Mean Number Of Roots Research Articles

Impact of Various IBA Concentrations on Rooting in Apical Shoot Cutting and Stem Cuttings of Callistemon lanceolatus (Sm.) Sweet in Polytunnel Conditions

The study was undertaken to identify the optimal concentration of IBA and type of cuttings suitable for propagation of Callistemon lanceolatus and it was conducted at the Vanavarayar Institute of Agriculture, Pollachi, Coimbatore, India from December 2022 to September 2023. The experiment addressed the challenges of seed propagation, such as their seed capsules need to be activated by fire and the unpredictable nature of seed germination, which results in high progeny diversity due to genetic recombination. The investigation focused on apical and stem cuttings subjected to fifteen different treatments with three replications each; the Tukey test was used for statistical analysis; the cuttings were treated with various concentrations of Indole-3-butyric acid (IBA) and placed under polytunnel conditions; observations were made after 45 days, measuring sprouting percentage, rooting percentage, mean number of leaves, shoot length, mean number of roots, and root length per cutting. In light of these issues, the study explored clonal propagation as an alternative for mass multiplication of elite trees. The results of the study depicted that the maximum rooting percentage and root length were observed in T6 (73.28±8.87%) and (12.54±0.00cm) due to the exogenous application of auxin. On the whole, the result of this investigation suggest that apical shoot cuttings with IBA at 6000ppm can be made to induce maximum rooting and to produce high quality planting stock material for clonal propagation in Callistemon lanceolatus.

Establishment of In Vitro Cultures and Genetic Fidelity Analysis in Valeriana jatamansi -a High Value Endangered Medicinal Herb

The present study was conducted in the Department of Biotechnology, College of Horticulture and Forestry, Neri, Hamirpur, Himachal Pradesh, India during 2018–2020 for 18 months to develop an efficient, rapid and reproducible protocol for in vitro establishment along with analysing antioxidant activity. In the present study, surface sterilization protocol was standardized using Sodium hypochlorite and mercuric chloride as sterilant. Maximum direct shoot induction frequency from nodal explant was achieved on Murashige and Skoog medium enriched with BAP (5.0 mg l-l), TDZ (3.0 mg l-l) and NAA (1.0 mg l-l) for 18 days. Microshoots inoculated on MS media supplemented with 4.0 mg l-l BAP, 2.0 mg l-l TDZ, 1.0 mg l-l NAA and 0.5 mg l-l GA3) showed maximum frequency of shoot multiplication with average shoot length (4.36±0.08 cm) and average shoot number (14.00±0.57). A 75.00±1.15% rooting with significantly high mean root number (8.00±0.57) and root length was achieved in full strength MS medium, supplemented with same concentration i.e. 4.0 mg l-l BAP, 2.0 mg l-l TDZ, 1.0 mg l-l NAA and 0.5 mg l-l GA3) combination. Maximum plantlets survive after 1 year of acclimatization. RAPD and ISSR markers were used to confirmed genetic stability of in vitro raised plants by showing 100% monomorphism. High multiplication rate associated with genetic stability ensure the efficacy of the present in vitro clonal propagation protocol of this important medicinal plant.

