Efficient Plant Regeneration via Indirect Organogenesis in Carnation (Dianthus Caryophyllus Semperflorens flore pleno) Cultivars

ABSTRACT Callus induction and plant regeneration are important steps of in vitro plant breeding of ornamental plants. In this study, the effects of different combinations of plant growth regulators (PGRs), promoters, and minerals on callus induction and plant regeneration in different carnation cultivars were studied in a completely randomized design with three replications. For callus induction, 16 different combinations of 2,4-dichlorophenoxyacetic acid (2,4-D), 6-benzylaminopurine (BA), 1-naphthaleneacetic acid (NAA), and casein hydrolysate (CH) were studied using in vitro leaf explants. The Murashige and Skoog (MS) medium supplemented with 0.2 mg·dm-3 of 2,4-D and 200 mg·dm-3 of CH showed the highest frequency of callus induction. Among the cultivars, ‘Noblesse’ showed the highest rate of callus induction (91.67%). Regarding regeneration, BA, NAA, silver nitrate (AgNO3), and adenine hemisulfate (As) were used in ten different combinations. The ‘Cameron’, ‘Tabasco’, and ‘Noblesse’ cultivars with 95.24% regeneration percentage showed the highest rate of plant regeneration. Generally, in most cultivars, the highest regeneration rate and shoot number per explant were found in the MS medium supplemented with 3 mg·dm-3 of BA, 0.6 mg·dm-3 of NAA, 5 mg·dm-3 of AgNO3, and 40 mg·dm-3 of As. According to the results, the highest regeneration frequency was obtained when 40 mg·dm-3 of As was added to the medium. Finally, the flow cytometry analysis indicated that there were no significant differences between in vitro regenerated and control plants in terms of DNA ratios.

Evaluating sources of variability in inflorescence number, flower number and the progression of flowering in Sauvignon blanc using a Bayesian modelling framework

The time of flowering is key to understanding the development of grapevines. Flowering coincides with inflorescence initiation and fruit set, important determinants of yield. This research aimed to determine between and within-vine variability in 4-cane-pruned Sauvignon blanc inflorescence number per shoot, number of flowers per inflorescence and flowering progression using an objective method of assessing flowering via image capture and statistical analysis using a Bayesian modelling framework. The inflorescence number and number of flowers per inflorescence were measured by taking images over the flowering period. Flowering progression was assessed by counting open and closed flowers for each image over two seasons. An ordinal multinomial generalised linear mixed-effects model (GLMM) was fitted for inflorescence number, a Poisson GLMM for flower counts and a binomial GLMM for flowering progression. All the models were fitted and interpreted within a Bayesian modelling framework. Shoots arising from cane node one had lower numbers of inflorescences compared to those at nodes 3, 5 and 7, which were similar. The number of flowers per inflorescence was greater for basal inflorescences on a shoot than apical ones. Flowering was earlier, by two weeks, and faster in 2017/18 when compared to 2018/19 reflecting seasonal temperature differences. The time and duration of flowering varied at each inflorescence position along the cane. While basal inflorescences flowered later and apical earlier at lower insertion points on the shoot, the variability in flowering at each position on the vine dominated the date and duration of flowering.This is the first study using a Bayesian modelling framework to assess variability inflorescence presence and flower number, as well as flowering progression via objective quantification of open and closed flower counts rather than the more subjective method of visual estimation in the field or via cuttings. Although flower number differed for apical and basal bunches, little difference in timing and progression of flowering by these categories was observed. The node insertion point along a shoot was more important. Overall, the results indicate individual inflorescence variation and season are the key factors driving flowering variability and are most likely to impact fruit set and yield.

In vitro propagation from nodal segments of Lippia origanoides (chemotype A)

ABSTRACT: This research described an efficient micropropagation protocol for Lippia origanoides (Verbenaceae). Sterile seeds were used to obtain germinated seedlings in Murashige and Skoog medium (MS) supplemented with sucrose and agar. The nodal segments obtained from seedlings were grown on MS medium supplemented with different concentrations of gibberellic acid (GA), benzylaminopurine (BAP) and 1-naphthalenacetic acid (NAA) with BAP. The callus induction, shoots length, shoots number and root length, were analyzed. The treatments showed high percentage of callus formation at 0.5 to 1.5 mg L-1 of BAP alone or in combination with NAA (0.1 mg L-1). The highest value of shoot number per nodal segments was obtained at 1.5 mg L-1 of BAP (4.3 ± 0.8). The obtained plantlets were better rooted in vitro in the absence of plant growth regulators (PGRs) and they showed acclimatization rate of 90%. We reported a protocol for in vitro propagation and acclimatization of L. origanoides for A chemotypes from Colombia.

