Substrates and container types on rooting of peach cv. Okinawa hardwood cuttings of different diameters

Propagation by hardwood cuttings is an important technique for producing plant of many species. The technique is usually easy and inexpensive and makes it possible to high plants. But it is not commonly used for peach propagation in Brazil. The objective of this study was to evaluate the effects of different substrates, container types and cutting diameters on rooting of peach hardwood cuttings. The cuttings (two size groups, diameters of 2-6 mm and 6-10 mm) were inserted in six substrates (carbonized rice husk, vermiculite, washed sand, carbonized rice husk + vermiculite, carbonized rice husk + washed sand and vermiculite + washed sand, each 1:1 v/v), each in three types of containers (plastic bag, plastic tray and polystyrene tray). The best rooting occurred with smaller cuttings (47%) and larger cuttings (38%) in plastic bag with vermiculite, and with smaller cuttings in plastic bag with washed sand + vermiculite (33%). While these rooting percentages were unsatisfactory for commercial propagation, the results are encouraging and suggest need for more investigation to improve rooting.

Quantification of free auxins in semi-hardwood plant cuttings and microshoots by dispersive liquid–liquid microextraction/microwave derivatization and GC/MS analysis

Several studies have suggested that differences in the natural rooting ability of plant cuttings could be attributed to differences in endogenous auxin levels. Hence, during rooting experiments, it is important to be able to routinely monitor the evolution of endogenous levels of plant hormones. This work reports the development of a new method for the quantification of free auxins in auxin-treated Olea europaea (L.) explants, using dispersive liquid–liquid microextraction (DLLME) and microwave assisted derivatization (MAD) followed by gas chromatography/mass spectrometry (GC/MS) analysis. Linear ranges of 0.5–500 ng mL 1 and 1–500 mg mL 1 were used for the quantification of indole-3-acetic acid (IAA) and indole-3-butyric acid (IBA), respectively. Determined by serial dilutions, the limits of detection (LOD) and quantification (LOQ) were 0.05 ng mL 1 and 0.25 ng mL 1, respectively for both compounds. When using the calibration curve for determination, the LOQ corresponded to 0.5 ng mL 1 (IAA) and 0.5 mg mL 1 (IBA). The proposed method proved to be substantially faster than other alternatives, and allowed free auxin quantification in real samples of semi-hardwood cuttings and microshoots of two olive cultivars. The concentrations found in the analyzed samples are in the range of 0.131–0.342 mg g 1 (IAA) and 20–264 mg g 1 (IBA).

Adventitious root formation in cuttings of Backhousia citriodora F. Muell: 1. Plant genotype, juvenility and characteristics of cuttings

Backhousia citriodora is a commercially valuable Australian woody species that has a reputation for being recalcitrant in forming adventitious roots from cuttings. A study was carried out to determine whether maturation and plant genotype influenced rooting. It also tried to establish whether genotypic differences in rooting ability were related to characteristics of the cutting material. The rooting of cuttings in B. citriodora declines after maturation and is strongly influenced by genotype. The cutting characteristics of actively growing axillary buds, wide stems and mature leaves are associated with rooting and survival but not related to genotype. Furthermore, the 8-24 weeks required by B. citriodora to form roots from cuttings makes it difficult to distinguish between the characteristics that increase rooting and those characteristics that enhance survival. A subsequent disbudding experiment demonstrated that axillary buds per se have an inhibitory effect on rooting. This suggests that the presence of actively growing axillary buds are an indication of overall growth and condition of the stock plant unrelated to the formation of adventitious rooting. The effects of other cutting characteristics on rooting are also discussed. (C) 2004 Elsevier B.V. All rights reserved.

