RESUMEN
During microspore embryogenesis, microspores are induced to develop into haploid embryos. In Brassica napus, microspore embryogenesis is induced by a heat shock (HS), which initially produces embryogenic structures with different cell wall architectures and compositions, and with different potentials to develop into embryos. The B. napus DH4079 and DH12075 genotypes have high and very low embryo yields, respectively. In DH12075, embryo yield is greatly increased by combining HS and the histone deacetylase (HDAC) inhibitor trichostatin A (TSA). However, we show that HS + TSA inhibits embryogenesis in the highly embryogenic DH4079 line. To ascertain why TSA has such different effects in these lines, we treated DH4079 and DH12075 microspore cultures with TSA and compared the cell wall structure and composition of the different embryogenic structures in both lines, specifically the in situ levels and distribution of callose, cellulose, arabinogalactan proteins and high and low methyl-esterified pectin. For both lines, HS + TSA led to the formation of cell walls unfavorable for embryogenesis progression, with reduced levels of arabinogalactan proteins, reduced cell adhesion of inner walls and altered pectin composition. Thus, TSA effects on cell walls cannot explain their different embryogenic response to TSA. We also applied TSA to DH4079 cultures at different times and concentrations before HS application, with no negative effects on embryogenic induction. These results indicate that DH4079 microspores are hypersensitive to combined TSA and HS treatments, and open up new hypotheses about the causes of such hypersensitivity.
Asunto(s)
Brassica napus , Pared Celular , Genotipo , Respuesta al Choque Térmico , Ácidos Hidroxámicos , Brassica napus/genética , Brassica napus/efectos de los fármacos , Brassica napus/fisiología , Pared Celular/metabolismo , Pared Celular/efectos de los fármacos , Ácidos Hidroxámicos/farmacología , Respuesta al Choque Térmico/efectos de los fármacos , Respuesta al Choque Térmico/genética , Polen/genética , Polen/efectos de los fármacos , Estrés FisiológicoRESUMEN
Yield in many crops is affected by abscission during the early stages of fruitlet development. The reasons for fruitlet abscission are often unclear but they may include genetic factors because, in some crops, self-pollinated fruitlets are more likely to abscise than cross-pollinated fruitlets. Pollen parentage can also affect final fruit size and fruit quality. Here, we aimed to understand the effects of pollen parentage on fruitlet retention and nut quality in orchards of macadamia (Macadamia integrifolia Maiden & Betche). We identified the pollen parent of macadamia 'cultivar '816' embryos by analysing single nucleotide polymorphisms (SNPs) in their DNA using customised MassARRAY and Single Allele Base Extension Reaction (SABER) methods. This allowed us to determine the proportions of self-fertilised and cross-fertilised progeny during premature fruit drop at 6 weeks and 10 weeks after peak anthesis, as well as at nut maturity. We determined how pollen parentage affected nut-in-shell (NIS) mass, kernel mass, kernel recovery, and oil concentration. Macadamia trees retained cross-fertilised fruitlets rather than self-fertilised fruitlets. The percentage of progeny that were cross-fertilised increased from 6% at 6 weeks after peak anthesis to 97% at nut maturity, with each tree producing on average 22 self-fertilised nuts and 881 cross-fertilised nuts. Three of the four cross-pollen parents provided fruit with significantly higher NIS mass, kernel mass, or kernel recovery than the few remaining self-fertilised fruit. Fruit that were cross-fertilised by '842', 'A4', or 'A203' had 16-29% higher NIS mass and 24-44% higher kernel mass than self-fertilised fruit. Nuts that were cross-fertilised by 'A4' or 'A203' also had 5% or 6% higher kernel recovery, worth approximately $US460-540 more per ton for growers than self-fertilised nuts. The highly selective abscission of self-fertilised fruitlets and the lower nut quality of self-fertilised fruit highlight the critical importance of cross-pollination for macadamia productivity.
