Isolation and selection of plant growth-promoting bacteria associated with sugarcane

v. 46, n. 2, p. 149-158, apr./jun. 2016.

Salicaceae Endophytes: Growth Promotion Potential in Rice (Oryza sativa L.) and Maize (Zea mays L.) and Bio-Control of Plant Pathogen

Thesis (Ph.D.)--University of Washington, 2016-08

Studies on the Catalytic Asymmetric Fischer Indolization

This work describes further developments and applications of the catalytic asymmetric Fischer indolization. In the first part of this thesis, the development of an organocatalytic asymmetric synthesis of helicenes via a Fischer indolization is discussed. The application of a novel SPINOL-derived phosphoric acid, featuring extended π-surfaces as 3,3‘-substituents which can potentially participate in π-interactions with the polyaromatic intermediate, afforded the corresponding products in high yields and enantioselectivities. The second part of this work describes the development of a catalytic asymmetric dearomatizing synthesis of 1,4-diketones via an interrupted Fischer indolization. Employing aryl hydrazines with α-substituents next to the hydrazine group prevents the rearomatization which takes place in common Fischer indole syntheses, thus enabling the hydrolysis of the generated diimine species. In the presence of STRIP as catalyst, a variety of different 1,4-diketones could be obtained in generally high yields, diastereo- and enantioselectivities. The last part of this thesis deals with the development of an organocatalytic asymmetric stereodivergent synthesis of novel 3H- and 2H-pyrroles, applying an interrupted Fischer indolization and for the 2H-pyrroles a subsequent stereospecific [1,5]-alkyl shift. Employing STRIP as catalyst afforded the corresponding products in good to excellent yields and enantioselectivities. Preliminary biological investigations of these novel structure motifs in cell-based assays, monitoring biological signal transduction pathways showed an inhibition of the Hedgehog signaling pathway in a μM range.

CHARACTERIZATION OF TOBACCO MOSAIC VIRUS DIRECTED REPROGRAMMING OF PHLOEM GENE EXPRESSION TO PROMOTE VIRAL PHLOEM LOADING AND SYSTMEIC MOVEMENT

Vascular phloem loading has long been recognized as an essential step in the establishment of a systemic virus infection. Yet little is known about this process and the mechanisms that control it. In this study, an interaction between the replication protein of Tobacco mosaic virus (TMV) and phloem specific auxin/indole acetic acid (Aux/IAA) transcriptional regulators was found to modulate virus phloem loading. Promoter expression studies show TMV 126/183 kDa interacting Aux/IAAs predominantly express and accumulate within the nuclei of phloem companion cells (CC). Furthermore, CC Aux/IAA nuclear localization is disrupted upon infection with an interacting virus but not during infection with a non-interacting virus. In situ analysis of virus spread shows the inability of TMV variants to disrupt Aux/IAA CC nuclear localization correlates with a reduced ability to load into the vascular tissue. Subsequent systemic movement assays also demonstrate that a virus capable of disrupting Aux/IAA localization is significantly more competitive at systemic movement than a non-interacting virus. Similarly, CC expression and over-accumulation of a degradation-resistant-interacting Aux/IAA protein was found to selectively inhibit TMV accumulation and phloem loading. Transcriptional expression studies demonstrate a role for interacting Aux/IAA proteins in the regulation of salicylic acid and jasmonic acid dependent host defense responses as well as virus specific movement factors including pectin methylesterase that are involved in regulating plasmodesmata size exclusion limits and promoting virus cell-to-cell movement. Further characterization of the phloem environment was done using two phloem specific promoters (pSUC2 and pSULTR2;2) to generate epitope-tagged polysomal-RNA complexes. Immuno-purification using the epitope tag allowed us to obtain mRNAs bound to polysomes (the translatome) specifically in phloem tissue. We found the phloem translatome is uniquely altered during TMV infection with 90% and 88% of genes down regulated in the pSUC2 and pSULTR2;2 phloem translatomes, compared to 31% of genes down regulated in the whole plant p35S translatome. Transcripts down regulated in phloem include genes involved in callose deposition at plasmodesmata, host defense responses, and RNA silencing. Combined, these findings indicate TMV reprograms gene expression within the vascular phloem as a means to enhance phloem loading and systemic spread.

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.

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.

