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.

In vivo and in vitro disappearance of energy and nutrients in novel carbohydrates and cereal grains by pigs

In vivo and in vitro experiments were conducted to determine digestibility of GE and nutrients, as well as DE and ME of carbohydrates fed to growing pigs. The objective of Exp. 1 was to determine the DE and ME of 4 novel carbohydrates fed to pigs. The 4 novel carbohydrates were 2 sources of resistant starch (RS 60 and RS 70), soluble corn fiber (SCF), and pullulan. These carbohydrates were produced to increase total dietary fiber (TDF) intake by humans. Maltodextrin (MD) was used as a highly digestible control carbohydrate. The DE and ME for RS 60 (1,779 and 1,903 kcal/kg, respectively), RS 75(1,784 and 1,677 kcal/kg, respectively), and SCF (1,936 and 1,712 kcal/kg, respectively) were less (P < 0.05) than for MD (3,465 and 3,344 kcal/kg, respectively) and pullulan (2,755 and 2,766 kcal/kg, respectively), and pullulan contained less (P < 0.05) DE and ME than MD. However, there was no difference in the DE and ME for RS 60, RS 75, and SCF. The varying degrees of small intestinal digestibility and differences in fermentability among these novel carbohydrates may explain the differences in the DE and ME among carbohydrates. Therefore, the objectives of Exp. 2 were to determine the effect of these 4 novel carbohydrates and cellulose on apparent ileal (AID) and apparent total tract (ATTD) disappearance, and hindgut disappearance (HGD) of GE, TDF, and nutrients when added to diets fed to ileal-cannulated pigs. The second objective was to measure the endogenous flow of TDF to be able to calculate the standardized ileal disappearance (SID) and standardized total tract (STTD) disappearance of TDF in the 4 novel fibers fed to pigs. Results of the experiment indicated that the AID of GE and DM in diets containing cellulose or the novel fibers was less (P < 0.05) than of the maltodextrin diet, but the ATTD of GE and DM was not different among diets. The addition of RS 60, RS 75, and SCF did not affect the AID of acid hydrolysed ether extract (AEE), CP, or ash, but the addition of cellulose and pullulan reduced (P < 0.01) the AID of CP. The average ileal and total tract endogenous losses of TDF were calculated to be 25.25 and 42.87 g/kg DMI, respectively. The SID of TDF in diets containing RS 60, SCF, and pullulan were greater (P < 0.01) than the SID of TDF in the cellulose diet, but the STTD of the SCF diet was greater (P < 0.05) than for the cellulose and pullulan diets. Results of this experiment indicate that the presence of TDF reduces small intestinal disappearance of total carbohydrates and energy which may reduce the DE and ME of diets and ingredients. Therefore, the objective of Exp. 3 was to determine the DE and ME in yellow dent corn, Nutridense corn, dehulled barley, dehulled oats, polished rice, rye, sorghum, and wheat fed to growing pigs and to determine the AID and ATTD of GE, OM, CP, AEE, starch, total carbohydrates, and TDF in these cereal grains fed to pigs. Results indicated that the AID of GE, OM, and total carbohydrates was greater (P < 0.001) in rice than in all other cereal grains. The AID of starch was also greater (P < 0.001) in rice than in yellow dent corn, dehulled barley, rye, and wheat. The ATTD of GE was greater (P < 0.001) in rice than in yellow dent corn, rye, sorghum, and wheat. With a few exceptions, the AID and ATTD of GE and nutrients in Nutridense corn was not different from the values for dehulled oats. Likewise, with a few exceptions, the AID, ATTD, and HGD of GE, OM, total carbohydrates, and TDF in yellow corn, sorghum, and wheat were not different from each other. The AID of GE and AEE in dehulled barley was greater (P < 0.001) than in rye. The ATTD of GE and most nutrients was greater (P < 0.001) in dehulled barley than in rye. Dehulled oats had the greatest (P < 0.001) ME (kcal/kg DM) whereas rye had the least ME (kcal/kg DM) among the cereal grains. Results of the experiment indicate that the presence of TDF and RS may reduce small intestinal digestibility of starch in cereal grains resulting in reduced DE and ME in these grains. Digestibility experiments involving animals are time consuming and expensive. Therefore, the objective of Exp. 4 was to correlate DM and OM digestibility obtained from 3 in vitro procedures with ATTD of GE and with the concentration of DE in 50 corn samples that were fed to growing pigs. The second objective was to develop a regression model that can predict the ATTD of GE or the concentration of DE in corn. The third objective was to evaluate the suitability of using the DaisyII incubator as an alternative to the traditional water bath when determining in vitro DM and OM digestibility. Results indicated that corn samples incubated with Viscozyme for 48 h in the DaisyII incubator improved (P < 0.001) the ability of the procedure to detect small differences in the ATTD of GE or to detect small differences in the concentration of DE in corn. Likewise, compared with using cellulase or fecal inoculum, the variability in the ATTD of GE and the variability in the DE in corn was better (R2 = 0.56; P < 0.05 and R2 = 0.53; P < 0.06, respectively) explained if Viscozyme was used than if cellulase or fecal inoculum was used. A validated regression model that predicted the DE in corn was developed using Viscozyme and with the corn samples incubated in the DaisyII incubator for a 48 h. In conclusion, this present work used the pig as a model for human gastrointestinal function and evaluates carbohydrates from 2 different nutritional perspectives – humans and animals. The addition of novel carbohydrates reduced the digestibility of energy in the diets without necessarily reducing the digestibility of other nutrients. Thus, supplementation of novel carbohydrates in the diets may be beneficial for the management of diabetes. Aside from diabetic management, cereal grains such as rye and sorghum, may also help in BW management because of there low caloric value, but for undernourished individuals, dehulled oats, dehulled barley, and rice are the ideal grains. From an animal nutrition standpoint, high concentration of dietary fiber is undesirable because it reduces feed efficiency. Therefore, the inclusion of feed ingredients that have a high concentration of dietary fiber is often limited in animal diets. Although in vivo determination is ideal, in vitro procedures are useful tools to determine caloric value of food and feed ingredients.