Asunto(s)
Frutas , Macadamia , Polimorfismo de Nucleótido Simple , Macadamia/genética , Frutas/genética , Frutas/crecimiento & desarrollo , Semillas/genética , Semillas/crecimiento & desarrollo , Autofecundación , Polen/genética , Polen/crecimiento & desarrollo , Polen/efectos de los fármacos , ADN de Plantas/genética , Nueces/genética , Nueces/crecimiento & desarrollo , PolinizaciónRESUMEN
Changing ambient temperature often impairs plant development and sexual reproduction, particularly pollen ontogenesis. However, mechanisms underlying cold stress-induced male sterility are not well understood. Here, we exposed Chinese cabbage (Brassica campestris) to different cold conditions during flowering and demonstrated that the tetrad stage was the most sensitive. After completion of pollen development at optimal conditions, transient cold stress at the tetrad stage still impacted auxin levels, starch and lipid accumulation, and pollen germination, ultimately resulting in partial male sterility. Transcriptome and metabolome analyses and histochemical staining indicated that the reduced pollen germination rate was due to the imbalance of energy metabolism during pollen maturation. The investigation of ß-glucuronidase (GUS)-overexpressing transgenic plants driven by the promoter of DR5 (DR5::GUS report system) combined with cell tissue staining and metabolome analysis further validated that cold stress during the tetrad stage reduced auxin levels in mature pollen grains. Low-concentration auxin treatment on floral buds at the tetrad stage before cold exposure improved the cold tolerance of mature pollen grains. Artificially changing the content of endogenous auxin during pollen maturation by spraying chemical reagents and loss-of-function investigation of the auxin biosynthesis gene YUCCA6 by artificial microRNA technology showed that starch overaccumulation severely reduced the pollen germination rate. In summary, we revealed that transient cold stress at the tetrad stage of pollen development in Chinese cabbage causes auxin-mediated starch-related energy metabolism imbalance that contributes to the decline in pollen germination rate and ultimately seed set.
Asunto(s)
Brassica , Metabolismo Energético , Ácidos Indolacéticos , Polen , Polen/efectos de los fármacos , Polen/genética , Polen/fisiología , Polen/crecimiento & desarrollo , Ácidos Indolacéticos/metabolismo , Metabolismo Energético/efectos de los fármacos , Brassica/genética , Brassica/fisiología , Brassica/metabolismo , Brassica/efectos de los fármacos , Respuesta al Choque por Frío/fisiología , Regulación de la Expresión Génica de las Plantas/efectos de los fármacos , Plantas Modificadas Genéticamente , Frío , Germinación/efectos de los fármacosRESUMEN
Prunus serotina Ehrh. is an alien invasive neophyte widespread in European forests. So far, no effective methods of its elimination have been developed. For this reason, the aim of our study was to determine how herbicides affect the morphological characteristics of pollen grains. This knowledge may be crucial to control this invasive species. The current study was carried out in a research area of 2.7 ha located in the Zielonka Forest near Poznan, Poland (N 52°31'58.016â³, E 17°05'55.588â³). We tested morphological differences among ten features of P. serotina pollen, based on the samples collected from 15 control trees compared to the 50 trees treated with five different herbicides. In total 1950 pollen grains were measured. We confirmed the adopted hypotheses of long-term herbicide influence on P. serotina pollen. Pollen grains from the control trees had a longer equatorial axis, were more elongated in shape and had the largest range of exine thickness compared to the pollen from the herbicide-treated samples. Exine thickness in the control sample was on average 0.74 µm, ranging from 0.42 to 1.19 µm. The average values and the ranges of this trait in the samples treated with herbicides were larger (e.g. average exine thickness was from 0.90 to 0.95 µm). There were differences in the P/E ranges of variability between the control and herbicide-treated samples. In the control sample the P/E ratio was 1.32-2.04 and elongated forms of pollen shapes prevailed, while in the herbicide-treated samples it ranged from 1.03 to 1.47. The share of deformed pollen grains in the herbicide-treated samples was lower than expected, ranging from 8.7 to 25.3%, while in the control samples it was 6%. Logo and Mustang turned out to be the most effective among the herbicides used in the described research. The two used application methods were found to have an effect on pollen quality.
Asunto(s)
Herbicidas , Especies Introducidas , Polen , Prunus avium , Control de Malezas , Herbicidas/toxicidad , Polen/efectos de los fármacos , Polen/ultraestructura , Prunus avium/anatomía & histología , Prunus avium/efectos de los fármacos , Bosques , Control de Malezas/métodos , MicroscopíaRESUMEN
The development of floral organs is coordinated by an elaborate network of homeotic genes, and gibberellin (GA) signaling is involved in floral organ development; however, the underlying molecular mechanisms remain elusive. In the present study, we found that MOS4-ASSOCIATED COMPLEX 5A (MAC5A), which is a protein containing an RNA-binding motif, was involved in the development of sepals, petals, and stamens; either the loss or gain of MAC5A function resulted in stamen malformation and a reduced seed set. The exogenous application of GA considerably exacerbated the defects in mac5a null mutants, including fewer stamens and male sterility. MAC5A was predominantly expressed in pollen grains and stamens, and overexpression of MAC5A affected the expression of homeotic genes such as APETALA1 (AP1), AP2, and AGAMOUS (AG). MAC5A may interact with RABBIT EARS (RBE), a repressor of AG expression in Arabidopsis flowers. The petal defect in rbe null mutants was at least partly rescued in mac5a rbe double mutants. These findings suggest that MAC5A is a novel factor that is required for the normal development of stamens and depends on the GA signaling pathway.