Adventitious root formation in cuttings of Backhousia citriodora F. Muell 2. Seasonal influences of temperature, rainfall, flowering and auxins on the stock plant

A 2-year study was carried out on established trees at two sites in southeastern Queensland, Australia, to identify environmental factors that influenced rooting of Backhousia citriodora from cuttings. Complex interactions of rainfall events above 20 mm from the preceding month and mean maximum temperature on stock plants resulted in a correlation with rooting success of r = 0.81 and 0.74 for sites at The University Of Queensland, Gatton Campus, and Cedar Glen, respectively. A more detailed investigation under controlled environmental conditions showed that maintaining stock plants at temperatures between 10 and 30degreesC had no direct effect on rooting capacity. However, temperature was correlated with growth, which may have an indirect effect on root formation. In spring floral initiation was found to only delay rooting and had no effect on the final rooting percentage. A series of seasonal experiments demonstrated that application of the auxins indole-3-acetic acid, indole-3-butyric acid and napthaleneacetic acid over a range of concentrations (1000-8000 mug/ml) did not significantly increase rooting compared to the control and there is no practical advantage in applying auxins. Seasonal results and the temperature experiment also suggest that under a glasshouse environment with higher temperatures in winter and an adequate supply of water, B. citriodora cuttings can be successfully rooted over the whole year. (C) 2004 Elsevier B.V. All rights reserved.

Starch, growth, and development in cuttings of rootstock "Paulsen 1103" from plants infested with ground pearl.

ABSTRACT The ground pearl, Eurhizococcus brasiliensis, is considered an important pest of vineyards in southern Brazil, with affected plants exhibiting leaf chlorosis, reduction in vigor, fading, and death. This study evaluated the quality of hardwood cuttings produced from plants infected (I) and not infected (NI) by ground pearl. ?Paulsen 1103? (Vitis berlandieri × Vitis rupestris) plants were grown for 29 months in brick-built raised beds either infested or not infested by ground pearl; then, 12 one-year-old branches with a maximum of 12 buds each were cut from each plant, subdivided into three portions (4 buds cutting-1), and subjected to destructive and nondestructive testing. Destructive testing comprised determining fresh and dry weight, length, internode diameters, and percentage of starch. Nondestructive testing comprised assessing the potential for bud sprouting and shoot development. Each mother plant in the I and NI beds was considered a replicate, with a total of 360 cuttings per treatment. It was observed that cuttings from infected plants had significantly lower (P<0.05) internode diameter, length, and fresh and dry weight than those of the uninfected plants. The percentage of starch content of the cuttings did not differ significantly. All cuttings showed the same percentage (100%) of bud breaking and no changes in growth and development of seedlings regardless of source. Given these results, it was concluded that vines of ?Paulsen 1103? infested with ground pearl produce smaller cuttings than those of uninfected plants but with no reduction in bud break percentage or seedling development. Key words: Margarodidae, Vitaceae, insect?plant interaction, carbohydrates. RESUMO A pérola-da-terra, Eurhizococcus brasiliensis, tem sido considerada uma importante praga dos vinhedos no sul do Brasil, sendo que as plantas atacadas manifestam clorose foliar, redução no vigor, definhamento e morte. Este trabalho teve por objetivo avaliar a qualidade de estacas lenhosas produzidas a partir do contraste de videiras infestadas (I) e não infestadas (NI) por pérola-da-terra. Após 29 meses de cultivo em canteiros de alvenaria, em presença ou ausência de pérola-da-terra, cada planta da variedade ?Paulsen 1103? (Vitis berlandieri × Vitis rupestris) foi submetida à retirada de 12 ramos de ano, com no máximo 12 gemas cada, sendo subdivididos em três porções 4 gemas estaca-1) e submetidos a avaliações destrutivas e não destrutivas. As destrutivas consistiram em determinar massas fresca e seca, comprimento, diâmetro de entrenós e percentual de amido. As avaliações não estrutivas consistiram em testar o potencial de brotação e desenvolvimento das estacas. Cada planta matriz dos canteiros I e NI foi considerada uma repetição, totalizando 360 estacas por tratamento. As estacas das plantas infestadas tiveram uma redução (P<0,05), em relação às não infestadas, em diâmetro, comprimento e massas fresca e seca. Não houve contraste significativo do percentual de amido avaliado das estacas. Quanto à brotação, destaca-se que todas as estacas apresentaram o mesmo percentual (100%), independente da origem, sem alterações no desenvolvimento e crescimento das mudas. Diante desses resultados, salienta-se que videiras ?Paulsen 1103? infestadas por pérola-da-terra produzem estacas menores, porém não há comprometimento no percentual de brotação e desenvolvimento das mudas, quando comparadas com plantas não infestadas. Palavras-chave: Margarodidae, Vitaceae, interação inseto-planta, carboidrato

