Concentrations Of Picloram Research Articles

First evidence of age-dependent decreases in non-persistent pesticide mixtures in nestlings of a farmland raptor

Intensive use of synthetic pesticides in conventional agriculture may harm non-target organisms through sublethal effects on life-history traits. Farmland birds are exposed throughout their life cycle, but the fate of non-persistent pesticide mixtures in wild birds remains unknown. In this study, we investigated changes in pesticide contamination levels in Montagu’s harrier (Circus pygargus) nestlings during their growth. In total, 35 chicks were sampled twice during the rearing period, and blood was tested for 116 pesticides to assess pesticide load through two proxies; the number of pesticides detected and the sum of pesticide concentrations. Body mass and tarsus length were also measured to estimate body condition. Across the two sampling times, nine herbicides, five insecticides and four fungicides were detected. Contamination levels decreased significantly with nestling age irrespective of sampling date, and there was no relationship between pesticide load and body condition. Moreover, concentrations of chlorpyrifos-methyl, fenpropidin, metamitron, picloram and S-metolachlor, all detected throughout the rearing period, were unrelated to any of the explanatory variables. However, ethofumesate concentrations decreased significantly with chick age. This study provides the first evidence that non-persistent pesticide mixtures can decrease with age in wild nestlings. This has implications for understanding how chicks are contaminated and provides new insights on pesticide fate within organisms.

Effect of gamma irradiation on proliferation and growth of friable embryogenic callus and in vitro nodal cuttings of ugandan cassava genotypes.

Cassava (Manihot esculenta Crantz) production and productivity in Africa is affected by two viral diseases; cassava mosaic disease (CMD) and cassava brown streak disease (CBSD). Induced mutagenesis of totipotent/embryogenic tissues or in vitro plant material can lead to the generation of CMD and/or CBSD tolerant mutants. To massively produce non-chimeric plants timely and with less labor, totipotent cells or tissues are a pre-requisite. This study aimed to determine the effect of gamma radiation on the proliferation and growth of friable embryogenic callus (FEC) and in vitro nodal cuttings respectively. To obtain FEC, 2-6 mm sized leaf lobes of nine cassava genotypes were plated on Murashige and Skoog (MS) basal media supplemented with varying levels (37, 50, 70, 100) μM of picloram for production of organized embryogenic structures (OES). The OES of five cassava genotypes (Alado, CV-60444, NASE 3, NASE 13 and TME 204) were crushed and plated in Gresshoff and Doy (GD) basal media in combination with the amino acid tyrosine in varying concentrations for FEC production. FEC from five cassava genotypes and in vitro nodal cuttings of nine genotypes were irradiated using five different gamma doses (0, 5, 10, 15, 20 and 25 Gy) at a dose rate of 81Gy/hr. The lethal dose (LD)50 was determined using the number of roots produced and flow cytometry was done to determine the ploidy status of plants. The highest production of OES was noted in Alado across varying picloram concentrations, while TME 204 obtained the highest amount of FEC. The irradiated FEC gradually died and by 28 days post irradiation, FEC from all five cassava genotypes were lost. Conversely, the irradiated in vitro nodal cuttings survived and some produced roots, while others produced callus. The LD50 based on number of roots varied from genotype to genotype, but plants remained diploid post-irradiation. Accordingly, the effect of gamma irradiation on Ugandan cassava genotypes (UCGs) was genotype-dependent. This information is foundational for the use of in vitro tissues as target material for cassava mutation breeding.

Optimized medium composition in Physalis alkekengi callus culture altered nitric oxide level for inducing antioxidant enzyme activities and secondary metabolites