Asunto(s)
Flores/efectos de los fármacos , Giberelinas/farmacología , Proteínas de Unión al ARN/genética , Proteínas de Unión al ARN/metabolismo , Flores/genética , Flores/metabolismo , Regulación de la Expresión Génica de las Plantas/efectos de los fármacos , Genes Homeobox/efectos de los fármacos , Genes Homeobox/genética , Genes de Plantas/efectos de los fármacos , Genes de Plantas/genética , Morfogénesis/efectos de los fármacos , Morfogénesis/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Polen/efectos de los fármacos , Polen/genética , Factores de Transcripción/genética , Factores de Transcripción/metabolismoRESUMEN
Pollen selection affects honeybee colony development and productivity. Considering that pollen is consumed by young in-hive bees, and not by foragers, we hypothesized that young bees learn pollen cues and adjust their preferences to the most suitable pollens. To assess whether young bees show preferences based on learning for highly or poorly suitable pollens, we measured consumption preferences for two pure monofloral pollens after the bees had experienced one of them adulterated with a deterrent (amygdalin or quinine) or a phagostimulant (linoleic acid). Preferences were obtained from nurse-aged bees confined in cages and from nurse bees in open colonies. Furthermore, we tested the bees' orientation in a Y-maze using a neutral odour (Linalool or Nonanal) that had been previously associated with an amygdalin-adulterated pollen. Consumption preferences of bees, both in cages and in colonies, were reduced for pollens that had been adulterated with deterrents and increased for pollens that had been supplemented with linoleic acid. In the Y-maze, individuals consistently avoided the odours that they had previously experienced paired with the deterrent-adulterated pollen. Results show that nurse-aged bees associate pollen-based or pollen-related cues with either a distasteful/malaise experience or a tasty/nutritious event, leading to memories that bias their pollen-mediated response.
Asunto(s)
Amigdalina/química , Abejas/fisiología , Conducta Alimentaria/fisiología , Aprendizaje , Ácido Linoleico/química , Néctar de las Plantas/fisiología , Polen/química , Animales , Contaminación de Alimentos/análisis , Polen/efectos de los fármacosRESUMEN
BACKGROUND: A mannitol stress treatment and a subsequent application of n-butanol, known as a microtubule-disrupting agent, enhance microspore embryogenesis (ME) induction and plant regeneration in bread wheat. To characterize changes in cortical (CMT) and endoplasmic (EMT) microtubules organization and dynamics, associated with ME induction treatments, immunocytochemistry studies complemented by confocal laser scanning microscopy (CLSM) were accomplished. This technique has allowed us to perform advanced 3- and 4D studies of MT architecture. The degree of MT fragmentation was examined by the relative fluorescence intensity quantification. RESULTS: In uni-nucleated mannitol-treated microspores, severe CMT and EMT fragmentation occurs, although a complex network of short EMT bundles protected the nucleus. Additional treatment with n-butanol resulted in further depolymerization of both CMT and EMT, simultaneously with the formation of MT aggregates in the perinuclear region. Some aggregates resembled a preprophase band. In addition, a portion of the microspores progressed to the first mitotic division during the treatments. Bi-nucleate pollen-like structures showed a high MT depolymerization after mannitol treatment and numerous EMT bundles around the vegetative and generative nuclei after n-butanol. Interestingly, bi-nucleate symmetric structures showed prominent stabilization of EMT. CONCLUSIONS: Fragmentation and stabilization of microtubules induced by mannitol- and n-butanol lead to new configurations essential for the induction of microspore embryogenesis in bread wheat. These results provide robust insight into MT dynamics during EM induction and open avenues to address newly targeted treatments to induce ME in recalcitrant species.
