RESUMO
The main difficulty for the cultivation and conservation of bromeliad species is the reduced number of propagules and slow growth of many of the species, resulting in a low propagation efficiency. Bromeliad plants are hardy and relatively easy to cultivate, with a high ornamental and ecological importance. Aiming at efficient micropropagation rates of V. hieroglyphica, a highly valued bromeliad, with very low propagation efficiency, a temporary immersion system was used and compared to semisolid and liquid static medium. Cultures obtained from in vitro germinated seeds were used as explants, maintaining their genetic diversity. Micropropagation with this simple temporary immersion system, composed of two autoclavable flasks, each with one opening for the attachment of 22 µm syringe filters, connected by a rubber stopper and an inner glass tube. In the bottom flask, an air valve is attached to the filter, which is subsequently connected to an aquarium pump and a timer and plugged to an outlet. This simple temporary immersion system showed improved micropropagation efficiency and is a method that can also be evaluated for other species.
Assuntos
Bromeliaceae , Imersão , Catéteres , Reprodução , SementesRESUMO
The use of temporary immersion systems (TIS) for plant micropropagation is an efficient technique for plant production, and we have applied it for the production of alstroemerias. This method involves the cultivation of explants such as rhizomes and axillary buds in a nutrient medium to stimulate shoot growth. TIS offer advantages such as accelerated multiplication processes, uniform production, and cost reduction. This process has shown promise in meeting the growing demand for alstroemeria plants in the market. This chapter describes a specific protocol for temporary immersion bioreactor micropropagation of the "Albatroz" cultivar, with the potential for large-scale automation.
Assuntos
Alstroemeria , Imersão , Automação , Reatores Biológicos , NutrientesRESUMO
KEY MESSAGE: Lastly, the bZIP gene family encompasses genes that have been reported to play a role in flower development, such as bZIP14 (FD). Notably, bZIP14 is essential for Flowering Locus T (FT) initiation of floral development in Arabidopsis (Abe et al. 2005). Cotton (Gossypium hirsutum L.) is the world's most extensively cultivated fiber crop. However, its reproductive development is poorly characterized at the molecular level. Thus, this study presents a detailed transcriptomic analysis of G. hirsutum at three different reproductive stages. We provide evidence that more than 64,000 genes are active in G. hirsutum during flower development, among which 94.33% have been assigned to functional terms and specific pathways. Gene set enrichment analysis (GSEA) revealed that the biological process categories of floral organ development, pollen exine formation, and stamen development were enriched among the genes expressed during the floral development of G. hirsutum. Furthermore, we identified putative Arabidopsis homologs involved in the G. hirsutum gene regulatory network (GRN) of pollen and flower development, including transcription factors such as WUSCHEL (WUS), INNER NO OUTER (INO), AGAMOUS-LIKE 66 (AGL66), SPOROCYTELESS/NOZZLE (SPL/NZZ), DYSFUNCTIONAL TAPETUM 1 (DYT1), ABORTED MICROSPORES (AMS), and ASH1-RELATED 3 (ASHR3), which are known crucial genes for plant reproductive success. The cotton MADS-box protein-protein interaction pattern resembles the previously described patterns for AGAMOUS (AG), SEEDSTICK (STK), SHATTERPROOF (SHP), and SEPALLATA3 (SEP3) homolog proteins from Arabidopsis. In addition to serving as a resource for comparative flower development studies, this work highlights the changes in gene expression profiles and molecular networks underlying stages that are valuable for cotton breeding improvement.
Assuntos
Flores , Regulação da Expressão Gênica de Plantas , Redes Reguladoras de Genes , Gossypium , Gossypium/genética , Gossypium/crescimento & desenvolvimento , Gossypium/fisiologia , Flores/genética , Flores/crescimento & desenvolvimento , Reprodução/genética , Transcriptoma , Perfilação da Expressão Gênica , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Arabidopsis/genética , Arabidopsis/crescimento & desenvolvimento , Arabidopsis/fisiologiaRESUMO
Barbacenia graminifolia is a Velloziaceae species endemic to the campos rupestres in Serra do Cipó, Minas Gerais state (Brazil). This biome is characterised by high irradiance and limited water conditions. Unlike other resurrection plants, B. graminifolia can maintain a high hydric status (>80%) after 28 days of water suppression before desiccation. We investigated the physiological and metabolic mechanisms associated with structural changes that allow B. graminifolia to maintain hydration under a prolonged water deficit and to recover after desiccation. After 30 days of water deficit, desiccated plants exhibited chlorophyll degradation, a 178.4% and 193.7% increase in total carotenoids and MDA, respectively, and twice the CAT and APX activity compared to hydrated plants. The metabolite profile showed increased amino acids, carbohydrates, saturated fatty acids and benzoic acids during dehydration, while trichloroacetic acid cycle acids were higher in hydrated and rehydrated plants. Anatomical and ultrastructural data corroborated the physiological and metabolic changes and revealed the presence of mucilaginous cells with high water retention capacity. Our data indicated that combined strategies of assimilatory metabolism shutdown, accumulation of compatible solutes and antioxidant compounds, increase in hydrophilic molecules, changes in the composition of membrane lipids and remodelling of cell organelles conditioned the efficiency of B. graminifolia in delaying water loss, tolerating further desiccation and quickly recovering after rehydration. These attributes evidence that this species is well adapted to cope with adverse environmental conditions, mainly directing the metabolism to an efficient antioxidant response and improving its capacity to retain water during the dry season.
