Callus Diameter Research Articles

Effect of Rootone-F concentration on the cutting-growth of three species of coffee (Coffea sp.)

Liberica, Robusta and Excelsa coffee are cross-pollinated so that their vegetative propagation uses cuttings. Plant growth regulators (PGR) such as Rootone-F could accelerate root growth on cuttings. The study aimed to determine the effect of Rootone-F concentration on the growth of one segment cuttings. The study was conducted at the Indonesian Industrial and Beverage Crops Research Institute (IIBCRI) from January to March 2020. The research design was a factorial, Completely Randomized Design, with three replications. The first factor was coffee species, i.e., Robusta (BP 936), Liberica (LIM 1 and LIM 2), and Excelsa, and the second factor was Rootone-F concentrations were 0, 100, 200, 300, 400, and 500 ppm. The characters observed were callus diameter, leaves number, shoots number, and leaf length. The results showed there was an interaction between coffee species and Rootone-F concentration on shoots number and leaf length. Every factor affected callus diameter and leaves number, respectively. Application without Rootone-F (0 ppm) gave a higher value of callus diameter and leaves number of LIM 1. Rootone-F concentration that could increase the shoots number and leaf length was 500 ppm for Lim 1, 400 ppm for Lim 2, 0 ppm for BP 936, and 100 ppm for Excelsa.

The effects of zinc oxide nanoparticles loaded with IAA and IBA on in vitro rooting ofapple microcuttings

In the micropropagation of difficult-to-root plants, the low rates of rooting cause high economic losses. The local apple (Malus domestica Borkh.) cultivar #67, which is highly resistant to fire blight disease, has been shown to have low rooting success. Nanoparticles loaded with auxins could increase the rooting success with their physicochemical properties. To increase rooting percentage, the effects of zinc oxide nanoparticles (nZnO), nZnO loaded with indole-3-acetic acid (IAA-nZnO) and indole-3-butyric acid (IBA-nZnO) at different concentrations (1.0, 2.0, 3.0, 4.0, 5.0, and 6.0 mg L-1) on in vitro rooting of microcuttings were investigated. The concentrations of auxin carried by the nanoparticles were 2.8, 5.7, 8.0, 10.8, 13.7, 16.0 μM IAA or 2.5, 4.4, 6.9, 9.3, 11.3, 13.8 μM IBA. The various concentrations of IAA (0.0, 2.8, 5.7, 11.4, 17.1 μM) and IBA (0.0, 2.5, 4.9, 9.8, 14.8 μM) were also examined. All substances as described previously were incorporated to half-strength Murashige and Skoog (½ MS) medium containing 2% sucrose at pH 5.8, and solidified with 0.7% agar. The microcuttings were cultured on these media for 5 days in the dark before being transferred to a 16 h photoperiod conditions at 25 ± 1 °C for 6 weeks. It was found that rooting did not occur at 0.0 μM IAA and IBA concentration and nZnO treatments. IBA-nZnO and IBA were more effective on the rooting of microcuttings. In the comparison of IBA-nZnO and IBA, the percentage of rooting was higher in IBA-nZnO (58.4%) than IBA (47.5%). In addition, 4.4 μM or 4.9 μM IBA, particularly, induced many favorable properties with high rooting percentage (62.7%) and root length (16.5 ± 3.5 mm) and a small callus diameter (5.3 ± 0.2 mm). In conclusion, IBA-nZnO loaded with 4.4 μM IBA could be a promising treatment for in vitro rooting of difficult-to-root apple cultivar #67.

Callus regeneration and polyploidy induction of Allium cepa L var. Bima Brebes using oryzalin

Polyploidy induction could increase shallot bulb-size to raise consumer preference and local shallot productivity. The research aimed to obtain an effective method of polyploidy induction on callus of onion (Allium cepa) var. Bima Brebes. The experiment was consisted of two experimental steps, which were callus induction of onion and polyploid induction of the callus. A 1×1 cm callus was treated by two drops of oryzalin with concentrations 0, 25, 50, 75, 100, and 120 μM. The ploidy level was identified based on morphological trait, stomatal analysis and DNA content using a flow cytometry. The results showed callus diameter, number of green spots, and number of shoots were decreased with increasing oryzalin concentration. The planlet leaves regenerated from oryzalin treated callus were darker than that of control. The flow cytometry analysis showed that planlets with 75 μM oryzalin was tetraploid, had longer and wider stomata than that of the control.