Asunto(s)
1-Butanol/farmacología , Manitol/farmacología , Microtúbulos/efectos de los fármacos , Polen/efectos de los fármacos , Triticum/efectos de los fármacos , Microscopía Confocal , Microtúbulos/ultraestructura , Desarrollo de la Planta , Triticum/embriología , Triticum/ultraestructuraRESUMEN
Bees are critical for crop pollination, but there is limited information on levels and sources of pesticide exposure in commercial agriculture. We collected pollen from foraging honey bees and bumble bees returning to colonies placed in blooming blueberry fields with different management approaches (conventional, organic, unmanaged) and located across different landscape settings to determine how these factors affect pesticide exposure. We also identified the pollen and analyzed whether pesticide exposure was correlated with corbicular load composition. Across 188 samples collected in 2 years, we detected 80 of the 259 pesticide active ingredients (AIs) screened for using a modified QuEChERS method. Detections included 28 fungicides, 26 insecticides, and 21 herbicides. All samples contained pesticides (mean = 22 AIs per pollen sample), with pollen collected from bees on conventional fields having significantly higher average concentrations (2019 mean = 882.0 ppb) than those on unmanaged fields (2019 mean = 279.6 ppb). Pollen collected by honey bees had more AIs than pollen collected by bumble bees (mean = 35 vs. 19 AIs detected at each farm, respectively), whereas samples from bumble bees had higher average concentrations, likely reflecting differences in foraging behavior. Blueberry pollen was more common in pollen samples collected by bumble bees (25.9% per sample) than honey bees (1.8%), though pesticide concentrations were only correlated with blueberry pollen for honey bees. Pollen collected at farms with more blueberry in the surrounding landscape had higher pesticide concentrations, mostly AIs applied for control of blueberry pathogens and pests during bloom. However, for honey bees, the majority of AIs detected at each farm are not registered for use on blueberry at any time (55.2% of AIs detected), including several highly toxic insecticides. These AIs therefore came from outside the fields and farms they are expected to pollinate. For bumble bees, the majority of AIs detected in their pollen are registered for use on blueberry during bloom (56.9% of AIs detected), though far fewer AIs were sprayed at the focal farm (16.7%). Our results highlight the need for integrated farm and landscape-scale stewardship of pesticides to reduce exposure to pollinators during crop pollination.
Asunto(s)
Abejas/fisiología , Arándanos Azules (Planta)/fisiología , Plaguicidas/toxicidad , Polen/fisiología , Animales , Arándanos Azules (Planta)/efectos de los fármacos , Polen/química , Polen/efectos de los fármacos , PolinizaciónRESUMEN
The current study was performed on eight years old peach (Prunus persica L. Batsch) trees cv. Florida prince to study the influence of spraying of commercial nano fertilizer on vegetative growth, pollen grain viability, yield, and fruit quality of the "Florida prince" peach cultivar. Furthermore, extracts from the nanofertilizer treated leaves were studied for their bioactivity as insecticidal or bactericidal activities against some stored grain insects and plant bacterial pathogens. Seventy uniform peach trees were sprayed three time as follow: before flowering; during full bloom, and one month later in addition using the water as a control. Commercial silver particales (Ag NPs) at 10, 12.5, and 15 mL/L and zinc particales (Zn NPs) at 2.5, 5 and 7.5 mL/L as recommended level in a randomized complete block design in ten replicates/trees. Spraying Ag NP at 15 mL/L increased shoot diameter, leaf area, total chlorophyll, flower percentage, fruit yield and fruit physical and chemical characteristics, followed by Ag NPs at 12.5 mL/L and Zn NPs at 7.5 mL/L. Moreover, Zn and Ag NPs caused a highly significant effect on pollen viability. Different type of pollen aberrations were detected by Zn NPs treatment. The commercial Ag NPs showed a high increase in pollen viability without any aberrations. The Ag NPs significantly increased the pollen size, and the spores also increased and separated in different localities, searching about the egg for pollination and fertilization. Peach leaves extract was examined for their insecticidal activity against rice weevil (Sitophilus oryzea L.) and the lesser grain borer (Rhyzopertha dominica, Fabricius) by fumigation method. The antibacterial activity of all treatments was also performed against molecularly identified bacteria. Ag NPs treated leaves extract at concentration 3000 µg/mL were moderate sufficient to inhibit all the bacterial isolates with inhibition zone (IZ) ranged 6-8.67 mm with high efficiency of acetone extracts from leaves treated with Ag NPs compared with Zn NPs. Also, S. oryzae was more susceptible to acetone extracts from leaves treated with both nanomaterials than R. dominica.
