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Article

Determining Optimal Mutation Induction of Philodendron billietiae Using Gamma Radiation and In Vitro Tissue Culture Techniques

by
Rattana Khamrit
1 and
Nakorn Jongrungklang
2,3,*
1
Faculty of Science and Technology, Loei Rajabhat University, Loei 42000, Thailand
2
Department of Agronomy, Faculty of Agriculture, Khon Kaen University, Khon Kaen 40002, Thailand
3
Northeast Thailand Cane and Sugar Research Center, Faculty of Agriculture, Khon Kaen University, Khon Kaen 40002, Thailand
*
Author to whom correspondence should be addressed.
Horticulturae 2024, 10(11), 1164; https://doi.org/10.3390/horticulturae10111164
Submission received: 27 September 2024 / Revised: 28 October 2024 / Accepted: 31 October 2024 / Published: 1 November 2024
(This article belongs to the Section Propagation and Seeds)
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Versions Notes

Abstract

:
The Philodendron billietiae is a popular ornamental plant, with mutated varieties in high demand and commanding premium prices. This research aims to identify the optimal medium for propagating Philodendron billietiae and to use gamma radiation to induce mutations. Shoot tips of Philodendron billietiae were cultured on Murashige and Skoog (MS) medium supplemented with various concentrations of 6-benzyladenine (BA), both alone and in combination with naphthalene acetic acid (NAA), to induce shoot formation. Different concentrations of NAA were then tested for root induction. Additionally, mutation induction was investigated using gamma radiation from cesium-137 as the primary radioactive source, with doses of 0, 10, 20, 30, 40, and 50 Gray. MS medium supplemented with 1.5 mg L−1 BA achieved the highest shoot induction, resulting in more shoots and leaves per shoot. The optimal medium for root induction was MS medium supplemented with 0.5 mg L−1 NAA. In the mutation induction experiment, increasing radiation doses resulted in decreased survival rates, fewer new shoots, and reduced leaf width and length. Gamma radiation at doses between 20 and 40 Gray induced morphological changes in the leaves of Philodendron billietiae. These findings provide valuable guidelines for the genetic improvement of Philodendron billietiae to enhance its commercial value.

1. Introduction

Philodendron, the second largest genus in the Araceae family, comprises over 500 species native to tropical and subtropical regions of America and the West Indies [1]. These plants are a prominent part of the native flora due to their abundance, climbing habit, and large, showy leaves. Philodendrons exhibit a variety of forms, with some species being vines and others having short, non-climbing stems. Their diverse leaf characteristics and attractive forms make them popular ornamental plants. They grow well in both partial shade and shade, making them suitable for decorating buildings and interior spaces. Their leaves can be used in vase arrangements or floral bouquets for various occasions, and they are used for ornamental desk plants, hanging baskets, totems, and potted plants [2]. This foliage plant is resistant to environmental conditions and maintains its beauty for a long time. It is easier to care for than flowering plants, making it highly desirable in the market, especially in Thailand. Due to their appealing foliage and ability to thrive indoors, Philodendron species are among the most popular foliage plants in Thailand [3].
Philodendron billietiae is highly sought after due to its rarity, high price, and distinctive orange-yellow petioles with wavy, ridged leaf edges [4]. Traditional propagation methods, such as seed propagation and stem cuttings, yield a limited number of plants and are time-consuming, resulting in an insufficient supply to meet demand. Plant tissue culture techniques offer a method of plant propagation that can produce a large number of disease-free plants in a short time. Conventional methods of propagation are slow and inconsistent with market demand. Micropropagation is the optimal solution to provide a consistent supply of high-quality plant materials at a desirable pace [5].
Currently, consumers are seeking ornamental plants with unique aspects of beauty, shapes, and colors. Therefore, methods to induce mutations in Philodendron billietiae are necessary. Since natural mutations occur at a low rate, various methods have been employed to increase mutation rates, such as radiation-induced mutations [6,7,8,9]. Gamma-ray mutagenesis offers several advantages compared to chemical and other mutagenesis methods. Notable benefits include a high frequency of mutation induction across a broad spectrum, significant penetrance, and a well-defined mutagenic process [10].
Gamma radiation is a popular method for inducing mutations in plants [11] due to its simplicity, safety (no residual radiation), and ability to induce a high mutation rate. This radiation has been applied in numerous plant species to induce mutations, including Torenia baillonii [12], Chrysanthemum [13], Orchid [14,15], Tulipa gesneriana L. [16]), Acalypha hispida [17], and Philodendron scandens [8]. This technique can produce various plant forms, leaf characteristics, and shapes different from the original [18]. For instance, leaf-color chimera mutants exhibited higher stability than leaf-shape chimeras, with stability dependent on the chimera type and the mutation’s location within the cell layers of the shoot apical meristem [19]. Moreover, higher doses of 40–50 Gray stimulated shoot multiplication but stunted growth in Curcuma heyneana. A high frequency of leaf mutations was observed, including deformation, split apexes, narrowing, stiffness, wrinkling, twisting, and variegation [20]. In plant mutational breeding, the primary gamma radiation sources for inducing mutations are the radioactive isotopes cobalt-60 and cesium-137. In Philodendron billietiae, Maikaeo et al. (2024) [6] reported that gamma radiation from cobalt-60 induced only a single specific mutation, resulting in pale leaves.
To date, there are no reports on using plant tissue culture techniques for propagating Philodendron billietiae combined with using gamma radiation from cesium-137 as the primary radioactive source to induce mutations. This approach can increase the quantity and potentially produce new and interesting characteristics for consumers. However, the appropriate medium for inducing shoot and root formation and the suitable intensity of gamma radiation to induce mutations must be established first. Therefore, this study aimed to determine appropriate in vitro tissue culture techniques for the propagation of Philodendron billietiae and gamma radiation to induce mutations to obtain Philodendron billietiae plants with novel characteristics. This approach provides a guideline for using in vitro tissue culture techniques for propagation and the use of gamma radiation to enhance novel ornamental plants.