Pemberian NAA (Naphthalene Acetic Acid) dan BAP (Benzil Amino Purine) dalam Inisiasi Petal Krisan (Chrysanthemum indicum L.) Terhadap Pertumbuhan Organogenesis Tunas Secara In Vitro pada Media MS (Murashige and Skoog)

The research was carried out at Laboratorium Pemuliaan Terpadu, precisely at Laboratorium Konservasi Balai Penelitian Tanaman Hias Kebun Percobaan Segunung, Pacet, Cianjur from March to June 2021. The Research Method uses experimental methods with non-parametric statistics with 12 treatments and 5 repeats. Perlakuan NAA dengan konsentrasi 0,1 ppm; 0,2 ppm; dan BAP yaitu konsentrasi 0,5 ppm; 1 ppm; dan 1,5 ppm. The results of the observations were analyzed descriptively using the Wallis Kruskal Test. Based on several observed variables, the results showed different results in the degree of significance between the explants in NAA and BAP administration, in callus diameter variables showed significant different results, while in variables the number and height of the shoots showed significantly different results. The results of the study showed NAA and BAP concentrations on A2B3 treatment (concentration 0.1 ppm NAA+1 ppm BAP) were able to produce the best shoots against the number of shoots. In A3B3 treatment (concentration 0.2 ppm NAA + 1 ppm BAP) is capable of producing the best shoot against the height of the shoot.

In vitro propagation of six selected sugarcane mutant clones through leaf explants

Six mutant clones of sugarcane with high productivity have been produced through tissue culture techniques combined with mutations using gamma-ray irradiation and Ethyl Methane Sulfonate. The six mutant clones have been tested for stability in the field. They are proven to have high productivity and yields, so that they are very potential to be developed as superior varieties. To support the planting material sufficiency of these clones, an efficient propagation method was needed. Media formulations with different physical properties and composition of growth regulators were tested to obtain high seedling propagation rates. The media formulation for callus induction was Murashige dan Skoog (MS) + 3 mg/l 2,4-D + 3 g/l casein hydrolysate + 3% sucrose and for shoot regeneration was MS + 0,5 mg/l BA + 0,1 mg/l IBA + 100 mg/l PVP and 2% sucrose. Shoot proliferation was carried out on MS liquid (1, ½) + (0.3; 0.5 mg/l) BA + 0.1 mg/l IBA + 1 mg/l Kinetin + (0; 0.5 mg/l) GA3+ sucrose 2%. The results showed that callus induction, callus regeneration, and shoot proliferation of sugarcane mutant clones were influenced by the genotype and medium composition. The fastest callus induction was obtained from the MSP-4 clone (5.82 days), and the longest was MSB-7 (8.82 days). The largest callus diameter was obtained from MSB-6 clone on MS medium containing 1 mg/l BA, 100 mg/l PVP, and 2% sucrose. The highest number of shoots was obtained from the MSB-6 clone, while the least number of shoots conducted from the MSB-8 clone. The MSB-8 clones were more difficult to regenerate compared to the others. The best media formulation for shoot proliferation was ½ MS containing 0.5 mg/l BA, 1 mg/l Kinetin, and 0.1 mg/l IBA, while the best formulation for rooting was ½ MS.