Extraction and purification of theobromine using ionic liquids

Theobromine is an alkaloid present in cocoa and it is used in the treatment of atherosclerosis, hypertension, angina, among others. Due to its importance, the aim of this work consists on the development of an efficient and sustainable technology for the extraction of theobromine from cocoa beans. For the development of a purification technique for theobromine extracted from cocoa, aqueous biphasic systems (ABS) composed of ionic liquids (ILs) were initially studied to infer on the most promising systems. Cholinium-based ILs, based on a non-toxic and biocompatible cation, were used combined with two polymers (PPG 400 and PEG 400) and an inorganic salt (K3PO4). The respective phase diagrams at 298 K and atmospheric pressure were determined, as well as their extraction efficiencies for theobromine. The results obtained indicate that K3PO4 has a greater ability to induce the formation of ABS compared to PEG 400 and PPG 400. ABS consisting of K3PO4 also have a high potential for the extraction of theobromine, with extraction efficiencies ranging between 96.4 and 99.9 %. Based on the most promising ILs for the purification step, they were further used in aqueous solution to extract theobromine from cocoa beans, with extraction yields ranging between 4.5% and 6.5 wt%. Finally, ABS were applied to the aqueous solutions containing theobromine from the cocoa extract, with extraction efficiencies ranging between 96.7 and 99.0%.

Transcriptome, carbohydrate, and phytohormone analysis of Petunia hybrida reveals a complex disturbance of plant functional integrity under mild chilling stress

Cultivation of chilling-tolerant ornamental crops at lower temperature could reduce the energy demands of heated greenhouses. To provide a better understanding of how sub-optimal temperatures (12 degrees C vs. 16 degrees C) affect growth of the sensitive Petunia hybrida cultivar 'SweetSunshine Williams', the transcriptome, carbohydrate metabolism, and phytohormone homeostasis were monitored in aerial plant parts over 4 weeks by use of a microarray, enzymatic assays and GC-MS/MS. The data revealed three consecutive phases of chilling response. The first days were marked by a strong accumulation of sugars, particularly in source leaves, preferential up-regulation of genes in the same tissue and down-regulation of several genes in the shoot apex, especially those involved in the abiotic stress response. The midterm phase featured a partial normalization of carbohydrate levels and gene expression. After 3 weeks of chilling exposure, a new stabilized balance was established. Reduced hexose levels in the shoot apex, reduced ratios of sugar levels between the apex and source leaves and a higher apical sucrose/hexose ratio, associated with decreased activity and expression of cell wall invertase, indicate that prolonged chilling induced sugar accumulation in source leaves at the expense of reduced sugar transport to and reduced sucrose utilization in the shoot. This was associated with reduced levels of indole-3-acetic acid and abscisic acid in the apex and high numbers of differentially, particularly up-regulated genes, especially in the source leaves, including those regulating histones, ethylene action, transcription factors, and a jasmonate-ZIM-domain protein. Transcripts of one Jumonji C domain containing protein and one expansin accumulated in source leaves throughout the chilling period. The results reveal a dynamic and complex disturbance of plant function in response to mild chilling, opening new perspectives for the comparative analysis of differently tolerant cultivars.

A transcriptome approach towards understanding the development of ripening capacity in 'Bartlett' pears (Pyrus communis L.)

Background: The capacity of European pear fruit (Pyrus communis L.) to ripen after harvest develops during the final stages of growth on the tree. The objective of this study was to characterize changes in 'Bartlett' pear fruit physico-chemical properties and transcription profiles during fruit maturation leading to attainment of ripening capacity. Results: The softening response of pear fruit held for 14days at 20°C after harvest depended on their maturity. We identified four maturity stages: S1-failed to soften and S2- displayed partial softening (with or without ET-ethylene treatment); S3 - able to soften following ET; and S4 - able to soften without ET. Illumina sequencing and Trinity assembly generated 68,010 unigenes (mean length of 911bp), of which 32.8% were annotated to the RefSeq plant database. Higher numbers of differentially expressed transcripts were recorded in the S3-S4 and S1-S2 transitions (2805 and 2505 unigenes, respectively) than in the S2-S3 transition (2037 unigenes). High expression of genes putatively encoding pectin degradation enzymes in the S1-S2 transition suggests pectic oligomers may be involved as early signals triggering the transition to responsiveness to ethylene in pear fruit. Moreover, the co-expression of these genes with Exps (Expansins) suggests their collaboration in modifying cell wall polysaccharide networks that are required for fruit growth. K-means cluster analysis revealed that auxin signaling associated transcripts were enriched in cluster K6 that showed the highest gene expression at S3. AP2/EREBP (APETALA 2/ethylene response element binding protein) and bHLH (basic helix-loop-helix) transcripts were enriched in all three transition S1-S2, S2-S3, and S3-S4. Several members of Aux/IAA (Auxin/indole-3-acetic acid), ARF (Auxin response factors), and WRKY appeared to play an important role in orchestrating the S2-S3 transition. Conclusions: We identified maturity stages associated with the development of ripening capacity in 'Bartlett' pear, and described the transcription profile of fruit at these stages. Our findings suggest that auxin is essential in regulating the transition of pear fruit from being ethylene-unresponsive (S2) to ethylene-responsive (S3), resulting in fruit softening. The transcriptome will be helpful for future studies about specific developmental pathways regulating the transition to ripening. © 2015 Nham et al.

