcleo y que muestran una unión específica al ADN con diferente afinidad a secuencias diana y presentan diversas capacidades de activación transcripcional en ensayos de protoplastos de Arabidopsis. El dominio C-terminal de AtCDF3 es esencial para esta localización y su capacidad activación, la delección de este dominio reduce la tolerancia a sequía en plantas transgénicas 35S::AtCDF3. Análisis por microarray revelan que el AtCDF3 regula un set de genes involucrados en el metabolismo del carbono y nitrógeno. Nuestros resultados demuestran que el gen AtCDF3 juega un doble papel en la regulación de la respuesta a estrés por sequía y bajas temperaturas y en el control del tiempo de floración. En el segundo capítulo de este trabajo se lleva a cabo la identificación de 34 genes Dof en tomate que se pueden clasificar en base a homología de secuencia en cuatro grupos A-D, similares a los descritos en Arabidopsis. Dentro del grupo D se han identificado cinco genes DOF que presentan características similares a los Cycling Dof Factors (CDFs) de Arabidopsis. Estos genes son considerados ortólogos de Arabidopsis CDF1-5, y han sido nombrados como Solanum lycopersicum CDFs o SlCDFs. Los SlCDF1-5 son proteínas nucleares que muestran una unión específica al ADN con diferente afinidad a secuencias diana y presentan diversas capacidades de activación transcripcional in vivo. Análisis de expresión de los genes SlCDF1-5 muestran diferentes patrones de expresión durante el día y son inducidos de forma diferente en respuesta a estrés osmótico, salino, y de altas y bajas temperaturas. Plantas de Arabidopsis que sobre-expresan SlCDF1 y SlCDF3 muestran un incremento de la tolerancia a la sequía y salinidad. Además, de la expresión de varios genes de respuesta estrés como AtCOR15, AtRD29A y AtERD10, son expresados de forma diferente en estas líneas. La sobre-expresión de SlCDF3 en Arabidopsis promueve un retardo en el tiempo de floración a través de la modulación de la expresión de genes que controlan la floración como CONSTANS (CO) y FLOWERING LOCUS T (FT). En general, nuestros datos demuestran que los SlCDFs están asociados a funciones aun no descritas, relacionadas con la tolerancia a estrés abiótico y el control del tiempo de floración a través de la regulación de genes específicos y a un aumento de metabolitos particulares. ABSTRACT Tomato (Solanum lycopersicum L.) is one of the horticultural crops of major economic importance in the Spanish territory. However, its production is being affected by adverse environmental conditions such as salinity, drought and extreme temperatures. To resolve the problems triggered by stress conditions, a number of agricultural techniques that reduce the negative effects of stress are being frequently applied. However, the development of stress tolerant varieties is of a great interest. In this direction, the technological progress in obtaining and analysis of transgenic plants facilitated the study and evaluation of selected genes in relation to stress tolerance. Recent studies have shown that a use of regulatory genes such as transcription factors (TFs) is a great tool to obtain new tomato varieties with greater tolerance to abiotic stresses. The DOF (DNA binding with One Finger) proteins form a family of plant-specific TFs (Yangisawa, 2002) that are involved in the regulation of particular plant processes such as nitrogen assimilation, photosynthetic carbon fixation, seed germination, secondary metabolism and flowering time bur their precise roles in abiotic stress tolerance are largely unknown. The work described in this thesis aims at the study of the DOF type regulatory genes to increase tolerance to abiotic stress in both model species and the tomato. In the first chapter of this thesis, we present molecular characterization of the Arabidopsis CDF3 gene as well as its role in the response to abiotic stress and in other developmental processes. AtCDF3 is highly induced by drought, extreme temperatures, salt and abscisic acid (ABA) treatments. The cdf3-1 T-DNA insertion mutant was more sensitive to drought and low temperature stresses, whereas the AtCDF3 overexpression enhanced the tolerance of transgenic plants to drought, cold and osmotic stress comparing to the wild-type (WT) plants. In addition, these plants exhibit increased photosynthesis rates and stomatal aperture. AtCDF3 is localized in the nuclear region, displays specific binding to the canonical DNA target sequences and has a transcriptional activation activity in Arabidopsis protoplast assays. In addition, the C-terminal domain of AtCDF3 is essential for its localization and activation capabilities and the deletion of this domain significantly reduces the tolerance to drought in transgenic 35S::AtCDF3 overexpressing plants. Microarray analysis revealed that AtCDF3 regulated a set of genes involved in nitrogen and carbon metabolism. Our results demonstrate that AtCDF3 plays dual roles in regulating plant responses to drought and low temperature stress and in control of flowering time in vegetative tissues. In the second chapter this work, we carried out to identification of 34 tomato DOF genes that were classified by sequence similarity into four groups A-D, similar to the situation in Arabidopsis. In the D group we have identified five DOF genes that show similar characteristics to the Cycling Dof Factors (CDFs) of Arabidopsis. These genes were considered orthologous to the Arabidopsis CDF1 - 5 and were named Solanum lycopersicum CDFs or SlCDFs. SlCDF1-5 are nuclear proteins that display specific binding to canonical DNA target sequences and have transcriptional activation capacities in vivo. Expression analysis of SlCDF1-5 genes showed distinct diurnal expression patterns and were differentially induced in response to osmotic, salt and low and high temperature stresses. Arabidopsis plants overexpressing SlCDF1 and SlCDF3 showed increased drought and salt tolerance. In addition, various stress-responsive genes, such as AtCOR15, AtRD29A and AtERD10, were expressed differently in these lines. The overexpression of SlCDF3 in Arabidopsis also results in the late flowering phenotype through the modulation of the expression of flowering control genes such CONSTANS (CO) and FLOWERING LOCUS T (FT). Overall, our data connet SlCDFs to undescribed functions related to abiotic stress tolerance and flowering time through the regulation of specific target genes and an increase in particular metabolites.