Effectiveness of Nail Dynamization in Delayed Union of Tibial Shaft Fractures: Relationship Between Fracture Morphology, Callus Diameter, and Union Rates

Dynamization has already been described as a secondary intervention for delay unions of tibial shaft fractures treated with intramedullary nailing. Although it's a common procedure, it is not widely supported in the literature. The purpose of this study was to determine the union rate of nail dynamization in cases of delayed union of diaphyseal tibial fractures, and assess the effect of fracture morphology on union rates. We retrospectively analyzed a series of 199 consecutive tibial shaft fractures. We recorded the dynamization rate, period from nailing to dynamization, nailing to the union, the fracture pattern (according to AO/ASIF and whether it was closed or open), the callus diameter before dynamization (fracture healing index; FHI) and union/failure rates. Out of a total of 199 fractures treated during the study period, 41 (20.6%) were dynamized. After applying inclusion and exclusion criteria, 39 patients with 39 fractures were included in the study. The mean time from nailing to dynamization was 18.4 ± 7.2weeks. The union rate was 92.3% (n = 36) over a mean time of 14.1 ± 5.6weeks as from dynamization. The overall failure rate was 6.7% (n = 3). There was no significant association between failure and AO/ASIF classification (p > 0.05) or fracture exposure (X 2 = 0.19; p = 0.66). The pre-dynamization FHI of ≥ 1.17 was significantly associated with consolidation (p < 0.05). In cases of delayed union of tibial fractures, dynamization offered a high union rate associated with pre-dynamization FHI, while fracture morphology did not affect the failure rate.

Elicitation of industrially promising vanillin type aromatic compound 2-hydroxy 4-methoxy benzaldehyde (MBAlD) yield in the in-vitro raised medicinal crop Hemidesmus indicus (L) R. Br. by methyl jasmonate and salicylic acid

Growth promoting and secondary metabolite enhancing roles of the elicitors, methyl jasmonate (MeJA) and salicylic acid (SA) were investigated in the popular medicinal crop Hemidesmus indicus (L) R. Br. or Indian Sarsaparilla grown in-vitro. In-planta elicitor treatment (MeJA; 25, 50, and 75 μM and SA; 0.75, 1.5, and 2 mg/mL) induced growth of nodular meristemoids, promoted overall growth and demonstrated enhanced bioactivity and accumulation of root specific alkaloid 2-hydroxy 4-methoxy benzaldehyde (MBAld). A fast and effective micropropagation protocol including both direct and indirect organogenesis and regeneration was standardized for H. indicus (HI-KOL). Murashige and Skoog (MS) media fortified with 6-benzylaminopurine (BAP) (1.5 mg/L) produced the highest (6.7) number of shoots/ explant and average shoot length was 11.7 cm. It was followed by thiodiazuron (TDZ) (1.5 mg/L) (avg. shoot/explant 6.5 and avg. shoot length 11.1 cm). The percentage of emergence of shoot initials from the light green-compact base callus was highest (100%) in MS media supplemented with TDZ (2 mg/L) followed by indole-3-butyric acid (IBA) or indole-3-acetic acid (IAA) (0.50 mg/L). Highest average callus diameter (3.7 cm) was noted in MS media fortified with TDZ (2.0 mg/L) + IBA (0.50 mg/L). For root initiation, IBA (1.5 mg/L) fortified MS medium demonstrated the best result (average root length 9.1 cm). Best responsive in-vitro raised plant materials also demonstrated significant total phenolics, flavonoid, tannin, and photosynthetic pigment contents when compared with other samples and in-vitro grown plantlets. Competitive antioxidant activity 1,1-diphenyl2-picrylhydrazyl (DPPH) and ferric reducing antioxidant power (FRAP) assays, activity of antioxidant enzymes [superoxide dismutase (SOD), ascorbate peroxidase (APX), and catalase (CAT)], and elicitor stimulated pancreatic lipase inhibitory activities were also noted. In addition, a rapid, validated, and sensitive densitometric protocol for high performance thin layer chromatography (HPTLC) quantification of MBAld was also standardized and it was noted that both MeJA and SA stimulated accumulation of MBAld in in-vitro samples but the effect was more prominent in MeJA treated plant derived root extracts with 75 μM MeJA treatment produced 3.41 ± 0.8 mg/g MBAld which was higher than the mother plant (MP) (3.30 ± 0.2 mg/g). HPTLC analyses also revealed phytochemical fidelity among the MPs and the treated and non-treated micropropagated plant samples.

In vitro Callus Induction under NaCl Salt Stress and Subsequent Plant Regeneration in Oats (Avena sativa L.)