Assuntos
Antioxidantes , Dessecação , Antioxidantes/metabolismo , Ácido Tricloroacético , Água/metabolismo , Plantas/metabolismo , Clorofila/metabolismo , Carotenoides , Carboidratos , Lipídeos de Membrana , Ácidos Graxos , Benzoatos , AminoácidosRESUMO
Protoplasts are an attractive explant source for biotechnological tools widely used on citrus genetic improvement, such as somatic hybridization and direct genetic transformation. These delicate and responsive materials are subjected to cell proliferation induction and differentiation of somatic embryos which further regenerate into entire plants. The isolation of viable protoplasts followed by regeneration of plants through somatic embryogenesis is an important methodology for breeding applications. The methods presented here can also be used as a reference for protoplast work in other species, followed by protocol optimization for different species/genotypes.
Assuntos
Citrus , Protoplastos , Citrus/genética , Desenvolvimento Embrionário , Melhoramento Vegetal , Técnicas de Embriogênese Somática de Plantas/métodosRESUMO
Somatic embryogenesis has been obtained in many citrus cultivars; however, the efficiency of the system is genotype dependent and culture synchronization is important to reach more efficient systems. In this chapter we present a detailed protocol of somatic embryogenesis induction from nucellar tissue and the use of an alternative method of callus sieving for culture synchronization and embryo production. This is a simple method which can also be evaluated for other species aiming at better culture efficiency and somatic embryo production.
Assuntos
Citrus , Citrus/genética , Desenvolvimento Embrionário , Técnicas de Embriogênese Somática de Plantas/métodosRESUMO
In situ hybridization with mRNA probes enables the detection and localization of gene expression in plant somatic embryogenesis samples. BbrizSERK is a gene that is expressed in embryogenic cells and tissues of Brachiaria. Here we describe methods used for in situ hybridization to localize BbrizSERK transcripts during somatic embryogenesis of Brachiaria brizantha according to the plant material and observations intended, using paraffin or butyl methyl methacrylate resin-embedded samples, as well as a method for whole-mount preparation applicable for the analysis of other genes involved in embryogenic processes, along with other in vitro processes.
Assuntos
Brachiaria , Brachiaria/genética , Desenvolvimento Embrionário , Hibridização In SituRESUMO
With its accumulation in upland rice, cadmium (Cd) can easily enter the human food chain, which poses a global health threat considering nearly half of the human population depends on rice as a staple food source. A study was conducted to (1) evaluate Cd accumulation by rice cultivars, grown in Cd-polluted Tropical Oxisols, with different levels of Cd tolerance; (2) quantify Cd transfer from soil to rice shoots and grain; and (3) estimate daily Cd intake by humans. Three rice cultivars, characterized by low (Cateto Seda-CS), medium (BRSMG Talento-BT), and high (BRSMG Caravera-BC) Cd uptake capacity, were investigated. Rice cultivars were exposed to increasing soil Cd concentrations (0.0, 0.7, 1.3, 3.9, 7.8, and 11.7 mg kg-1). Analysis was performed on soil, shoots, and grain. Shoot biomass and grain yield decreased with increasing Cd supply, suggesting the following Cd tolerance: CS > BT > BC. Cadmium concentrations in shoots and grain increased when exposed to Cd. Only CS did not exceed the maximum Cd limit permitted in food (0.40 mg kg-1), when rates up to 1.3 mg kg-1 of Cd were applied to soil. Considering daily rice consumption levels in Brazil, Cd intake often exceeds maximum tolerable levels. Continuous monitoring of soil Cd concentrations is a pivotal step in avoiding hazards to humans. Such monitoring is important on a global scale since outside of Asia, Brazil is the leading rice-producing and rice-consuming country.
