Plant-specific environmental and developmental signals regulate the mismatch repair protein MSH6 in Arabidopsis thaliana.

The DNA mismatch repair (MMR) is a postreplicative system that guarantees genomic stability by correcting mispaired and unpaired nucleotides. In eukaryotic nuclei, MMR is initiated by the binding of heterodimeric MutS homologue (MSH) complexes to the DNA error or lesion. Among these proteins, MSH2-MSH6 is the most abundant heterodimer. Even though the MMR mechanism and proteins are highly conserved throughout evolution, physiological differences between species can lead to different regulatory features. Here, we investigated how light, sugar, and/or hormones modulate Arabidopsis thaliana MSH6 expression pattern. We first characterized the promoter region of MSH6. Phylogenetic shadowing revealed three highly conserved regions. These regions were analyzed by the generation of deletion constructs of the MSH6 full-length promoter fused to the ß-glucuronidase (GUS) gene. Combined, our in silico and genetic analyses revealed that a 121-bp promoter fragment was necessary for MSH6 expression and contained potential cis-acting elements involved in light- and hormone-responsive gene expression. Accordingly, light exposure or sugar treatment of four-day old A. thaliana seedlings triggered an upregulation of MSH6 in shoot and root apical meristems. Appropriately, MSH6 was also induced by the stem cell inducer WUSCHEL. Further, the stimulatory effect of light was dependent on the presence of phyA. In addition, treatment of seedlings with auxin or cytokinin also caused an upregulation of MSH6 under darkness. Consistent with auxin signals, MSH6 expression was suppressed in the GATA23 RNAi line compared with the wild type. Our results provide evidence that endogenous factors and environmental signals controlling plant growth and development regulate the MSH6 protein in A. thaliana.

Composite coatings of beeswax + naphthalene acetic acid preserved fruit quality and antioxidants in stored lemon fruits.

Lemon fruits are well recognized for their richness in antioxidants. The present study was conducted to maintain the antioxidant properties of lemon fruits under long term cold storage. Fruits were given different treatments of naphthalene acetic acid (NAA) @ 50, 100 and 150 ppm plus beeswax (BW) @ 2% and were stored at 6-8 °C and 90-95% RH for 60 days. At the end of storage, fruits coated with NAA (50 ppm) + BW (2%) retained 42.14 and 34.61% antioxidants, 62.72 and 56.54% phenolic content and 17.72 and 13.80% hydroxyl radical scavenging capacity in peel and pulp, respectively as compared to the control. This treatment also resulted in lesser weight loss (5.27%), higher ascorbic acid content (46.31 mg 100 ml-1 juice) and titratable acidity (5.23%). Hence, NAA + BW coatings were promising for the maintenance of the postharvest antioxidant quality of stored lemons. Supplementary Information: The online version contains supplementary material available at 10.1007/s10068-023-01364-4.

Transcriptome and metabolome profiling unveils the mechanisms of naphthalene acetic acid in promoting cordycepin synthesis in Cordyceps militaris.

Cordycepin, an important active substance in Cordyceps militaris, possesses antiviral and other beneficial activities. In addition, it has been reported to effectively promote the comprehensive treatment of COVID-19 and thus has become a research hotspot. The addition of naphthalene acetic acid (NAA) is known to significantly improve the yield of cordycepin; however, its related molecular mechanism remains unclear. We conducted a preliminary study on C. militaris with different concentrations of NAA. We found that treatment with different concentrations of NAA inhibited the growth of C. militaris, and an increase in its concentration significantly improved the cordycepin content. In addition, we conducted a transcriptome and metabolomics association analysis on C. militaris treated with NAA to understand the relevant metabolic pathway of cordycepin synthesis under NAA treatment and elucidate the relevant regulatory network of cordycepin synthesis. Weighted gene co-expression network analysis (WGCNA), transcriptome, and metabolome association analysis revealed that genes and metabolites encoding cordycepin synthesis in the purine metabolic pathway varied significantly with the concentration of NAA. Finally, we proposed a metabolic pathway by analyzing the relationship between gene-gene and gene-metabolite regulatory networks, including the interaction of cordycepin synthesis key genes; key metabolites; purine metabolism; TCA cycle; pentose phosphate pathway; alanine, aspartate, and glutamate metabolism; and histidine metabolism. In addition, we found the ABC transporter pathway to be significantly enriched. The ABC transporters are known to transport numerous amino acids, such as L-glutamate, and participate in the amino acid metabolism that affects the synthesis of cordycepin. Altogether, multiple channels work together to double the cordycepin yield, thereby providing an important reference for the molecular network relationship between the transcription and metabolism of cordycepin synthesis.