Asunto(s)
Prunus persica/efectos de los fármacos , Prunus persica/metabolismo , Antibacterianos/farmacología , Antiinfecciosos/farmacología , Clorofila/metabolismo , Fertilizantes , Florida , Insecticidas/farmacología , Nanopartículas del Metal , Extractos Vegetales/metabolismo , Hojas de la Planta/efectos de los fármacos , Polen/efectos de los fármacos , Polinización/efectos de los fármacos , Plata , Árboles/efectos de los fármacos , ZincRESUMEN
Pollinators and other insects are experiencing an ongoing worldwide decline. While various environmental stressors have been implicated, including pesticide exposure, the causes of these declines are complex and highly debated. Fungicides may constitute a particularly prevalent threat to pollinator health due to their application on many crops during bloom, and because pollinators such as bees may consume fungicide-tainted pollen or nectar. In a previous study, consumption of pollen containing the fungicide Pristine® at field-relevant concentrations by honey bee colonies increased pollen foraging, caused earlier foraging, lowered worker survival, and reduced colony population size. Because most pollen is consumed by young adults, we hypothesized that Pristine® (25.2% boscalid, 12.8% pyraclostrobin) in pollen exerts its negative effects on honey bee colonies primarily on the adult stage. To rigorously test this hypothesis, we used a cross-fostering experimental design, with bees reared in colonies provided Pristine® incorporated into pollen patties at a supra-field concentration (230 mg/kg), only in the larvae, only in the adult, or both stages. In contrast to our predictions, exposure to Pristine® in either the larval or adult stage reduced survival relative to control bees not exposed to Pristine®, and exposure to the fungicide at both larval and adult stages further reduced survival. Adult exposure caused precocious foraging, while larval exposure increased the tendency to forage for pollen. These results demonstrate that pollen containing Pristine® can induce significant negative effects on both larvae and adults in a hive, though the magnitude of such effects may be smaller at field-realistic doses. To further test the potential negative effects of direct consumption of Pristine® on larvae, we reared them in vitro on food containing Pristine® at a range of concentrations. Consumption of Pristine® reduced survival rates of larvae at all concentrations tested. Larval and adult weights were only reduced at a supra-field concentration. We conclude that consumption of pollen containing Pristine® by field honey bee colonies likely exerts impacts on colony population size and foraging behavior by affecting both larvae and adults.
Asunto(s)
Abejas/fisiología , Compuestos de Bifenilo/toxicidad , Fungicidas Industriales/toxicidad , Niacinamida/análogos & derivados , Estrobilurinas/toxicidad , Animales , Fungicidas Industriales/farmacología , Insectos , Larva/efectos de los fármacos , Niacinamida/toxicidad , Plaguicidas/toxicidad , Néctar de las Plantas , Polen/efectos de los fármacos , PolinizaciónRESUMEN
Insect pollinators are threatened by multiple environmental stressors, including pesticide exposure. Despite being important pollinators, solitary ground-nesting bees are inadequately represented by pesticide risk assessments reliant almost exclusively on honeybee ecotoxicology. Here we evaluate the effects of realistic exposure via squash crops treated with systemic insecticides (Admire-imidacloprid soil application, FarMore FI400-thiamethoxam seed-coating, or Coragen-chlorantraniliprole foliar spray) for a ground-nesting bee species (Hoary squash bee, Eucera pruinosa) in a 3-year semi-field experiment. Hoary squash bees provide essential pollination services to pumpkin and squash crops and commonly nest within cropping areas increasing their risk of pesticide exposure from soil, nectar, and pollen. When exposed to a crop treated at planting with soil-applied imidacloprid, these bees initiated 85% fewer nests, left 5.3 times more pollen unharvested, and produced 89% fewer offspring than untreated controls. No measurable impacts on bees from exposure to squash treated with thiamethoxam as a seed-coating or foliage sprayed with chlorantraniliprole were found. Our results demonstrate important sublethal effects of field-realistic exposure to a soil-applied neonicotinoid (imidacloprid) on bee behaviour and reproductive success. Soil must be considered a potential route of pesticide exposure in risk assessments, and restrictions on soil-applied insecticides may be justified, to mitigate impacts on ground-nesting solitary bee populations and the crop pollination services they provide.
Asunto(s)
Abejas/efectos de los fármacos , Insecticidas/farmacología , Neonicotinoides/farmacología , Densidad de Población , Animales , Canadá , Productos Agrícolas , Ambiente , Polen/efectos de los fármacosRESUMEN
Fern spores and pollen are haploid plant germplasm of microscopic nature that can be used to regenerate full plants through germination (fern spores) or to fertilize seed-bearing plants through breeding programs (pollen). Due to their short life span in conventional storage (i.e., dry at -20 °C), the use of cryopreservation has been indicated for long-term ex situ conservation. While fern spores of most species and pollen from many seeded plants tolerate desiccation and can be stored dry at liquid nitrogen temperatures, some pollen is desiccation sensitive, and cryopreservation protocols require controlled drying and cooling and some level of cryoprotection. In this chapter we describe the cryopreservation process for fern spores used in the Millennium Seed Bank of Royal Botanic Gardens, Kew, including some details of the fern spores harvest and cleaning methods. In addition, two protocols for pollen cryopreservation are described, one generic for desiccation-tolerant pollen that can be used for multiple species and one specific for a desiccation sensitive pollen (Zea mays).
