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 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.

Araucaria clonal forestry: types of cuttings and mother tree sex in field survival and growth.

Araucaria angustifolia é uma espécie nativa potencial para a silvicultura brasileira. No entanto, uma série de desafios e limitações técnicas ainda persistem, dificultando sua expansão silvicultural, dentre os quais se destaca a falta de tecnologias de clonagem de materiais genéticos superiores, bem como sua avaliação em condições de campo. Assim, objetivou-se avaliar a potencialidade da utilização de mudas de araucária oriundas de estaquia e de sementes para produção madeireira, por meio da avaliação da sobrevivência e crescimento a campo. Clones provenientes de matrizes masculinas e femininas, de diferentes tipos de estacas e mudas de sementes foram plantadas em espaçamento 3 x 3 m. O experimento foi conduzido em delineamento inteiramente casualizado, com três tratamentos e parcelas de uma planta (one tree plot). Clones do sexo feminino e de estacas contendo o ápice apresentaram maior crescimento em diâmetro à altura do peito (6,4 cm) e altura total (3,6 m) aos 74 meses após o plantio, seguidas das de sementes e demais clones, com resultados similares. Conclui-se que a estaquia é uma técnica potencial de produção de mudas de araucária para fins madeireiros e é favorecida pela utilização de estacas proveniente de matrizes femininas e com ápice.

Influência da fonte material e do tempo de cura na propagação vegetativa da pitaya vermelha (Hylocereus undatus Haw)

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

Análises bioquímicas e hsitológicas na micropropagação de abacaxizeiro 'Gomo de Mel' submetido a reguladores vegetais

Pós-graduação em Agronomia (Horticultura) - FCA

Enraizamento de mini-estacas de jabuticabeiras, pitangueira, araçazeiro amarelo e sete capoteiro

The most native fruit trees are belonging to Myrtaceae family, which it have as main marketing potential their fruit. Despite the wide acceptance of the fruits of these native fruit cultura, the establishment of commercial orchards is still necessary, because if it prevails extraction in the forest. To start the cultivo in the orchard, the first point is on the mother plant choice, which should provide superior characteristics when compared to other genotypes. Then, it is necessary to choose the method to can produce satisfactory amount of seedlings and preferably without it to lose the mother plant characteristics. For this, it adopts the asexual thechniques, with option for grafting, cuttings and air layering. These techniques when tested with native fruits tree, it proved limiting in theses results, with this, it should to test other it to recommend its use, especially, those fruit native of higher potential as jabuticaba tree, pitanga tree, sete capote tree and araça amarelo tree. The aim of this study was to test the use of asexual propagation through mini-cuttings in these native fruit trees, according to the time of collection, the mini-cutting length and concentration of IBA, as well as, it to relate the results of rooting with tryptophan extracted at certain times. The work was carried out at Universidade Tecnológica Federal do Paraná – Câmpus Dois Vizinhos, Brazil. The samples were collected each two months. The mini-cutting were prepared with 6 or 8 cm, with a pair of leaves reduced to 25% of the original size. The mini-cuttings had their base immersed in liquid solution of indole-butyric acid (IBA) in the concentrations of 0, 3000 and 6000 mg L-1 and then were placed in tubes containing commercial substrate. The experimental design was completely randomized with factorial 2 x 3 x 6 (mini-cutting length x IBA concentration x time of collection), with four replications, it being each plot varied according to the amount of shoots obtained by period time. After 120 days, the rooting and callus formation (%), average number of roots per mini-cutting and the average length of the roots were evaluated. After 60 days of these evaluations, the survival of mini-cuttings rooted after transplant was evaluated. It was evaluated also the production of mini-cuttings of each size in each period time. At the end of the experiment it was evaluated the percentage of survival of mother plantlets. For analysis of tryptophan was used materials branches, leaves and twigs with leaves, taken from the materials used for the production of mini-cutting. It was recommended for hybrid jabuticaba tree the use mini-cutting with eight cm, treated with 6000 mg L-1 of IBA and collected in June. For jabuticaba tree of cabinho and araça amarelo tree the period for propagation by mini-cuttings should be in August, regardless of IBA concentration and length of the mini-cutting. In the jabuticaba tree sabara and sete capote tree is important to obtain more satisfactory results realized the collect in October or December, with the same independence of other levels tested in other factors. However, for sete capote tree should test other techniques to increase the efficiency of propagation. And with pitanga tree recommended to the collection in June, but with 6cm the application of 3000 mg L-1 of IBA and 8 cm with 6000 mg L-1 of IBA.