In vitro study on the effect of cytokines and auxins addition to growth medium on the micropropagation and rooting of Paulownia species (Paulownia hybrid and Paulownia tomentosa).

This study represents an efficient preliminary protocol for in vitro mass production of two Paulownia species (Paulownia hybrid and Paulownia tomentosa) seedlings by using seed explant. Different concentrations of benzyladenine (BA) or Kinetin (Kin) (0.0, 2.0, 4.0, 6.0, 8.0 and 10.0 mg/L) were tested during multiplication stage. The number of shoots/explants was significantly increased with increasing either BA or Kin concentration; however, the shoot length significantly decreased. Data show that media fortified by BA (10 mg/L) combined with indole butyric acid (IBA) at 1.0 or 1.5 mg/L recorded the highest number of shoots/explant (9.13 and 9.25, respectively). After six weeks during the multiplication stage, data cleared that media fortified by benzyladenine (10 mg/L) combined with IBA at 0.5 mg/L recorded the highest shoot length (3.23 cm). The inclusion of indole butyric acid (IBA) or naphthalene acetic acid (NAA) at 1.0-1.5 mg/L to the medium significantly increased the number of roots/plantlets and the highest root length. The results indicated that IBA supplementation was more effective than NAA for in vitro rooting of both Paulownia species. The best treatment for multiplication was 10 mg/L and 8.0-10 mg/L BA for P. hybrid and P. tomentosa, respectively. Peat moss and sand (1:1, v/v) or peat moss and sand (1:2, v/v) were investigated as soil mixture during the adaptation stage. The results referred that Paulownia species plantlets were successfully survived (100 %) in soil mixture contained peat moss: sand (1:2, v/v). This mixture recorded the highest values of plantlet height and number of leaves/plantlets.

Alleviation of salinity stress effects on agro-physiological traits of wheat by auxin, glycine betaine, and soil additives.

Soil salinity is a major constraint to wheat production; it causes a severe reduction in wheat growth and yield. Alleviation of salinity effects on physiological, biochemical, and yield of wheat cultivars; Sids 14 and Misr 3 using some soil additions (control, Molasses and Humic acid), compatible solutes, and growth regulators (water as control, Naphthalene acetic acid, and Glycine betaine) were investigated in salt-affected soils. Results indicated that Misr 3 was superior to Sids 14 in all studied characteristics except flag leaf area, relative water content, plant height and recorded lower and desirable value of leaf temperature. The addition of Molasses (24 L ha-1) or Humic acid (12 L ha-1) significantly increased physiological and biochemical characteristics. At the same time, flag leaf temperature, proline, and malondialdehyde (MDA) content were decreased, yield and its attributes also increased except No. kernel spike-1. Foliar spray of Naphthalene acetic acid (NAA) at 30 mg L-1. or glycine betaine (GB 100 mM) also positively affected the studied characteristics, where Glycine betaine recorded the highest Relative water content and Fv/Fm. In contrast, NAA recorded the most increased Catalase (CAT) activity, and the Number of spikes m-2 and insignificant differences were observed between them in grain yield. It could be recommended the cultivation of Misr 3 with Molasses and GB under saline soils.

Clonal Propagation of Valeriana jatamansi Retains the Essential Oil Profile of Mother Plants: An Approach Toward Generating Homogenous Grade of Essential Oil for Industrial Use.

Valeriana jatamansi Jones (Syn. V. wallichii DC.) is an aromatic, medicinal herb used as a tranquilizer and in treating sleep disorders. Rhizome is mainly used to extract essential oil (EO) and valepotriates. High quality and economic yield of rhizomes are available in the third year of growth. Therefore, the cultivation of V. jatamansi is not picking up, and over-exploitation of this plant from wild habitats to meet the increasing demand of the pharmaceutical industry is the cause of threat to the genetic diversity of the species. Further, collections from the wild are heterogeneous, resulting in variable produce. The development of clonal lines can ensure uniform quality and yield of rhizome biomass. An effective clonal propagation method was standardized using different hormonal concentrations of naphthalene acetic acid (NAA) on apical shoot cuttings from the selected clone CSIR-IHBT-VJ-05 for different time durations and raised over various planting media. NAA treatment of 50 ppm concentration for 30 min was found optimum for root induction in apical shoots of V. jatamansi. Variations for EO composition within the clone were non-significant, while samples of the control population were variable. The best quality EO (patchouli alcohol ∼62%) was available during the third year of plant growth. A propagation technique for large-scale quality plant material (QPM) production has been standardized to reduce the stress over natural resources and promote V. jatamansi for use in the aromatic and pharmaceutical industry.