Asunto(s)
Técnicas de Cultivo de Célula/métodos , Criopreservación/métodos , Crioprotectores/farmacología , Helechos/citología , Polen/citología , Esporas/citología , Venas Umbilicales/citología , Proliferación Celular , Células Cultivadas , Helechos/efectos de los fármacos , Polen/efectos de los fármacos , Esporas/efectos de los fármacos , Venas Umbilicales/efectos de los fármacosRESUMEN
Pollen grain was explored as a new carrier for enzyme immobilization. After being modified with boric acid-functionalized titania, the pollen grain was able to covalently immobilize glycosylated enzymes by boronate affinity interaction under very mild experimental conditions (e.g., pH 7.0, ambient temperature and free of organic solvent). The glucose oxidase and horse radish peroxidase-immobilized pollen grain became a bienzyme system. The pollen grain also worked as an indicator of the cascade reaction by changing its color. A rapid, simple and cost-effective approach for the visual detection of glucose was then developed. When the glucose concentration exceeded 0.5 mM, the color change was observable by the naked eye. The assay of glucose in body fluid samples exhibited its great potential for practical application.
Asunto(s)
Bioensayo/métodos , Enzimas Inmovilizadas/química , Glucosa Oxidasa/química , Glucosa/análisis , Peroxidasa de Rábano Silvestre/química , Polen/química , Bioensayo/instrumentación , Glucemia/análisis , Ácidos Bóricos/química , Color , Humanos , Concentración de Iones de Hidrógeno , Cinética , Microscopía Electrónica de Rastreo , Polen/efectos de los fármacos , Polen/ultraestructura , Solventes/química , Temperatura , Titanio/químicaRESUMEN
A biological assessment of environmental quality was performed using the tropical plant species Tradescantia pallida (Rose) D.R. Hunt. var. purpurea exposed to different levels of air contamination in urban intersections with high volume of vehicle traffic. Air quality (average daily levels of particulate material in the PM1, 2.5, 10 fractions) and traffic volume in crossing intersections were monitored for 30 days before the collection of plants. Frequency of micronuclei and pollen abortivity in inflorescences collected at different intersections with gradual levels of traffic volume were evaluated as biomarkers of genotoxicity. In addition, the concentrations of bioaccumulated heavy metals in the leaves of the collected plants were also investigated. The proposed biological assessment model found a positive association between the environmental variables (traffic volume; concentration of particulate material) and biological effects (leaf concentration of Cr and Cd; micronucleus frequencies and pollen abortivity).
Asunto(s)
Contaminantes Atmosféricos/toxicidad , Daño del ADN , Tradescantia/genética , Contaminación por Tráfico Vehicular/efectos adversos , Salud Urbana , Contaminación del Aire/análisis , Monitoreo del Ambiente/métodos , Metales Pesados/farmacología , Pruebas de Micronúcleos , Pruebas de Mutagenicidad , Hojas de la Planta/química , Hojas de la Planta/efectos de los fármacos , Polen/efectos de los fármacosRESUMEN
Self-incompatibility (SI) is used by many angiosperms to prevent self-fertilization and inbreeding. In common poppy (Papaver rhoeas), interaction of cognate pollen and pistil S-determinants triggers programmed cell death (PCD) of incompatible pollen. We previously identified that reactive oxygen species (ROS) signal to SI-PCD. ROS-induced oxidative posttranslational modifications (oxPTMs) can regulate protein structure and function. Here, we have identified and mapped oxPTMs triggered by SI in incompatible pollen. Notably, SI-induced pollen had numerous irreversible oxidative modifications, while untreated pollen had virtually none. Our data provide a valuable analysis of the protein targets of ROS in the context of SI-induction and comprise a benchmark because currently there are few reports of irreversible oxPTMs in plants. Strikingly, cytoskeletal proteins and enzymes involved in energy metabolism are a prominent target of ROS. Oxidative modifications to a phosphomimic form of a pyrophosphatase result in a reduction of its activity. Therefore, our results demonstrate irreversible oxidation of pollen proteins during SI and provide evidence that this modification can affect protein function. We suggest that this reduction in cellular activity could lead to PCD.