Physalis alkekengi L. is a valuable medicinal plant from the Solanaceae family and has multiple therapeutic applications. This study aimed to develop an optimized protocol for callogenesis in P. alkekengi to obtain friable calluses with high biomass. The effect of different concentrations of picloram, casein hydrolysate (CH), basal media (Murashige and Skoog (MS) and Gamborg (B5)), and static magnetic field (SMF) were investigated on the callus induction and growth, signaling molecules, and enzymatic and non-enzymatic antioxidants. Results showed that CH (200 mgL−1) and SMF4 mT for 90 min increased callus induction and fresh weight in P. alkekengi, while different concentrations of picloram reduced callogenesis. Hypocotyl explants showed various callogenesis and metabolic responses depending on the basal medium type. The 2B5 medium supplied with CH 200 (mgL−1) induced friable and cream calluses with high biomass (0.62 g) compared to the MS medium (control). The maximum activity of superoxide dismutase and catalase activities was identified in the 2B5 medium and peroxidase in the 2MS medium. The highest total phenolic (129.44 µg g−1DW) content and phenylalanine-ammonia lyase activity were obtained in the 2MS medium, and total withanolides (49.86 µg g-1DW) and DPPH radical scavenging activity were observed in the 2B5 medium. The 2MS medium boosted the hydrogen peroxide and nitric oxide levels, while their contents alleviated in the 2B5 medium, although these parameters were higher than the control. The findings of this study suggest that an effective protocol for successful callogenesis in P. alkekengi and the nutrient composition of culture medium by affecting the level of signaling molecules can control the antioxidant defense system and callus growth.

Analysis of secondary metabolites through callus cell suspension culture of black cumin (Nigella sativa L.) with PGR combination on MS media

This study aims to resolve the effect of using ZPT picloram and BAP and a combination of MS media on cell suspension culture and secondary metabolite compounds contained in black cumin (Nigella sativa L.) callus. The enquiry was performed at the Tissue Culture Laboratory, Faculty of Agriculture and Phytochemical Laboratory, Faculty of Pharmacy, Universitas Sumatera Utara, from February 2023 to July 2023. This study used a Totally Randomized Design (RAL) method amid 2 treatment elements and each treatment had 3 replications. The first treatment factor was ZPT concentration in MS medium (Z): Z1 (Picloram 0 ppm + BAP 1 ppm), Z2 (Picloram 1 ppm + BAP 1 ppm), Z3 (Picloram 1 ppm + BAP 2 ppm), Z4 (Picloram 2 ppm + BAP 2 ppm). The second factor is the composition of the MS (M) media: M1 (½ MS) and M2 (MS). Parameters observed were callus morphology, growth index, SCV (Settled Cell Volume), PCV (Peacked Cell Volume) and analysis of secondary metabolite content. The outcomes exposed that the arrangement of MS media and the PGR concentration of picloram and BAP had no expressive impact on all parameters. Z4 and M2 concentrations provided the most optimal growth.

Effect of Picloram and Desiccation on the Somatic Embryogenesis of Lycium barbarum L.

An efficient and reproducible in vitro method for indirect somatic embryogenesis was optimized by culturing leaf and leaf with petiole explants of Lycium barbarum L. Murashige and Skoog (MS) medium, supplemented with various concentrations of Picloram and 2,4-Dichlorophenoxyacetic acid (2,4-D), individually and in combinations, were tested. Picloram (1.0 µM) showed a better response compared to 2,4-D and results indicate it to be a better auxin for induction of somatic embryos for Goji berry. It was seen that the leaf explants were more responsive in callus and somatic embryo induction than the leaf with petiole explant when incubated in the dark for 5 weeks. Embryogenic callus, after being transferred to MS medium containing Benzyl amino purine (BAP) in 1.0 µM, 5.0 µM and 10.0 µM, began to differentiate in light after one week. MS medium with 1.0 µM Picloram + 10 µM BAP resulted as the most favorable treatment for somatic embryogenesis in Lycium barbarum L. Removal of plant growth regulators from MS medium and culturing induced calluses under 16 h photoperiod resulted in globular, heart, torpedo, cotyledons, and further development into plantlets. Well-developed plants have been obtained and are capable of acclimatizing in ex vitro conditions. In addition, the effects of desiccation treatments (0, 1, 3, 6, 9 h, and 12 h) on embryogenic callus for somatic embryo induction were found to be directly proportional to the length of desiccation treatment at room temperature. After 9 h and 12 h of desiccation treatments, 60% and 90% of plated calluses resulted in somatic embryos, respectively. In a L. barbarum callus mass, Acetocarmine and Evans blue double staining differentiated between embryogenic and non-embryogenic callus. These findings will help Goji berry improvement by elite clone production, ex situ conservation projects, scaling up plant production, and agronomy for the commercial production of this superfruit in the future.