Potassium: A key modulator for cell homeostasis.

Potassium (K) is the most vital and abundant macro element for the overall growth of plants and its deficiency or, excess concentration results in many diseases in plants. It is involved in regulation of many crucial roles in plant development. Depending on soil-root interactions, complex soil dynamics often results in unpredictable availability of the elements. Based on the importance index, K is considered to be the second only to nitrogen for the overall growth of plants. More than 60 enzymes within the plant system depend on K for its activation, in which K act as a key regulator. K helps plants to resist several abiotic and biotic stresses in the environment. We have reviewed the research progress about K's role in plants covering various important considerations of K highlighting the effects of microbes on soil K+; K and its contribution to adsorbed dose in plants; the importance of K+ deficiency; physiological functions of K+ transporters and channels; and interference of abiotic stressor in the regulatory role of K. This review further highlights the scope of future research regarding K.

Indirect somatic embryogenesis of purple coneflower (Echinacea purpurea (L.) Moench): a medicinal-ornamental plant: evaluation of antioxidant enzymes activity and histological study.

Purple coneflower (Echinacea purpurea (L.) Moench) is a widely used medicinal and ornamental plant. In the present study, the callus embryogenesis was examined using benzyl adenine (BA) at three levels (3, 4, 5 mg L-1), 1-Naphthalene acetic acid (NAA) at three levels (0.1, 0.2 and 0.5 mg L-1) with or without activated charcoal (1 g L-1), coconut milk (50 ml L-1) and casein hydrolysate (50 mg L-1) in the MS (Murashige and Skoog 1962) medium. The embryogenesis indirectly occurred with the production of callus. The calli were observed in three forms: undifferentiated, embryogenic and organogenic. The embryogenic calli were dark green and coherent with a faster growth rate. The highest embryogenesis (100%) and embryonic regeneration (plantlet production) were obtained in the combined BA + NAA treatments with the activated charcoal, coconut milk and casein hydrolysate. However, the combined treatments of growth regulators failed to produce somatic embryos without the use of coconut milk and casein hydrolysate. The maximum amount of protein, peroxidase and catalase activity of embryogenic calli (2.02, 1.79 and 6.62ΔOD/Min/mg.protein, respectively), and highest percentage of acclimatization success (29.3% of plants) were obtained in the combined treatment of 5 mg L-1 BA + 0.5 mg L-1 NAA + activated charcoal + coconut milk + casein hydrolysate. The highest amount of chlorophyll content (33.3 SPAD value) and growth characteristics of acclimatized plantlets were observed in the media containing 3 mg L-1 BA + 0.1 and 0.2 mg L-1 NAA + 1 g. L-1 combined activated charcoal, coconut milk, casein hydrolysate. The histological studies confirmed the somatic embryogenesis in purple coneflower. Generally, it was found that the somatic embryogenesis of E. purpurea occurs at high levels of BA and low levels of NAA with the addition of coconut milk and casein hydrolysate.

Feasible regeneration and agro bacterium-mediated transformation of Brassica juncea with Euonymus alatus diacylglycerol acetyltransferase (EaDAcT) gene.

In the present study an effort has been made to optimize the in vitro regeneration protocol for Agrobacterium-mediated transformation of Brassica juncea, because of its importance as oilseed crops. The highest callus induction frequency of 87% was observed on MS (Murashige and Skoog, 1962) medium supplemented with 4 µM 6-benzyladenine (BA) after four weeks of culture period. Subculturing of organogenic calli in MS media with a similar hormonal composition resulted in shoot organogenesis after six weeks of culture cultivation. The highest shoot induction frequency (92%) was recorded on MS medium containing 4 µM BA in combination with 1 µM of α-naphthalene acetic acid (NAA). Further, well-developed roots were formed in MS media augmented with 6 µM of Indole acetic acid (IAA) in combination with 1 µM Kinetin (Kn). Cotyledon explants were exploited in vitro for the successful transformation of B. juncea. A binary vector comprised of the Euonymus alatus diacylglycerol acetyltransferase (EaDAcT) gene under the transcriptional control of a glycinin promoter and with a basta selection marker was introduced into A. tumefaciens strain GV3101 via electroporation. EaDAcT gene is responsible for unusual triacylglycerol's production where the sn-3 position is esterified with acetate instead of the long-chain fatty acid found in the triacylglycerol's. The highest regeneration frequency (100%) of transgenic shoots was observed on MS medium supplemented with 4 µM BA plus 1 µM NAA in the presence of 25 mg l-1 basta and 160 mg l-1 timintin. The efficiency of stable transformation was found to be approximately 7% in the transgenic plants. Moreover, the transformed regenerated shoots were confirmed by PCR analysis using EaDAcT gene-specific primers.

