Concentrations Of BAP Research Articles

WORTHY MICROPROPAGATION PROTOCOLS OF SEVEN CULTIVARS OF POMEGRANATE (PUNICA GRANATUM L.) CULTIVARS IN THE PROVINCE OF DUHOK, KURD ISTAN RIGION OF IRAQ.

An efficient attempt was conducted to increase the mass production of seven local cultivars of pomegranate (Punica granatum L.) Masafik, Melisse, Radisho, Armishte, Diala, Halapja, and Dwarf pomegranate cultured in the Kurdistan region of Iraq from October 2021 to February 2023. Micropropagation technique was applied using eight combinations of plant growth regulators (PGRs) added to two media (MS and WPM). The results indicated that immersing explants in 70% Ethyl alcohol (EtOH) for (2 minutes) followed by immersion in (2.5%) Sodium Hypochlorite (NaOCl) plus 3 drops of tween 20 for 13 mins. was the more suitable combination for explant sterilization. MS and WPM media salts supplemented together with eight combinations of PGRs were carried out to evaluate the more suitable combination for the initiation stage. The obtained data clarified that the ½ strength of the MS medium (1/2 MS) supplemented with 0.5 BAP plus 0.5 mg/l Kinetin increases the pomegranate explant initiation response percentage. WPM basal medium was the preferred media to be used in the multiplication stage. The impact of different levels and protocols of Cytokinins, Auxins, and Gibberellins were investigated by eight protocols including using BAP, GA3, and adenine sulfate. The outcomes indicated that adding both concentrations of BAP (0.5 and 1.0 mg/l) separately supported by GA3 and adenine sulfate in amounts of (0.3 and 30 mg/l) respectively again were the most suitable protocol that affected the multiplication of regenerated shoots when compared to using BAP alone. Valuable results were found in rooting response after 6 weeks by using ½ strength of MS media along with two levels of Auxins (NAA, and IBA). Moreover, the visual observation records that around 85-90% of rooted cultures were successfully acclimatized by transferring the cultures from in vitro to the ex-vitro environment. Finally, Dwarf cultivars exceed all the other cultivars in studded parameters.

In vitro Culture Optimization of Pomelo Seeds (Citrus maxima (Burm.) Merr.): A South Sulawesi Orange.

<b>Background and Objective:</b> Indonesia boasts a variety of delicious tropical fruits, including pomelo, mainly grown in Pangkep Regency, South Sulawesi Province. However, in this region, some challenges hinder such as inadequate care, aging trees and limited seed supply hinder productivity in this region. <i>In vitro</i> culture methods present a solution by rapidly producing high quality, disease-free pomelo seeds. This study aims to determine the optimal concentration of the BAP added to the culture medium to induce shoots from pomelo seeds. <b>Materials and Methods:</b> The seeds were planted on MS media with the addition of BAP hormone (0.5, 1, 1.5, 2 and 2.5 ppm) and 0 ppm as the control. The experimental units were arranged in a CRD and analyzed using SPSS 20.0 software, employing the Shapiro-Wilk normality test and Levene's Statistic for homogeneity. If the data met the normality and homogeneity assumptions, ANOVA was applied, followed by the DMRT for a parametric test. Otherwise, a non-parametric test namely the Kruskal-Wallis was conducted and differences were further analyzed using the Mann-Whitney test at a 5% significance level. <b>Results:</b> The application of the BAP accelerated shoot emergence, with the most rapid development occurring on the 10th day after planting (DAP), at a BAP concentration of 2.5 ppm for red pomelo. For white pomelo and sweet pomelo, shoots appeared on the 19th and 20th days, respectively at a 2 ppm BAP. Interestingly, root development was fastest between the 4th and 6th DAP in 0 ppm BAP (control). <b>Conclusion:</b> The addition of the BAP at a concentration of 1.5 ppm in the culture medium promotes faster shoot emergence and has a significant impact on the number of shoots in red pomelo.