Control of flowering time in temperate cereals: genes, domestication, and sustainable productivity

The control of flowering is central to reproductive success in plants, and has a major impact on grain yield in crop species. The global importance of temperate cereal crops such as wheat and barley has meant emphasis has long been placed on understanding the genetics of flowering in order to enhance yield. Leads gained from the dissection of the molecular genetics of model species have combined with comparative genetic approaches, recently resulting in the isolation of the first flowering time genes in wheat and barley. This paper reviews the genetics and genes involved in cereal flowering pathways and the current understanding of how two of the principal genes, Vrn and Ppd, have been involved in domestication and adaptation to local environments, and the implications for future breeding programmes are discussed.

Ecophysiology of seed dormancy and the control of germination in early spring-flowering Galanthus nivalis and Narcissus pseudonarcissus (Amaryllidaceae)

Seed dormancy induction and alleviation in the winter-flowering moist temperate woodland species Galanthus nivalis and Narcissus pseudonarcissus are complex and poorly understood. Temperature, light and desiccation were investigated to elucidate their role in the germination ecophysiology of these species. Outdoor and laboratory experiments simulating different seasonal temperatures, seasonal durations, and temperature fluctuations; the presence of light during different seasons; and intermittent drying (during the summer period) over several ‘years’ investigated the importance of these factors in germination. Warm summer-like temperatures (20°C) were necessary for germination at subsequent cooler autumn-like temperatures (greatest at 15°C in G. nivalis and 10°C in N. pseudonarcissus). As the warm temperature duration increased so did germination at subsequent cooler temperatures; further germination occurred in subsequent ‘years’ at cooler temperatures following a second, and also third, warm period. Germination was significantly greater in darkness, particularly in G. nivalis. Dormancy increased with seed maturation period in G. nivalis, because seeds extracted from green capsules germinated more readily than those from yellow. Desiccation increased dormancy in an increasing proportion of N. pseudonarcissus seeds the later they were dried in ‘summer’. Seed viability was only slightly reduced by desiccation in N. pseudonarcissus but was poor and variable in G. nivalis. Shoot formation occurred both at the temperature at which germination was greatest and also if 5°C cooler. In summary, continuous hydration of seeds of both species during warm summer-like temperatures results in the gradual release of seed dormancy; thereafter, darkness and cooler temperatures promote germination. Cold temperatures, increased seed maturity (G. nivalis), and desiccation (N. pseudonarcissus) increase dormancy while light inhibits germination.