Effects of light intensity and plant growth regulators on callus proliferation and shoot regeneration in the ornamental succulent Haworthia.

BACKGROUND: Haworthia are desert succulents belonging to the Asphodelaceae family. Haworthia species are cultivated commercially as ornamentals and some rare species are quite valuable at retail market but growth slowly and difficult to propagation. However, an efficient micropropagation protocol was remained insufficient. RESULTS: The organogenic cultures obtained from inflorescence explants were cultured on Murashige and Skoog (MS) medium supplemented with various combinations of 6-benzylaminopurine (BA) and α-naphthalene acetic acid (NAA) under a light intensity of 10 µmol m-2 s-1 or 45 µmol m-2 s-1. The highest callus proliferation index (93.15%) with 1.0 mg L-1 BA + 0.1 mg L-1 NAA under a light intensity of 10 µmol m-2 s-1. The best shoot proliferation rates were on media with either 1 mg L-1 BA + 0-0.4 mg L-1 NAA (65.57-81.01%) under a light intensity of 45 µmol m-2 s-1. The highest root length (15.57 mm) and the highest rooting frequency (17 roots per shoot) were obtained when adventitious shoots were inoculated on MS medium with 0.4 mg L-1 NAA + 0.4 mg L-1 IBA. The survival rate of the transplanted plantlets was about 100%. The efficient micropropagation protocol proliferated Haworthia regenerate plants from inflorescence within 11 weeks. CONCLUSIONS: The present study determined the best combination of light intensity and plant growth regulators (PGRs) for improved organogenesis of Haworthia during propagation by tissue culture. This optimized protocol showed light intensity is an important factor for efficient callus or shoot regeneration. These results indicate that it will be useful to optimize the light conditions for future commercial cultivation, germplasm conservation, genetic engineering and molecular biology research of this ornamental plant.

Development of rapid, sensitive and selective fluorimetric method for determination of 1-naphthalene acetic acid in cucumber by using magnetite-molecularly imprinted polymer.

In this study, a novel method based on the determination of 1-naphthalene acetic acid with the usage of magnetite-molecularly imprinted polymer prior to fluorimetric detection has been developed. Magnetite-molecularly imprinted polymer has been used for the first time as selective adsorbent for the determination of 1-naphthalene acetic acid. The adsorption capacity of the synthesized polymer was found to be 2.18 ±â€¯0.36 mg g-1 (n = 3). Limit of detection (LOD) and limit of quantification (LOQ) of the method were found to be 0.75 and 2.50 µg L-1, respectively. Linearity of the calibration graph for the proposed method was observed within the range of 20-700 µg L-1. The proposed method seems to be rapid where the detection procedure for 1-naphthalene acetic acid can be completed within a total time of 1 h. The same imprinted polymer can be used for the determination of 1-naphthalene acetic acid with quantitative sorption and recovery values repeatedly for at least ten times. The effects of some potential organic interferences were investigated. Proposed method has been successfully applied to determine 1-naphthalene acetic acid in cucumber, where the recoveries of the spiked samples were found to be in the range of 93.7-104.5%. Characterization of the synthesized polymer was also evaluated. By combining the high capacity, cheapness, reusability and selectivity of the magnetic adsorbent with the dynamic calibration range, rapidity, simplicity, and sensitivity of fluorimetry, the proposed method seems to be an ideal method for the determination of trace levels of 1-naphthalene acetic acid.

Plant Growth Regulators Improve the Production of Volatile Organic Compounds in Two Rose Varieties.