Production of cuttings in response to stock plant temperature in the subtropical eucalypts, Corymbia citriodora and Eucalyptus dunnii

Propagation of subtropical eucalypts is often limited by low production of rooted cuttings in winter. This study tested whether changing the temperature of Corymbia citriodora and Eucalyptus dunnii stock plants from 28/23A degrees C (day/night) to 18/13A degrees C, 23/18A degrees C or 33/28A degrees C affected the production of cuttings by stock plants, the concentrations of Ca and other nutrients in cuttings, and the subsequent percentages of cuttings that formed roots. Optimal temperatures for shoot production were 33/28A degrees C and 28/23A degrees C, with lower temperatures reducing the number of harvested cuttings. Stock plant temperature regulated production of rooted cuttings, firstly by controlling shoot production and, secondly, by affecting the ensuing rooting percentage. Shoot production was the primary factor regulating rooted cutting production by C. citriodora, but both shoot production and root production were key determinants of rooted cutting production in E. dunnii. Effects of lower stock plant temperatures on rooting were not the result of reduced Ca concentration, but consistent relationships were found between adventitious root formation and B concentration. Average rooting percentages were low (1-15% for C. citriodora and 2-22% for E. dunnii) but rooted cutting production per stock plant (e.g. 25 for C. citriodora and 52 for E. dunnii over 14 weeks at 33/28A degrees C) was sufficient to establish clonal field tests for plantation forestry.

The effects of photoperiod and light spectrum on stock plant growth and rooting of cuttings of Cotinus coggygria 'Royal Purple'

Extending the season of production and improving the scheduling of ornamental crops are key commercial objectives for nurserymen. In some woody species, the period in which cuttings can be rooted successfully is transient, thus limiting the opportunities for scheduled production. Optimum rooting often occurs in early- to mid-summer coinciding with periods of active shoot growth. The relationship between this shoot activity and root initiation was investigated in Cotinus coggygria 'Royal Purple'. Shoot growth on stock plants was manipulated by altering the photoperiod or light quality. Results indicated there were seasonal effects on rooting, but the importance of shoot activity varied with harvest time. Cuttings harvested in August had high rooting percentages, irrespective of photoperiod, and despite shoot growth terminating in response to the short-day treatment. In contrast, by September, rooting percentage was highest in cuttings from plants under long-days, which had maintained greatest shoot growth activity. Cotinus shoots grown in vitro under 16 h days showed reduced shoot growth and increased rooting competence compared with shoots grown under 8 h days. Growing stock plants under polythene films, which altered the amount and quality of the incident light, influenced the rooting of cuttings harvested in August, but no consistent relationship with shoot activity was apparent. From a practical viewpoint, maintaining shoot activity late in the season may prolong the period for propagation by cuttings; but, from a scientific viewpoint, processes associated with an active shoot apex do not provide a complete explanation of seasonal variation in rooting.

Rooting cuttings of Syringa vulgaris cv. Charles Joly and Corylus avellana cv. Aurea: the influence of stock plant pruning and shoot growth

The relationship between shoot growth and rooting was examined in two, 'difficult-to root' amenity trees, Syringa vulgaris L. cv. Charles Joly and Corylus avellana L. cv. Aurea. A range of treatments reflecting severity of pruning was imposed on field-grown stock prior to bud break. To minimise variation due to the numbers of buds that developed under different treatments, bud number was restricted to 30 per plant. Leafy cuttings were harvested at different stages of the active growth phase of each species. With Syringa, rooting decreased with later harvests, but loss of rooting potential was delayed in cuttings collected from the most severe pruning treatment. Rooting potential was associated with the extent of post-excision shoot growth on the cutting but regression analyses indicated that this relationship could not entirely explain the loss of rooting with time, nor the effects due to pruning. Similarly, in Corylus rooting was promoted by severe pruning, but the relationship between apical growth on the cutting and rooting was weaker than in Syringa, and only at the last harvest did growth play a critical role in determining rooting. Another unusual factor of the last harvest of Corylus was a bimodal distribution of roots per cutting, with very few rooted cuttings having less than five roots. This implies that, for this harvest at least, the potential of an individual cutting to root is probably not limited by the number of potential rooting sites.