Optimizing Western Flower Thrips Management on French Beans by Combined Use of Beneficials and Imidacloprid

Western flower thrips (WFT), Frankliniella occidentalis (Pergande), is an important pest of vegetable crops worldwide and has developed resistance to many insecticides. The predatory mites Neoseiulus (=Amblyseius) cucumeris (Oudemans), the entomopathogenic fungus Metarhizium anisopliae (Metsch.), and an insecticide (imidacloprid) were tested for their efficacy to reduce WFT population density and damage to French bean (Phaseolus vulgaris L.) pods under field conditions in two planting periods. Metarhizium anisopliae was applied as a foliar spray weekly at a rate of one litre spray volume per plot while imidacloprid was applied as a soil drench every two weeks at a rate of two litres of a mixture of water and imidacloprid per m(2). Neoseiulus cucumeris was released every two weeks on plant foliage at a rate of three mites per plant. Single and combined treatment applications reduced WFT population density by at least three times and WFT damage to French bean pods by at least 1.7 times compared with untreated plots. The benefit-cost ratios in management of WFT were profitable with highest returns realized on imidacloprid treated plots. The results indicate that M. anisopliae, N. cucumeris, and imidacloprid have the potential for use in developing an integrated pest management program against WFT on French beans.

Avaliação da potencial migração de Helicoverpa armigera (Lepidoptera: Noctuidae) por massas de ar para áreas produtoras de cultivos hospedeiros do Estado de São Paulo.

