Jatropha curcas L. as a Plant Model for Studies on Vegetative Propagation of Native Forest Plants.

Even though it is a forest native plant, there are already several studies evaluating the small genome of Jatropha curcas L., which belongs to the Euphorbiaceae family, and may be an excellent representative model for the other plants from the same family. Jatropha curcas L. plant has fast growth, precocity, and great adaptability, facilitating silvicultural studies, allowing important information to be obtained quickly, and reducing labor costs. This information justifies the use of the species as a model plant in studies involving the reproduction of native plants. This study aimed to evaluate the possibility of using Jatropha curcas L. as a model plant for studies involving native forest plants and establish possible recommendations for the vegetative propagation of the species using hardwood cuttings. The information collected can be helpful to other native forest plant species, similar to Jatropha curcas L. To this end, the effects of hardwood cutting length (10, 20, and 30 cm) and the part of the hardwood cuttings (basal, middle, and apex) were evaluated. Moreover, the influence of immersing the hardwood cuttings in solutions containing micronutrients (boron or zinc) or plant regulators (2,4-D, GA3) and a biostimulant composed of kinetin (0.09 g L-1), gibberellic acid (0.05 g L-1), and 4-indole-3-butyric acid (0.05 g L-1). The experiments were carried out in duplicates. In one duplicate, sand was used as the substrate, and rooting evaluations were made 77 days after planting. In another duplicate, a substrate composed of 50% soil, 40% poultry litter, and 10% sand was used, and the evaluations of the saplings were performed 120 days after planting. The GA3 solutions inhibited the roots' and sprouts' emissions, while immersion in 2,4-D solution increased the number of primary roots at 77 days after planting. The hardwood cuttings from the basal part of the branch had the best results for producing saplings.

Biomass accumulation of Panax vietnamensis in cell suspension cultures varies with addition of plant growth regulators and organic additives.

OBJECTIVE: To evaluate the impact of plant growth regulators including kinetin (KN), benzyl adenine and naphthalene acetic acid, yeast extract and casein hydrolyzate on biomass accumulation of Vietnamese ginseng Panax vietnamensis (P. vietnamensis) in cell suspension culture. METHODS: Cell suspension cultures were established from friable calluses derived from leaves and petioles of 3-year-old in-vitro P. vietnamensis plants. The cell suspension cultures were grown in Murashige and Skoog basal media supplemented with various concentrations of KN, benzyl adenine, naphthalene acetic acid, and yeast extract and casein hydrolyzate. RESULTS: All tested factors generated an increase in the cell biomass of P. vietnamensis in suspension culture, but the impact of each varies depended on the factor type, concentration, and incubation period. Addition of 2.0 mg/L KN resulted in the largest biomass increase after 24 d, (57.0 ± 0.9) and (3.1 ± 0.1) mg/mL fresh and dry weight, respectively, whereas addition of benzyl adenine or naphthalene acetic acid produced optimum levels of Panax cell biomass at 1.0 and 1.5 mg/L, respectively. Addition of the elicitor yeast extract led to a 1.4-2.4 fold increase in biomass of P. vietnamensis, while addition of casein hydrolyzate enhanced biomass accumulation 1.8-2.6 fold. CONCLUSIONS: The addition of each factor causes significant changes in biomass accumulation of P. vietnamensis. The largest biomass accumulation is from cultures grown in MS media containing 2.0 mg/L KN for 24 d. The outcome of the present study provides new insights into the optimal suspension culture conditions for studies on the in vitro cell biomass production of P. vietnamensis.

MicroRNA and Transcription Factor: Key Players in Plant Regulatory Network.

Recent achievements in plant microRNA (miRNA), a large class of small and non-coding RNAs, are very exciting. A wide array of techniques involving forward genetic, molecular cloning, bioinformatic analysis, and the latest technology, deep sequencing have greatly advanced miRNA discovery. A tiny miRNA sequence has the ability to target single/multiple mRNA targets. Most of the miRNA targets are transcription factors (TFs) which have paramount importance in regulating the plant growth and development. Various families of TFs, which have regulated a range of regulatory networks, may assist plants to grow under normal and stress environmental conditions. This present review focuses on the regulatory relationships between miRNAs and different families of TFs like; NF-Y, MYB, AP2, TCP, WRKY, NAC, GRF, and SPL. For instance NF-Y play important role during drought tolerance and flower development, MYB are involved in signal transduction and biosynthesis of secondary metabolites, AP2 regulate the floral development and nodule formation, TCP direct leaf development and growth hormones signaling. WRKY have known roles in multiple stress tolerances, NAC regulate lateral root formation, GRF are involved in root growth, flower, and seed development, and SPL regulate plant transition from juvenile to adult. We also studied the relation between miRNAs and TFs by consolidating the research findings from different plant species which will help plant scientists in understanding the mechanism of action and interaction between these regulators in the plant growth and development under normal and stress environmental conditions.