An effort was made to identify the in vitro responsiveness for callus induction in NaCl (salt stress) and subsequent plant regeneration of oats (Avena sativa L.). Callus induction was tested in different concentrations (0, 5, 10, 15 and 20 mg/l) of NaCl salt added in MS. The experiment was laid out in CRD considering 2,4-D growth regulator and NaCl salt as factors. The best response in callus induction was observed on MS containing 3 mg/l 2,4- D and 5 mg/l NaCl where the frequency of callus induction was 68.57%, callus weight was 0.36 g and callus diameter was 7.63 mm. During regeneration, the maximum frequency of regeneration (41.67%), regenerated shoots per callus (14.80), shoot length (12.14 cm), root number (10.90) and root length (3.76 cm) was observed from the callus derived from MS containing 3 mg/l 2,4-D and 5 mg/l NaCl. Plantlets that were regenerated from callus induced in MS with 3 mg/l 2,4-D and 5 mg/l NaCl showed higher rate of ex vitro survivability. Therefore, MS with 3 mg/l 2,4-D and 5 mg/l NaCl found better for in vitro callus induction and subsequent plant regeneration from mature seeds of oat.&#x0D; Plant Tissue Cult. &amp; Biotech. 30(2): 253-266, 2020 (December)

The Effect of Strontium Ranelate on Fracture Healing: An Animal Study.

Background Strontium ranelate (StR) is an antiosteoporotic agent previously utilized for the enhancement of fracture union. We investigated the effects of StR on fracture healing using a rabbit model. Methods Forty adult female rabbits were included in the study and were divided in 2 equal groups, according to StR treatment or untreated controls. All animals were subjected to osteotomy of the ulna, while the contralateral ulna remained intact and served as a control for the biomechanical assessment of fracture healing. Animals in the study group received 600 mg/kg/day of StR orally. All animals received ordinary food. At 2 and 4 weeks, all animals were euthanatized and the osteotomy sites were evaluated for healing through radiological, biomechanical, and histopathological studies. Results The treatment group presented statistically significant higher callus diameter, total callus area, percentage of fibrous tissue (p < 0.001), vessels/mm2, number of total vessels, and lower osteoclast number/mm2 (p < 0.05) than the control group at 2 weeks. Additionally, the treatment group presented significantly higher percentages of new trabecular bone, vessels/mm2, osteoclast number/mm2, and lower values for callus diameter, as well as total callus area (p < 0.05), than the control group at 4 weeks. At 4 weeks, in the treatment group, force applied (p = 0.003), energy at failure (p = 0.004), and load at failure (p = 0.003) were all significantly higher in the forearm specimens with the osteotomized ulnae compared to those without. Radiological bone union was demonstrated for animals receiving StR at 4 weeks compared with controls (p = 0.045). Conclusion StR appears to enhance fracture healing but further studies are warranted in order to better elucidate the mechanisms and benefits of StR treatment.

Effects of a novel biodegredable implant system on a rat tibia fracture model.

This study aimed to determine the effects of a novel biodegradable implant releasing platelet-derived growth factor (PDGF) at the fracture site on fracture healing in a rat tibia fracture model. In this study, 35 male Sprague-Dawley rats weighing between 300 and 350 g were used. The rats were divided into four groups: Group A (control group without any treatment, n=10), Group B (spacer without PDGF Group, n=10), Group C (spacer with PDGF group, n=10), and Group D (healthy rat Group, n=5). Standardized fractures were created in the right tibias of rats, and then biodegradable implants made of poly-β-hydroxybutyrate-co-3-hydroxy valerate were implanted at the fracture sites in Groups B and C. In Group C, implants were loaded with 600 ng of PDGF. Animals were sacrificed 30 days after the operation, and fracture healing in each group was assessed radiologically based on the Goldberg score. Furthermore, the anteroposterior (AP) and mediolateral (ML) callus diameters were measured macroscopically, and fracture sites were mechanically tested. In the radiological assessment, Group C showed higher fracture healing rate than Groups A and B (p=0.001), whereas no significant difference was found between group C and Group D (p>0.05). In the macroscopic assessment, while Group C exhibited the thickest AP callus diameter (p=0.02), no significant differences in ML callus diameters existed among the groups (p>0.05). Mechanical testing revealed that Group C had higher torsional strength (p=0.001) and stiffness than Groups A and B (p=0.001) while there was no significant difference between Groups C and D (p>0.05). Biodegradable implant releasing PDGF may have positive effects on fracture healing.