2. Materials and Methods

2.1. Studying the Appropriate Medium for Shoot Induction from Shoot Tips of Philodendron billietiae

Shoot tips were prepared by removing leaf sheaths until small buds were obtained. They were washed with dishwashing liquid for 5 min, then rinsed with tap water until the foam was gone. The explants were soaked in 70% ethanol for 5 min, then surface-sterilized with 20% Clorox solution containing 1–2 drops of Tween-20 for 10 min with agitation. The explants were then transferred to 15% Clorox solution with 1–2 drops of Tween-20 for 10 min. Finally, they were rinsed with sterile distilled water 5 times, for 5 min each. Shoot tips of 0.5 cm were cultured on MS (Murashige and Skoog) [21] medium supplemented with 6-benzyladenine (BA) at concentrations of 0.5, 1, 1.5, and 2 mg L−1 alone, and BA at concentrations of 0.5, 1, 1.5, and 2 mg L−1 in combination with 0.5 mg L−1 naphthalene acetic acid (NAA). A control treatment with 0 mg L−1 BA and 0 mg L−1 NAA was also included. All media were supplemented with 30 g L−1 sucrose and 1.8 g L−1 Gellan Gum, with pH adjusted to 5.8. Cultures were maintained in a growth room with 3000 lux light intensity, a 16 h photoperiod, and a temperature of 25 ± 2 °C for 12 weeks. The percentage of shoot formation, number of shoots per explant, and number of leaves were recorded.

2.2. Examining Suitable Media for Root Induction in Philodendron billietiae Shoots

Philodendron billietiae shoots approximately 0.5 cm in height were cultured on MS medium supplemented with NAA at concentrations of 0, 0.5, 1, 1.5, and 2 mg L−1. All media were supplemented with 30 g L−1 sucrose and 1.8 g L−1 Gellan Gum, with pH adjusted to 5.8. Cultures were maintained in a growth room with 3000 lux light intensity, a 16 h photoperiod, and a temperature of 25 ± 2 °C for 8 weeks. The percentage of root formation, number of roots per shoot, and root length were recorded.