Development of An Efficient Protocol for Rapid Propagation of Curculigo orchioides Using Leaf Explant Culture

Curculigo orchioides is one of the most common medicinal plants used by diverse cultures and tribal groups. The roots of the plant are used medicinally in Asian countries. Curculigo orchioides have the ability to regenerate through seeds and tubers, but the regeneration rate is low. Plant tissue culture method was believed to have potential for rapid multiplication of this medicinal plant. An efficient protocol for rapid propagation of Curculigo orchioides, of the family Amaryllidaceae, was developed using leaf explants culture. The leaf explants (1 cm x 1 cm squares) cultured on Murashige and Skoog (MS) basal medium were supplemented with various concentrations and combinations of auxins and cytokinins with temperature 25 ± 2°C, relative humidity 85-90% and photoperiod of 12 hours light (2000-3000 lux). Callus induction was obtained within 4 weeks, 2,4-D at 3 mg/l formed profuse callus and the degree was found to be the highest (+++) among all the treatments. The best response to shoot induction, with maximum shoot number 5.33 (mean number of shoots per explant) was obtained using 1.0 mg/l 6-benzyl aminopurine (BAP) in combination with 1.0 mg/l Kinetin. In vitro shoots were induced for rooting on 0.5 mg/l of NAA supplemented medium. In order for seedlings propagated in vitro to adapt to natural conditions, plants were growned on a substrate coir: husk ash: sand (with the ratio of 0.5: 0.5: 1) in a greenhouse (humidity: 70%, temperature: 28-300C) gave 88.33% survival rate after 8 weeks of culture. With the results received, this is an effective approach to propagating Curculigo orchioides.

Determination of the Effect of Magnesium Oxide Nanoparticles (MgO-NP) on in Vitro Culture of Cowpea (Vigna Unguiculata L. Walp)

This study was carried out to determine the effect of different doses of MgO nanoparticles (MgO-NP's) on in vitro regeneration of cowpea. MgO NPs used in the study were synthesized using walnut shell extract by green synthesis method. MgO nanoparticles with 35-40 nm size were used in this research. In the study, the effect of different doses of MgO NP applied to cowpea plant on all in vitro parameters was found to be significant. Considering the parameters examined, the best results in morphogenesis were 185 mg/L, 370 mg/L and 555 mg/L MgO-NPs applications, the highest shoot formation rate was obtained 82.50% with 555 mg/L(MgO-NPs) and 72.50% with 370 mg/L (MgO-NPs). The highest mean values of shoot number, shoot number per explant and shoot length were observed in the application of 370 mg/L MgO-NPs with 61.25, 17.50, and 2.075 (unit) respectively. The best root formation rate was obtained from control and 370 mg/L MgO application at a rate of 27.50%. The highest values were 6.75 (pieces) and 370 mg/L MgO-NPs in the number of roots per explant, 555 mg/L MgO-NPs with 10% in callus formation, and 1.575 (cm) and 370 mg/L MgO-NPs in root length. When all the examined parameters were evaluated, it was determined that the application of 370 mg/L MgO-NPs gave the best results in terms of in vitro parameters.

Impact of Phytomediated Zinc Oxide Nanoparticles on Growth and Oxidative Stress Response of In Vitro Raised Shoots of Ochradenus arabicus

Biogenic nanoparticles have potential roles in the growth and development of plants and animals as they are ecofriendly and free of chemical contaminants. In this study, we assessed the effects of phytomediated zinc oxide nanoparticles (ZnONPs) on shoot growth, biochemical markers, and antioxidant system response in Ochradenus arabicus, which is a medicinal plant. The shoot length and fresh and dry weights were found to be higher in groups with 5 and 10 mg/L ZnONPs than in the control. At high concentrations of ZnONPs (50, 100, and 300 mg/L), biomass was decreased in a concentration-dependent manner. The shoot number was observed to be highest at 50 mg/L among all applied concentrations of ZnONPs. The levels of the stress markers proline and TBARS were found to be higher in shoots treated with 100 and 300 mg/L ZnONPs than in the control as well as NP-treated shoots. The levels of antioxidant enzymes were significantly increased at high concentrations of nanoparticles compared with the control. Thus, synthesized phytomediated ZnONPs from shoots of O. arabicus and their application to the same organ of O. arabicus in vitro were found to be effective as a low concentration of nanoparticles promoted shoot growth, resulting in high biomass accumulation. Thus, using green nanotechnology, such endemic plants could be conserved in vitro and multiple shoots could be produced by reducing the phytohormone concentration for multiple uses, such as the production of potential secondary metabolites.

Salinity stress affects growth and physiology of mulberry (Morus sp.)

Mulberry (Morus sp.) plant is used to feed silkworms, and the leaves contain compounds with medicinal properties of secondary metabolites. However, the content of these compounds tends to increase under stress conditions, for instance, salt stress. This study, therefore, aimed to determine the accessions of mulberry with tolerance for salt stress. The stem cuttings of seven accessions from 5 regions, Bogor, Pati, Situbondo, Bali, and Gowa, were planted following a factorial randomized block design with 3 replications. Subsequently, the first factor using the accessions, and the second factor using NaCl solution (0.0%, 0.2%, 0.3%, and 0.4% concentrations) were performed. The variables observed were growth (leaves number, plant height, and shoots number), photosynthesis rate, total chlorophyll, and proline content. The results showed that the M6 accession exhibited tolerance under high salt stress, based on the leaves number, plant height, shoot number, photosynthesis rate, and proline content. Furthermore, an increase in salt concentration was discovered to cause a decrease in growth, photosynthesis rate, and total chlorophyll content. Also, proline accumulation stimulated by high salt stress possibly plays an important role in salinity tolerance.