Evaluation of fungicide applications to sweet orange at different flowering stages for control of postbloom fruit drop caused by Colletotrichum acutatum

Postbloom fruit drop (PFD), caused by Colletotrichum acutatum, produces blossom blight, fruit abscission and persistent calyces. in groves of Pera-Rio and Natal sweet orange located in Santa Cruz do Rio Pardo and Rincao, São Paulo, Brazil, four experiments were carried out to evaluate the effectiveness of fungicides sprayed alone or as mixtures, at different flowering stages for the control of PFD of citrus. The number of symptomatic flowers, the percentage of fruit set (FS), and the relationship between persistent calyces and total fruit weight per plant were evaluated. The fungicides carbendazim and folpet were sprayed at 0.50 ml and 1.25 ga.i. l(-1) of water, respectively, were superior by all the criteria to the other treatments. Carbendazim and folpet fungicides performed best when they were applied at the green bud through hollow ball stages. Difenoconazole, independent of application timing, was less effective by all criteria used. Application of mancozeb at 1.60 ga.i. l(-1) at the green bud stage followed by application of mancozeb in a tank mix with carbendazim or folpet at 1.0 ml and 1.25 g a.i. l(-1), respectively, during green bud bloom and hollow ball stages were effective for disease control. Carbendazim combined with 0.25% KNO3, reduced the number of persistent calyces and increased fruit production significantly. Applications must be made between green bud and hollow ball stages for best control. Applications only at hollow ball or open flower stages did not provide effective disease control. (C)2007 Elsevier Ltd. All rights reserved.

Photoperiod Control of Gibberellin Levels and Flowering in Sorghum1

Regulation of rhythmic peaks in levels of endogenous gibberellins (GAs) by photoperiod was studied in the short-day monocot sorghum (Sorghum bicolor [L.] Moench). Comparisons were made between three maturity (Ma) genotypes: 58M (Ma1Ma1, Ma2Ma2, phyB-1phyB-1, and Ma4Ma4 [a phytochrome B null mutant]); 90M (Ma1Ma1, Ma2Ma2, phyB-2phyB-2, and Ma4Ma4); and 100M (Ma1Ma1, Ma2Ma2, PHYBPHYB, and Ma4Ma4). Plants were grown for 14 d under 10-, 14-, 16-, 18-, and 20-h photoperiods, and GA levels were assayed by gas chromatography-mass spectrometry every 3 h for 24 h. Under inductive 10-h photoperiods, the peak of GA20 and GA1 levels in 90M and 100M was shifted from midday, observed earlier with 12-h photoperiods, to an early morning peak, and flowering was hastened. In addition, the early morning peaks in levels of GA20 and GA1 in 58M under conditions allowing early flowering (10-, 12-, and 14-h photoperiods) were shifted to midday by noninductive (18- and 20-h) photoperiods, and flowering was delayed. These results are consistent with the possibility that the diurnal rhythm of GA levels plays a role in floral initiation and may be one way by which the absence of phytochrome B causes early flowering in 58M under most photoperiods.

The pattern and control of reproductive development in non-astringent persimmon (Diospyros kaki L.): a review

Non-astringent persimmon is rapidly expanding as a new fruit crop in warm subtropical regions of the world, Most research and development of this fruit crop has occurred in Japan, where there is a considerable amount of published literature on its performance. Much of this information is not readily accessible to other countries and needs to be interpreted and modified for other climatic regions. This paper reviews reproductive events from floral initiation to the completion of fruit growth. The timing and significance of these events is described in relation to the phenological cycle. Method of improving flowering, reducing fruit drop and altering the fruit maturity period are discussed. (C) 1997 Elsevier Science B.V.

