Crotalaria ochroleuca (Fabaceae) is rich in bioactive compounds used for nematode control. Transformed root culture allows the production of a large amount of hairy root biomass, conditioned by auxin biosynthesis-related genes integrated via the process of Agrobacterium rhizogenes infection, which is known to cause a hairy root phenotype. We transformed A. rhizogenes to increase the biomass of hairy roots and optimized the process of production of bioactive compounds. The transformed nature of hairy roots was confirmed by polymerase chain reaction, which confers kanamycin resistance. The transformed roots were cultured in full-strength in liquid media to realize the growth of hairy roots and the production of bioactive compounds. The transformed roots were grown in a culture medium supplemented with elicitors to produce total phenols, and flavonoids. Chromatographic analysis of transformed roots revealed the presence of flavonoids apigenin-6,8-C-diglucoside and luteolin-6-C-glucoside. The results were obtained by conducting physiological and biochemical studies with the Flavonoids and studying the pathways that led to the production of large amounts of bioactive compounds from the hairy roots of C. ochroleuca. It was observed that the extraction of the compounds significantly affected nematodes and insect larvae, resulting in significantly high levels of economic damage to crops.

Abstract The aim of this work was to evaluate the effect of five light qualities [fluorescent lamps (FL); white (WL), red (RL), blue (BL), or red/blue LEDs (RBL)] and two types of flask sealing (with or without porous membranes) in the in vitro germination and growth of two Capsicum frutescens cultivars (Espaguetinho and Cayenne). For this, two seeds were inoculated per flask (350 mL) containing 50 mL of MS medium with 100 mg L-1 myo-inositol, 30 g L-1 sucrose, 6.0 g L-1 agar, and pH 5.8 ± 0.1. The plants were kept under culture room conditions (16-h photoperiod, 25 ± 2°C and irradiance of 40 μmol m-2 s-1). The experiment was arranged in CRD, 2x5x2 factorial scheme (Cultivar X Light qualities X Sealing type). At 42 days, the following traits were evaluated: percentage of germination; root and shoot length; number of green leaves, senescent leaves and nodal segments; and fresh and dry weight. Plants grown under RL and BRL, showed greater number of green leaves, and longer shoots and roots. The use of porous membranes remarkably reduced the number of senescent leaves. Thus, the use of porous membranes, RL and BRL are recommended since they improve growth and development of pepper in vitro.

Abstract Intraspecies or interspecies crossings transfer relevant alleles between plants. However, some interspecies crossings involving Passiflora species impede ovule fertilization and the viable development of seeds. Thus, the purpose of this study was to verify the viability of interspecific crossings and monitor pollen tube development. The experiment had six species of Passiflora in the reciprocal crossings. Histochemical tests aimed to evaluate the percentage of intraspecies or interspecies crossings that resulted in fruit development and pollen tube development. Ovule fertilization and fruit development occurred in determined directions of crossings when controlling the female or male genitor, but only one case of reciprocal crossing had success. In crossings with no fruit development, histological analysis showed that some callus developed in the stigma and style, confirming unilateral and interspecies incompatibility in the genus Passiflora to some species and some directions of crossings.

Abstract: Somatic embryogenesis from explants from hermaphrodite papaya mother plants is an alternative for the production of true-to-type plants without the need for sexing. This study aimed to analyze hormonal and osmotic inducers in different somatic embryogenesis stages in the commercial hermaphrodite hybrid papaya UENF/Caliman 01. Leaf disks from in vitro shoots originated from ex vitro hermaphrodite plants were cultured in induction medium supplemented with different concentrations of 2,4-D (6, 9, 12, 15, and 18 μM) and 4-CPA (19, 22, 25, 28, and 31 μM). After 90 days, the formation of somatic embryos was verified. The 2,4-D induced the formation of light brown calli with low frequency (20%) of somatic embryogenesis. However, 4-CPA (25 μM) induced 96% of embryogenic calli, which were transferred to maturation medium (MM) and cultured for 30 days. The MM contained ABA (0.5 μM) and AC (15 g L-1) and produced 36.6 somatic embryos callus-1, mainly on cotyledonary stage. Cotyledonary embryos were transferred to germination medium supplemented with gibberellic acid (GA3) (0.0, 1.44, 2.88, and 4.32 µM), and the conversion into plantlets was enhanced with GA3 at 2.88 µM.

Abstract This study evaluated the effect of osmoregulators and carbohydrates on the maturation and germination of somatic embryos of papaya ‘Golden THB’. Cotyledon explants from papaya seedlings germinated in vitro on basal MS medium were cultured on somatic embryogenesis induction medium (IM) containing MS salts, myo-inositol, sucrose, agar and p-chlorophenoxyacetic acid. After 50 days, embryogenic calli were transferred onto maturation media (MM) for 45 additional days. For experiment 1, a MS-based medium supplemented with abscisic acid, activated charcoal and concentrations of PEG 6000 (0; 40; 50; 60 and 70 g L-1) was used, whereas for experiment 2 malt extract concentrations (0; 0.1; 0.2; 0.3 and 0.4 g L-1) were assessed. The normal cotyledonary somatic embryos produced in experiment 2 were transferred to the germination medium (GM). The GM consisted of full-strength MS medium, sucrose, agar and was supplemented with myo-inositol at varying concentrations (0; 0.275; 0.55 and 0.825 mM). The PEG concentrations tested impaired the maturation of ‘Golden THB’ papaya somatic embryos. The MM, supplemented with malt extract at 0.153 g L-1, promoted the greatest development of normal somatic embryos (18.28 SE calli-1), that is, two cotyledonary leaves produced 36.56 SE calli-1. The supplementation with 0.45 mM myo-inositol provided the highest germination percentage (47.42%) and conversion to emblings.