Biomass accumulation of Panax vietnamensis in cell suspension cultures varies with addition of plant growth regulators and organic additives

Objective To evaluate the impact of plant growth regulators including kinetin (KN), benzyl adenine and naphthalene acetic acid, yeast extract and casein hydrolyzate on biomass accumulation of Vietnamese ginseng Panax vietnamensis (P. vietnamensis) in cell suspension culture. Methods Cell suspension cultures were established from friable calluses derived from leaves and petioles of 3-year-old in-vitro P. vietnamensis plants. The cell suspension cultures were grown in Murashige and Skoog basal media supplemented with various concentrations of KN, benzyl adenine, naphthalene acetic acid, and yeast extract and casein hydrolyzate. Results All tested factors generated an increase in the cell biomass of P. vietnamensis in suspension culture, but the impact of each varies depended on the factor type, concentration, and incubation period. Addition of 2.0 mg/L KN resulted in the largest biomass increase after 24 d, (57.0 ± 0.9) and (3.1 ± 0.1) mg/mL fresh and dry weight, respectively, whereas addition of benzyl adenine or naphthalene acetic acid produced optimum levels of Panax cell biomass at 1.0 and 1.5 mg/L, respectively. Addition of the elicitor yeast extract led to a 1.4–2.4 fold increase in biomass of P. vietnamensis, while addition of casein hydrolyzate enhanced biomass accumulation 1.8–2.6 fold. Conclusions The addition of each factor causes significant changes in biomass accumulation of P. vietnamensis. The largest biomass accumulation is from cultures grown in MS media containing 2.0 mg/L KN for 24 d. The outcome of the present study provides new insights into the optimal suspension culture conditions for studies on the in vitro cell biomass production of P. vietnamensis.

Biomass accumulation of Panax vietnamensis in cell suspension cultures varies with addition of plant growth regulators and organic additives

OBJECTIVE@#To evaluate the impact of plant growth regulators including kinetin (KN), benzyl adenine and naphthalene acetic acid, yeast extract and casein hydrolyzate on biomass accumulation of Vietnamese ginseng Panax vietnamensis (P. vietnamensis) in cell suspension culture.@*METHODS@#Cell suspension cultures were established from friable calluses derived from leaves and petioles of 3-year-old in-vitro P. vietnamensis plants. The cell suspension cultures were grown in Murashige and Skoog basal media supplemented with various concentrations of KN, benzyl adenine, naphthalene acetic acid, and yeast extract and casein hydrolyzate.@*RESULTS@#All tested factors generated an increase in the cell biomass of P. vietnamensis in suspension culture, but the impact of each varies depended on the factor type, concentration, and incubation period. Addition of 2.0 mg/L KN resulted in the largest biomass increase after 24 d, (57.0 ± 0.9) and (3.1 ± 0.1) mg/mL fresh and dry weight, respectively, whereas addition of benzyl adenine or naphthalene acetic acid produced optimum levels of Panax cell biomass at 1.0 and 1.5 mg/L, respectively. Addition of the elicitor yeast extract led to a 1.4-2.4 fold increase in biomass of P. vietnamensis, while addition of casein hydrolyzate enhanced biomass accumulation 1.8-2.6 fold.@*CONCLUSIONS@#The addition of each factor causes significant changes in biomass accumulation of P. vietnamensis. The largest biomass accumulation is from cultures grown in MS media containing 2.0 mg/L KN for 24 d. The outcome of the present study provides new insights into the optimal suspension culture conditions for studies on the in vitro cell biomass production of P. vietnamensis.