In vitro multiplication of Eugenia calycina Cambess. (Myrtaceae) under the influence of BAP and NAA

Eugenia calycina Cambess. (Myrtaceae) is a fruit species native to Brazil indicated for the recovery of degraded areas. Generally its propagation occurs by seeds, but leads to an increase in the juvenile period of the plants, problems in maintaining seed viability and pathogen incidence. However, with the application of tissue culture techniques it is possible to produce seedlings in large scale, uniformity of the plants obtained and reduction in the period of seed germination. Thus, the objective of this research was to define the best medium for in vitro multiplication of E. calycina. Nodal segments of 1 cm, extracted from E. calycina plants already established in vitro, were used as explants. The treatments consisted of different concentrations of BAP (0; 0.5; 1 and 2 mg L-1) and NAA (0; 0.5; 1 and 2 mg L-1). The experimental design was completely randomized, in a 4x4 factorial scheme, with four concentrations of BAP and four concentrations of NAA, with four replications and three tubes per replication. After 90 days, shoot number, length and fresh mass of the aerial part, production and fresh mass of callus were evaluated. The MS medium plus 2.0 mg L-1 of BAP is ideal for in vitro multiplication of E. calycina.

Induksi Tunas Karamunting (Rhodomyrtus tomentosa) secara Invitro

Tissue culture techniques provide an alternative to vegetative plant propagation efforts in conservation efforts for Karamunting, the existence of which is becoming increasingly rare. The explant as the material used is the result of in vitro germination using several concentrations of GA3. This research was structured in the form of a factorial experiment with a completely randomized factorial design (RAL) with 2 factors. The first factor is the BAP concentration with three levels, namely B1 = BAP 0.5 B2 = BAP1 ppm and B3 = BAP 1.5 ppm. The second factor is the concentration of TDZ with three levels, namely T1 = TDZ 0 ppm, T2 = TDZ 0.25 ppm and T3 = TDZ 0.5 ppm. The results of the research showed that there was an interaction between the growth regulators BAP and TDZ on the percentage of live explants and the height of karamunting explants. In terms of the percentage of live explants, giving BAP 0.50 ppm without TDZ is the best concentration with a live percentage of 100%. Treatment of 1 ppm BAP and 0.25 ppm TDZ produced the highest shoots, namely 2.83 mm. The time of emergence of shoots and the number of shoots per explant had no interaction and the influence of a single factor, several concentrations of BAP and TDZ.

Respon Pertumbuhan Tanaman Porang (Amorphophallus muelleri Blume.) Dengan Penambahan NAA (Naphthalene Acetic Acid) dan BAP (6-Benzyl Amino Purine) Pada Perbanyakan Secara In Vitro

Porang is one of the export crops that are much hunted in several countries. The increasing number of needs has resulted in limited porang seeds so that there is a need for propagation of porang plants with good quality, one of which is by propagation through tissue culture or in vitro. In order to propagate porang plants in vitro, this research tries to discover the ideal concentration of NAA and BAP. From August to December 2022, this study was conducted in the Plant Tissue Culture Laboratory of Jember State Polytechnic. The experiment employed a parametric statistical design with 2 treatments: 0.5 mg/L NAA + 1.5 mg/L BAP (P1) and 0.5 mg/L + 2.5 mg/L BAP (P2), with each treatment being repeated 10 times. According to the findings, the parameters of callus emergence time (46 HST) and callus diameter (1.51 cm) were best achieved by administering NAA and BAP at a dosage of 0.5 mg/L NAA + 2.5 mg/L BAP. According to the study aims, the optimal concentration for explant growth and development is believed to be 0.5 mg/l NAA + 2.5 mg/l BAP.

Pertumbuhan Tanaman Anggrek Cattleya (Cattleya eximia) secara In-Vitro pada Media MS dengan Subtitusi NAA dan BAP

Orchid plants are one of good prospects commodities for the floriculture business because they have high aesthetic value. Cattleya orchid has distinctive characteristics with large, beautiful, brightly colored and fragrant flowers. In vitro propagation is one way to meet the increasing demand for orchids. In vitro culture techniques require a lot of culture media to multiply plants in large quantities, so an alternative is needed to increase the effectiveness of using media such as the addition of plant growth regulators. Therefore, research was carried out on the growth of in vitro cattleya orchids on MS media with NAA and BAP substitutions. This study aimed to determine the effect of adding NAA and BAP to MS media on the growth of in vitro cattleya orchid explants. This research was conducted from June to September 2022 at the Jember State Polytechnic Tissue Culture Laboratory. This study used completely randomized design factorial. The first factor was the NAA concentration consisted of 3 levels, namely 0, 1 and 2 ppm. The second factor was the BAP concentration consisted of 3 levels, namely 0, 2 and 4 ppm. Observational variables included plant height, the number of shoots, leaves and roots. Data were analyzed using analysis of variance (ANOVA) and Duncan's Multiple Range Test (DMRT) at 5% level. Giving NAA 2 ppm + BAP 2 ppm was the best treatment for the number of leaves while giving NAA 2 ppm resulted in the best root growth.