Somatic embryo production and GFP genetic transformation in elite Ugandan cassava genotypes

Friable embryogenic callus (FEC) production is a prerequisite for the genetic transformation of cassava (Manihot esculenta Crantz). This process necessitates the development of organized embryogenic structures (OES) as a precursor. However, the production of OES and FECs in cassava is dependent on the genotype. This study aimed to optimize somatic embryo production in elite Ugandan cassava genotypes (NASE13, NASE19, and NAROCASS1) and demonstrate genetic transformation using the enhanced green fluorescent protein (eGFP) gene. OES were induced by initiating leaf-lobe explants on Murashige and Skoog (MS) media at various concentrations of 24D and picloram followed by friable embryogenic callus (FEC) production and regeneration. There were highly significant differences among the different media compositions in the percentage response to OES formation (p < 0.001). NASE 13 exhibited the highest regeneration frequency of 38.4%; while NAROCASS1 had the lowest regeneration frequency of 10.5% recorded. The presence of GFP gene in NAROCASS1 was confirmed by polymerase chain reaction (PCR) analysis of the transformed lines. Compared to existing studies, this research pioneers genotype-specific optimization for somatic embryo production and successful genetic transformation in Ugandan cassava variaties. The utilization of these genotypes, already embraced by farmers, presents promising opportunities to enhance them with desirable traits, further augmenting cassava cultivation.

Embryogenic callus induction of Indonesian Cassava (Menti and Adira 4) on different picloram concentrations

Cassava (Manihot esculenta Crantz) is an important staple crop in tropic countries. A reproducible somatic embryogenic callus production will be useful in the biotechnology breeding program for plants including cassava. Menti and Adira-4 are two types of high yield cassava genetic resources in Indonesia. Immature leaf lobes from cassava in vitro were used as explants on MS basal medium with five level concentrations of Picloram: 0 mg L−1, 3 mg L−1, 5 mg L−1, 10 mg L−1, and 12 mg L−1. Initial embryogenic callus were seen on the explants in 2 weeks after treatment. Explants were then subcultured to the same medium every three weeks until three cycles of subculture to proliferate pro-embryos. Result showed that MS basal medium containing picloram 3 – 12 mg L−1 could induce embryogenic callus formation in Menti and Adira 4 immature leaf lobe explants. The highest percentage of explants forming embryogenic callus was on D medium (MS + 3 mg L−1 Picloram). These findings would suggest that at all picloram concentrations tested both varieties of cassava showed a high embryogenic callus production. High frequency on embryogenic callus formation is a key step in cassava regeneration system specifically to support the modern breeding program.

Somatic embryogenesis and plant regeneration from transverse thin cell layers of adult peach palm (Bactris gasipaes) lateral offshoots.

In this work, we report a successful protocol to obtain in vitro peach palm (Bactris gasipaes Kunth) “Diamantes 10” plants through somatic embryogenesis from transverse thin cell layer (TCL) explants, dissected from three sections (basal, medial, and apical) of lateral offshoots of adult plants cultured on different concentrations of 4-amino-3,5,6-trichloropicolonic acid (picloram). After swelling and development of primary callus in all treatments, without any strong effect of explant origin or picloram concentration, it was possible to observe the formation of embryogenic structures and the exact point from where they developed. Browning was also observed and correlated to the induction treatments, although it was not an impairment for the production of embryogenic structures. Subsequent maturation and conversion of somatic embryos into plantlets allowed their acclimatization 17 months after culture initiation (ACI), which was quicker than previous reports with juvenile tissues (from embryos or seed-germinated plantlets). To the best of our knowledge, this is the first report on peach palm regeneration through somatic embryogenesis from TCL explants from adult plants and could constitute, after fine-tuning the acclimatization stage, a tool for mass clonal propagation of elite genotypes of this open-pollinated crop, as well as for the establishment of conservation strategies of in situ gene bank plant accessions endangered due to aging and other threats.

An Optimized Protocol for Indirect Organogenesis from Root Explants of Agapanthus praecox subsp. orientalis ‘Big Blue’