Optimization of an in vitro regeneration system for colombian indica rice varieties / Optimización de un sistema in vitro de regeneración para variedades colombianas de arroz indica

In order to optimize an in vitro regeneration system for colombian rice varieties, the amount of plant regulators used for each step of the process was modified, together with other factors such as seed position and time of exposition to plant regulators. Modification of 2,4-D concentration during callus induction and coculture has a negative effect on regeneration potential. However, it is possible to increase the amount of produced calli by modifying the position in which the seed is placed on the culture medium. Moreover, a longer callus induction time decreased the regeneration percentage. During the regeneration step, for FEDEARROZ 2000 variety, a combination of 1,0 mg/l g/l mg/l of naftalenacetic acid + 4,0 mg/l g/l mg/l of kinetin increases the regeneration percentage, without increasing the amount of abnormal shoots and without decreasing their size. For FEDEARROZ 50 variety, the best combination of plant regulators is 0,5 mg/l g/l mg/l of naftalenacetic acid + 2,0 mg/l g/l mg/l of kinetin.

Para optimizar un sistema de regeneración in vitro de variedades colombianas de arroz, se modificaron las concentraciones de los reguladores utilizados a lo largo de cada etapa del proceso de cultivo, junto con otros factores como la posición de siembra de la semilla y el tiempo de exposición a los reguladores. Se encontró que la modificación de la concentración de 2,4-D para las etapas de inducción de callo y cocultivo repercute de forma negativa en la regeneración. Por otra parte, es posible incrementar el porcentaje de formación de callo cambiando la posición de siembra de la semilla. Adicionalmente, un mayor tiempo de formación de callo provocó una disminución del porcentaje de regeneración. En la etapa de regeneración, para la variedad FEDEARROZ 2000 se encontró que una combinación de 1,0 mg/l g/l mg/l de ácido naftalenacético + 4,0 mg/l g/l mg/l de kinetina aumenta el porcentaje de regeneración, sin incrementar la cantidad de brotes anormales y sin disminuir de forma considerable el tamaño de los brotes. Para la variedad FEDEARROZ 50, la mejor combinación de reguladores durante la regeneración es de 0,5 mg/l g/l mg/l de ácido natfalenacético + 2,0 mg/l g/l mg/l de kinetina.

Giberelina, citocinina e auxina na qualidade química de bagas de uva 'Superior Seedless' / Gibberellin, citocinin and auxin on chemical quality of 'Superior Seedless' grape berries

Os experimentos foram conduzidos no município de Juazeiro/BA com a finalidade de avaliar o efeito de reguladores vegetais compostos por giberelina, citocinina e auxina na qualidade química de bagas de uva do cultivar Superior Seedless. No primeiro ensaio estudaram-se os efeitos de Stimulate (biorregulador) e X-Cyte (citocinina) associado a uma nova formulação comercial de giberelina (N-Large) e ao Pro-Gibb, produto usualmente utilizado como fonte de giberelina, aplicados via pulverização na fase de desenvolvimento das bagas (18, 21, 51 e 56 dias após a poda de produção). Para tanto se empregou os seguintes tratamentos: T1: Pro-Gibb; T2: Stimulate (Dose 1); T3: Stimulate (Dose 2); T4: Stimulate (Dose 3); T5: Pro-Gibb + X-Cyte (Dose Baixa - DB); T6: Pro-Gibb + X-Cyte (Dose Média - DM); T7: Pro-Gibb + X-Cyte (Dose Alta - DA); T8: N-Large; T9: N-Large + X-Cyte (DB); T10: N-Large + X-Cyte (DM); T11: N-Large + X-Cyte (DA). O segundo ensaio teve por objetivo verificar o efeito da nova formulação de giberelina (N-Large) associada ou não a citocinina (X-Cyte) também aplicados via pulverização direcionada aos cachos na fase de desenvolvimento das bagas (17, 55 e 66 dias após a poda de produção). Assim, os seguintes tratamentos foram empregados: T1: Pro-Gibb; T2: N-Large (DB); T3: N-Large (DM); T4: N-Large (DA); T5: N-Large (DB) + X-Cyte (DB); T6: N-Large (DB) + X-Cyte (DM); T7: N-Large (DB) + X-Cyte (DA); T8: N-Large (DM) + X-Cyte (DB); T9: N-Large (DM) + X-Cyte (DM); T10: N-Large (DM) + X-Cyte (DA); T11: N-Large (DA) + X-Cyte (DB); T12: N-Large (DA) + X-Cyte (DM); T13: N-Large (DA) + X-Cyte (DA). O delineamento experimental foi em blocos ao acaso com quatro repetições, onde se considerou cada parcela uma repetição com três plantas úteis na mesma linha de plantas. Por ocasião da colheita, momento em que a média dos cachos apresentava teor de sólidos solúveis superior a 15 ºBrix, foram retiradas bagas para as análises de sólidos solúveis, pH, acidez titulável e, cálculo do ratio (SS/AT). Em ambos os ensaios os reguladores vegetais avaliados não proporcionaram alterações significativas nas características químicas das bagas de uva 'Superior Seedless'. Assim, a ausência de diferença de resposta entre o produto comercialmente empregado (Pro-Gibb) e os demais testados (Stimulate, X-Cyte e N-Large) os torna promissores no cultivo da uva 'Superior Seedless', fornecendo maior número de alternativas aos produtores da região do Vale do São Francisco.