In vitro micropropagation of Aloe elegans Tod. using offshoot cuttings

ObjectiveAloe elegans Tod. is an ecologically, environmentally, medicinally, and commercially useful aloe species in Ethiopia and Eritrea. Unfortunately, it is highly threatened due to industrial and urban expansion and traditional mining and agricultural activities. As a consequence, it is included in the IUCN List of Threatened Species since 2013. The plant is getting thinly populated in many parts of the Tigrai floristic region since it is being exploited for traditional and commercial purposes. Therefore, this study was aimed to develop a reproducible, large-scale micropropagation protocol using offshoot cuttings in Murashige and Skoog (MS) media enriched with plant growth regulators (PGRs).ResultsSterilized explants cultured in full-strength MS media enriched with 0.25 mg/L benzyl amino purine (BAP) and 0.10 mg/L naphthaleneacetic acid (NAA) resulted in 100% healthy and live (i.e., initiated) explants after four weeks of initiation study. Unsupplemented initiation media (control) yielded only 14.3% initiated explants. The initiated explants were tested for their shooting response to produce microshoots by incubating in different concentrations and combinations of BAP and NAA for four weeks. Fewer days to shooting (13.0 ± 1.0 days), higher mean shoot number (5.0 ± 1.0), and higher mean shoot length (9.20 ± 2.35 cm) were observed with 1.0/0.50, 1.0/0.25, and 1.0 /0.50 mg/L BAP/NAA combinations, respectively. The rooting responses of the microshoots toward producing plantlets were also tested by incubating them in half-strength MS media enriched with different concentrations of NAA and indole-3-butyric acid (IBA) for four weeks. Fewer mean days to rooting (12.0 ± 1.0 days), higher mean root number (8.0 ± 4.0), and higher mean root length (7.53 ± 3.03 cm) were observed in MS media enriched with 0.75, 0.75, and 1.25 mg/L IBA, respectively. The responses of A. elegans plantlets to primary (in greenhouse) and secondary (in nursery shade and direct sunlight) acclimatization in coco peat, composted soil, and manured soil media were high – with survival percentages of 87.5–97.8% in three to four weeks.

Protocolo de enraizamento in vitro de porta-enxertos de Pyrus comunnis

ABSTRACT: Effective protocols for in vitro rooting for woody fruit trees are still a challenge for in vitro seedling production, especially when there is a need to insert new cultivars or rootstocks. These protocols are essential to accelerate studies in plant breeding programs and for seedling distribution. This study evaluated the use of 6-Benzylaminopurine (IBA) in in vitro rooting of Pyruscomunnis rootstocks, clones ‘OHxF87’ and Pyrodwarf. Explant exposure times (0, 24, 48, 72, and 96 hours) to 20 mg L-1 IBA were tested for in vitro rooting. The exposure to IBA resulted in rooting rates above 80%, surpassing some results reported in the literature. The 24-hour treatment provided 81,81% survival, leading to an average growth of five roots with 19 mm length, for ‘OHxF87’ rootstock. The same exposure time resulted in the highest survival rate (75%) and the highest mean root number, seven roots per plant with 10 mm length, for ‘PDW’ rootstock. Root formation did not occur in the absence of synthetic auxin. Therefore, it can be concluded that a 24-hour exposure at 20 mg L-1 IBA was sufficient to promote in vitro rooting in ‘OHxF87’ and Pyrodwarf rootstocks’.

In vitro direct and indirect regeneration of plants from nodal and petiole explants in Pelargonium odoratissimum (L.) Herit.

Nodal and petiole explants were employed to study the direct and indirect regeneration from Pelargonium odoratissimum in vitro. Direct shoots regeneration of nodal segments was tried in MS medium containing 1 and 2 mg l-1 BAP and 0.5 mg l-1 IBA. The highest mean shoots number and the greatest shoots per explant number were obtained in the medium containing 2 mg l-1 BAP. Nodal segments were the source of petiole explants and the resulting petioles were cultured in ½ MS medium supplemented by 1, 1.5, 2 and 4.5 mg l-1 BAP enriched with 0.1, 1 and 1.5 mg l-1 NAA. With the petiole explants, the lowest browning percentage, the highest callus induction and also, the top number of shoots per explant were recorded in 2 mg l-1 BAP + 0.1 mg l-1 NAA medium. The medium supplemented with 0.2 mg l-1 NAA exhibited the desired effect on rooting percentage and mean root number and length. The rooted young plants were transferred to the pots containing peat-moss and perlite (1:1) and the acclimatization was successful since, more than 90 % of plants adapted-well in the greenhouse conditions. This in-vitro propagation methodology would be advisable to the plant production systems and to whom wish to produce the clonal homogenous plants for the commercial ideas and for the detailed molecular studies.