ABSTRACT The aim of this study was to evaluate somatic embryogenesis in juvenile explants of the THB papaya cultivar. Apical shoots and cotyledonary leaves were inoculated in an induction medium composed of different concentrations of 2,4-D (6, 9, 12, 15 and 18 µM) or 4-CPA (19, 22, 25, 28 and 31 µM). The embryogenic calluses were transferred to a maturation medium for 30 days. Histological analysis were done during the induction and scanning electron microscopy after maturing. For both types of auxin, embryogenesis was achieved at higher frequencies with cotyledonary leaves incubated in induction medium than with apical shoots; except for callogenesis. The early-stage embryos (e.g., globular or heart-shape) predominated. Among the auxins, best results were observed in cotyledonary leaves induced with 4-CPA (25 µM). Histological analyses of the cotyledonary leaf-derived calluses confirmed that the somatic embryos (SEs) formed from parenchyma cells, predominantly differentiated via indirect and multicellular origin and infrequently via synchronized embryogenesis. The secondary embryogenesis was observed during induction and maturation phases in papaya THB cultivar. The combination of ABA (0.5 µM) and AC (15 g L-1) in maturation medium resulted in the highest somatic embryogenesis induction frequency (70 SEs callus-1) and the lowest percentage of early germination (4%).

Foi avaliado o comportamento de segmentos nodais lenhosos e herbáceos de plantas adultas e, os efeitos da caseína hidrolisada (CH 500 mg L-1), da cinetina (KIN; 6-furfurilaminopurina 0,46 µM e 4,65 µM) e do carvão ativado (CA 1,5 g L-1) na indução e na posterior multiplicação de brotos, visando estabelecer um sistema de propagação in vitro a partir de plantas adultas de Ginkgo biloba. Segmentos nodais lenhosos não emitiram brotos axilares e apresentaram 100% de contaminação fúngica e bacteriana durante o cultivo. Segmentos nodais de ramos herbáceos foram desinfestados com sucesso e apresentaram alta capacidade regenerativa in vitro. A CH foi essencial para a indução de brotos axilares e também para a posterior multiplicação destes, estimulando a formação de brotos em 85% dos segmentos nodais e múltiplos brotos em 35% destes na fase de estabelecimento das culturas. Na fase de multiplicação, 66,6% dos propágulos multiplicaram formando novos brotos e, 33,3% formaram múltiplos brotos quando cultivados com CH. A KIN e o CA provocaram efeitos inibitórios no processo organogênico de ginkgo. Foram observados dois padrões distintos de desenvolvimento de brotos in vitro, à semelhança do que ocorre in vivo: 1) brotos curtos, com segmentos internodais muito próximos, expansão de algumas folhas e de crescimento lento e; 2) brotos longos, com nós separados por segmentos internodais distintos e com maior crescimento apical que os anteriores. Este é o primeiro relato da formação in vitro de múltiplos brotos em segmentos nodais de plantas adultas de Ginkgo biloba.

The in vitro performance of herbaceous and woody nodal segments from adult plants and the effect of hydrolyzed casein (HC 500 mg L-1), kinetin (KIN; 6-furfurylaminopurine 0.46 and 4.65 µM) and activated charcoal (AC 1.5 g L-1) were evaluated upon new shoots induction and development, and to establish a system of in vitro propagation from adult plants of Ginkgo biloba. Woody nodal segments did not produce axillary shoots and presented 100% of bacterial and fungal contamination in culture. However, nodal segments from herbaceous shoots were successfully disinfected and displayed high in vitro morphogenic capacity. The HC was essential for the axillary shoots induction and further multiplication, stimulating shoot formation in 85% of the cultured nodal segments and multiple shoots induction in 35% of them at establishment stage. During the multiplication stage, 66.6% of propagules formed new shoots and 33.3% of them formed multiple shoots when cultured with HC. The KIN and AC inhibited the organogenic process in ginkgo. Two distinct patterns of sprouts development were observed in vitro, similar to what occurs in vivo: 1) short shoots with crowded internodes and expansion of only a few leaves and slow growth; 2) long shoots with separated nodes and marked apical growth. This is the first report of multiple shoots in vitro formation in nodal segments obtained from adult plants of Ginkgo biloba.