Micropropagation of Three Endemic Begonias Using Various Hormones Concentration and Culture Media Application

Three species of Begonias endemic to Java and Sumatra, namely Begonia leuserensis, Begonia atricha and Begonia scottii, were conserved in Cibodas Botanic Gardens as sources of germplasm for ornamental plant and/or medicines. However, the information on efficient hormones concentration and their culture media application through an in vitro propagation effort is still limited. Therefore, this study aimed to explain the growth response of three species of Begonias using various hormones concentrations and culture media through in vitro propagation. The culture media using Murashige & Skoog (MS) media that combinedwith 6-Benzyladenine (BA) dan Thidiazuron (TDZ) hormones in different concentrations i.e. 0.5 mg/L, 1 mg/L, 2 mg/L, and 3 mg/L. Observation parameter included shoot number, plantlets height, and leaves number. The data were analyzed using analysis of variance (ANOVA) with the F test at a 5% significance level. The results showed that three species of Begonias were observed to have different growth responses in the combination of MS+BA and MS+TDZ media. The combination of MS+TDZ media produces more shoots number, while the combination of MS+BA media influenced higher in leaves number. A concentration of 0.5 mg/L of hormone showed a good regeneration, therefore were recommended for in vitro propagation of Begonia species.

Protocol Optimization for Rapid Multiplication of Cassava (Manihot esculenta Crantz) using the Nodal Segment

Background: Cassava is a vital food security crop for millions of people, especially in sub-Saharan Africa. Since the crop produces a reasonable yield on marginal soils, it could help alleviate world hunger. Consequently, the increase in cassava production and its quality characteristics are significant. However, the low multiplication rate of this main crop has delayed the spread of improved varieties among farmers. As a result, tissue culture techniques can be a viable solution to overcome these challenges. Methods: The study used a nodal segment as an explant to evaluate different concentrations of BAP and NAA for an efficient, cost-effective in vitro mass multiplication of the AWC-1 cassava variety. CuSO4, commercial bleach and ethanol had used to sterilize nodal explants taken from greenhouse-grown plants at various time intervals. Result: The best medium for micro shoots induction had found to be medium without growth regulators. Among different treatments used for shoot multiplication purposes, the maximum shoot number has been recorded on M.S. medium supplemented with 0.75 mg/l BAP and 0.2 mg/l NAA. Medium with 0.5 mg/l NAA concentration was the best for rooting induction. A survival rate of 86% has obtained in the greenhouse and the plantlets appeared to be morphologically normal.

Germination and seedling growth response of mango (Mangifera Indica L.) cultivars to different nursery potting media

Background Mango is an important cash crop greatly contributing for food security of fruit growers in northwestern parts of Tigray region, northern Ethiopia. However, information was critically lacking with respect to germination and growth response of mango rootstock to varying compositions of growing media in the region. Thus, an experiment was conducted during 2018 to evaluate germination and seedling growth response of mango cultivars to different combinations of nursery potting media in a net house at Indasellassie. Seeds of Local, Dodo and Keitt cultivars of mango were sown in full top soil (M1); top soil: sawdust: sand (3:2:1)(M2); top soil: FYM (Farmyard manure): sand (3:2:1)(M3); and top soil: FYM: sawdust: (3:2:1)(M4). Randomized complete block design in a factorial arrangement with three replications were used. Data on germination and growth parameters were collected and analyzed using GenStat software. Results The findings of the experiment revealed that mango cultivar had significantly affected most of the germination and growth parameters except shoot number. Moreover, potting media affected most of the parameters except days to germination, germination percentage, stem diameter, root number, and shoot number. Interaction effects of cultivar and growing media significantly affected all parameters except internode length, shoot number and stem diameter. Accordingly, interactions of local mango cultivar and M3 media combination gave the highest for most of the parameters with the order of M3 = M1 = M4 > M2 for germination percentage, M3 = M1 > M4 = M2 for plant height, M3 > M1 > M2 = M4 for leaf number, M3 > M1 > M4 > M2 for leaf area, M3 = M1 > M2 = M4 for stem diameter, M3 = M1 > M2 > M4 for root number, M3 > M2 > M1 = M4 for root length, M3 > M1 > M2 = M4 for fresh weight, M3 > M1 = M2 = M4 for dry weight and Vigorosity II of mango. Conclusion Considering better mango performance on seed germination, seedling growth and establishment in the study area, mango growers should use Local mango cultivar with soil potting media combinations of top soil: FYM: sand in the ratio of 3:2:1 for improving productivity and food security.