Ortet Age Effect, Anatomy and Physiology of Adventitious Rooting in Tilia mandshurica Softwood Cuttings

Tilia mandshurica is recognized as one of the most valuable timber, ornamental and nectariferous tree species, and its cutting propagation is very difficult. To evaluate the influence of ortet age on the rooting of T. mandshurica cuttings and the relationship between the variations of nutrients, enzyme activities, endogenous hormones and the formation of adventitious roots, a series of softwood cuttings of T. mandshurica were collected from 2-, 7-, 15- and 20-year-old healthy ortets, respectively. The rooting percentage, mean root number and total root length were investigated after 60 days of cultivation. Phenotypic and anatomical changes and the variations of organic nutrients, enzyme activities and endogenous hormones were measured during adventitious root formation. The results showed that ortet age effect existed in T. mandshurica softwood cuttings. The adventitious rooting of 2-year-old ortets was superior to that of 7-, 15- and 20-year-old ortets. No root primordium was visible in T. mandshurica softwood cuttings before cultivation, and the root primordia were induced after planting. The adventitious roots derived from vascular cambium and callus in cuttings. Soluble sugar increased during root primordium induction and decreased in adventitious root elongation. Soluble protein decreased during root primordium induction and increased in adventitious root elongation. Polyphenol oxidase (PPO) activity increasing and peroxidase (POD) and 3-indoleacetic acid oxidase (IAAO) activities decreasing were conducive to root primordium induction. High 3-indoleacetic acid (IAA) and gibberellin (GA3) level could promote root primordium induction and low IAA and GA3 level was beneficial to adventitious root elongation. Abscisic acid (ABA) and zeatin riboside (ZR) inhibited root primordium induction and adventitious root elongation. IAA, ABA and ZR coordinated with each other in the process of root primordium induction and adventitious root formation. IAA/ABA increasing could promote root primordium induction, and IAA/ZR increasing was beneficial to root primordium induction and adventitious root elongation. The soluble sugar content, PPO activity, IAA/ABA and IAA/ZR of softwood cuttings from 2-year-old ortets were relatively high, which may be the reason for the better adventitious rooting. The present study provides a reference to build a propagation by cuttings technology of linden trees.

Influence of basal vitamins, growth regulators, and explants on in vitro organogenesis from synthetic seeds of Citrus jambhiri Lush.

An efficient shoot and root organogenesis protocol has been developed from synseeds derived from various explants of Citrus jambhiri Lush. Optimum synseeds were developed using sodium alginate (0.5-0.75%) in 1.0% CaCl2 solution. Shoot organogenesis was examined under various basal vitamin mediums (MS Nitsch and MSB5) supplemented with various concentrations of adenine sulfate (ADS) and 6-benzyl amino purine (BAP) from different explants such as cotyledonary junction, shoot tip, and nodal explants. The synseed regeneration response ranged between 60-100% among the vitamins, cytokinins and explants used. The number of shoots per synseeds was higher (13.4) in MSB5-BAP (1.5 mg L-1), followed by 12.8 in MSN-ADS (2.0 mg L-1), 11.2 in MSN-ADS (1.0 mg L-1), and 10.8 in MSB5-ADS (1.0 mg L-1) from the synseeds developed using 0.75% sodium alginate. The mean number of roots per explant was higher (4.2) in ½MSN+IAA (1.0 mg L-1). Similarly, the mean root length was higher (5.2 cm) in ½MSN+IAA (0.5 mg L-1) followed by 4.2 cm in ½MSN+IAA (1.0 mg L-1). Regenerants derived from synseeds have shown no somaclonal variations, confirming that the plantlets are true-to-type to their parental progenies. The encapsulated plantlets showed >90% survivability while transferred at Kachai village, Manipur, India. The results of the present study encourage the use of various vitamin mediums and explants for large-scale propagation of C. jambhiri through synseeds.