Indoor exposure to polycyclic aromatic hydrocarbons associated with solid fuel use in rural China.

Polycyclic aromatic hydrocarbons (PAHs) are widespread environmental contaminants associated with various health risks including lung cancer. Indoor exposure to PAHs, particularly from the indoor burning of fuels, is significant; however, long-term large-scale assessments of indoor PAHs are hampered by high costs and time-consuming in field sampling and laboratory experiments. A simple fuel-based approach and statistical regression models were developed as a trial to predict indoor BaP, as a typical PAH, in China, and consequently spatiotemporal variations in indoor BaP and indoor exposure contributions were discussed. The results show that the national population-weighted indoor BaP concentration has decreased substantially from 46.1ng/m3 in 1992 to 6.60ng/m3 in 2017, primarily due to the increased use of clean energies for cooking and heating. Indoor BaP exposure contributed to > 70% of the total inhalation exposure in most cities, particularly in regions where solid fuels are widely utilized. With limited experimental observation data in building statistical models, quantitative results of the study are associated with high uncertainties; however, the study undoubtedly supports effective countermeasures on indoor PAHs from solid fuel use and the importance of promoting clean household energy usage to improve household air quality.

Application of BAP and IBA for In Vitro Callus Formation of Biduri (Calotropis gigantea)

A herbaceous plant called biduri (Calotropis gigantea) is useful as a medicine. The leaves, roots, and flowers are just a few of the organs that contain secondary metabolites. A callus is an accumulation of secondary metabolite-producing amorphous cells. Through the induction of plant growth regulators, callus formation can be induced. The goal of the study was to find the BAP and IBA concentration that would promote callus formation in biduri. The initialstem segments of biduri, which were 1.5 months old and 2 cm long, were used as explants. A factorial design using two components and three repetitions of each was employed in a fully randomized design. BAP concentrations (0, 1, 2, 3, 4 ppm) and IBA concentrations were the first and second factors, respectively (0, 1, 2, 3, 4 ppm). The callus induction time, callus morphology (color and texture), and callus appearance percentage were the factors that were observed. The results demonstrated that a green, compact callus could be produced at a concentration of 4 ppm BAP and 1 ppm IBA. The proportion of callus manifestation rose and the callus induction time decreased with an increase in BAP concentration of up to 4 ppm.

The Effect of PVP (Polivinil Pirolidon) and BAP (6-benzylamino purine) on Shoots Induction of Teak Plus Perhutani (Tectona grandis)

The goal of this study is to see how BAP and PVP concentrations affect in vitro teak shoot induction. The research was carried out at Diponegoro University’s Tissue Culture Laboratory, Faculty of Animal Husbandry and Agriculture, between April and June 2022. The research method was a 4x4 factorial design based on a completely randomized design (CRD) with three replications. The first factor was the concentration of PVP (mg.L-1): 0, 110, 220, and 330, and the second was the concentration of BAP (ppm): 0, 1.3, 2.6, and 3.9. Duncan’s Multiple Range Test (DMRT) and orthogonal polynomials were used to test the data for variance. The interaction between the 0 mg.L-1 PVP treatment and the 110, 220, and 330 mg.L-1 PVP treatments after the addition of 2.6 ppm BAP had the quickest callus emergence time, with an average of 7 days. PVP treatments of 110 mg.L-1 and 220 mg.L-1 had a significant effect on the shoot length parameter. BAP concentrations of 1.3 and 2.6 ppm were found to increase the shoot length, number of shoots, and number of leaves response parameters. The 1.3 ppm BAP treatment also reduced browning by 17% in the explants. There was no effect of PVP or BAP treatments on shoot emergence time, leaf emergence time, or the percentage of explants that formed shoots.