Propagação de Camellia sinensis: efeito do genótipo, estaca, substrato, recipiente e ácido indolbutírico

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Enraizamento de estacas de Trichilia catigua A. Juss (catigua) em diferentes estações do ano

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

2,6-di-hidroxiacetofenona no enraizamento de estacas semilenhosas de pessegueiro

A propagação do pessegueiro no Brasil baseia-se na enxertia de cultivares-copa em porta-enxertos propagados por sementes, e uma alternativa de propagação para esta frutífera poderia ser a estaquia. O objetivo deste trabalho foi avaliar o potencial de enraizamento de estacas semilenhosas de cultivares de pessegueiro através da aplicação de 2,6-di-hidroxiacetofonona (2,6-DHAP) antes do tratamento com AIB (ácido indolbutírico). As estacas foram preparadas a partir de ramos coletados das cultivares Delicioso Precoce, Jóia 1 e Okinawa, em dezembro de 2001, para serem tratadas na base com 2,6-DHAP (0 e 300mg L-1), por 4h, em aeração e depois com AIB (200mg L-1 e 2500mg L-1). As estacas foram plantadas em bandejas de poliestireno expandido com vermiculita fina e colocadas em casa de nebulização, por 45 dias. A aplicação de 300mg L-1 de 2,6-DHAP antes da aplicação de 200mg L-1 de AIB em estacas de 'Okinawa' proporcionou aumentos nos resultados das características de maior relevância para a propagação por estacas e pode ser uma técnica interessante para a estaquia em pessegueiro.

Enraizamento de estacas lenhosas de pessegueiro cv. Okinawa em diferentes diâmetros de ramos, substratos e recipientes

O trabalho foi desenvolvido no Departamento de Produção Vegetal - Setor Horticultura da Faculdade de Ciências Agronômicas (FCA) da Universidade Estadual Paulista Júlio de Mesquita Filho (UNESP), Campus de Botucatu (SP) com o objetivo de avaliar a influência de diferentes substratos e tipos de recipientes no potencial de enraizamento de estacas lenhosas de pessegueiro cultivar Okinawa com diferentes diâmetros. As estacas foram tratadas com 2,5g L-1 de ácido indol-butírico. O período de permanência das estacas na casa de vegetação foi de 50 dias. Os tratamentos consistiram de seis substratos: areia, casca de arroz carbonizada, vermiculita, areia + casca de arroz carbonizada, areia + vermiculita e casca de arroz carbonizada + vermiculita, com as misturas na proporção 1:1v/v, três tipos de recipientes: sacos plásticos, bandejas de poliestireno expandido e bandejas plásticas e dois grupos de estacas com diâmetros diferentes: 2 a 6mm e 6 a 10mm. O melhor resultado de enraizamento foi obtido em sacos plásticos com vermiculita independente do diâmetro das estacas.

2,6-Di-hidroxiacetofenona e tipo de corte basal no enraizamento de estacas semi-lenhosas de pessegueiro Okinawa

A propagação do pessegueiro (Prunus persica) no Brasil é baseada na enxertia sobre porta-enxertos oriundos de sementes. Outros métodos de propagação de frutíferas poderiam ser utilizados para o pessegueiro visando obtenção de materiais de melhor qualidade. Neste trabalho objetivou-se avaliar o efeito do 2,6-di-hidroxiacetofenona aplicado previamente ao ácido indol-butírico em estacas semi-lenhosas de pessegueiro da cultivar Okinawa preparadas com diferentes tipos de corte basal. Foram coletados ramos do porta-enxerto Okinawa em dezembro de 2001 para o preparo das estacas sem folhas, com 12cm de comprimento, 7mm de diâmetro, quatro gemas e diferentes tipos de cortes basais (corte longitudinal, corte lateral e corte da casca) tratado-as na base com 0 e 300mg L-1 de 2,6-di-hidroxiacetofenona por 4h em aeração (oxigenação) e depois com 2500mg L-1 ácido indol-butírico por 5s. As estacas foram plantadas em bandejas de poliestireno expandido (72 células) com vermiculita fina e colocadas em casa de nebulização por 45 dias. A aplicação de 300mg L-1 de 2,6-DHAP e a realização de lesões nas bases das estacas foram eficientes para aumentar o enraizamento das estacas semi-lenhosas do pessegueiro Okinawa , demonstrando que estas técnicas podem ser utilizadas para a propagação de pessegueiro por estaquia.