The tolerance of steers (Bos taurus) to sorghum ergot (Claviceps africana) in a feedlot during the cooler months in sub-tropical Queensland

Two experiments tested the tolerance of steers (Bos taurus) to sorghum ergot (Claviceps africana) during cooler months in south-east Queensland. Sorghum grain containing 2.8% ergot and 28 mg/kg ergot alkaloids (84% dihydroergosine, 10% dihydroelymoclavine, 6% festuclavine) was incorporated into feedlot rations. In a previous study in summer–autumn, ergot (1.1–4.4 mg alkaloids/kg ration) severely reduced performance in steers when the temperature–humidity index (THI; dry bulb temperature °C + 0.36 dew-point temperature °C + 41.2) was ~70, whereas a THI of ~79 was tolerated by steers fed ergot-free rations. Experiment 1 was conducted in winter–spring, with rations containing 0, 2.8, 5.6, 8.2 or 11.2 mg ergot alkaloids/kg ration. All ergot inclusions depressed feed intake (14% average reduction) and growth rate (34% average reduction), even when the weekly average daily THI was less than 65. Rectal temperatures were occasionally elevated in ergot-fed steers (P < 0.05), primarily when the THI exceeded ~65. All ergot inclusions depressed plasma prolactin concentrations in steers. Experiment 2 was predominantly carried out in winter, with weekly average daily THI <65 throughout the experiment. Rations containing 0, 0.28, 0.55 or 1.1 mg ergot alkaloids/kg were fed for 4 weeks but produced no significant effect on feed intakes and growth rates of steers. Alkaloid concentrations were then changed to 0, 2.1, 4.3 and 1.1 mg/kg, respectively. Subsequently, feed intakes declined by 17.5% (P < 0.05), and growth rates by 28% (P > 0.05) in the group receiving 4.3 mg/kg alkaloid, compared with Controls. Plasma prolactin concentrations were depressed, relative to the Controls, by dietary alkaloid inclusion greater than 1.1 mg/kg, with alkaloid intake of 4.3 mg/kg causing the greatest reduction (P < 0.05). Cattle performance in these studies shows steers can tolerate up to ~2 mg ergot alkaloid/kg (0.2% ergot) in feedlot rations under low THI conditions (< ~60–65), but previous findings indicate a much lower threshold will apply at higher THI (>65).

The tolerance of steers (Bos taurus) to sorghum ergot (Claviceps africana) in a feedlot during the cooler months in sub-tropical Queensland