Manipulating Some Culturing Conditions Enhances Production of Solanine in Microshoots, Callus and Cell suspension Cultures of Solanum nigrum L.: A Wild Medicinal Plant

Solanum nigrum L. is a medicinal plant of solanaceae family with distinguished therapeutic properties. Traditionally, S. nigrum. had been used as an anti-tumorgenic, antioxidant, hepatoprotective, diuretic, and antipyretic agent. The most important alkaloid member in this plant is solanine. Therefore, this study was conducted to utilize tissue culture techniques for the enhancement of solanine production in the in vitro grown cultures of this promising neglected plant. For callus growth and development experimental part, the highest callus growth parameters (callus diameter (21.4 mm) and callus fresh weight (2202.4 mg) were obtained in callus grown on Murashige &amp; Skoog MS media supplemented with 2,4-Dichlorophenoxyacetic acid (2.0 mg·L-1) plus 1.5 mg·L-1 Thidiazuron. Similar trend was also obtained in cell suspension culture experiment, as maximum growth was recorded at similar hormone combination. Moreover, High-performance liquid chromatography analysis revealed that, solanine was affected by growth regulator type and concentration. The highest solanine levels were obtained when the explants were treated with 6-benzylaminopurine at level of 2.0 mg·L-1, as solanine content reached up to (2.61, 1.53 mg.g-1) for callus and cell suspension, respectively, while, microshoot contained the highest solanine (4.52 mg.g-1 DW) at 6-benzylaminopurine level of 1.6 mg.L-1. Additionally, carbon source had positively affected solanine level, where 0.2 M sucrose resulted in production of the highest amounts (3.13, 2.03 and 1.20 mg.g-1 DW) of solanine in microshoots, callus and cell suspension, respectively. Also, exposing microshoots and callus to light intensity of (100 µmol.m -2 s-1) yielded the highest solanine content (4.03 and 1.26 mg.g-1 DW, respectively),while the lowest solanine levels (1.50 and 0.48 mg.g-1 DW) were observed in plant material exposed to the lowest light intensity treatment (25 µmol.m -2 s-1) .

Effects of Phytohormone Compatibility on Callus Growth of Rosa rugosa tunb

To obtain superior quality callus and to regenerate root system of Rosa rugosa tunb and improve culture efficiency, this experiment used a multi-factor orthogonal test method to establish a sterile system with young leaves of R. rugosa tunb as experimental materialsand different concentrations of hormones in each stage. Explore the effects of plant callus growth. The results showed that the optimal disinfection method of explants was 70% alcohol 30s+0.1% HgCl2 8min. At this time, the browning rate was 46.70%, and the infection rate was 10.00%, which was 14.40% and 72.67% lower than that before disinfection. [Primary medium]: MS+1.5 mg/L 6-BA+3.0 mg/L NAA+3% sucrose+0.9% agar, the induction rate at this time is 75.00%, an increase of 62.50% compared with no added hormone. [Proliferation medium] MS+2.0mg/L 6-BA+0.2mg/L NAA+3% sucrose+0.9% agar, the callus diameter reached 2.49cm after proliferation, the growth was strong, and the callus was highly available. [Rooting medium] MS+0.4mg/L IBA+0.5g/L AC+3% sucrose+ 0.9% agar, the rooting rate is 75.00%, the root system is long and the growth is robust.

In vitro callus initiation from Indigofera (Indigofera zollingeriana Miq.)