Agapanthus praecox has become a burgeoning variety in the flower market due to its high ornamental value with unique large blue-purple inflorescence. For rapid entering into the market, tissue culture technology or organogenesis has an attractive application over the conventional reproduction approach. In this study, a highly efficient protocol based on indirect organogenesis has been successfully established for A. praecox subsp. orientalis ‘Big Blue’. Two types of explants, root tips versus root segments, were compared for callus induction frequency in response to the induction culture media. The induction media contain Murashige and Skoog’s (MS) Basal Salt supplemented with various concentrations of picloram (PIC), 2,4-Dichlorophenoxyacetic acid (2,4-D), thidiazuron (TDZ), kinetin (KT) and naphthalene acetic acid (NAA). Of the two types of explants, root tips were found to be more effective for callus induction than root segments. Among the induction media tested, the highest callus induction rate (100.00%) was achieved when cultured on MS supplemented with 2.0 mg/L PIC, 1.5 mg/L KT and 0.1 mg/L NAA, which was probably accredited to higher endogenous phytohormone contents, especially of 3-indoleacetic (IAA). The optimal medium for callus proliferation was MS + 1.0 mg/L PIC + 1.0 mg/L 6-BA + 0.4 mg/L NAA, and the fresh weight increased by 72.74%. After being transferred onto the adventitious bud induction medium for 25 days, shoots were dedifferentiated from the surface of the flourishing callus, which then developed to the plantlet with roots in 90 days. The plantlets were transplanted in a greenhouse with a survival rate of 92.86%. This study innovatively established an indirect organogenesis tissue culture system of A. praecox with roots as explants, which provided a practical reference in its application.

Determination of genotoxic damages of picloram and dicamba with comet assay in Allium cepa rooted in tissue culture and distilled water.

Many genotoxicity tests allow us to understand the mechanism of damages on genetic material occurring in living organisms against various physical and chemical agents. One of them is the Comet test. The current study aimed to evaluate genotoxic caused by picloram and dicamba to root meristems of Allium cepa utilizing comet assay. Two different protocols were used for rooting and auxin/pesticide application. (i) A. cepa bulbs were rooted in MS medium and then treated with Murashige and Skoog (MS) medium (control) and 0.67, 1.34, 2.01, 2.68, 3.35, 4.02, and 8.04mg/L of picloram and dicamba using aseptic tissue culture techniques. (ii) A. cepa bulbs were then rooted in bidistilled water and treated with 0 (control), 0.67, 1.34, 2.01, 2.68, 3.35, 4.02, and 8.04mg/L of picloram and dicamba in distilled water. The A. cepa root tip cells in both treatment groups were examined using comet test to find the possible DNA damaging effects of picloram and dicamba. The results obtained at all the concentrations were statistically compared with their control groups. Almost at all the concentrations of Picloram and dicamba increased comet tail intensity (%) and tail moment in roots treated in MS medium. Two highest concentrations revealed toxic effect. On the other hand, DNA damaging effect of both auxins was only noted on the highest (> 4.02mg/L) in roots treated in distilled water. This study approve and confirm genotoxic effects of how growth regulators on plants. These findings give an evidence of DNA damage in A. cepa. Therefore, both picloram and dicamba should only be used in appropriate and recommended concentrations in agriculture to conserve ecosystem and to pose minimum threat to life.

In vitro embryo formation and plantlet regeneration from unpollinated ovaries of Lilium L

AbstractThe in vitro culture of unpollinated ovaries is one of the techniques used to induce gynogenesis. To explore the mechanism of lily gynogenesis and obtain haploids to shorten the breeding cycle, the in vitro effects of flower bud sampling time, exogenous plant growth regulators (PGRs), and different concentrations of picloram on embryo formation in unpollinated ovaries of Lilium L. ‘Sorbonne’ (oriental hybrids) were studied. The results indicated that flower buds ≥8 cm in length were most suitable. The PGRs picloram and 2,4‐dichlorophenoxyacetic acid (2,4‐D) with 6‐benzylaminopurine (6‐BA) or kinetin were useful in inducing ovule expansion in vitro. However, most ovules did not survive on the media containing 2,4‐D, whereas those grown on media containing picloram developed normally. In particular, a lower concentration of picloram (0.5 mg L−1) was found to be beneficial to the development of ovules into embryogenic shoots, whereas higher concentrations were more favorable to the development of shoots from the ovary wall. In total, 87 regenerated plants were obtained from unpollinated ovaries of Sorbonne. The chromosome ploidy of 20 of these was determined by counting root tip chromosomes, and eight of the regenerated plants were found to be haploids.