Methodology of Micropropagation of Elite Genotype in Lotus (Nelumbo nucifera) Genotype Lakshmi

The lotus (Nelumbo nucifera Gaertn.) is an aquatic plant grown extensively throughout Asia, mostly for commercial and ornamental purposes. It is mostly propagated via rhizomes. A potent and reproducible plant regeneration system for lotus has been developed in this work employing rhizomes, shoot tips from the mother plant, and mature and immature embryos collected from eighteen and twenty-six-day-old aseptically fertilized seed. Shoot induction, number of shoots, and length of roots were induced on Murashige and Skoog's (MS) basal medium supplemented with various concentrations of 6-Benzylaminopurine (BAP) mgL-1 (0.5, 1, 2, 3, 4). The response of the explant varied with different BAP concentrations. The most favorable outcomes were achieved using MS medium treated with 0.5 mgL-1 6-Benzylaminopurine (T2).

In vitro multiplication of “Kepok” banana (Musa acuminata x Musa balbisiana) using different concentration of BAP and NAA in MS media

This study aims to determine the effect of different composition of BAP and NAA concentration added to Murashige and Skoog (MS) basic media on the in vitro multiplication of Indonesian “Kepok” banana variety. The research was conducted at the Tissue Culture Laboratory of Bonto-Bonto Horticultural Plant Seed Center. A completely randomized design (CRD) was employed with two factors tested. Four concentrations of each BAP and NAA used were 0 ppm, 4.5 ppm, 5 ppm, and 5.5 ppm for BAP, and 0 ppm, 2 ppm, 2.5 ppm, and 3 ppm for NAA resulted in 15 composition of BAP:NAA in addition to control (without the hormones). The results show no interaction between the BAP and NAA treatments, BAP concentration of 5 ppm gave the best result on parameter time to shoots emergence (7.50 DAP). NAA concentration of 2.5 ppm gave the best results on root emergence time (8.61 DAP), time to plantlet formation (11.72 DAP), number of roots (4.81 roots) and root length (5.93 cm).

BAP and Kinetin Application for In Vitro Bud Induction of Vanilla (Vanilla planifolia Andrews.) from Stem Node Explants in the Initiation Stage

Due to the scarcity of seedlings, vanilla farming in Indonesia is not at its best. The goal of this study was to comprehend how BAP and Kinetin affected the introduction of vanilla buds as a sustainable seedling strategy. The study is carried out at Salatiga, Central Java, Indonesia, at the Plant Tissue Laboratory of the Plantation Seeds Center and the Plant Tissue Laboratory of Diponegoro University’s Physiology and Plant Breeding Laboratory. A completely randomized design (CRD) 5 x 3 with two replications was utilized in the study. The first factor is BAP concentration: B0 = 0 mg.L−1, B1 = 0.6 mg.L−1, B2 = 1.2 mg.L−1, B3 = 1.8 mg.L−1, and B4 = 2.4 mg.L−1. The second factor is Kinetin concentration: K0 = 0 mg.L−1, K1 = 1 mg.L−1, and K2 = 2 mg.L−1. The result shows addition of BAP is able to shorten the day of bud emergence at 8 days. The addition of BAP and Kinetin did not significantly affect the number of buds. Application of BAP 1.44 mg.L−1 was able to increase bud length. There is interaction between Kinetin 2 mg.L−1 and the addition of BAP 2.4 mg.L−1 resulting in the largest bud diameter with a linear regression form. There is interaction between Kinetin 0 mg.L−1 and BAP 1.2 mg.L−1 resulting in the highest number of roots.

Different effects of continuous and pulsed Benzo[a]pyrene exposure on metabolism and antioxidant defense of large yellow croaker: Depend on exposure duration

This study aims to compare differential effects of continuous and pulsed BaP exposures on metabolism and antioxidant defense in the liver of large yellow croaker. Fish were subjected to BaP for 4 days and 36 days in three exposure regimes with the same time-averaged concentration of BaP: 4 μg/L BaP continuously, 8 μg/L BaP for 24 h every other day or 16 μg/L BaP for 24 h every 4 days. Our results showed that compared to pulsed BaP exposures, continuous BaP exposure reduced BaP metabolism (CYP1A, CYP3A and AHR transcriptional expressions, GSH content, GSH/GSSG ratio, EROD and GST activities) and antioxidant defense (T-SOD activity) on day 4, resulting to the increases in MDA and PC contents, indicating that continuous BaP exposure induced more severe oxidative damage during the early stage of exposure. But continuous BaP exposure reduced MDA and PC contents by improving BaP metabolism and antioxidant defense during the late stage of exposure. CYP1B transcriptional expression and CAT activity were unsuitable biomarkers of both continuous and pulsed BaP exposures. In conclusion, our results demonstrated differential effects of continuous and pulsed exposures on BaP metabolism and antioxidant responses, which were depend on exposure duration.