Effective Methods for Adventitious Root Regeneration on Weeping Fig Stems

When transplanting mature Ficus trees, the large root balls are expensive to treat, handle, and move. This study aimed to identify the optimal wounding method and auxin treatment for regenerating adventitious roots (ARs) from weeping fig (Ficus benjamina L.) stems to uptake additional water and to compensate for fewer absorption roots in the smaller root balls at transplantation. We adopted a two-factorial experiment involving the wounding methods (three-line cut (3LC) and rectangular peel (RP)) and auxin treatments (2000 mg·L−1 Indole-3-butyric acid (IBA), 2000 mg·L−1 IBA + 2000 mg·L−1 1-naphthaleneacetic acid (NAA), and 4000 mg·L−1 IBA). The rooting rate of each treatment, the mean root number, the length of the three longest ARs, and the dry weight of ARs in each wound were evaluated. The treatment combination using 4000 mg·L−1 IBA with RP13 (rectangular peel 1/3 the perimeter of the stem) consistently exhibited the best rooting results in 2019 and 2020. It had a 100% rooting rate, a mean of 18.5 roots, a 16.8 cm root length, and a 1640 mg dry weight in the wounds. All auxin treatments demonstrated a superior rooting ability as compared to water treatments. The RP method regenerated more roots than the 3LC method. Doubling the RP length to be 2/3 of the perimeter improved the rooting ability. The locations of ARs varied under different treatment combinations, with 4000 mg·L−1 IBA on RP13 demonstrating the most diversified distribution on four edges of the wounds. Thus, it is recommended to regenerate ARs from stems of F. benjamina trees.

Influence of spectral quality on the rooting of Corymbia and Eucalyptus spp. minicuttings

The pursuit of better adaptation in clonal plants seedling production processes based on the minicutting technique has expanded the use of species and hybrid combinations of genera Corymbia and Eucalyptus in the composition of commercial crops. The aim of the work was to evaluate the effect of spectral quality on the rooting of Eucalyptus andrewsii, E. saligna, E. microcorys, E. cloeziana, E. pilularis, E. grandis, E. grandis × E. urophylla and Corymbia torelliana minicuttings to help better understanding the production of clonal plants. E. grandis × E. urophylla and C. torelliana root anatomy was analyzed. The effects of spectral quality on the rooting of minicuttings were evaluated based on three sources (fluorescent, red and blue). Survival (SUR), callogenesis (CAL), oxidation (OXI) and rooting (RO) percentage; length (RL) and diameter of the largest root (ROD); mean number of roots per minicutting (NRM), root epidermis thickness (RET), root cortex diameter (RCD), diameter of the root vascular cylinder (DRVC) and root diameter (RD) were evaluated at 30 days. Based on the results, wavelength specificity was a useful technology to optimize the large-scale production of clonal plants of Eucalyptus. Fluorescent spectral quality was the most appropriate source in the rooting of E. saligna (68.7%), E. microcorys (43.7%), E. pilularis (75.0%) and C. torelliana (75.0%) minicuttings; blue spectral quality was the most appropriate for E. andrewsii (55.5%), E. grandis (75.0%) and E. grandis × E. urophylla (81.3%); and red spectral quality was the most appropriate for E. cloeziana (56.2%).

Mapping of quantitative trait loci for root elongation in rice (Oryza sativa L.) treated with various nitrogen sources and concentrations