Ácido indol-butírico, 2,6-di-hidroxiacetofenona e gene ARRO-1 no enraizamento de estacas de pessegueiro

Pós-graduação em Agronomia (Horticultura) - FCA

Seed and vegetative propagation methods for the rare Florida native species Chionanthus pygmaeus (pygmy fringetree)

Chionanthus pygmaeus Small (pygmy fringetree) (Oleaceae) is an endemic and rare Florida species, which has an attractive, small habit giving it great potential for use in managed landscapes. Members of the genus Chionanthus are difficult to propagate via cuttings and possess complex seed dormancies that are not well understood. Conservation of pygmy fringetree and its potential for commercial propagation for use in managed landscapes is contingent on a better understanding of its complex seed dormancy and enhancement of its propagation. I conducted two experiments to assess sexual and asexual propagation methods for pygmy fringetree. The first experiment was conducted to determine what factors are involved in overcoming seed dormancy. Various scarification treatments, which mimicked conditions seeds are exposed to in the wild, were investigated to determine their effects on germination of 20-year-old seeds originally collected from the species’ native range. Treatments included endocarp removal, sulfuric acid, boiling-water, and smoke-water treatments. Prior to treatment initiation, seed viability was estimated to be 12%. Treated seeds went through two cold- and two warm-stratification periods of 4°C and 25°C, respectively, in a dark growth chamber. After 180 days, none of the treatments induced early germination. Seeds were then tested for viability, which was 11%. Seed dormancy of the species is apparently complex, allowing some of the seeds to retain some degree of viability, but without dormancy requirements satisfied. The second experiment was conducted to assess if pygmy fringetree could be successfully propagated via hardwood or root cuttings if the appropriate combination of environmental conditions and hormones were applied. Hardwood and root cuttings were treated with either 1000 ppm IBA talc, 8000 ppm IBA talc, or inert talc. All cuttings were placed on a mist bench in a greenhouse for 9 weeks. Hardwood cuttings were supplemented with bottom heat at 24 °C. No treatments were successful in inducing adventitious root formation. I conclude that pygmy fringetree seeds possess complex dormancy that was not able to be overcome by the treatments utilized. However, this result is confounded by the age of the seeds used in the experiment. I also conclude that vegetative propagation of pygmy fringetree is highly dependent on the time of year cuttings are harvested. Further research of both seed and asexual propagation methods need to be explored before pygmy fringetree can be propagated on a commercial scale.

Understanding the role of auxins and oxidative enzymes on adventitious root formation in olive (olea europaea L.) cultivars