Effect of Adding NAA and BAP on the Growth of Kepok Kuning Banana (Musa paradisiaca L.) Planlet on MS Media in Vitro

The availability of Kepok Kuning Banana (Musa paradisiaca L.) seeds is increasingly limited due to physiological diseases such as leaf wilt. Therefore, tissue culture techniques were carried out as a solution for propagating Kepok Kuning banana seeds which disease free. This study's purpose is to determine the effect of adding NAA and BAP to MS media in vitro on the growth of Kepok Kuning Banana plantlets. This study used a completely randomized design with one treatment factor, the combination of NAA and BAP concentrations with five treatments; A (4ppm NAA + 8ppm BAP), B (5ppm NAA + 7ppm BAP), C (6ppm NAA + 6ppm BAP), D (7ppm NAA + 5ppm BAP), E (8ppm NAA + 4ppm BAP). Plantlet growth data including number of shoots, leaves, roots, and plantlet height were analyzed using one-way ANOVA test followed by Duncan's test at 5% level. The results showed that the addition of NAA and BAP affected the growth of Kepok Kuning banana plantlets on MS media in vitro. This is shown in the optimal number of shoots and leaves in treatment A, treatment B, and treatment C. Meanwhile, the number of roots and plantlet height is optimal in treatment D, and treatment E.

Polycyclic aromatic hydrocarbons (PAHs) in ambient air of Guangzhou city: Exposure levels, health effects and cytotoxicity

Polycyclic aromatic hydrocarbons (PAHs) in PM2.5 pose potentially serious threats to human health. In this study, the distribution characteristics of 16 priority controlled, fine PM (PM2.5)-bound PAHs in the ambient air of Guangzhou city were analysed from 2016 to 2019. Four high-molecular-weight PAHs with the highest annual average concentrations were benzo[ghi]perylene (BghiP; 0.757 ng/m3), indeno(1,2,3-cd)pyrene (IcdP; 0.627 ng/m3), benzo[b]fluoranthene (BbF, 0.519 ng/m3) and 3,4-benzopyrene (BaP; 0.426 ng/m3). Increasing concentrations of BghiP, IcdP, BbF and BaP were associated with increasing numbers of outpatient visits for respiratory diseases, indicating that exposure to these PAHs potentially causes acute respiratory injury in residents. Acute exposure of the human bronchial epithelial cell line BEAS-2B cells to BghiP, IcdP, BbF and BaP in vitro resulted in acute inflammation, DNA damage and apoptosis. Further bioinformatic analysis indicated that nuclear receptor subfamily 1 group D member 1 (NR1D1) may be a key target gene involved in mediating the toxic effects of BghiP. Collectively, our results suggest that BghiP and the other PAHs represented by it can damage the respiratory system and induce lung cancer. This study provides valuable evidence regarding the potential health risks posed by local ambient PAHs pollution.

OPTIMIZATION OF Talinum paniculatum Gaertn. ROOT INDUCTION AND THE EFFECT OF PHOSPHATE CONCENTRATIONS AND AMMONIUM:NITRATE RATIO ON BIOMASS OF ADVENTITIOUS ROOTS IN IN VITRO CULTURE

Java ginseng (Talinum paniculatum Gaertn.) is a medicinal plant, the roots of which are commonly used in traditional medicine. In its natural habitat, the roots grow very slowly, requiring two to three years to produce 100 g of roots per plant. Plant tissue culture could therefore provide an alternative means of accelerating root growth. This research aimed to optimize root induction and determine the effect of phosphate (KH2PO4) concentration and the ratio between ammonium and nitrate (ammonium:nitrate) on the biomass of Java ginseng adventitious roots in in vitro culture. Stem and leaf were used as explants and various combinations and concentrations of IBA and BAP, kinetin, and TDZ were used as growth regulators. Leaf explants were grown in Murashige and Skoog (MS) media supplemented with IBA 2 mg/L and various concentrations of phosphate (170; 212.5; 255; 297.5; 340; 382.5; 425; 467.5; 510 mg/L) and various ammonium:nitrate ratios (21:19 mM as the control, 0:30 mM, 10:20 mM, 15:15 mM, 20:10 mM, 30:0 mM). Cultures were maintained for 6 weeks. The observed parameters were fresh weight, dry weight, the duration of root formation, and the number and length of adventitious roots. The data were analyzed using Analysis of Variance. The results showed that the concentration of phosphate and the ammonium:nitrate ratio significantly influenced the amount, length, fresh weight, and dry weight of Java ginseng adventitious root. The highest fresh weight (37.47 mg) and dry weight (5.53 mg) were achieved in the treatment of double phosphate concentration (KH2PO4 340 mg/L), while an ammonium:nitrate ratio of 10:20 mM was the optimum treatment to produce the highest biomass (fresh weight 73.6 mg and dry weight 8.2 mg).