Effects of different factors on friable callus induction and establishment of cell suspension culture of Hovenia dulcis (Rhamnaceae)

Abstract Medicinal plants are an important therapeutic option for a large share of the world’s population. To establish an in vitro culture system for the production of secondary metabolites from Hovenia dulcis, we studied the effect of auxins, cytokinins, absence of light, and silver nitrate on the development of friable callus. Callus cultures were established for the first time and used to obtain cell suspension cultures. Supplementation with KIN (Kinetin) produced calli with both compact and friable areas, while the addition of TDZ (Thidiazuron) only produced compact callus. The maintenance of cultures in the dark induced a slight enhancement on friability when the auxin PIC (Picloram) was present in the culture medium. The addition of silver nitrate promoted the formation of friable calli. Dry weight analysis showed no significant differences in biomass growth, and, therefore, 2.0 mg.L-1 was considered the most suitable treatment. The presence of silver nitrate was not required for the establishment of cell suspension cultures. Dry weight analysis of cell suspensions showed higher biomass production in the absence of silver nitrate. PIC promoted 100% of cell suspension culture formation in the absence of silver nitrate, and higher biomass production was observed with the lowest concentration (0.625 mg.L-1). No morphological differences were observed among the different concentrations of PIC. Phytochemical screening showed the presence of saponins, flavonoids, flavonols and catechins in the extracts obtained from H. dulcis calli. These results show that the cell cultures herein established are potential sources for the production of H. dulcis secondary metabolites of medicinal interest.

Induction, Multiplication, and Evaluation of Antioxidant Activity of Polyalthia bullata Callus, a Woody Medicinal Plant.

Polyalthia bullata is an endangered medicinal plant species. Hence, establishment of P. bullata callus culture is hoped to assist in mass production of secondary metabolites. Leaf and midrib were explants for callus induction. Both of them were cultured on Murashige and Skoog (MS) and Woody Plant Medium (WPM) containing different types and concentrations of auxins (2,4-dichlorophenoxyacetic acid (2,4-D), α-naphthaleneacetic acid (NAA), picloram, and dicamba). The callus produced was further multiplied on MS and WPM supplemented with different concentrations of 2,4-D, NAA, picloram, dicamba, indole-3-acetic acid (IAA), and indole-3-butyric acid (IBA) media. The quantification of total phenolic content (TPC), total flavonoid content (TFC) and antioxidant capacity was further carried out on P. bullata callus, and the results were subjected to correlation analysis. Among the media, the WPM + 16.56 µM picloram (53.33 ± 22.06%) was the best for callus induction while MS + 30 µM dicamba was the best for callus multiplication. The TPC, TFC, and EC50 of DPPH scavenging activity were determined at 0.657 ± 0.07 mg GAE/g FW, 0.491 ± 0.03 mg QE/g, and 85.59 ± 6.09 µg/mL in P. bullata callus, respectively. The positive correlation between DPPH scavenging activity with TPC was determined at r = 0.869, and that of TFC was at r = 0.904. Hence, the P. bullata callus has an ability to accumulate antioxidants. It therefore can be a medium for secondary metabolites production.

In Vitro Induction of Callus and Flowers in Immature Oil Palm Inflorescences

This work aimed to investigate the effects of different combinations of auxin (0, 225 and 450 µM Picloram) and Murashige and Skoog salts (50% and 100% MS salt concentrations, respectively named as ½ MS and MS) in mediate the induction of callus and flowers from immature inflorescences (developmental stages I-9, I-12 and I-15) of the oil palm BRS – Manicoré hybrid. In I-9 inflorescence explants, the rate of callus induction was 17% higher in ½ MS than in MS; while in both I-12 and I-15 inflorescence explants, callus induction was over than 95% regardless of the MS salt concentration. The higher rate of callus induction in I-9 inflorescences was mediated by 450 µM Picloram, while the callus induction rates for either I-12 or I-15 inflorescences were higher than 92% regardless of Picloram concentration. Floral induction was lower in I-9 inflorescence explants (45% floral induction rate) than in both I-12 and I-15 inflorescence explants (floral induction rate higher than 90%). Our results suggest that Picloram (450 µM) and MS salts concentrations (½ MS) led to the highest rates of calluses and flowers induction in younger inflorescence explants (I-9); however, the medium composition is indifferent on callus and flower induction in I-12 and I-15 inflorescences.

Induction of Callogenesis, Organogenesis, and Embryogenesis in Non-Meristematic Explants of Bleeding Heart and Evaluation of Chemical Diversity of Key Metabolites from Callus.