2.3. Inducing Mutations Using Acute Gamma Irradiation

Philodendron billietiae plants were cultured on MS medium for 2 weeks. Plants of uniform size were then subjected to acute gamma irradiation at the Nuclear Technology Research Center, Kasetsart University, Bangkok, Thailand, using a MARK 1-30 gamma irradiator (Serial No. 1116) loaded with 4500 Curies of Cs-137, manufactured by J.L. Shepherd & Associates, San Fernando, CA, USA. The radiation doses used were 0, 10, 20, 30, 40, and 50 Gray. After irradiation, the plants were transferred to MS medium supplemented with 1.5 mg L−1 BA. Cultures were maintained in a growth room with 3000 lux light intensity, a 16 h photoperiod, and a temperature of 25 ± 2 °C for 22 weeks, with subculturing every 4 weeks. Survival rates were recorded, and the LD50 (lethal dose causing 50% mortality) or Critical Dose was calculated using regression analysis after 6 weeks of culture. The number of new shoots, leaf width, leaf length, and any abnormal characteristics were recorded after 16 weeks of culture. Subculturing of gamma-irradiated Philodendron billietiae shoots (0, 10, 20, 30, and 40 Gray) was performed four times at four-week intervals.

2.4. Statistical Analysis

The collected data were subjected to one-way analysis of variance according to a completely randomized design (CRD). Means were compared with Tukey test at a 95% confidence level. Statistical analysis was conducted using SPSS version 23.

3. Results

3.1. Determination of the Appropriate Medium for Shoot Induction of Philodendron billietiae

The culture of shoot tips on MS medium supplemented with BA at concentrations of 0.5, 1.0, 1.5, and 2.0 mg L−1 alone, and BA at concentrations of 0.5, 1.0, 1.5, and 2 mg L−1 in combination with NAA at 0.5 mg L−1, for 12 weeks, resulted in the enlargement of Philodendron billietiae shoot tips and a change in tissue color from white to green (Figure 1). Shoot formation was observed in MS medium supplemented with plant growth regulators, with shoot induction rates varying from 40% to 100% across formulations (Table 1). The MS medium supplemented with 1.5 mg L−1 BA induced the highest shoot formation rate at 100%, with an average of 3.00 shoots per explant and an average of 3.37 leaves per shoot. Moreover, BA at a concentration of 1.5 mg L−1, combined with 0.5 mg L−1 NAA, resulted in a 100% shoot formation rate, with an average of 2.00 shoots per explant and 2.82 leaves per shoot. In contrast, MS medium without plant growth regulators resulted in shoot tips turning brown and failing to develop into shoots.

3.2. Determination of Suitable Media for Root Induction in Philodendron billietiae Shoots

Root induction of Philodendron billietiae from shoots varied with NAA doses (Figure 2). Philodendron billietiae shoots cultured on medium with 0.5 mg L−1 NAA demonstrated the best rooting performance, achieving a 100% rooting induction percentage, an average of 5.70 roots per shoot, and an average root length of 2.53 cm. Shoots cultured with 1.0 mg L−1 NAA also achieved 100% rooting induction, but produced an average of 3.70 roots per shoot and a root length of 2.41 cm. In contrast, shoots without NAA (0 mg L−1) showed a 70% rooting induction percentage, with an average of 0.8 roots per shoot but a longer average root length of 3.39 cm (Table 2).

3.3. Inducing Mutations Using Acute Gamma Irradiation

The survival rate of Philodendron billietiae shoots was assessed six weeks after gamma irradiation. As the gamma radiation dose increased, the number of surviving shoots decreased. Shoots irradiated with 10, 20, 30, and 40 Gray in particular had survival rates of 96.67%, 80.00%, 53.33%, and 26.67%, respectively, compared to a 100% survival rate in the non-irradiated control group. No shoots survived at the 50 Gray dose (Figure 3 and Table 3). The relationship between radiation dose and survival rate was analyzed to determine the LD50 value after 6 weeks using regression analysis. The equation y = −2.371x + 112.06 (R2 = 0.816) was derived, indicating that a dose of 26.17 Gray resulted in 50% mortality of Philodendron billietiae shoots (Figure 4).
There were no significant differences in the number of new shoots and leaf width among the experimental groups exposed to 0–20 Gray. Additionally, the leaf length did not significantly differ among the groups exposed to 0–10 Gray. However, across the 0–50 Gray range, these values varied (p < 0.05) from those of the irradiated plants. As the radiation dose increased, new shoot formation, leaf width, and leaf length all decreased (Table 4).
Morphological changes in the irradiated Philodendron billietiae plants were observed, with new shoots exhibiting three distinct leaf characteristics: (1) small, light green leaves, (2) small, pale-yellow leaves, and (3) small, yellow-variegated green leaves. Plants irradiated with 10 Gray produced 8 new shoots with small, light green leaves. Plants exposed to 20 Gray of radiation produced two new plants with small, pale-yellow leaves. At 30 Gray, two new plants exhibited small, pale-yellow leaves, while two others displayed yellow-green variegated leaves. At 40 Gray, one new plant had small, pale-yellow leaves, and another showed yellow-green variegated leaves (Figure 5).