ABSTRACT Roots are important plant organs that absorb water and nutrients. Root length responses are influenced by the plants’ sources of nitrogen (N), such as ammonium (NH4 +) and nitrate (NO3 −) ions and their concentrations. Here, to determine the genetic resources with the potential to improve the root system of rice (Oryza sativa L.), the root development traits of a core set of 334 introgression lines (ILs) were assessed in plants grown in hydroponic culture medium containing low (5 μM) or high (500 μM) concentrations of N in the form of NH4 + or NO3 −. ILs were classified into three clusters. The mean value of maximum root length (MRL) in cluster I was significantly higher than that in the other two clusters under all conditions. Root number (RN) differed significantly between clusters IIa and IIb, with the mean RN being higher in cluster IIa. In addition, the order of the mean total root length of the clusters was cluster I > IIa > IIb. Therefore, ILs in cluster I are considered genetic resources for improving root development traits under all tested N conditions. Among the ILs, YTH187 – which is derived from YP5 as a donor – had a longer MRL under all conditions. Quantitative trait loci (QTL) analyses, performed using recombinant inbred lines derived from a cross between IR 64 and YTH187, revealed that qRL5.3-YP5 and qRL6.5-YP5 were detected under all N conditions. In contrast, qRL4.1-YP5 was detected only under low KNO3 concentrations, whereas qRL8.1-YP5 was detected under high concentrations of NH4Cl and KNO3. YP5 alleles at these QTLs increased MRL. QTLs for root length in rice grown in hydroponic culture have not previously been detected in the vicinity of qRL8.1-YP5; thus, this may be a novel QTL that controls root length in rice. Moreover, qRL8.1-YP5 and qRL4.1-YP5 could be useful QTLs to determine the mechanism of root elongation under KNO3 conditions because such QTLs have not yet been detected under these conditions in rice. In future studies, further fine-mapping and characterization of qRL8.1-YP5 could reveal the mechanism of root elongation under high KNO3 concentration conditions.

Rooting Responses of Female and Male Cuttings of Leucadendron elimense E. Phillips subsp. elimense on the Type and Concentration of Auxin

Abstract Female and male plants of difficult-to-root species Leucadendron elimense subsp. elimense were investigated for rooting potential with three rooting hormones: indole-3-acetic acid (IAA), 1-naphthaleneacetic acid (NAA), and indole-3-butyric acid (IBA) at three different concentrations (2000 ppm, 4000 ppm, and 6000 ppm). The experiment was made under essential misting, bottom heat, and a naturally ventilated greenhouse. After 18 weeks the experiment terminated and callusing and rooting percentage, root number, and root length were determined. Female cuttings rooted in the highest percentage (80%), root number (>30), and root length (close to 100 mm) after treatment with IAA at 4000 ppm. Under the above treatment male cuttings rooted in 70%, with a mean root number of 24 and root length of 90 mm. The efficient rooting results could aid in saving the species in its natural habitat and supporting restoration ecology, as well as introducing this species into the flower market.

Effect of Phytohormones on rooting behaviour in air layers of Phoebe cooperiana, an economically important tree species of Arunachal Pradesh, India

This study reports the response of air layers of Phoebe cooperiana (Common name: Tapil) to different concentrations of rooting hormones in mid hills of Arunachal Pradesh. Ten trees between the ages of 7 and 10 were selected at Boleng village of Siang district, Arunachal Pradesh, India for the experiment. Eight treatments comprising of IBA and NAA at concentrations of 1000ppm, 2000ppm and 3000ppm each, Rootex hormone for semi-hardwood and control were applied to a total of 240 layers during April 2020. After 60 days, rooting percentage, mean number of roots and length of the longest root were recorded. Significant variation (p<0.05) among treatments was observed for all root parameters. Among treatments, the highest rooting percentage and mean a number of roots were obtained in layers treated with NAA 3000ppm (80% and 21.33 respectively). The length of the longest root was the highest in layers treated with NBA 2000ppm (4.23cm). The highest survival percentage of layers two months after transplanting was observed in those treated with NAA 3000ppm (54.16%) and the least in control (23.07%). The technique offers an easy and cheap method of propagation for farmers as well as for government and private agencies to enhance seedling production of the species.