Asunto(s)
Papaver/fisiología , Proteínas de Plantas/metabolismo , Polen/fisiología , Autoincompatibilidad en las Plantas con Flores/fisiología , Actinas/metabolismo , Secuencia de Aminoácidos , Aminoácidos/metabolismo , Proteínas del Citoesqueleto/metabolismo , Peróxido de Hidrógeno/toxicidad , Pirofosfatasa Inorgánica/metabolismo , Nitrosación , Oxidación-Reducción , Papaver/efectos de los fármacos , Péptido Hidrolasas/metabolismo , Péptidos/metabolismo , Proteínas de Plantas/química , Polen/efectos de los fármacos , Tubo Polínico/efectos de los fármacos , Tubo Polínico/metabolismo , Complejo de la Endopetidasa Proteasomal/metabolismo , Procesamiento Proteico-Postraduccional/efectos de los fármacos , Autoincompatibilidad en las Plantas con Flores/efectos de los fármacos , SolubilidadRESUMEN
KEY MESSAGE: After cryopreservation, the occurrence of apoptosis-like programmed cell death events induced by the accumulation of ROS reduces pollen viability. Cryopreservation, as a biotechnological means for long-term preservation of pollen, has been applied to many species. However, after cryopreservation, the viability of pollen significantly decreases via a mechanism that is not completely clear. In this study, the pollen of Paeonia lactiflora 'Zi Feng Chao Yang', which exhibits significantly reduced viability after liquid nitrogen (LN2) storage, was used to study the relationship among pollen viability, programmed cell death (PCD) and reactive oxygen species (ROS). The apoptosis rate was increased significantly in pollen with decreased viability after cryopreservation, and the changes in ROS generation and hydrogen peroxide (H2O2) were consistent with the apoptosis rate. Correlation analysis results showed that the apoptosis rate is positively correlated with ROS generation and H2O2 content. In addition, ascorbic acid (AsA), glutathione (GSH) and ascorbic acid reductase (APX) levels were significantly correlated with ROS and H2O2. After LN2 preservation for 8 months, the exogenous antioxidants AsA and GSH at appropriate concentrations significantly decreased H2O2 content, inhibited PCD indicator levels, and increased cryopreserved pollen viability. These observations suggest that PCD occurred in pollen during LN2 preservation for 1-8 months and was induced by the accumulation of ROS in pollen after cryopreservation, thus explaining the main reasons for the reduction in pollen viability after cryopreservation in LN2.
Asunto(s)
Apoptosis , Criopreservación , Paeonia/citología , Paeonia/fisiología , Polen/citología , Polen/fisiología , Especies Reactivas de Oxígeno/metabolismo , Supervivencia Tisular , Antioxidantes/metabolismo , Apoptosis/efectos de los fármacos , Ácido Ascórbico/farmacología , Glutatión/farmacología , Humedad , Estrés Oxidativo/efectos de los fármacos , Paeonia/efectos de los fármacos , Polen/efectos de los fármacos , Supervivencia Tisular/efectos de los fármacosRESUMEN
Although exogenous melatonin can enhance the drought tolerance of plants, reports on the role of melatonin in drought tolerance in male reproductive organs are limited. To explore this, a pot experiment was conducted with cotton cultivar Yuzaomian 9110 to study the effects of exogenous melatonin (100, 200, and 1000 µM) on male fertility and related carbohydrate metabolism in anther under drought. Results showed that drought inhibited the translocation of carbon assimilates to anthers, however, melatonin application (100 and 200 µM) significantly improved the translocation of carbon assimilates to drought-stressed anthers. Drought reduced the deposition of starch, the hydrolysis of sucrose into hexoses, the generation of adenosine triphosphate (ATP) in anthers, restricting pollen viability and germination. Nevertheless, the appropriate melatonin concentrations (100 and 200 µM) increased the starch accumulation by enhancing ADP-glucose pyrophosphorylase and soluble starch synthases activities and accelerated the hydrolysis of sucrose by increasing sucrose synthase, acid and alkaline invertases activities in drought-stressed anthers. Appropriate melatonin concentrations (100 and 200 µM) also could help to generate more ATP for reproductive activities of drought-stressed anthers, finally increasing the pollen viability and germination of drought-stressed plants. These findings suggest that drought inhibited male fertility of cotton, but a precise melatonin application could regulate the carbohydrate balance of drought-stressed anthers to improve male fertility. This is the first report demonstrating the important role of exogenous melatonin in improving male fertility under drought conditions by regulating the carbohydrate metabolism in the male part of cotton.