The study focused on the influence of the plant growth regulators (PGRs) benzyladenine (BA) and naphthalene acetic acid (NAA) on the production of volatile organic compounds (VOCs) from the flowers of two modern rose varieties, Hybrid Tea and Floribunda. Thirty-six plants of Hybrid Tea and Floribunda were tested. Benzyladenine and naphthalene acetic acid were applied at 0, 100 and 200 mg/L to both rose varieties. Gas chromatography, coupled with flame ionization detection and mass spectrometry, was used to analyze and identify the volatile organic compounds from the flowers. A three-phase fiber 50/30 µm divinylbenzene/carboxen/polydimethylsiloxane was used to capture VOCs, at 2, 4 and 8 weeks, and 4 weeks was selected as it had the highest peak area. In total, 81 and 76 VOCs were detected after treatment of both rose varieties with BA and NAA, respectively. In addition, 20 compounds, which had significant differences between different treatments, were identified from both rose varieties. The majority of VOCs were extracted after the application of 200 mg (BA and NAA) /L of formulation, and four important compounds, cis-muurola-4(141)5-diene, y-candinene, y-muurolene and prenyl acetate, increased significantly compared to the controls. These compounds are commercially important aroma chemicals. This study used the rapid and solvent-free SPME method to show that BA and NAA treatments can result in significant VOC production in the flowers of two rose varieties, enhancing the aromatic value of the flowers. This method has the potential to be applied to other valuable aromatic floricultural plant species.

Differential Production of Phenylpropanoid Metabolites in Callus Cultures of Ocimum basilicum L. with Distinct In Vitro Antioxidant Activities and In Vivo Protective Effects against UV stress.

Ocimum basilicum L. (Purple basil) is a source of biologically active antioxidant compounds, particularly phenolic acids and anthocyanins. In this study, we have developed a valuable protocol for the establishment of in vitro callus cultures of O. basilicum and culture conditions for the enhanced production of distinct classes of phenylpropanoid metabolites such as hydroxycinnamic acid derivatives (caffeic acid, chicoric acid, rosmarinic acid) and anthocyanins (cyanidin and peonidin). Callus cultures were established by culturing leaf explants on Murashige and Skoog medium augmented with different concentrations of plant growth regulators (PGRs) [thidiazuron (TDZ), α-naphthalene acetic acid (NAA), and 6-benzyl amino purine (BAP)] either alone or in combination with 1.0 mg/L NAA. Among all the above-mentioned PGRs, NAA at 2.5 mg/L led to the highest biomass accumulation (23.2 g/L DW), along with total phenolic (TPP; 210.7 mg/L) and flavonoid (TFP; 196.4 mg/L) production, respectively. HPLC analysis confirmed the differential accumulation of phenolic acid [caffeic acid (44.67 mg/g DW), rosmarinic acid (52.22 mg/g DW), and chicoric acid (43.89 mg/g DW)] and anthocyanins [cyanidin (16.39 mg/g DW) and peonidin (10.77 mg/g DW)] as a function of the PGRs treatment. The highest in vitro antioxidant activity was determined with the ORAC assay as compared to the FRAP assay, suggesting the prominence of the HAT over the ET-based mechanism for the antioxidant action of callus extracts. Furthermore, in vivo results illustrated the protective action of the callus extract to limit the deleterious effects of UV-induced oxidative stress, ROS/RNS production, and membrane integrity in yeast cell culture. Altogether, these results clearly demonstrated the great potential of in vitro callus of O. basilicum as a source of human health-promoting antioxidant phytochemicals.

Effects of an Auxin and a brassinosteroid on physical, chemical and biochemical attributes of 'Galaxy' apples / Ação da auxina e do brassinosteroide nos atributos físico-químicos e bioquímicos de maçãs 'Galaxy'

ABSTRACT: The aim of this study was to study the effect of the auxin naphthalene acetic acid (NAA) and the brassinosteroid 28-homocastasterone (28-HCS) applied every 15 days (harvest 2015/16) or every 21 days (harvest 2016/17) after full bloom, on the physical, chemical and biochemical attributes of the 'Galaxy' apples. The study was performed at a commercial orchard at Vacaria county, Rio Grande do Sul State, Brazil. The following treatments were applied: water (control), NAA (0.1%), 28-HCS (10‒6 M) and NAA (0.1%) + 28-HCS (10‒6 M). For this purpose, 300 μL of the plant growth regulator solutions were applied to the fruit peduncles. Fruits treated with NAA every 21 days presented higher starch-iodine index and lower total soluble solids (TSS), titratable acidity (TA), lightness index (L) and hue angle (h°) than 28-HCS-treated fruits. Fruits treated every 15 days with NAA presented reduced skin rupture force (SRF), total antioxidant activities (TAA) of the skin and flesh, and hydrogen peroxide content compared to the control fruits. In addition, NAA application every 15 and 21 days resulted in increased anthocyanin content in the skin. Application of NAA + 28-HCS reduced superoxide dismutase activity. 28-HCS applications increased TSS, color attributes (C and h°) of the skin, total antioxidant activity of the skin and flesh and peroxidase activity compared to control fruits. These results are expected to help to understand how plant growth regulators affect apple quality. In addition, results described here are also expected to help on the development of strategies to reduce post harvest losses and to increase fruit shelf life.