Resumo Helicoverpa armigera (Hübner, 1808) (Lepidoptera: Noctuidae) é um inseto exótico, altamente polífago, que foi detectado no Brasil em 2013 causando sérios danos em cultivos de soja, milho e algodão, localizados em áreas estaduais específicas das regiões Nordeste e Centro-Oeste com a presença do bioma Cerrado. Após o período de um ano o inseto foi identificado em várias regiões do País. A literatura indica que o inseto adulto apresenta características físicas que favorecem sua grande capacidade de voo, tanto em distância (100-1000 km) quanto a elevadas altitudes (maiores que 1,5 km acima do nível do solo), aumentando o seu potencial para dispersão de longo alcance, principalmente por voos migratórios das áreas já atacadas. Por essa razão, as massas de ar brasileiras podem atuar como potenciais facilitadoras à atividade migratória do inseto a longas distâncias para as áreas de maior concentração de seus cultivos hospedeiros preferenciais, resultante da atração natural do inseto aos fatores químicos delas decorrentes. Desse modo, devem ser consideradas em avaliações de áreas mais propícias à presença de H. armigera para orientar monitoramentos e estratégias de manejo locais. Este trabalho avaliou a potencial migração de H. armigera por massas de ar para áreas produtoras de cultivos hospedeiros do estado de São Paulo, considerando seus municípios produtores de algodão, soja, milho, tomate, feijão, café e laranja; as principais massas de ar brasileiras; a presença de grandes áreas com cultivos de eucalipto e pinus (outras potenciais barreiras físicas ao inseto); e a localização do bioma Cerrado paulista. Informações das áreas produtoras e das áreas plantadas por município de 2008, 2012 e 2014, disponibilizadas pelo IBGE, foram utilizadas em planos de informação georreferenciados, elaborados em ArcGis 10.3, para identificar a localização de suas respectivas grandes áreas em cada ano. O cruzamento georreferenciado também resultou mapas das áreas plantadas em 2014 com todos os cultivos citados, indicando as de maior potencial de infestação/reinfestações de H. armigera no estado de São Paulo, para orientar monitoramentos e estratégias de Manejo Integrado de Pragas - MIP locais. Abstract Helicoverpa armigera (Hübner, 1808) (Lepidoptera: Noctuidae) is an exotic insect highly polyphagous, which was detected in Brazil in 2013 causing serious damage to soybean, corn and cotton crops located in specific state areas in the northeast and center-west regions with the presence of Cerrado Biome (similar to the Savannah). After a oneyear period, the insect was detected in several regions of the country. Foreign literature indicates that the adult phase of the insect shows certain physical characteristics which favor its great flight capacity, even related to reach long distances (100-1000 Km) as many as to reach high altitudes (overtaken 1.5 Km above the ground level), increasing its potential of farthest dispersion, mainly by migratory flights from the areas already attacked. Therefore, the Brazilian air masses may act as potentially enhance the insect's farthest migratory activity to areas of greater concentration of its preferential host crops due to the insect's natural attraction to their chemical factors. Thus, migration potential of H. armigera by air masses toward preferential host crop areas must be considered in evaluations of most favorable areas to the presence of the insect to guide monitoring and local management strategies. The present work evaluated the H. armigera migration potential by air masses toward producing areas of host crops in São Paulo state, Brazil, considering its municipalities producing cotton, soybean, corn, tomato, bean, coffee, orange, eucalyptus and pine trees, as well as the main Brazilian air masses and the presence of Cerrado Biome in the state. Information of producing and of planting areas with the annual crops by municipalities in 2008, 2012 and 2014, provided by IBGE, were used in cross-maps made using ArcGis 10.3, in order to identify the location of their respective greater areas in each year. Cross-maps also provided results indicating the planting areas in 2014 with all crops already mentioned, as well as the potential areas to infestation/reinfestations of H. armigera in São Paulo State, in order to guide monitoring and local strategies of IPM.

The legitimation of risk and Bt cotton: a case study of Bantala village in Warangal, Andhra Pradesh, India