Asunto(s)
Metabolismo de los Hidratos de Carbono , Sequías , Gossypium , Melatonina , Polen , Metabolismo de los Hidratos de Carbono/efectos de los fármacos , Fertilidad/efectos de los fármacos , Gossypium/efectos de los fármacos , Melatonina/farmacología , Polen/efectos de los fármacosRESUMEN
Graphene related materials (GRMs) are currently being used in products and devices of everyday life and this strongly increases the possibility of their ultimate release into the environment as waste items. GRMs have several effects on plants, and graphene oxide (GO) in particular, can affect pollen germination and tube growth due to its acidic properties. Despite the socio-economic importance of sexual reproduction in seed plants, the effect of GRMs on this process is still largely unknown. Here, Corylus avellana L. (common Hazel) pollen was germinated in-vitro with and without 1-100⯵gâ¯mL-1 few-layer graphene (FLG), GO and reduced GO (rGO) to identify GRMs effects alternative to the acidification damage caused by GO. At 100⯵gâ¯mL-1 both FLG and GO decreased pollen germination, however only GO negatively affected pollen tube growth. Furthermore, GO adsorbed about 10 % of the initial Ca2+ from germination media accounting for a further decrease in germination of 13 % at the pH created by GO. In addition, both FLG and GO altered the normal tip-focused reactive oxygen species (ROS) distribution along the pollen tube. The results provided here help to understand GRMs effect on the sexual reproduction of seed plants and to address future in-vivo studies.
Asunto(s)
Corylus/efectos de los fármacos , Grafito/toxicidad , Reproducción/efectos de los fármacos , Calcio/metabolismo , Supervivencia Celular/efectos de los fármacos , Flores/efectos de los fármacos , Concentración de Iones de Hidrógeno , Polen/efectos de los fármacos , Tubo Polínico/efectos de los fármacos , Especies Reactivas de Oxígeno/metabolismoRESUMEN
Effective mutagenesis is critical for connecting traits of interest to specific plant genes. The development of site-directed mutagenesis and sequenced-indexed genetics resources in maize allows for targeted analysis of individual genes. These reverse genetics approaches have the potential for confirmation bias by only studying candidate genes for association with traits of interest. Genetic screens of induced, random mutations are important for identifying novel loci as well as interacting factors for known mutant loci. Chemical mutagenesis provides very high mutation rates and can be used for a variety of screen designs. This chapter provides an updated protocol for ethyl methanesulfonate (EMS) mutagenesis of maize pollen using paraffin or mineral oil. Mutagenesis occurs in mature pollen causing nonconcordant endosperm and embryo genotypes as well as sectored M1 plants. Considerations for these factors in genetic screens are discussed.
Asunto(s)
Metanosulfonato de Etilo/farmacología , Mutagénesis/efectos de los fármacos , Mutágenos/farmacología , Polen/efectos de los fármacos , Zea mays/efectos de los fármacos , Endospermo/efectos de los fármacos , Endospermo/genética , Genes de Plantas/efectos de los fármacos , Mutación/efectos de los fármacos , Tasa de Mutación , Polen/genética , Zea mays/genéticaRESUMEN
Increasing use of silver nanoparticles (AgNPs) in myriad applications including electronics, medicines and agriculture has led to serious concerns regarding its release to plant ecosystems. Over the years, numerous studies have demonstrated the toxic impact of AgNPs in a variety of cell and tissue systems involved in vegetative growth across a wide range of plant species. However, assessing their impact on haploid phase of plant life cycle was restricted only to a study with Kiwifruit. In this study, in vitro pollen performance of Peltophorum pterocarpum at two endpoints i.e., germination and tube growth was assessed to evaluate the impact of nanoparticulate or ionic form of silver. Increasing concentrations of AgNO3/AgNPs significantly reduced the pollen germination and retarded the tube growth. The EC 50 values indicated a more potent toxic effect of AgNPs than AgNO3 on pollen germination as well as tube growth. Impairment of pollen performance was more pronounced at the stage of emergence of pollen tube. Extensive alterations in the muri and lumen of exine as revealed through SEM analysis and subsequent blockage of germpore might disrupt the emergence of pollen tube. The dynamics of pollen tube growth was analyzed with polynomial models of different degrees. A high degree of polynomial, the quintic model was able to approximate the real data points with highest coefficient of determination and smallest RMSE, compared to other models. An oscillating pattern of tube growth was portrayed with the passage of time in all the treatments that fits well with the established mechanistic oscillatory model of tube growth. It appears that exposure to AgNO3/AgNPs inhibited pollen germination and retarded tube growth without affecting the oscillatory behavior of tip-growth.