Assuntos
Oryza , Poluentes do Solo , Cádmio/análise , Monitoramento Ambiental , Humanos , Solo , Poluentes do Solo/análiseRESUMO
MAIN CONCLUSION: Characterization of anther and ovule developmental programs and expression analyses of stage-specific floral marker genes in Gossypium hirsutum allowed to build a comprehensive portrait of cotton flower development before fiber initiation. Gossypium hirsutum is the most important cotton species that is cultivated worldwide. Although cotton reproductive development is important for fiber production, since fiber is formed on the epidermis of mature ovules, cotton floral development remains poorly understood. Therefore, this work aims to characterize the cotton floral morphoanatomy by performing a detailed description of anther and ovule developmental programs and identifying stage-specific floral marker genes in G. hirsutum. Using light microscopy and scanning electron microscopy, we analyzed anther and ovule development during 11 stages of flower development. To better characterize the ovule development in cotton, we performed histochemical analyses to evaluate the accumulation of phenolic compounds, pectin, and sugar in ovule tissues. After identification of major hallmarks of floral development, three key stages were established in G. hirsutum floral development: in stage 1 (early-EF), sepal, petal, and stamen primordia were observed; in stage 2 (intermediate-IF), primordial ovules and anthers are present, and the differentiating archesporial cells were observed, marking the beginning of microsporogenesis; and in stage 6 (late-LF), flower buds presented initial anther tapetum degeneration and microspore were released from the tetrad, and nucellus and both inner and outer integuments are developing. We used transcriptome data of cotton EF, IF and LF stages to identify floral marker genes and evaluated their expression by real-time quantitative PCR (qPCR). Twelve marker genes were preferentially expressed in a stage-specific manner, including the putative homologs for AtLEAFY, AtAPETALA 3, AtAGAMOUS-LIKE 19 and AtMALE STERILITY 1, which are crucial for several aspects of reproductive development, such as flower organogenesis and anther and petal development. We also evaluated the expression profile of B-class MADS-box genes in G. hirsutum floral transcriptome (EF, IF, and LF). In addition, we performed a comparative analysis of developmental programs between Arabidopsis thaliana and G. hirsutum that considered major morphoanatomical and molecular processes of flower, anther, and ovule development. Our findings provide the first detailed analysis of cotton flower development.
Assuntos
Flores , Regulação da Expressão Gênica de Plantas , Gossypium , Flores/anatomia & histologia , Flores/genética , Perfilação da Expressão Gênica , Gossypium/genética , Gossypium/crescimento & desenvolvimento , Óvulo Vegetal/genéticaRESUMO
MAIN CONCLUSION: In Brachiaria brizantha BbrizSERK1, BbrizSERK2 and BbrizSERK3 were identified. SERK expression marks somatic embryogenesis, sexual MMC, and sexual and apomictic PMC. BbrizSERK3 might have a regulatory role in reproductive development. Somatic embryogenesis receptor-like kinase (SERK) consists of plasma membrane receptor genes that have been characterized in various species, associated with several aspects of plant development, including reproduction. SERK genes are involved in anther development and in early embryo development in sexual and asexual seed formation. To comprehend the complexity of the SERK genes and their function in Brachiaria reproduction, we performed a homology-based search in a genomic database of a sexual B. brizantha and identified sequences of three SERK genes, BbrizSERK1, BbrizSERK2, and BbrizSERK3. RNASeq data showed equivalent abundance of BbrizSERK1 and BbrizSERK2 transcripts in ovaries at early megasporogenesis of sexuals and apomicts, while BbrizSERK3 transcripts were more abundant in ovaries of sexuals than in apomicts. BbrizSERK3 results in three coding sequences due to alternative splicing, among them Variant 1 results in a protein with all the predicted domains of a SERK. BbrizSERK transcripts were detected in male reproductive tissues of both sexual and apomictic plants, suggesting a role in controlling anther development. BbrizSERK transcripts were detected early in ovule development, in the integuments, and in the megaspore mother cell of the sexual plant, but not in the cells that give rise to apomictic embryo sacs, suggesting a role in female reproductive development of sexuals. This paper provides evidences that SERK genes plays a role in the onset and establishment of somatic embryogenesis and in the reproductive development of B. brizantha and suggests a distinct role of BbrizSERK in apomixis initiation.