The ecdysteroid 20-hydroxyecdysone (20E) is a steroid hormone found in insects and few plants. To analyze its temporal and spatial distribution in Pfaffia glomerata (Amaranthaceae), we selected among 71 accessions, the accession 13 which showed the highest root 20E total quantity (1.48 g/plant root). The 20E was constantly detected in flowers, leaves, stems, and roots, but its percentage was variable throughout its development. The highest 20E percentage was found in flowers (0.82%), roots (0.66%), leaves (0.60%), and stems (0.24%). While stems showed the least variable 20E percentage (0.13% - 0.24%), followed by roots (0.42% - 0.66%), and flowers (0.47% - 0.82%). Leaves showed the greatest 20E percentage variation (0.21% - 0.60%). The stem and leaves 20E total amount remained constant, showing a little variation while root 20E total amount increased over time, suggesting that 20E may be accumulated in roots. These findings suggest that 20E has tissue specific functions in plant and may have ecological significance since uncoordinated pulses of 20E are often lethal to insects.

Anualmente, as plantas produzem aproximadamente 180 bilhões de toneladas de celulose, sendo o maior reservatório de carbono orgânico no planeta. A celulose é um homopolímero linear composto por resíduos de glicose unidos por meio de ligações beta(1-4). A síntese coordenada das cadeias de glicose é orquestrada por complexos específicos ligados à membrana plasmática (CelS). Postula-se que o CelS é composto por aproximadamente 36 subunidades da sintase da celulose (CESA). Cada CelS sintetiza 36 cadeias de glicose dispostas lado a lado antes de serem organizadas em microfibrilas, que são, posteriormente, associadas com outros polímeros da parede celular. As 36 cadeias de glicose presentes em uma microfibrila são estabilizadas por pontes de hidrogênio intra e inter-cadeias, conferindo grande estabilidade às microfibrilas. As microfibrilas elementares são dispostas lado a lado, permitindo a formação das macrofibrilas. Várias isoformas da CESA podem estar envolvidas no processo de biossíntese de celulose e, no mínimo, três tipos de isoformas da CESA podem ser necessárias para a organização funcional de cada CelS em plantas superiores.

Annually, plants produce about 180 billion tons of cellulose making it the largest reservoir of organic carbon on Earth. Cellulose is a linear homopolymer of beta(1-4)-linked glucose residues. The coordinated synthesis of glucose chains is orchestrated by specific plasma membrane-bound cellulose synthase complexes (CelS). The CelS is postulated to be composed of approximately 36 cellulose synthase (CESA) subunits. The CelS synthesizes 36 glucose chains in close proximity before they are further organized into microfibrils that are further associated with other cell wall polymers. The 36 glucose chains in a microfibril are stabilized by intra- and inter-hydrogen bonding which confer great stability on microfibrils. Several elementary microfibrils come together to form macrofibrils. Many CESA isoforms appear to be involved in the cellulose biosynthetic process and at least three types of CESA isoforms appear to be necessary for the functional organization of CelS in higher plants.

Dezesseis plantas transgênicas (R0) de maracujá-amarelo (Passiflora spp.) foram obtidas expressando um RNA não-traduzível correspondente à região 3' do gene NIb e 5' do gene CP de um isolado brasileiro do CABMV. As plantas R0 foram propagadas vegetativamente por estaquia e inoculadas com os isolados CABMV-MG1 e CABMV-PE1. Uma das plantas (TE5-10) foi resistente ao isolado CABMV-MG1, porém suscetível ao isolado CABMV-PE1. As demais plantas transgênicas foram suscetíveis a ambos os isolados, apresentando sintomas sistêmicos semelhantes ao de plantas não-transformadas. A ausência de replicação viral nas plantas TE5-10 foi confirmada por ELISA indireto. A análise da transcrição do transgene nas plantas TE5-10 demonstrou que estas não acumulam o mRNA transgênico, mesmo antes da inoculação com o vírus. Após a inoculação, o RNA viral foi detectado apenas em plantas inoculadas com o isolado CABMV-PE1. Esses resultados confirmam que a planta TE5-10 é resistente ao isolado CABMV-MG1, e sugerem que o mecanismo da resistência é baseado em silenciamento gênico pós-transcricional, o qual já se encontra ativado nas plantas antes da inoculação com o vírus.

Sixteen transgenic yellow passionfruit (Passiflora spp.) plants (R0) were obtained which express a non-translatable transgenic RNA corresponding to the 3' region of the NIb gene and the 5' region of the CP gene, derived from the genome of a Brazilian isolate of Cowpea aphid-borne mosaic virus (CABMV). The transgenic plants were propagated by stem cuttings and challenged by sap inoculation with isolates CABMV-MG1 and CABMV-PE1. One transgenic plant (TE5-10) was resistant to the isolate CABMV-MG1, but susceptible to CABMV-PE1. The remaining transgenic plants developed systemic symptoms, equal to non-transformed plants, when inoculated with either isolate. The absence of virus in TE5-10 plants was confirmed by indirect ELISA. Transcription analysis of the transgene demonstrated that the TE5-10 plant did not accumulate transgenic mRNA, even before inoculation. After inoculation, viral RNA was only detected in plants inoculated with CABMV-PE1. These results confirm that the transgenic plant TE5-10 is resistant to isolate CABMV-MG1, and suggest that the resistance mechanism is post-transcriptional gene silencing, which is already activated in the transgenic plants before virus inoculation.