Role of homobrassinolide, abscisic acid, and 6-benzylamino purine on delaying flower senescence in Gladiolus grandiflora

A study was conducted to investigate the effects of homobrassinolide (HBL), abscisic acid (ABA) and 6-benzylamino purine (BAP) on the post-harvest life of 2 Gladiolus grandiflora cultivars, snow princess and nova lux. Different concentrations of HBL, ABA and BAP were applied to cut flowers. The experimental results showed that ABA and BAP treatments significantly increased post-harvest life in snow princess and nova lux cultivars compared to untreated flowers. However, HBL treatment was unsuccessful in delaying senescence in gladiolus. Furthermore, ABA and BAP treatments were more effective in delaying senescence in the nova lux cultivar than in the snow princess cultivar. Vase solutions containing BAP (500 M) and ABA (10 M) were the most efficient in extending the life of the cut floral spike of the nova lux variety (10 and 9 days respectively) followed by the snow princess variety (8 and 7 days respectively). Increasing fresh weight of flowers, vase solution uptake and membrane integrity along with a decrease in pH, malondialdehyde content, and lipoxygenase (LOX) activity prolonged flowers' post-harvest life. In terms of the post-harvest life of G. grandiflora, BAP outperformed ABA in improving the flower longevity of Gladiolus by maintaining higher physiological and biochemical stability in petals.

In vitro propagation of oil-bearing Rosa damascena using phloroglucinol: A protocol for rapid and high-quality shoot multiplication and rooting

Damask rose (Rosa damascena Mill.), of the family Rosaceae, is an important ornamental-medicinal, essential oil-bearing species. Due to its economic value, the aim of this study was to establish a new protocol for the in vitro propagation of R. damascena ‘Isfahan' using phloroglucinol. Axillary bud explants, Murashige and Skoog (MS) medium, and combinations of various plant growth regulators (PGRs) were used for the development of an effective method for rapid and high-quality shoot multiplication and in vitro rooting of Damask rose ‘Isfahan’. In the establishment step, 0.1 mg·L–1 phloroglucinol (PG), 0.2 mg·L–1 gibberellic acid (GA3), 0.1 mg·L–1 indole-3-butyric acid (IBA) and 1 mg·L–1 6-benzylaminopurine (BAP) were used. Secondary explants (nodal segments containing an axillary bud), obtained after 30 days of culture, were transferred to the proliferation media enriched with different concentrations of PG (0, 0.1, 0.2 and 0.3 mg·L–1), BAP (0, 1, 2 and 3 mg·L–1) and IBA (0, 0.1, 0.2 and 0.3 mg·L–1). A total of 64 combinations of these PGRs were investigated as a factorial experiment on the basis of a completely randomized design. Results showed that the shortest time until bud break (4.3 days), the highest number of green leaves (13.3) and lowest number of yellow leaves (1.3) were obtained in the explants grown on the medium containing 0.3 mg·L–1 PG + 2 mg·L–1 BAP + 0.2 mg·L–1 IBA. The largest number of shoots (4.8), and the highest fresh (5.9 g) and dry (1.2 g) weight of roots per explant were produced in the medium supplemented with 0.2 mg·L–1 PG + 2 mg·L–1 BAP + 0.3 mg·L–1 IBA. The highest regeneration coefficient (2.53), stem length (6.47 cm), fresh (2.32 g) and dry (0.51 g) weight of stem were obtained in the medium with 0.2 mg·L–1 PG + 2 mg·L–1 BAP + 0.2 mg·L–1 IBA. On the other hand, the highest number of roots (4.7) per explant were produced in the media supplemented with 0.3 mg·L–1 PG + 2 mg·L–1 BAP + 0.3 mg·L–1 IBA. None of the shoots regenerated roots in the absence of IBA. In most measured traits, the lowest values of the measured traits were obtained in the control medium without PGRs. Rooted plantlets were transferred to pots containing perlite and peat moss in 2:1 proportion and acclimatized in a greenhouse with a mean 82% survival rate. The positive role of PG in micropropagation was confirmed. The obtained results are more efficient than the previously established ones. Therefore, this cost-effective protocol can be used for the in vitro mass propagation of R. damascena Mill.