Trials were carried out in Juazeiro, Bahia State, Brazil, aiming to test plant regulators composed by gibberellin, cytokine and auxin effects on chemical quality of Superior Seedless grape berries. The first trial studied the effects of Stimulate (bio regulator) and X-Cyte (cytokine) associated to a new gibberellin formulation (N-Large ) and associated to Pro-Gibb , which is a product used as source of gibberellin. Products were sprayed at berries development phase (18, 21, 51 and 56 days after spur-pruning). Treatments were: T1: Pro-Gibb ; T2: Stimulate (Dose 1); T3: Stimulate (Dose 2); T4: Stimulate (Dose 3); T5: Pro-Gibb + X-Cyte (Low Dose - DB); T6: Pro-Gibb + X-Cyte (Intermediate Dose - DM); T7: Pro-Gibb + X-Cyte (High Dose - DA); T8: N-Large ; T9: N-Large + X-Cyte (DB); T10: N-Large + X-Cyte (DM); T11: N-Large + X-Cyte (DA). The second trial aimed to assess the effect of the new gibberellin formulation (N-Large ) associated or not with cytokine (X-Cyte ) also sprayed straight over the bunches at berries development phase (17, 55 e 66 days after spur-pruning). Treatments were: T1: Pro-Gibb - blank; T2: N-Large (DB); T3: N-Large (DM); T4: N-Large (DA); T5: N-Large (DB) + X-Cyte (DB); T6: N-Large (DB) + X-Cyte (DM); T7: N-Large (DB) + X-Cyte (DA); T8: N-Large (DM) + X-Cyte (DB); T9: N-Large (DM) + X-Cyte (DM); T10: N-Large (DM) + X-Cyte (DA); T11: N-Large (DA) + X-Cyte (DB); T12: N-Large (DA) + X-Cyte (DM); T13: N-Large (DA) + X-Cyte (DA). Experimental design was random blocks with four repetitions with each repetition/parcel having three useful plants in the same row. At harvest, when bunches average had soluble solids over 15 ºBrix, berries were collected for soluble solids, pH, titratable acidity analysis as well as (SS/AT) ratio calculation. In both trials, plant regulators evaluated did not provide significant changes on chemical quality of 'Superior Seedless' grape berries. Therefore, the lack of differences on response between the commercially used product (Pro-Gibb) and the other

Desenvolvimento vegetativo e produção de óleo essencial de patchouli (Pogostemon cablin (Blanco) Benth. ) após a aplicação de ácido giberélico e extrato de alga marinha / Development and essential oil production of Pogostemon cablin (Blanco) Benth. patchouli after applying gibberellic acid and seaweed extract