Synergistic interaction of exogenous auxin and reduced level of stockplant irradiance on the rooting of cola acuminata (Pal. De Beauv.) stem cuttings: applications for agroforestry trees domestication

ABSTRACT The establishment of an efficient propagation protocol is one of the most important steps in the domestication of agroforestry trees. Using Cola acuminata as a biological model, the present study investigated the interaction between exogenous auxin and reduced levels of stockplant irradiance for its effect on the rooting of stem cuttings. In a 3 × 4 factorial experiment, three levels of stockplant irradiance and four concentrations of indole 3-butyric acid (IBA) were tested for their effect on the rooting potential of leafy stem cuttings. Relationship between flavonoid content of stockplants and rooting potential of stem cuttings was also investigated. Results showed that percentage of cuttings rooted significantly increased, as did the mean number of roots, with decreasing levels of stockplant irradiance. Likewise, rooting was enhanced by all applications of IBA. 100% rooting was achieved under the combination of 50% and 25% ambient irradiance together with 2% and 4% IBA. The mean number of roots was greatest (11.4 ± 0.2) when 4% IBA was combined with 25% ambient sunlight. Flavonoid content of plant extracts decreased with stockplant irradiance. There were significant negative correlations between flavonoid content of plant extracts and the rooting parameters of cuttings.

Simulating root distribution of plant individual with a three-dimensional model

AimsSoil resources distribute heterogeneously in natural ecosystems, which is known as soil heterogeneity. Responses of plant roots to soil heterogeneity are important for their adaptation to the changes of environments. However, few observations of root systems in soil are recorded, and there is a lack of comprehensive theory for roots responses to soil heterogeneity. MethodsSoil heterogeneity including configurational heterogeneity and qualitative heterogeneity is developed in three dimensions via extending a stochastic spatially explicit lattice-based habitat model in two dimensions. Root dynamics of plants growing in such soil heterogeneity in three dimensions are simulated. In these simulations, three parameters (i.e. ratio of root distributed in resource-rich patches, mean root length and root number of plants) are measured. ResultsComparing with the previous two dimensional model of heterogeneous pattern formation, this three-dimensional model is time-saving and more efficiency. The spatial pattern of habitats, as well as the adaptability of plants to soil heterogeneity, determines how plants place their roots into resource-rich and resource-poor patches. The plasticity of root architecture plays an important role in the root response to soil heterogeneity. Specifically, plants with the ability to respond to resources would adopt a large number of roots with short root length as the ratio of resource-rich patches is relatively small, while they would adopt a small number of roots with long root length as the ratio of resource-rich patches is relatively large. ConclusionsSuch realistic three-dimensional model allows us to better explore the effects of soil heterogeneity on plant root dynamics. Applying this model in root dynamics improves our understanding of root distribution in spatial habitat and of the formation mechanism of root morphology.

Effect of Medium Composition on In vitro Seed Germination and Plant Development in Kentucky Bluegrass (Poa pratensis L. cv. Evora)

In the current study, the effects of De Greef & Jacobs (DG), Linsmaier & Skoog (LS), Murashige & Skoog (MS), and Schenk & Hildebrandt (SH) media were tested on seed germination and plant development in Poa pratensis cv. Evora. The highest germination rate (83±2.74%) was found on SH medium, whereas LS medium gave the lowest (46±4.18%) germination rate. The statistically same leaf numbers were recorded from SH (2.10±0.27) and DG (2.12±0.18) media. SH and DG media gave 4.28±0.28 cm and 4.16±0.31 cm mean leaf lengths, respectively. SH medium gave the maximum mean root number (3.09±0.26). However, the LS medium gave the lowest mean root number (1.84±0.10). The longest roots (1.43±0.19 cm) were observed in the plants grown in SH medium. However, DG medium had the minimum mean root length (0.81±0.08 cm). In conclusion, SH medium should be preferred over the other medium tested in in vitro tissue culture studies on the species to increase the biomass production yield. The efficacy of SH medium in the in vitro propagation of P. pratensis could be increased using growth promoters.