The aimed of this research was to callus initiation of I. zollingeriana Miq using a various concentration of 2,4-D and source of explants. The source of explants was the first factor which consisted of 2 types of explants, leaf (A1) and stem (A2) and concentration of 2,4-D was the second factor with several levels D0 (MS0), D1 (0.0 mg/L), D2 (1.0 mg/L), D3 (2.0 mg/L), D4 (3.0 mg/L), and D5 (4.0 mg/L). The results showed that source of explants and plant growth regulator of 2,4-D afford give a significant effect on the time of callus induction and very significant effect for the weight of callus, but no interaction between source of explants and 2,4-D for a time of callus induction and weight of callus. The fastest of callus initiation on MS media added by 2,4-D with a concentration of 2.0 and 3.0 mg/L (3.55 weeks after planting). The largest callus diameter showed on MS media without 2,4-D. The biggest weight of callus showed on MS media added by 2,4-D with a concentration of 3.0 mg/L. Callus had friable textured and creamy white-colored. Ultimately, the fastest of callus initiation from the source of explants was stem with 3.51 weeks after planting.

Induction of kopyor coconut embryogenic callus using 2.4-D and TDZ

This research was conducted to obtain an effective method to produce true to type coconut kopyor seeds and determine the effect of 2,4-D with or without TDZ in inducing callus to produce somatic embryos. This research was conducted in the Lampung State Polytechnic laboratory with concentrations of 2,4-D 0, 2.5, 5, 7.5, and 10 mg.L-1 with or without the addition of 0.5 mg.L-1. Further analysis of the data used the 5% LSD test. The parameters observed in this study were: percentage of callus explants, callus diameter, callus weight. The results showed that increasing the concentration of 2,4-D and 7.5 mg. L-1 could increase the percentage of embryogenic callus, callus diameter, and callus weight and the addition of 0.5 TDZ concentration to the 2,4-D medium resulted in a decrease in parameters. The best 2,4-D concentration to induce callus embryogenic in kopyor embryos is 7.5 mg. L-1. The use of 2,4-D for embryogenic callus formation can accelerate callus growth and obtain kopyor coconut seeds true to type.

Induction and Growth of Endosperm Cali of Rimau Gerga Lebong (RGL) Citrus on Several Media Compositions

Rimau Gerga Lebong (RGL) variety is one of the main orange fruit commodities in Lebong Regency of Bengkulu Province, which has a competitive advantage and has good market potential. However, high number of seed characteristic makes this orange fruit becomes less popular. Triploid genotype formation through endosperm culture in vitro is an alternative solution to develop seedless orange fruit. The objective of this study was to determine the best composition medium for callus induction of endosperm of RGL orange seeds as the foremost step of in vitro triploid plant development. The research was conducted from August 2017 until December 2018 at the Plant Tissue Culture Laboratory, Department of Agronomy, Faculty of Agriculture, University of Bengkulu. The experiment was arranged in a completely randomized design, with eight treatment combinations i.e. G1 (MT + 5 ppm BAP + 2 ppm 2,4-D), G2 (MT + 5 ppm BAP + 2 ppm 2,4-D + 500 ppm CH), G3 (MT + 5 ppm BAP + 2 ppm 2,4-D + 500 ppm + 0.5 ppm Kinetin), G4 (MT + 5 ppm BAP + 2 ppm 2,4-D + 500 ppm CH + 0.5 ppm to 500 ppm Kinetin + ME) , G5 (MS + 5 ppm BAP + 2 ppm 2,4-D), G6 (MS + 5 ppm BAP + 2 ppm 2,4-D + 500 ppm CH), G7 (MS + 5 ppm BAP + 2 ppm 2,4- D + 500 ppm CH + 0.5 ppm Kinetin), G8 (MS + 5 ppm BAP + 2 ppm 2,4-D + 500 ppm CH + 0.5 ppm to 500 ppm Kinetin + ME), with three replications. The experimental unit was five culture bottles containing three explants per bottle. Observations were conducted on days to callus formation, rate of callus formation, callus weight, callus diameter, callus color and texture. The results showed that media of MS + 5 ppm BAP + 2 ppm 2,4-D + 500 ppm CH was the best media for callus induction, and MT + 5 ppm BAP + 2 ppm 2,4-D + 500 ppm CH was the best media for callus development.Keyword: RGL citrus, calli, endosperm, kinetin, BAP