Pogostemon cablin (Blanco) Benth., comumente conhecido como patchouli, é uma espécie aromática utilizada pela indústria de perfumes devido a fragrância amadeirada e propriedade fixadora do óleo essencial. Fatores que afetam o teor dos constituintes do óleo essencial devem ser avaliados visando obter matéria prima de melhor qualidade. O experimento foi conduzido em casa de vegetação com delineamento inteiramente casualizado em esquema fatorial 2x3 e 4 repetições, sendo dois níveis de giberelina (0 e 200 mg L-1) e três níveis de extrato de alga marinha (0, 15 e 30 mg L-1). A aplicação dos reguladores vegetais foi realizada 30 dias após o plantio das mudas, e a avaliação do experimento deu-se 45 dias após a aplicação dos tratamentos. Não houve diferença significativa para a altura das plantas, a concentração de extrato de alga marinha de 15 mg L-1 promoveu aumento no número de folhas em relação à testemunha e a concentração de 30 mg L-1 promoveu decréscimo. Em relação à área foliar e ao número de folhas houve relação inversa para os níveis de extrato de alga marinha, onde o tratamento com maior concentração do regulador resultou em aumento da área foliar e redução do número de folhas. O teor de óleo essencial foi superior após a aplicação de 15 mg L-1 de extrato alga marinha isoladamente ou combinado com GA3. A produtividade do óleo essencial também aumentou com a aplicação 15 mg L-1 de extrato de alga marinha e quando utilizado somente GA3. A utilização de extrato de alga marinha aumentou a concentração de patchoulol no óleo essencial.

Pogostemon cablin (Blanco) Benth., commonly known as patchouli, is an aromatic species used by the perfume industry due to its woody fragrance and the fixative characteristic of its essential oil. Factors affecting the content of the constituents of essential oils should be evaluated in order to obtain raw materials with better quality. The experiment was carried out in a greenhouse in a completely randomized 2x3 factorial arrangement and four replications, with two levels of gibberellin (0 and 200 mg L-1) and three levels of seaweed extract (0, 15 and 30 mg L-1). Plant regulators were applied thirty days after planting the seedlings; the experiment was evaluated forty-five days after the treatments. There was no significant difference in plant height; the 15 mg L-1 concentration of seaweed extract promoted an increase in the number of leaves in relation to the control; and the 30 mg L-1 concentration promoted a decrease. In relation to leaf area and leaf number, there was an inverse relationship for the seaweed extract levels, in which the treatment with higher a concentration of the regulator resulted in an increased leaf area, reducing the number of leaves. The essential oil content was higher after applying 15 mg L-1 of seaweed extract alone or combined with GA3. The yield of essential oil also increased by applying 15 mg L-1 of seaweed extract and when only GA3 was used. The use of seaweed extract increased the patchoulol concentration in the essential oil.

Atividade das enzimas invertases e acúmulo de sacarose em cana-de-açúcar sob efeito do nitrato de potássio, etefon e etil-trinexapac1 / Invertases enzymes activity and sucrose accumulation in sugarcane (Saccharum officinarum L.) under effect the potassium nitrate, ethephon and ethyl-trinexapac

Conduziu-se este trabalho, com o objetivo de avaliar a atividade das enzimas invertases ácida e neutra e sua influência no processo de regulação do acúmulo de sacarose nos colmos da cana-de-açúcar, cultivar RB855453, sob efeito do nitrato de potássio comparado aos maturadores, da classe dos retardantes do crescimento, etefon e etil-trinexapac, buscando contribuir para o entendimento da ação desse composto químico. Os tratamentos constituíram-se da aplicação de quatro maturadores da classe dos retardantes do crescimento, cujos ingredientes ativos são etefon, etil-trinexapac, nitrato de potássio e nitrato de potássio + boro, e a maturação natural como testemunha, em delineamento experimental em blocos casualizados com cinco repetições. Os níveis enzimáticos das invertases ácida e neutra foram afetados de forma e intensidade distintas em função do princípio ativo utilizado como maturador e das condições climáticas; contudo, de forma geral, os níveis da invertase ácida manifestaram-se superiores aos da invertase neutra sem comprometer o acúmulo de sacarose nos colmos da cana-de-açúcar cultivar RB855453. O nitrato de potássio apresentou efeito maturador na cultura da cana-de-açúcar possibilitando o acúmulo de sacarose nos colmos, contudo as condições climáticas afetaram sua eficiência agronômica, tendo em vista que atua como indutor do processo de maturação.

The objective of this work was to evaluate the acid and neutral invertases enzymes activity and its influence in process of regulation sucrose accumulation in stems of RB855453 variety sugarcane under effect the potassium nitrate in compared with ripeners, of class of retainers growth, ethephon and ethyl-trinexapac, for to contribute the understanding of act this chemical compost. The treatments consisted of four ripeners of class of growth retainers, which active ingredient are ethefon, ethyl-trinexapac, potassium nitrate and potassium nitrate + boron and natural ripener as control, in a randomized block design with five replicates. The acid and neutral invertases enzymes levels were affected of form and intensity differents due active ingredient used as ripening and weather conditions; however, in general, the acid invertase levels were greatest the neutral invertase whitout to compromise the sucrose accumulation in stems of RB855453 cultivar sugarcane. The Potassium nitrate had effect ripener in sugarcane allowing sucrose accumulation in stems, however the weather conditions affected its agronomical efficiency, therefore act as inducers of the ripening process.