Lamprocapnos spectabilis (L.) Fukuhara is a perennial plant species valued in the horticultural, cosmetic, and pharmaceutical markets. To date, however, there were no studies on tissue culture systems in this species when adjusted from non-meristematic explants. The aim of this study is to induce callogenesis, organogenesis, and somatic embryogenesis in non-meristematic explants of Lamprocapnos spectabilis ‘Alba’ cultured in various media and to analyze the chemical diversity of the produced callus. Leaf, petiole, and internode explants were cultured on the modified Murashige and Skoog (MS) medium fortified with various combinations and concentrations of 6-benzyladenine (BA), indole-3-acetic acid (IAA), 1-naphthaleneacetic acid (NAA), 2,4-dichlorphenoxyacetic acid (2,4-D), and picloram (PIC). After 10 weeks of culturing, the morphogenetic response of explants was evaluated and the concentration of chlorophylls, carotenoids, anthocyanins, and polyphenols in callus was analyzed. There was no influence of explant type on the callogenesis efficiency (62.1–65.3%). The highest fresh weight of callus was produced on leaf explants in the presence of 2,4-D or PIC. In contrast, the highest share of dry weight was found in internode-derived calli and cultured on IAA-supplemented medium (up to 30.8%). Only 2.5% of all explants regenerated adventitious shoots, while rhizogenesis was reported in 4.5% of explants. Somatic embryos were produced indirectly by 0% to 100% of explants, depending on the culture medium and explant type. The highest mean number of embryos (11.4 per explant) was found on petioles cultured in the MS medium with 0.5 mg·L−1 BA and 1.0 mg·L−1 PIC. Calli cultured in media with NAA usually contained a higher content of primary and secondary metabolites. There was also a significant impact of explant type on the content of anthocyanins, polyphenols, and carotenoids in callus. Further studies should focus on the elicitation of metabolites production in callus culture systems of the bleeding heart.

High-efficiency somatic embryogenesis techniques for different hybrids of cut lilies

Somatic embryogenesis (SE) is a unique method of in vitro regeneration for plant reproduction, germplasm resource preservation, and molecular-assisted breeding. Although lily is an internationally valued ornamental plant, its SE technology is still incomplete. Here, the effects of different explants, picloram (PIC) concentrations, and light conditions on somatic embryo induction, development, germination and seedling formation were studied in five Lilium varieties belonging to different hybrids. The embryo histomorphology, development and origins were observed, and a broadly efficient embryogenesis system was firstly established in Lilium. With leaf explants, all varieties tested underwent indirect somatic embryogenesis (SE). With scale explants, the Lilium Asiatic ‘Strawberry and Cream’, Lilium Oriental ‘Double Surprise’, and Lilium longiflorum Thunb. ‘White Heaven’ underwent indirect SE and the Lilium longiflorum × Asiatic hybrid ‘Eyeliner’ and Lilium longiflorum × Oriental ‘Triumphator’ underwent direct SE. Somatic embryogenesis in the above varieties involved four stages: globular, heart-shaped, torpedo-shaped, and cotyledon embryos. Two original SE pathways (endogenous and exogenous) were observed, and their appearance times differed. In indirect SE, exogenous embryos appeared first, followed by endogenous embryos; whereas in direct SE, the order was reversed. The different somatic embryo origins and the proportion of internal and external origins were closely related to the occurrence of somatic embryos by direct or indirect pathways and significantly affected the induction cycle of somatic embryos. Dark culture promoted lily somatic embryo induction, while 16 h/day light culture promoted germination, and the germination rates of all five lily varieties exceeded 90%. Murashige Skoog (MS) medium + (1.5–2.5 mg L−1) PIC + 0.2 mg L−1 1-Naphthaleneacetic acid (NAA) was suitable for SE from lily leaves and scales with an induction efficiency of 88–96%. Our research developed a widely applicable regeneration procedure for different hybrids of lily via somatic embryogenesis. Morphohistological analysis confirmed the origins, developmental stages and dynamic changes of cells during in vitro morphogenesis. This study will provide a valuable reference for the propagation of Lilium.

Production of Flavonoids in Callus Cultures of Sophora flavescens Aiton.