Hormone physiology of pea mutants prevented from flowering by mutations gi or veg1

The veg1 (vegetative) mutant in pea (Pisum sativum L.) does not flower under any circumstances and gi (gigas) mutants remain vegetative under certain conditions. gi plants are deficient in production of floral stimulus, whereas veg1 plants lack a response to floral stimulus. During long days in particular, these non-flowering mutant plants eventually enter a stable compact phase characterised by a large reduction in internode length, small leaves and growth of lateral shoots from the upper-stem (aerial) nodes. The first-order laterals in turn produce second-order laterals and so on in a reiterative pattern. The apical bud is reduced in size but continues active growth. Endogenous hormone measurements and gibberellin application studies with gi-1, gi-2 and veg1 plants indicate that a reduction in gibberellin and perhaps indole-3-acetic acid level may account, at least partially, for the compact aerial shoot phenotype. In the gi-1 mutant, the compact phenotype is rescued by transfer from a 24- to an 8-h photoperiod. We propose that in plants where flowering is prevented by a lack of floral stimulus or an inability to respond, the large reduction in photoperiod gene activity during long days may lead to a reduction in apical sink strength that is manifest in an altered hormone profile and weak apical dominance.

Inheritance of late flowering in natural variants of soybean cultivars under short-day conditions

The objective of this work was to determine the inheritance of the long juvenile period trait in natural variants of the Doko, BR 9 (Savana), Davis, Embrapa 1 (IAS 5RC), and BR 16 soybean cultivars. Complete diallel crosses were made between the Doko and BR 16 cultivars and their variants. A 3:1 segregation ratio was observed in the F2 populations of the 'Doko' x Doko-18T, 'Doko' x Doko-Milionária, 'Davis' x São Carlos, and 'BR 9 (Savana)' x MABR92-836 (Savanão) crosses, indicating that the long juvenile period trait is controlled by a pair of recessive genes. The difference in late flowering between the Doko cultivar and both of its variants was caused by a recessive spontaneous mutation at the same genetic locus. However, the variants Doko-18T and Doko-Milionária are identical mutants that share a pair of genes that control the long juvenile period under short-day conditions. These mutants can be used in breeding programs to develop cultivars adapted to low-latitude tropical regions.

Control of tomato bacterial wilt through the incorporation of aerial part of pigeon pea and crotalaria to soil

The use of organic matter that improves the physical, chemical and biological soil properties has been studied as an inducer of suppressiveness to soilborne plant pathogens. The objective of this work was to evaluate the effect of different sources and concentrations of organic matter on tomato bacterial wilt control. Two commercially available organic composts and freshly cut aerial parts of pigeon pea (Cajanus cajan) and crotalaria (Crotalaria juncea) were incorporated, in concentrations of 10, 20 and 30 % (v/v), into soil infested with Ralstonia solanacearum. The soil with the fresh organic matter of pigeon pea and crotalaria was incubated for 30 and 60 days before planting. Tomato seedlings of cv. Santa Clara were transplanted into polyethylene bags with 3 kg of the planting substrate (infested soil + organic matter). The wilting symptoms and percentage of flowering plants were evaluated for 45 days. All evaluated concentrations with incorporation and incubation for 30 days of aerial parts of pigeon pea and crotalaria controlled 100% tomato bacterial wilt. With 60 days of incubation, only the 10 % concentration of pigeon pea and crotalaria did not control the disease. These results suggest that soil incorporation of fresh aerial parts of pigeon pea and crotalaria is an effective method for bacterial wilt control.

Weather requirements and rain forecast to time fungicide application for Fusarium head blight control in wheat

The aim of this study was to determine the minimum conditions of wetness duration and mean temperature required for Fusarium head blight infection in wheat. The weather model developed by Zoldan (2008) was tested in field experiments for two wheat cultivars grown in 2005 (five sowing dates) and 2006 (six sowing dates) in 10 m² plots with three replicates. The disease was assessed according to head incidence (HI), spikelet incidence (SI), and the interaction between these two methods was called head blight severity (HBS). Starting at the beginning of anthesis, air temperature and head wetness duration were daily recorded with an automatic weather station. With the combination of these two factors, a weather favorability table was built for the disease occurrence. Starting on the day of flowering beginning (1 - 5% fully exserted anthers), the sum of daily values for infection favorability (SDVIF) was calculated by means of a computer program, according to Zoldan (2008) table. The initial symptoms of the disease were observed at 3.7% spikelet incidence, corresponding to 2.6 SVDFI. The infection occurs in wheat due to rainfall which results in spike wetting of > 61.4 h duration. Rainfall events forecast can help time fungicide application to control FHB. The name of this alert system is proposed as UPF-scab alert.