RESUMO: O objetivo deste estudo foi avaliar a ação do ácido naftaleno acético (ANA) e do brassinosteroide (catasterona), aplicados a cada 15 (safra 2015/16) ou 21 (safra 2016/17) dias a partir dos 40 dias após a plena floração, nos atributos físico-químicos e bioquímicos de maçãs 'Galaxy'. O estudo foi realizado em um pomar comercial de macieiras 'Galaxy' no município de Vacaria, RS. Os tratamentos avaliados foram: água (controle), ANA (0,1%), catasterona (10‒6 M) e ANA (0,1%) + catasterona (10‒6 M). As aplicações foram realizadas no pedúnculo dos frutos, utilizando 300 µL de solução por pedúnculo. Frutos tratados com ANA a cada 21 dias tiveram maior índice de iodo-amido e menores valores de sólidos solúveis (SS), acidez titulável e dos atributos de cor L e ângulo hue (h°) do que os frutos tratados com catasterona. Aplicações de ANA a cada 15 dias reduziu a força de ruptura da casca (FRC), atividade antioxidante (AAT) da casca, da polpa e conteúdo de peróxido de hidrogênio. Além disso, ANA aplicada a cada 15 ou 21 dias aumentaram o teor de antocianinas na casca. ANA + catasterona reduziu a atividade da enzima superóxido dismutase. Entretanto, aplicações de catasterona a cada 15 dias aumentou o teor de SS, atributos de cor (C e h°) na casca, atividade antioxidante da casca e da polpa, compostos fenólicos totais (CFT) e atividade da enzima peroxidase dos frutos de maçãs. Portanto, estudos adicionais devem ser realizados para confirmar a ação desses fitorreguladores na manutenção da qualidade e redução das perdas pós-colheitas de maçãs 'Galaxy'.

High frequency regeneration of plants from cotyledon and hypocotyl cultures in Brassica oleracea cv. Pride of India.

Morphogenic potential of cabbage cv. Pride of India, for multiple shoot induction was tested under in vitro conditions using cotyledon and hypocotyl explants. Aseptically grown seven to nine days old seedlings of cabbage were used as source of explants for reproducible plant regeneration studies. Forty different concentrations and combinations of TDZ (alone), TDZ with adenine, TDZ with NAA and TDZ with IAA were tried. Maximum shoot regeneration response from cotyledon explants (91.11%) and hypocotyl (94.40%) was obtained on MS medium containing 0.330 mg/l TDZ + 79.70 mg/l Adenine and 0.220 mg/l TDZ + 0.088 mg/l IAA, respectively. Rooting was achieved within two to three weeks on all the rooting media, but MS medium containing 0.10 mg/l NAA produced the maximum number of strong and healthy roots (100%). The regenerated complete plantlets with healthy roots and shoot system were transferred to pots containing sterilized cocopeat and successfully acclimatized and no phenotypic variations were observed among regenerated plants. Highly efficient, reproducible plant regeneration protocol has been standardized in cabbage cv. Pride of India, which would be valuable for Agrobacterium-mediated gene transfer studies in cabbage.

Multiplication, rooting in vitro, and acclimatization of Brassavola tuberculata Hook. (Orchidaceae), an orchid endemic to the brazilian atlantic rainforest / Multiplicação, enraizamento in vitro e aclimatização de Brassavola tuberculata Hook (Orchidaceae), uma orquídea endêmica da mata atlântica brasileira

The objective of this study was to promote the establishment of an in vitro culture of Brassavola tuberculata, testing different concentrations of naphthalene acetic acid (NAA) and 6-benzylaminopurine (BAP) on multiplication and rooting, evaluating different substrates during acclimatization, as well as the effect of in vitro treatments. After germination, the seedlings of B. tuberculata were subjected to culture on MS medium supplemented with different concentrations of NAA and BAP, and multiplication and rooting were assessed. During acclimatization, different substrates were tested: S1, Plantmax® and vermiculite (1: 1); S2, Plantmax® and grit (1: 1); and S3, dust fern. Also the effect of the in vitro culture treatments was evaluated: T1, control; T5, (2.5 µM NAA +5 µM BAP); and T7, (5 µM NAA + 0 µM BAP). The favorable balance of cytokinins promoted by treatment T5 yielded the largest number of shoots and leaves in B. tuberculata. The greatest length of leaves and roots, and highest root number were observed in the treatment T7, favored by the presence of auxin. This treatment had a positive effect with respect to plant acclimatization: T7 associated with substrate S1 provided the most suitable conditions for acclimatization of seedlings of B. tuberculata, providing greater number and length of leaves, and high survival rate.