Olive (Olea europaea L.), one of the main crops in the Mediterranean basin, is mainly propagated by cuttings, a classical propagation method that relies on the ability of the cuttings to form adventitious roots. While some cultivars are easily propagated by this technique, some of the most interesting olive cultivars are considered difficult-to-root which poses a challenge for their preservation and commercialization. Therefore, increasing the current knowledge on adventitious root formation is extremely important for species like olive. This research focuses on evaluating the role of free auxins and oxidative enzymes on adventitious root formation of two olive cultivars with different rooting ability - ‘Galega vulgar’ (difficult-to-root) and ‘Cobrançosa’ (easy-to-root). In this context, free auxin levels and enzyme activities were determined in in vitro-cultured ‘Galega vulgar’ microshoots and in semi-hardwood cuttings of cvs. ‘Galega vulgar’ and ‘Cobrançosa’. To attain this goal, an analytical method for the quantification of free indole-3-acetic acid (IAA) and indole-3-butyric acid (IBA) was developed, which is based on dispersive liquid-liquid microextraction followed by microwave derivatization (DLLME-MAD) and gas chromatography-mass spectrometry (GC/MS) analysis. The developed method was validated in terms of linearity, recovery, limit of detection (LOD) and limit of quantification (LOQ) and proved to be useful in the analysis of two very different types of plant tissues. The results from auxin quantification in olive samples point at a relationship between free auxin levels and rooting ability of both microshoots and semihardwood cuttings. A defective IBA-IAA conversion, resulting in a peak of free IAA during initiation phase, seems to be associated with low rooting ability. Likewise, differences in the activity of oxidative enzymes also appear to be related with rooting ability. Higher polyphenol oxidases (PPO) activity is likely related with an easyto- root behavior, while the opposite is true for peroxidases (POX) (including IAA oxidase (IAAox)) activity. A possible hypothesis for adventitious root formation in olive microcuttings is presented herein for the first time. Free auxins, oxidative enzymes, alternative oxidase (AOX) and reactive oxygen species (ROS) are some of the factors that may be involved in this highly complex physiological process. Interestingly, while temporal changes in auxin levels were similar between microshoots and semihardwood cuttings, the conclusions obtained from enzyme activity results in microshoots didn’t translate to semi-hardwood tissues, showing the emerging need for adaptation of classical agronomical research studies to modern techniques; Resumo: Procurando compreender o papel das auxinas e enzimas oxidativas na formação de raízes adventícias em cultivares de oliveira (Olea europaea L.) A oliveira (Olea europaea L.) é uma das principais culturas da bacia Mediterrânica e é propagada maioritariamente por estacaria, um processo altamente dependente da capacidade das estacas para formar raízes adventícias. Enquanto algumas cultivares são fáceis de propagar desta forma, algumas das cultivares de oliveira mais interessantes são consideradas difíceis de enraizar, o que dificulta a sua preservação e comercialização e torna extremamente importante aprofundar o conhecimento sobre o enraizamento adventício desta espécie. Este trabalho foca-se na avaliação do papel das auxinas livres e das enzimas oxidativas na formação de raízes adventícias em duas cultivares de oliveira com diferente capacidade de enraizamento - ‘Galega vulgar’ (difícil de enraizar) e ‘Cobrançosa’ (fácil de enraizar). Neste contexto, determinaram-se os níveis de auxinas livres e as actividades de enzimas oxidativas em microestacas de ‘Galega vulgar’ cultivadas in vitro bem como em estacas semi-lenhosas das cvs. ‘Galega vulgar’ e ‘Cobrançosa’. Para tal foi necessário desenvolver uma metodologia analítica para a quantificação de ácido indol-3-acético (IAA) e ácido indol-3-butírico (IBA), baseada em microextracção dispersiva líquido-líquido (DLLME) seguida de derivatização em microondas (MAD) e análise por cromatografia gasosa acoplada a espectrometria de massa (GC/MS). O método desenvolvido foi validado em termos de linearidade, recuperação, limite de detecção (LOD) e limite de quantificação (LOQ), e mostrou-se eficaz na análise de dois tipos de tecidos vegetais bastante diferentes. Os resultados da análise de auxinas em amostras de oliveira apontam para uma possível relação entre os níveis de auxinas livres e a capacidade de enraizamento, tanto em microestacas como em estacas semi-lenhosas. Uma conversão IBA-IAA deficiente, que resulta num pico de IAA durante a fase de iniciação, parece estar associada à baixa capacidade de enraizamento. Por outro lado, a capacidade de enraizamento também parece estar relacionada com diferenças na actividade de enzimas oxidativas. Comportamentos fáceis de enraizar estão associados a actividade mais elevada das polifenoloxidases (PPO), enquanto o oposto é verdade para a actividade das peroxidases (POX) (incluindo a IAA oxidase (IAAox)). Neste trabalho propõe-se pela primeira vez uma possível explicação para o enraizamento adventício em microestacas de oliveira. Auxinas livres, enzimas oxidativas, oxidase alternativa (AOX) e espécies reactivas de oxigénio (ROS) são alguns dos factores envolvidos neste processo fisiológico altamente complexo. Curiosamente, enquanto as alterações temporais nos níveis de auxinas foram semelhantes entre microestacas e estacas semi-lenhosas, o mesmo não se observou relativamente à actividade enzimática, o que mostra a necessidade de adaptação dos estudos agronómicos tradicionais às técnicas correntes.