Morphogenesis features of <i>Hedysarum alpinum</i> L. in <i>in vitro</i> culture

The plants of Hedysarum alpinum L. in the North are characterized by a long pregenerative period, the laboriousness of seed propagation, low field germination of seeds, rare self-sowing, and the impossibility of vegetative propagation. In connection with this, it seemed relevant to introduce this rare species (included in the Red Book of the Komi Republic) into aseptic culture to preserve and maintain the gene pool, as well as to further develop the technology of its cultivation to obtain medicinal raw materials. The morphogenesis was studied, and the peculiarities of the regeneration of adventitious shoots of Hedysarum alpinum in in vitro culture were revealed. During three subcultivations, the influence of two cytokinin growth regulators, 6-benzylaminopurine (BAP) and kinetin (KIN), at concentrations from 0,5 to 1,0 mg/l, on the regeneration potential of different explants of Hedysarum alpinum in in vitro culture was determined on the WPM environment. A high frequency of regeneration of shoots from single-node segments and cotyledon nodes was obtained 79% and 65%, respectively. In the second and third passages, there was an increase in the regenerative activity of the cotyledon node explants and an increase in the multiplication factor to 5 shoots per explant. It was shown that relatively high concentrations (1,0 mg/l) of BAP and CIN increase the number of shoots per explant but inhibit their growth and development. For the reproduction stage proper, the medium containing BAP 0,5 + CIN 0,5 + IAA 0,1 mg/l is optimal. It was shown that the initiation of rhizogenesis actively took place on the WPM medium supplemented with IAA 0,5 and IBA 0,5 mg/l, on which the proportion of rhizogene explants was 77%. The maximum values for the length and number of roots were obtained on the medium with IAA 1,0 mg/l and with the combined use of two auxins (IAA with IBA at 0,5 mg/l). The regenerated plants were transplanted into non-sterile conditions, and their survival rate under ex vitro conditions was 50%.

Direct and indirect shoot regeneration from leaf explants of Centratherum punctatum Cass., A wild ornamental plant

Centratherum punctatum Cass. is an important wild ornamental medicinal plant belonging to Asteraceae family. It has anti-inflammatory, anti-cancerous, anti-hypersensitive, anti-bacterial and anti-fungal properties and is commonly used in hair care preparations. In this study we describe a protocol for high frequency direct and indirect shoot regeneration from leaf explants. In vitro derived leaves were cultured on MS medium supplemented with different concentrations of TDZ, ZN or BAP (0.1–2.0 mg/L) alone or in combination with NAA (0.1–0.7 mg/L) for shoot regeneration and varying concentrations of 2, 4-D or NAA (0.2–2.0 mg/L) for callus induction. Optimum direct shoot regeneration response was obtained from in vitro derived leaves in MS medium supplemented with 0.5 mg/L TDZ (61.4 ± 3.90% response; 27.4 ± 1.99 shoots per explant; mean shoot length of 4.8 ± 0.81 cm). Out of the 7, 14, 21 and 28-day-old leaves, 14-day-old leaves resulted in maximum shoot induction indicating the influence of explant age. Lamina with petiole exhibited a significant increase in shoot regeneration compared to that without petiole. Pretreating the explants with 1.0 mg/L TDZ for 24 h prior to culture produced the highest response of 92 ± 1.84% with a mean number of 38.7 ± 1.24 shoots per explant. 2, 4-D was found favourable for callus induction than NAA for callus induction from leaf explants of C. punctatum. The 21-day-old leaf showed the optimum callus induction. The optimum shoot regeneration from callus was obtained in 1.5 mg/L TDZ (84.2 ± 4.09% response with 58.2 ± 2.46 shoots per explant). Well-developed shoots with 3.0–4.0 cm length were rooted on half strength MS medium containing 0.5 mg/L IBA. The rooted shoots were transplanted to soil and acclimatized with 96.8 ± 3.26% success. Analysis of clonal fidelity of the micropropagated plants through SCoT marker analysis revealed the genetic similarity of mother plant and in vitro derived plants.