Objetivou-se, com este trabalho, promover o estabelecimento do cultivo in vitro de Brassavola tuberculata, testando diferentes concentrações de ANA e BAP na multiplicação e no enraizamento, e avaliar diferentes substratos e o efeito dos tratamentos de cultivo in vitro na aclimatização. Após a germinação das sementes, as plântulas de B. tuberculata foram submetidas ao cultivo em meio MS suplementado com diferentes concentrações de ácido naftaleno acético (ANA) e 6-benzilaminopurina (BAP), sendo avaliados a multiplicação e o enraizamento. Foram testados diferentes substratos: S1 (Plantmax e vermiculita (1:1)); S2 (Plantmax e areia (1:1)) e S3 (pó de xaxim) na aclimatização e, posteriormente, o efeito dos tratamentos do cultivo in vitro: T1 (controle), T5 (2,5 ANA + 5 BAP) e T7 (5 ANA + 0 BAP), na aclimatização. O balanço favorável às citocininas promovido pelo tratamento T5 (2,5 µM ANA + 5 µM BAP) promoveu maior número de brotos e de folhas em B. tuberculata. O maior comprimento das folhas, das raízes e maior número de raízes foi observado no tratamento T7 (5 µM ANA e 0 µM BAP), favorável a auxina. Este tratamento apresentou efeito positivo com relação a aclimatização das plantas: T7 associado ao substrato S1, Plantmax e vermiculita (1:1) proporcionou melhores condições para a aclimatização das plântulas de B. tuberculata, propiciando maior número e comprimento das folhas, e elevada taxa de sobrevivência.

Effects of naphthalene acetic acid (NAA) on the plant growth and sugars effects on the cut flowers Mokara chark kuan orchid / Efeitos do ácido naftaleno acético (naa) sobre o crescimento das plantas e de açúcares sobre flores cortadas da orquídea Mokara chark kuan

A study was carried out to investigate the effects of Naphthalene Acetic Acid (NAA) and sugars on the plants and flowers quality of Mokara Chark Kuan orchid respectively. Parameters were monitored between November 2014 to March 2015, with the application of three concentrations of NAA at 25, 50 and 100 mg/L in the field. The application of 25 mg/L NAA significantly increases the plant height, number of roots and total soluble solid content of Mokara Chark Kuan orchid plants. But, there was no significant effect on the leaves number, chlorophyll content and number of flowers. In case of storage experiment, the cut Mokara Chark Kuan flowers were treated with 3 % and 6 % glucose and sucrose. Physiological parameters: fresh weight, water uptake, petal thickness, discoloration and vase life were evaluated. It can be concluded that spraying with 25 mg/L NAA enhanced the plant growth and development of Mokara Chark Kuan orchid. The study also showed that 6 % sucrose was the best treatment for maintaining the post-harvest quality as well as vase life of cut Mokara Chark Kuan orchid flowers.

Um estudo foi realizado para investigar os efeitos do ácido naftaleno ácido acético (NAA) no crescimento de e açúcares no plantas e na qualidade de flores da orquídea Mokara Chark Kuan . Os parâmetros foram monitorados entre novembro de 2014 e março 2015, com a aplicação de três concentrações de ANA em 25, 50 e 100 mg / L sob condições de campo. A aplicação de 25 mg / L de ANA aumentou significativamente a altura da planta, o número de raízes e o teor de sólidos solúveis totais de plantas de orquídeas Mokara Chark Kuan. Todavia, não houve nenhum efeito significativo sobre o número de folhas, o teor de clorofila e o número de flores. No caso do experimento de armazenamento, flores cortadas de Mokara Chark Kuan foram tratadas com 3% e 6% de glicose e de sacarose. Os parâmetros fisiológicos: peso fresco, absorção de água, espessura pétala, descoloração e vida de vaso foram avaliados. Pode concluir-se que a pulverização com 25 mg / L de NAA aumentou o crescimento e o desenvolvimento das plantas de orquídea Mokara Chark Kuan. O estudo também mostrou que 6% de sacarose foi o melhor tratamento para a manutenção da qualidade pós-colheita, bem como vida de vaso em flores de corte da orquídea Mokara Chark Kuan .