In vitro propagation and DNA barcoding of the rare medicinal plant Aerva Lanata (L.) Juss. ex Schult grown in Gebel Elba

Aerva Lanata (L.) Juss. ex Schult is an important medicinal multipurpose plant that belongs to the family Amaranthaceae. An efficient protocol for In vitro propagation of A. Lanata was established. Nodal segment explants collected from mature shrubs from the Gebel Elba, South Egypt. Explants were cultured on Murashige and Skoog (MS) medium supplemented with different concentrations of 6-benzyl adenine (BA) (2.22, 4.44, 8.90 and 17.80 M) and Kinetin (Kin) (2.32, 4.60, 9.30, and 18.6M) individually for shoot induction. The highest shoot induction percentage (93.33%) with maximum number for shoots (4.07 shoots per explant) was noticed on MS medium supplemented with 4.44 M BA. The maximum number of multiple shoots per explants (9.13) was obtained in the combination BA (4.44 M) and NAA (2.14 M). For root induction, the highest rooting percentage (80%) with mean root number (4.33 roots per shoot) and length (3.83 cm) of roots were noticed on half- strength MS medium augmented with 4.90 M IBA. Rooted plantlets were transferred into plastic pots containing sand, peat moss and garden soil mixture at equal volumes, and successfully acclimatized in the greenhouse with a survival rate of 73.3%. Plant identification is a crucial aspect to understand and conserve plant diversity from extinction. DNA barcode analysis of A. Lanata was carried out using. The present in vitro propagation protocol would facilitate an alternative method for rapid and large-scal production of this important medicinal plant. We therefore, used the rbcL gene to identify A. lanata. Maximum likelihood tree analysis was performed to evaluate the discriminatory power of the rbcL gene. Our findings showed that using rbcL gene sequences enabled identifying the majority of the samples (90%) to the genus and species level.

High Efficiency In vitro Regeneration and Genetic Stability of Corallocarpus epigaeus - An Endangered Medicinal Plant

Indirect regeneration of plantlets from multiple shoot induction of Corallocarpus epigaeus was obtained from leaf and nodal explants on MS with different concentrations of BAP in combination with IAA/IBA or IBA alone. Among all the combinations, BAP and IBA exhibited maximum regeneration. High frequency of multiple shoots (89%) was obtained on BAP (2.0 mg/l) and IBA (1.5 mg/l) in nodal explants. Maximum mean shoot length of 6.8 ± 0.33 cm was obtained in nodal explants cultured on BAP (1.0 mg/l) + IBA (0.5 mg/l), followed by leaf explants with 6.7 ± 0.47 cm on BAP (3.0 mg/l) + IAA (2.5 mg/l). The highest frequency of rooting (88.3%) was obtained on NAA (1.0 mg/l) and IBA (2.0 mg/l) with 21.83 ± 0.57 mean number of roots. The well-rooted healthy plantlets were acclimatized with a survival rate of 80%. Inter simple sequence repeat (ISSR) analysis revealed the genetic similarity of in vitro raised plants with the mother plant.
 Plant Tissue Cult. & Biotech. 30(2): 219-229, 2020 (December)

Highly efficient rapid micropropagation and assessment of genetic fidelity of regenerants by ISSR and SCoT markers of Solanum khasianum Clarke

An efficient method of rapid micropropagation of Solanum khasianum Clarke was successfully established from the leaf, petiole, and nodal explants. The morphogenetic response of different concentrations of TDZ and BAP individually or in combination with auxins (IAA/IBA/2,4-D) was tested. Friable callus was obtained on different concentrations of BAP alone or in combination with IAA/IBA/2,4-D. Rapid multiple shoot induction was achieved from friable callus on MS medium supplemented with varying concentrations of TDZ and IBA. The leaf explants exhibited a high frequency of multiple shoots than petiole and nodal explants with an optimal percentage of response (92.73%), mean shoot number (53.5 ± 0.47), and shoot length (11.2 ± 0.53 cm) on MS medium augmented with TDZ (1.5 mg l−1) and IBA (1.5 mg l−1). Maximum rooting efficiency was achieved on MS medium with 1.5 mg l−1 IBA with 12.8 ± 0.36 mean number of roots. The in vitro rooted plants were acclimatized with a survival rate of 80%. The genetic fidelity of the regenerants assayed by the ISSR and the SCoT markers showed no genetic variation. The study examined the micropropagation responses of S. khasianum in the presence of various growth regulators and provided a simple and more suitable protocol adapted for the mass propagation of clones in this species. We have established a highly efficient micropropagation system for large scale production in Solanum khasianum. Evaluation of clonal fidelity by using ISSR and SCoT markers detected no somaclonal variations. The present study helps to the enhancement of potential alkaloids (solasodine) with the help of biotechnological tools.