Rooting of healthy and CVC-affected 'Valência' sweet orange stem cuttings, through the use of plant regulators

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

Effect of plant growth regulators in the rooting of Pinus cuttings

O trabalho avaliou o enraizamento de estacas de Pinus caribaea var. hondurensis Morelet sob a ação de diferentes níveis de reguladores vegetais. As estacas foram feitas de brotações de 4 a 6cm de comprimento de mudas de P. caribaea var. hondurensis Morelet com corte bisel na base sendo as acículas basais eliminadas. A base das estacas foram submetidas aos tratamentos por 2 segundos com os seguintes tratamentos: 1- NAA 2000mg L-1; 2- NAA 4000mg L-1; 3- NAA 6000mg L-1; 4- NAA 2000mg L-1 + PBZ 100mg L-1; 5- NAA 4000mg L-1 + PBZ; 6- NAA 6000mg L-1 + PBZ; 7- IBA 2000mg L-1; 8- IBA 4000mg L-1; 9- IBA 6000mg L-1; 10-IBA 2000mg L-1 + PBZ; 11- IBA 4000mg L-1 + PBZ; 12- IBA 6000mg L-1 + PBZ e testemunha. Após os tratamentos as estacas foram plantadas em tubetes contendo 50% de palha de arroz carbonizada e 50% de vermiculita. As avaliações realizadas aos 60 dias após o plantio mostraram que estacas de P. caribaea tratadas com IBA levaram a maior porcentagem de estacas enraizadas que aquelas tratadas com NAA, sendo o mais efetivo, IBA a 4000mg L-1 em conjunto com 100mg L-1 de paclobutrazol.

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.

Post Entry Plant Quarantine

The seeds of the majority of commercial crops must be grown for one generation in post-entry plant quarantine on arrival in Australia. Live plants and cuttings must also undergo quarantine screening on arrival, and spend a minimum of three months in quarantine.

Multiplication of hybrid aspen (Populus tremula L. x P. tremuloides Michx.) from cuttings

The primary aim of the present study was to find an efficient and simple method of vegetative propagation for producing large numbers of hybrid aspen (Populus tremuloides L. x P. tremula Michx.) plants for forest plantations. The key objectives were to investigate the main physiological factors that affect the ability of cuttings to regenerate and to determine whether these factors could be manipulated by different growth conditions. In addition, clonal variation in traits related to propagation success was examined. According to our results, with the stem cutting method, depending on the clone, it is possible to obtain only 1−8 plants from one stock plant per year. With the root cutting method the corresponding values for two-year-old stock plants are 81−207 plants. The difference in number of cuttings between one- and two-year-old stock plants is so pronounced that it is economically feasible to grow stock plants for two years. There is no reason to use much older stock plants as a source of cuttings, as it has been observed that rooting ability diminishes as root diameter increases. Clonal variation is the most important individual factor in propagation of hybrid aspen. The fact that the efficiently sprouted clones also rooted best facilitates the selection of clones for large-scale propagation. In practice, root cuttings taken from all parts of the root system of hybrid aspen were capable of producing new shoots and roots. However, for efficient rooting it is important to use roots smaller than one centimeter in diameter. Both rooting and sprouting, as well as sprouting rate, were increased by high soil temperature; in our studies the highest temperature tested (30ºC) was the best. Light accelerated the sprouting of root cuttings, but they rooted best in dark conditions. Rooting is essential because without roots the sprouted cutting cannot survive long. For aspen the criteria for clone selection are primarily fiber qualities and growth rate, but ability to regenerate efficiently is also essential. For large-scale propagation it is very important to find clones from which many cuttings per stock plant can be obtained. In light of production costs, however, it is even more important that the regeneration ability of the produced cuttings be high.

Stem anatomy and adventitious root formation in cuttings of Angophora, Corymbia and Eucalyptus

Many plantation eucalypts are difficult to propagate from cuttings, and their rooted cuttings often possess very few adventitious roots. We microscopically examined the stem anatomy of cuttings from 12 species of eucalypts and we determined whether adventitious root formation in auxin-treated cuttings of four species was limited to particular positions around the vascular tissue. Most species contained a central pith that was arranged in a four-pointed stellate pattern. The surrounding vascular tissue was also arranged in a stellate pattern near the shoot apex but it developed a more rectangular shape at the outer phloem as the stems enlarged radially. Adventitious roots formed at, or slightly peripheral to, the vascular cambium, and they formed at both the corners and the sides of the rectangular-shaped vascular tissue. The study highlighted that auxin-treated eucalypt cuttings can produce roots at multiple positions around the vascular tissue and so propagation methods can aim to produce more than four adventitious roots per rooted cutting. Higher numbers of adventitious roots could improve the root system symmetry, stability, survival and growth rate of clonal eucalypt trees. © 2015 by the authors; licensee MDPI, Basel, Switzerland.