The Effect of BAP and Kinetin Concentrations for shoot induction on Teak (Tectona grandis L.) with In Vitro method

The purpose of this research was to examine interaction effect between the different concentration of BAP hormone (Benzylaminopurine) and Kinetin hormone on the shoot induction of Jati (Tectona grandis L.) with in vitro method. This research used a completely randomized factorial design with 4x3 factorial pattern. The first factor was the concentration of BAP (B0 = 0 ppm; B1 = 1 ppm; B2 = 2 ppm dan B3 = 3 ppm). The second factor was concentration of Kinetin (K0 = 0 ppm; K2 = 0,75 ppm; dan K3 = 1,5 ppm). The parameters observed were time of callus appearance, callus length, time of shoot emergance, number of shoots, length of shoots, time of leaf development, number of leaves, percentage of growth, and percentage of contamination. The parameters obtained were analysed descriptively because the percentage of total growth was under 50%. The result showed that the concentration 1 ppm BAP and 0,75 ppm kinetin was better for stimulating the appearance of callus, shoots, leaves, and number of shoots. Callus diameter was better at concentration of BAP 3 ppm. The callus texture that was formed was included in the compact texture. The percentage of explants life was 33,30 – 66,60%. No contamination occurred in all explant samples.

Efficient plant regeneration and callus induction from nodal and hypocotyl explants of goji berry (Lycium barbarum L.) and comparison of phenolic profiles in calli formed under different combinations of plant growth regulators

Goji berry (Lycium barbarum L.) is a fruit that has many beneficial effects (such as antidiabetic, antioxidant, anticancer, antidepressant, and immunomodulatory) on human health because of several dietary constituents such as phenolics, vitamins, carotenoids, and polysaccharides. In order to develop in vitro culture protocols for callus induction and plant regeneration from different explants of Goji plants and to compare the phenolic composition in calli of different origin, various combinations of plant growth regulators (PGRs) were applied. Various types of explants (leaf, petiole, root, hypocotyl, and node) were cultured on MS medium containing 28 different concentrations and combinations of PGRs [thidiazuron (TDZ) and naphthalene acetic acid (NAA), TDZ and indole-3-acetic acid (IAA), benzyl adenine (BA) and NAA, alone TDZ, and alone BA]. The highest mean number of shoots (23.33 ± 1.86) and percentage of shoot formation (100%) were obtained from nodal explants on medium containing 0.5 mg/l BA alone. The highest mean callus diameter was obtained from hypocotyl explants on medium containing 0.25 mg/l TDZ and 0.1 mg/l IAA (21.40 ± 0.71 mm). The amounts of selected phenolic substances were significantly different in the callus obtained from different PGRs or combinations, individually. The combination of BA/NAA significantly increased the production and accumulation of chlorogenic acid and caffeic acid. The TDZ/IAA combination, TDZ alone, and TDZ/NAA combination significantly increased vanillic acid and rutin, gallic acid, and quercetin synthesis, respectively. These outcomes indicate that different PGRs lead to the production of different kinds of secondary metabolites and affect/accelerate accumulation in the callus of goji berry. The culture protocol described in this paper could be employed for the development of novel methods for the commercial production of goji secondary metabolites.

Short-term perioperative parecoxib is not detrimental to shaft fracture healing in a rat model.