Two experiments tested the tolerance of steers (Bos taurus) to sorghum ergot (Claviceps africana) during cooler months in south-east Queensland. Sorghum grain containing 2.8% ergot and 28 mg/kg ergot alkaloids (84% dihydroergosine, 10% dihydroelymoclavine, 6% festuclavine) was incorporated into feedlot rations. In a previous study in summer–autumn, ergot (1.1–4.4 mg alkaloids/kg ration) severely reduced performance in steers when the temperature–humidity index (THI; dry bulb temperature °C + 0.36 dew-point temperature °C + 41.2) was ~70, whereas a THI of ~79 was tolerated by steers fed ergot-free rations. Experiment 1 was conducted in winter–spring, with rations containing 0, 2.8, 5.6, 8.2 or 11.2 mg ergot alkaloids/kg ration. All ergot inclusions depressed feed intake (14% average reduction) and growth rate (34% average reduction), even when the weekly average daily THI was less than 65. Rectal temperatures were occasionally elevated in ergot-fed steers (P < 0.05), primarily when the THI exceeded ~65. All ergot inclusions depressed plasma prolactin concentrations in steers. Experiment 2 was predominantly carried out in winter, with weekly average daily THI <65 throughout the experiment. Rations containing 0, 0.28, 0.55 or 1.1 mg ergot alkaloids/kg were fed for 4 weeks but produced no significant effect on feed intakes and growth rates of steers. Alkaloid concentrations were then changed to 0, 2.1, 4.3 and 1.1 mg/kg, respectively. Subsequently, feed intakes declined by 17.5% (P < 0.05), and growth rates by 28% (P > 0.05) in the group receiving 4.3 mg/kg alkaloid, compared with Controls. Plasma prolactin concentrations were depressed, relative to the Controls, by dietary alkaloid inclusion greater than 1.1 mg/kg, with alkaloid intake of 4.3 mg/kg causing the greatest reduction (P < 0.05). Cattle performance in these studies shows steers can tolerate up to ~2 mg ergot alkaloid/kg (0.2% ergot) in feedlot rations under low THI conditions (< ~60–65), but previous findings indicate a much lower threshold will apply at higher THI (>65).

Estudo da influência da morfologia de nanopartículas de sílica mesoporosa na liberação do anticancerígeno indol-3-carbinol

Dissertação (mestrado)—Universidade de Brasília, Faculdade de Ceilândia, Programa de Pós-graduação em Ciências e Tecnologias em Saúde, 2015.

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.

Hyperspectral sensing to detect the impact of herbicide drift on cotton growth and yield

Yield loss in crops is often associated with plant disease or external factors such as environment, water supply and nutrient availability. Improper agricultural practices can also introduce risks into the equation. Herbicide drift can be a combination of improper practices and environmental conditions which can create a potential yield loss. As traditional assessment of plant damage is often imprecise and time consuming, the ability of remote and proximal sensing techniques to monitor various bio-chemical alterations in the plant may offer a faster, non-destructive and reliable approach to predict yield loss caused by herbicide drift. This paper examines the prediction capabilities of partial least squares regression (PLS-R) models for estimating yield. Models were constructed with hyperspectral data of a cotton crop sprayed with three simulated doses of the phenoxy herbicide 2,4-D at three different growth stages. Fibre quality, photosynthesis, conductance, and two main hormones, indole acetic acid (IAA) and abscisic acid (ABA) were also analysed. Except for fibre quality and ABA, Spearman correlations have shown that these variables were highly affected by the chemical. Four PLS-R models for predicting yield were developed according to four timings of data collection: 2, 7, 14 and 28 days after the exposure (DAE). As indicated by the model performance, the analysis revealed that 7 DAE was the best time for data collection purposes (RMSEP = 2.6 and R2 = 0.88), followed by 28 DAE (RMSEP = 3.2 and R2 = 0.84). In summary, the results of this study show that it is possible to accurately predict yield after a simulated herbicide drift of 2,4-D on a cotton crop, through the analysis of hyperspectral data, thereby providing a reliable, effective and non-destructive alternative based on the internal response of the cotton leaves.

Utilização da berberina na identificação dos mastócitos do molusco Anomalocardia brasiliana

Berberine is an alkaloid used as a fluorochrome in the identification of heparin and DNA. Enerback, 1974, described the technique used until today to study granules rich in heparin of vertebrate mast cells. Santos et al., 2003, studied mast cells of the mollusk Anomalocardia brasiliana using biochemical and histological analysis. This work used the fluorescent dye berberine technique to improve characterization of these cells. Mollusk organs (ctenidium and mantle) were processed with routine histological techniques. Tissue sections were treated with berberine 0,02% in redistilled water acidified to pH 4, by the addition of citric acid for 20 minutes. The visualization was made through fluorescence microscopy with ultraviolet region emission. The mast cell fluorescence had a strong yellow color, where cell nuclei appeared more greenish. This result was very similar to the ones reported before. Mast cells are location at the epithelium surface is the same in both organs, mantle and ctenidium. The fluorescence was easily observed in the granules. Therefore, this technique showed to be good and sensitive to study mast cell of invertebrates