Flavonoids, including maackiain (Maac) from Sophora flavescens Aiton roots, have many pharmacological properties, such as antitumor, antimicrobial, and antifungal activities. This research aimed to develop an in vitro plant and callus culture system for S. flavescens for the purpose of generating an alternative production system for enhancing Maac production, as Maac is usually present in very small amounts in S. flavescens’ roots. We arranged the optimal conditions of different tissues of S. flavescens and supplemented the medium with various plant growth regulators (PGRs). The highest induction and proliferation rates of callus was shown in combination treatments of all concentrations of thidiazuron (TDZ) and picloram. In addition, calli induced with leaf explants cultured on 2.0 mg/L picloram and 0.5 mg/L 6-benzyladenine (BA) in Murashige and Skoog (MS) medium had the highest accumulation of the active metabolite Maac. In vitro shoots were regenerated on medium containing combinations of TDZ and α-Naphthalene acetic acid (NAA). A reliable protocol for the mass production of secondary metabolites using a callus culture of S. flavescens was successfully established.

Herbicide determination in Brazilian propolis using high pressure liquid chromatography

ABSTRACT Propolis is a widely used medicinal product sourced by bees from vegetation that may be frequently irrigated with herbicides. Exposure to herbicides can affect bees’ health and the quality of commercial propolis. The objective of this study was to calculate the concentrations of glyphosate, aminomethylphosphonic acid (AMPA), picloram and atrazine in different types of propolis from Brazil using high-performance liquid chromatography (HPLC). Four types of propolis (brown, green, red, and yellow) were evaluated for a total of 19 samples. Of these types of propolis, 47% tested positive for the herbicides atrazine (5 to 17.4 µg/g) and AMPA (10.2 to 11.3 µg/g). No samples were reported to be positive for glyphosate; however, the presence of AMPA indicates its existence. The concentrations observed in this study are less than international maximum-residue-level standards.

Growth of Elaeocarpus grandiflorus callus cultures in ms medium with various concentrations of growth regulators

Rejasa contains bioactive compounds.Production of bioactive compounds can be done through callus cultures. This study will examine rejasa callus the growth of in various types and concentrations of growth regulators. The independent variables of this study are the types and concentrations of growth regulators (2,4-dichlorophenoxyacetic acid (2,4-D) and Picloram). Dependent variable is growth of callus. Callus growth was determined from the percentage of explants that produced callus, time of callus formation, and callus morphology. The explants used were young petioles grown on Murashige& Skoog solid medium with the addition of 2,4-D and Picloram at various concentrations. The results showed that the lowest percentage of callus formation observed in explants grown on medium with addition of 3.5 ppm 2,4-D (27%). The explants grown on medium with the addition of 3.5 ppm picloram showed the highest callus growth percentage (93%). Explant grown on MS medium supplemented with3.5 picloram showed the best average time of the callus induction, 29.9 days. Callus that is formed mostly brown, and in some treatments produce green callus. Based on the results of this study, the best medium for induction of rejasa callus is MS medium with the addition of 3.5 ppmpicloram.

Explant type and hormone regime influences somatic embryogenesis and regeneration of cassava

Cassava (Manihot esculenta Crantz) is a tropical root crop that serves as a food staple and a vital income source to resource deprived farmers in the tropics. Despite its importance, cassava production and consumption is faced by a myriad of biotic/abiotic constraints. Genetic transformation which can be applied to mitigate these challenges however has as a prerequisite the availability of robust regeneration systems. This study evaluated the effect of explant type and hormone regime on somatic embryogenesis and regeneration of Kenyan cassava cultivars. The embryogenic competence of immature leaf lobe and stem explants of three cassava cultivars was determined by culturing them in MS medium supplemented with picloram (4, 6, 8 and 10 mg/l). The optimum media composition for embryo maturation, germination and plant recovery was assessed by culturing embryos in media supplemented with varying concentrations of 6-Benzylaminopurine (BAP), α–Naphthalene acetic acid (NAA) and Gibberellic acid (GA3). Somatic embryo formation frequencies in leaf explants were significantly higher (P≤0.05) than in stem explants. Embryo formation rates were found to increase with increasing concentrations of picloram. Cultivar 08/080 gave the least response to embryogenesis while embryogenesis rates for cultivar 08/274 were similar to the control cultivar TMS 60444. Maturation and plant recovery rates differed significantly with regard to the ratios of BAP, NAA and GA3 in the maturation media. The optimal medium for embryo germination and plant recovery was supplemented with 1 mg/l BAP, 0.02 mg/l NAA and 1.5 mg/l GA3. This optimized regeneration protocol can be coupled with mutation breeding or genetic transformation to improve the cassava germplasm. Key words: Cassava, explant, regeneration, somatic embryogenesis, plant growth regulator.