Naphthalene Acetic Acid Potassium Salt (NAA-K+) Affects Conidial Germination, Sporulation, Mycelial Growth, Cell Surface Morphology, and Viability of Fusarium oxysporum f. sp. radici-lycopersici and F. oxysporum f. sp. cubense in Vitro.

The response to exogenous addition of naphthalene acetic acid potassium salt (NAA-K+) to Fusarium oxysporum f. sp radici-lycopersici ATCC 60095 and F. oxysporum f. sp. cubense isolated from Michoacan Mexico soil is reported. The in vitro study showed that NAA-K+ might be effective in the control of Fusarium oxysporum. Exogenous application of NAA-K+ affected both spores and mycelium stages of the fungi. Viability testing using acridine orange and propidium iodide showed that NAA-K+ possesses fungal killing properties, doing it effectively in the destruction of conidia of this phytopathogenic fungi. Analysis of treated spores by scanning electron microscopy showed changes in the shape factor and fractal dimension. Moreover, NAA-K+ repressed the expression of brlA and fluG genes. The results disclosed here give evidence of the use of this synthetic growth factor as a substance of biocontrol that presents advantages, and the methods of application in situ should be explored.

Development of an efficient callus proliferation system for Rheum coreanum Nakai, a rare medicinal plant growing in Democratic People's Republic of Korea.

A clonal mass propagation to obtain mountainous sources of Rheum coreanum Nakai, a rare medicinal plant in Democratic People's Republic of Korea was established by rhizome tissue culture. Whole plants were selected and collected as a vigorous individual free from blights and harmful insects among wild plants of R. coreanum grown on the top of Mt. Langrim (1.540 m above the sea) situated at the northern extremity of Democratic People's Republic of Korea. Induction of the callus was determined using four organs separated from the whole plant and different plant growth regulators. The callus was successfully induced from rhizome explant on MS medium containing 2.4-D (0.2-0.3 mg/l). In the MS medium supplemented with a combination of BAP (2 mg/l) and NAA (0.2 mg/l), single NAA (0.5 mg/l), or IBA (0.5 mg/l), a higher number of shoot, root and plantlets was achieved. The survival rate on the mountainous region of the plantlets successfully acclimatized (100%) in greenhouse reached 95%, and yields of crude drug and contents of active principles were higher than those obtained by sexual and vegetative propagation. This first report of R. coreanum tissue culture provides an opportunity to control extinction threats and an efficient callus proliferation system for growing resources rapidly on a large scale.

Elicitation of Medicinally Important Antioxidant Secondary Metabolites with Silver and Gold Nanoparticles in Callus Cultures of Prunella vulgaris L.

Prunella vulgaris L. (P. vulgaris) is an important medicinal plant with a wide range of antiviral properties. Traditionally, it is known as self-heal because of its faster effects on wound healing. It is commonly known as a natural antiseptic due to the presence of various polyphenols. There is lack of research efforts on its propagation and production of bioactive compounds under field and in vitro conditions. In this study, the effects of different ratios (1:2, 1:3, 2:1, and 3:1) of silver (Ag) and gold (Au) nanoparticles (NPs) alone or in combination with naphthalene acetic acid (NAA) were investigated for callus culture development and production of secondary metabolites. The Ag (30 µg l-1), AgAu (1:2), and AgAu (2:1) NPs in combination with NAA (2.0 mg l-1) enhanced callus proliferation (100 %) as compared to the control (95 %). Among the different NPs tested, AuNPs with or without NAA produced higher biomass in log phases (35-42 days) of growth kinetics. Furthermore, AgAu (1:3) and AuNPs alone enhanced total protein content (855 µg-BSAE/mg-fresh weight (FW)), superoxide dismutase (0.54 nM/min/mg-FW), and peroxidase (0.39 nM/min/mg-FW) enzymes in callus cultures. The AgAuNPs (1:3) in combination with NAA induced maximum accumulation of phenolics (TPC 9.57 mg/g-dry weight (DW)) and flavonoid (6.71 mg/g-DW) content. Moreover, AgAuNPs (3:1) without NAA enhanced antioxidant activity (87.85 %). This study provides the first evidence of NP effect on callus culture development and production of natural antioxidants in P. vulgaris.