This article explores Ulrich Beck’s theorisation of risk society through focusing on the way in which the risk of Bt cotton is legitimated by six cultivators in Bantala, a village in Warangal, Andhra Pradesh, in India. The fieldwork for this study was conducted between June 2010 and March 2011, a duration chosen to coincide with a cotton season. The study explores the experience of the cultivators using the ‘categories of legitimation’ defined by Van Leeuwen. These are authorisation, moral evaluation, rationalisation and mythopoesis. As well as permitting an exploration of the legitimation of Bt cotton by cultivators themselves within the high-risk context of the Indian agrarian crisis, the categories also serve as an analytical framework with which to structure a discourse analysis of participant perspectives. The study examines the complex trade-off, which Renn argues the legitimation of ambiguous risk, such as that associated with Bt technology, entails. The research explores the way in which legitimation of the technology is informed by wider normative conceptualisations of development. This highlights that, in a context where indebtedness is strongly linked to farmer suicides, the potential of Bt cotton for poverty alleviation is traded against the uncertainty associated with the technology’s risks, which include its purported links to animal deaths. The study highlights the way in which the wider legitimation of a neoliberal approach to development in Andhra Pradesh serves to reinforce the choice of Bt cotton, and results in a depoliticisation of risk in Bantala. The research indicates, however, that this trade-off is subject to change over time, as economic benefits wane and risks accumulate. It also highlights the need for caution in relation to the proposed extension of Bt technology to food crops, such as Bt brinjal (aubergine).

Differences between root traits of early- and late-successional trees influence below-ground competition and seedling establishment.

The competitive influence of the root system of the exotic grass Urochloa brizantha and the widespread forb Leonotis nepetifolia on the emergence, survival and early growth of the seedlings of eight tropical heliophilous herbaceous species, six early-successional woody species and five late-successional woody species from Brazil, grown in 3500-cm3 pots and in greenhouse without light restriction were assessed. The density of fine-root systems produced by the forb and the grass in pots were 6.8 cm cm-3 soil and 48.1 cm cm-3 soil, respectively. Seedlings survival of the heliophilous herbaceous, early- and late-successional woody species were 86%, 70% and 100% in presence of the forb root system and 12%, 14% and 100% in competition with grass root system, respectively. The competitive pressure applied by the grass root system on seedling growth of the heliophilous herbaceous, early- and late-successional woody species were 2.4, 1.9 and 1.4 times greater than the forb root system. Total root length of the heliophilous herbaceous, early- and late-successional woody species grown without competitors were 13, 33 and 5 times greater than in competition with forb, and were 66, 54 and 6 times greater than in competition with grass root system, respectively. The averages of fine-root diameter of plants grown without competitors were 209 microm for the heliophilous herbaceous, 281 microm for early-successional trees and 382 microm for late-successional trees. The root system of the forb did not avoid seedling establishment of most plant species, but the grass root system hampered more the establishment of heliophilous herbaceous and early-successional woody species than the seedling establishment of late-successional woody species. The different density of root systems produced in soil by the forb and the grass, and the distinct root traits (e.g. root diameter and root tissue density) of the early- and late-successional plant species can explain the differences in the establishment of seedlings of plant species belonging to different groups of tropical succession when exposed to below-ground competition.

The Effect of Feed and Gonadotrophin Hormone on The Litter Size and Birth Weight of Local Goat

A study was conducted to determine the effect of ration and gonadotrophin hormone on the reproductive performance of breeding goat. Twenty-three does and six bucks having a similar age (1.5-2.0 years old) were exposed in this experiment. An amount of 200 gram of concentrate consisting of yellow corn (30%), rice brand (50%) and commercial concentrate CP 235 (20%) has been given to each experimental animal. The experimental animals were divided into two groups. The group was fed (12 animals) with M1 (roughage + concentrate) and the other group was fed (11 animals) with M2 (roughage). Moreover, each group were divided into four treatment namely: T1 (PGF2α), T2 (PGF2α + PMSG), T3 (PGF2α + HCG) and T4 (PGF2α + PMSG + HCG). The same treatment was applied to the other group. The data such as litter size and birth weight were collected from each experimental goat. Those data were analyzed using a 2 x 4 factorial experiment with unequal repetition. A composition between to treatment group was done using Least Significant Design (LSD). The result of the study showed that the effect of ration and gonadotrophin hormone on litter size and birth weight didn’t show any significant difference (P>0.05). The average of litter size and birth weight from roughage + concentrate treated animal (goat) were 1.92 and 1.51 kilogram respectively. However, the average of litter size and birth weight from roughage treated group were 1.58 and 1.48 kilogram respectively. (Animal Production 5(1): 5-10 (2003) Key words : Gonadotrophin, PGF2α, PMSG, Hormone, Goat