Comportamento do algodoeiro cultivar delta opal sob estresse hídrico com e sem aplicação de bioestimulante / Cotton plant cultivar delta opal behavior under water stress with or without biostimulant

Objetivou-se, com esse trabalho, avaliar o efeito do bioestimulante Stimulate®, aplicado ou não em diferentes fases do desenvolvimento do algodoeiro cv. Delta Opal, (Gossypium hirsutum L.) cultivado em casa de vegetação e submetido ou não ao estresse hídrico. Foram aplicados 25, 60 e 100 por cento do volume total de poros (VTP) preenchidos com água e o Stimulate® aplicado na semente, na semente e na fase de botão floral e sem a aplicação do bioestimulante, aos 69 dias após a semeadura. A duração do estresse hídrico foi de 15, 30 e 45 dias após a aplicação do estresse hídrico. O algodoeiro cv Delta Opal não tolerou deficiência hídrica de 25 por cento VTP iniciado na fase de botão floral, sendo observadas as menores alturas, diâmetros de colo, número de folhas, e comprometendo também a formação de estruturas reprodutivas. O bioestimulante na dose e na forma aplicada não proporcionou melhoras no desenvolvimento das plantas quando submetidas à falta de água, já em plantas submetidas ao excesso de água, houve aumento do diâmetro do colo das plantas.

This work was aimed at evaluating the effect of biostimulant Stimulate® applied or not at different development stages of cotton plant cv Delta Opal, (Gossypium hirsutum L.) cultivated in greenhouse, subjected or not to water stress. Applications of 25, 60 and 100 percent of total volume of pores (VTP) filled up with water were conducted, and then Stimulate® was sprinkled over the seed and on the flower bud stage, without biostimulant application 69 days after sowing. The water stress time period was that of 15, 30 and 45 days after water stress application. The cotton plant cv Delta Opal did not tolerate hydric deficit of 25 percent VTP on the flower bud set off, where smaller heights as well as smaller colon diameters and flower number were observed, all of which also endanger reproductive structure formation. The use of biostimulant on the dosage and the type of application did not render any improvement on plant development when submitted to lack of water; nevertheless there was a colon diameter increase on plants subjected to water excess.

Soluções conservantes em sorvetão pós-colheita / Preservative solutions in beehive ginger post-harvest

O objetivo deste trabalho foi estudar alguns aspectos da fisiologia pós-colheita de inflorescências de sorvetão cultivadas no Submédio São Francisco. Hastes florais recém-colhidas foram submetidas a diferentes tratamentos (água destilada; 75mg L-1 de nitrato de prata - AgNO3; 1000mg L-1 de cloreto de cobalto - CoCl2; 5mg L-1 de ácido giberélico - GA3 - Progibb® e 10mg L-1 de 6-Benzilaminopurina - BAP), em ambiente com temperatura e umidade controlada por 15 dias. A vida pós-colheita foi acompanhada a partir da escala de notas, da massa da matéria fresca e do consumo da solução conservante. O tratamento com AgNO3 em hastes de sorvetão, foi o mais eficiente na manutenção da vida de vaso de sorvetão, porém, devido a sua toxidez, recomenda-se o uso de GA3.

The objective of this research was to study some physiological post-harvest aspect of beehive ginger inflorescences grown in the lower middle San Francisco river basin. Flower stems just harvested were submitted to different treatments (distilled water; 75mg L-1 of silver nitrate - AgNO3; 1000mg L-1 of cobalt chloride - CoCl2; 5mg L-1 de GA3 - Progibb® and 10mg L-1 of 6-benzylamino purine - BAP), in an environment with controlled temperature and humidity, for 15 days. The post-harvest life was monitored from grading scale, fresh weight, and consumption of the preservative solution. The treatment with silver nitrate was the most efficient to keep the vase life of beehive ginger, however due to its toxity, the use of GA3 is recommended.