In vitro Regeneration of Saussurea heteromalla (D. Don.) Hand-Mazz Using Auxins and Current Status in Galiyat Abbottabad

Current Status of Saussurea heteromalla was investigated in Galiyat areas of District Abbottabad, viz., Jahaffar, Seri, Beeran Gali, Banj, Haryala, Daryala Gali, Sarbhanna, Barriyan, Akhreela and Broangiala. Saussurea heteromalla was found common in Seri, Sarbhanna and Barriyan; endangered in Haryala, Jahaffar, Banj, Daryala Gali and Beeran Gali and absent in Akhreela and Broangiala. In vitro regeneration of Saussurea heteromalla (D. Don.) Hand-Mazz on MS media was conducted and shoots were developed on full strength MS medium supplemented with 1 mgL-1 GA3. The developed shoots were transferred for root induction to half strength MS medium fortified with various concentrations of Indole butyric acid (IBA) and α-Naphthalene acetic acid (NAA) i.e. T1 (control), T2 (2 mgL-1 IBA), T3 (3 mgL-1 IBA), T4 (4 mgL-1 IBA), T5 (1 mgL-1 NAA), T6 (2 mgL-1 NAA) and T7 (3 mgL-1 NAA). Maximum mean shoot length (6.3 cm), mean number of leaves (7), mean number of nodes (5.25); highest root emergence percentage (71%), means root length (1.5 cm), mean number of roots (3) and highest survival rate (100%) was recorded in treatment T4. However, treatments T5 and T6 also seem to be effective for in vitro rooting of Saussurea heteromalla. Treatments T2, T3 and T7 showed minimum root growth. It was concluded that IBA at higher concentration is more effective for in vitro rooting and better shoot growth of Saussurea heteromalla whereas NAA also initiate rooting but at lower concentration.

The Effect of Different Applications on In vitro Bulb Development of an Endemic Hyacinth Plant (Hyacinthus orientalis L. subsp. chionophyllus Wendelbo) Grown in Turkey

In this study the effects of different sucrose concentrations, and the combinations of jasmonic acid (JA) with auxins (IAA or NAA) or with cytokinin (2iP) on the bulb induction and rooting of in vitro plantlets of Hyacinthus orientalis subsp. chionophyllus Wendelbo, which is endemic in Turkey, were investigated. The effect of four different sucrose concentrations (30, 45, 60 and 90 g L-1) on bulb formation in tissue culture was investigated. These plantlets were cultured on MS medium supplemented with several concentrations and combinations of JA (0.0, 1.0, 2.0 mg L-1) and 2iP (0.0, 0.25 and 0.50 mg L-1), IAA or NAA (0.5, 1.0 mg L-1). In JA- 2iP treatment, the highest number of bulblets (13.7 number/explant) was obtained by the combinations of JA 1.0 mg L-1 + 2iP 0.25 mg L-1. Also, the largest bulblets with the mean diameter of 7.9 mm were found on MS medium supplemented with JA 2.0 mg L-1. In JA – Auxin treatment, the mean root number per bulblet was highest (17.9 number/explant) and root formation rate was maximum (81.14%) on MS medium supplemented with IAA 1.0 mg L-1 + JA 2.0 mg L-1.