ObjectivesExperimental studies indicate that non-steroidal anti-inflammatory drugs (NSAIDs) may have negative effects on fracture healing. This study aimed to assess the effect of immediate and delayed short-term administration of clinically relevant parecoxib doses and timing on fracture healing using an established animal fracture model.MethodsA standardized closed tibia shaft fracture was induced and stabilized by reamed intramedullary nailing in 66 Wistar rats. A ‘parecoxib immediate’ (Pi) group received parecoxib (3.2 mg/kg bodyweight twice per day) on days 0, 1, and 2. A ‘parecoxib delayed’ (Pd) group received the same dose of parecoxib on days 3, 4, and 5. A control group received saline only. Fracture healing was evaluated by biomechanical tests, histomorphometry, and dual-energy x-ray absorptiometry (DXA) at four weeks.ResultsFor ultimate bending moment, the median ratio between fractured and non-fractured tibia was 0.61 (interquartile range (IQR) 0.45 to 0.82) in the Pi group, 0.44 (IQR 0.42 to 0.52) in the Pd group, and 0.50 (IQR 0.41 to 0.75) in the control group (n = 44; p = 0.068). There were no differences between the groups for stiffness, energy, deflection, callus diameter, DXA measurements (n = 64), histomorphometrically osteoid/bone ratio, or callus area (n = 20).ConclusionThis study demonstrates no negative effect of immediate or delayed short-term administration of parecoxib on diaphyseal fracture healing in rats.Cite this article: G. A. Hjorthaug, E. Søreide, L. Nordsletten, J. E. Madsen, F. P. Reinholt, S. Niratisairak, S. Dimmen. Short-term perioperative parecoxib is not detrimental to shaft fracture healing in a rat model. Bone Joint Res 2019;8:472–480. DOI: 10.1302/2046-3758.810.BJR-2018-0341.R1.

The effects of cryopreserved human amniotic membrane on fracture healing: Animal study

ObjectiveThe aim of this study was to investigate the effects of human amniotic membrane (HAM) on fracture healing in an animal model. MethodsStandard tibial diaphysial fractures were created in twenty-eight Wistar-Albino rats and treated with intramedullary Kirschner wire (K-wire) and HAM (HAM (+) group) or K-wire only (HAM (–) group). Fracture healing was evaluated by histological analysis, radiologic X-ray views and callus diameter measurements at 3rd and 6th weeks postoperatively. ResultsFracture healing was histologically better in the HAM (+) group and the difference was statistically significant at both 3rd and 6th weeks postoperatively (p < 0.05). The highest histologic scores and entire woven bone formation (Huo Stage 8–9) were obtained at 6th weeks postoperatively in the HAM (+) group. Histological examination also revealed predominant fibrous tissue and partial cartilage formation (Huo Stage 2) at the postoperative 3rd week in the HAM (-) group. Equal amounts of woven bone and cartilage formation (Huo Stage 6–7) were observed at 3rd weeks postoperatively in the HAM (+) group and at 6th weeks postoperatively in the HAM (-) group. The callus diameters were greater in the HAM (+) group and the difference was statistically significant (p < 0.05) at 3rd and 6th weeks postoperatively. Although there was only a statistically significant difference (p < 0.05) at the postoperative 3rd week, radiological scores tended to be higher in the HAM (+) group at both the 3rd and 6th weeks postoperatively. ConclusionHAM is a cheap and easily accessible alternative biological material. HAM may be used to support surgical treatment of fractures, particularly where bone healing is expected to last longer.

Callus and shoot induction of leaf culture Lilium longiflorum with NAA and BAP

Abstract. Lestari NKD, Deswiniyanti NW, Astarini IA, Arpiwi LM. 2019. Callus and shoot induction of leaf culture Lilium longiflorum with NAA and BAP. Nusantara Bioscience 11: 162-165. Lilium longiflorum Thunb., an Easter lily, is a common ornamental plant used as potted plant, cut flower, cosmetic and medicine. The research on tissue culture technique to induce shoots and callus is expected to increase the yield of lilies quickly and efficiently. This study aims to determine the effect of plant growth regulator combination of Naphthalene Acetic Acid (NAA) and Benzyl Amino Purine (BAP) on the development of leaf culture and the best concentration for callus induction and shoots of lily plants. This study was conducted in a completely randomized design using leaf explants in Vacin and Went medium, with combination of NAA and BAP (0; 0.5; 1 mg..L-1 ) as the treatments. The result of the eight-week observations shows that the plant growth regulators significantly affected either in days initiation callus, days initiation of shoot, percentage of callus, percentage of shoot, diameter of callus, no of shoot and length of shoot. The combination of 1 mg.L-1 NAA and BAP had the best effect in the percentage of explants forming shoots (100%), means number of shoots (5.8), and means length of shoot (11.6 cm).