Propagação de Camellia sinensis: efeito do genótipo, estaca, substrato, recipiente e ácido indolbutírico

Tem sido relatado que as estacas de Camellia sinensis possuem baixa capacidade de emitir raízes, motivando assim a realização de estudos básicos para otimização do processo de propagação por estacas. Assim sendo, o presente trabalho objetivou quantificar o potencial rizogênico de diferentes genótipos e o efeito da posição da estaca no ramo e incisão na base, do substrato, tamanho do recipiente e ácido indolbutírico no enraizamento de estacas semi-lenhosas dessa espécie. Para tal, foram coletados ramos dos genótipos IAC 259, F15 e Comum, em Pariquera-Açu-SP, no inverno de 2010. em seguida, preparadas as estacas, contendo uma gema e uma folha, foram mantidas em viveiro com 70% de sombreamento. Estacas da posição basal e mediana dos ramos são as mais adequadas para estaquia devido a menor mortalidade e maior enraizamento. A injúria na base da estaca não afeta a mortalidade e o enraizamento das estacas, porém induz à formação de calo. Também não houve diferenças na mortalidade e no enraizamento das estacas quando as mesmas foram mantidas em recipiente de 50, 90 e 120 cm³. Comparado com vermiculita, areia e casca de arroz carbonizada, o solo foi o melhor substrato para estaquia, que na presença do ferimento, juntamente com o tratamento das estacas com 10 g L-1 de AIB promoveu a maior porcentagem de enraizamento. Todavia, ainda nessa condição a mortalidade média das estacas foi de 42%. O potencial de enraizamento do genótipo Comum foi superior ao do IAC 259 e F15.

Tracking biochemical changes during adventitious root formation in olive (Olea europaea L.)

The activity of oxidative enzymes and the levels of free auxins were determined during adventitious root formation in olive explants. Rooting trials were performed both with in vitro-cultured micro shoots of the cultivar ‘Galega Vulgar’, treated with indole-3-butyric acid (IBA) and with salicylhydroxamic acid(SHAM) + IBA, as well as with semi-hardwood cuttings of the cultivars ‘Galega Vulgar’ (difficult-to-root)and ‘Cobrançosa’ (easy-to-root), treated with IBA. The auxin (IBA) was used in all experiments as a rooting promoter, while SHAM was used in micropropagation trials as rooting inhibitor, providing a negative control. Free indole-3-acetic acid (IAA) and IBA concentrations were determined in microshoots, as well as in semi-hardwood cuttings, throughout the rooting period at pre-established time-points. At the sametime-points, the enzymatic activity of polyphenol oxidases (PPO), peroxidases (POX), and IAA oxidase(IAAox) was evaluated in the microshoots. Microshoots treated with SHAM + IBA revealed higher POX and IAAox activity, as well as lower PPO activity, than those treated only with IBA. IAA levels were higher in IBA-treated microshoots during induction phase, but lower during early initiation phase. Incontrast, free IBA levels were higher in microshoots treated with SHAM + IBA during induction, but lower during initiation. A similar pattern of free auxin levels was observed in semi-hardwood cuttings of the two contrasting cultivars under evaluation. The similarities found on the auxin patterns of microshoots treated with SHAM and those of semi-hardwood cuttings of the difficult-to-root olive cultivar allow considering SHAM a reliable control for when simulation of a difficult-to-root behavior is necessary. The inhibitory effect of SHAM in root formation could be related with 1) the inhibition of alternative oxidase(AOX), leading to a down regulation of phenylpropanoid biosynthetic pathways, which would decrease the concentration of phenolic substrates for PPO; 2) an increase in IAAox activity resulting in lower free IAA levels or; 3) a defective conversion of IBA into IAA.