4. Discussion

4.1. Shoot Induction of Philodendron billietiae

BA, a cytokinin-type plant growth regulator, effectively stimulates cell division in various plant parts [22]. Cytokinin influence protein synthesis, facilitating rapid cell division, starting from the G1 phase of the cell cycle, the preparatory phase for DNA synthesis, progressing to the DNA synthesis phase (S-phase), where synthesized proteins and enzymes are utilized in mitosis. This process shortens the G2 phase, accelerating entry into mitosis [23]. The study also found that combining BA with NAA did not increase shoot and leaf numbers compared to using BA alone (Figure 1 and Table 1). This may be due to hormonal imbalance, leading to the inhibition of shoot formation, leaf development, growth, and plant development [24,25,26].
Our findings align with previous studies that utilized cytokinin alone. For example, Han and Park [27] reported that shoot tips of Philodendron cannifolium cultured on MS medium with 3 mg L−1 6-benzylaminopurine (BAP) produced a maximum of 2.0 shoots per explant. Similarly, Klanrit et al. [3] found that protocorm-like bodies of Philodendron erubescens ‘Pink Princess’ cultured on MS medium with 1 mg L−1 BAP generated up to 7.7 shoots per explant and a maximum of 4.1 leaves. In addition, Chen et al. [28] investigated the propagation of three Philodendron cultivars using nodal segments cultured on MS medium with cytokinins (TDZ, kinetin, and BA) at concentrations of 0.5 and 1 mg L−1. Their findings indicated that ‘Imperial Green’ achieved the highest shoot formation percentage (88.8%) with 0.5 mg L−1 kinetin, ‘Imperial Red’ exhibited the highest (80.6%) with 0.5 mg L−1 BA, and ‘Imperial Rainbow’ showed the highest (63.9%) with 1 mg L−1 kinetin. However, this study agrees with Alawaadh et al. [29], who reported that shoot tips cultured on MS medium with 1 mg L−1 BAP and 0.5 mg L−1 IBA produced up to 10.9 shoots per explant, demonstrating the effectiveness of combining auxins with cytokinins for shoot induction in Philodendron bipinnatifidum. Therefore, various Philodendron species can be effectively propagated in vitro using MS medium supplemented with BA, either alone or in combination with auxins. The optimal concentration of plant growth regulators in Philodendron tissue culture can vary significantly depending on factors such as species, cultivar, and explant type used [5,30,31,32,33].

4.2. Root Induction in Philodendron billietiae Shoots

NAA, an auxin, promotes protein and nucleic acid synthesis, cell division, and root induction [34]. The optimal auxin concentration in tissue culture media varies by Philodendron species and cultivar. For instance, Alawaadh et al. [29] reported that MS medium supplemented with 2 mg/L NAA induced 100% rooting and an average of 13.1 roots per shoot in Philodendron bipinnatifidum Schott ex Endl. Similarly, Han and Park [27] found that 0.5 mg L−1 NAA produced a maximum of 11.7 roots per shoot, with the highest root length measuring 4.4 cm.
In this study, Philodendron billietiae shoots cultured on MS medium without plant growth regulators produced the longest roots (Table 2). Increasing NAA concentrations led to a decrease in both the number and length of roots, likely due to the inhibitory effects of excessive exogenous NAA on root formation and growth. These findings agreed with research on Cryptocoryne wendtii by Klaocheed et al. [35], Tupistra albiflora by Palee [36], and Chrysanthemum morifolium by Chae [37]. MS medium containing 1 mg L−1 NAA induced 100% root formation in Tupistra albiflora, producing an average of 6.6 roots per explant [36]. The highest rooting success was achieved with 1.0 mg L−1 NAA, resulting in 100% rooting, an average of 36 roots per plantlet, and an average root length of 26.02 mm for Cryptocoryne wendtii [35]. In Chrysanthemum morifolium, the greatest root length of 36.2 ± 3.3 mm was achieved using SH medium with mg L−1 IAA, while fewer roots and shorter root lengths were observed with NAA [37].

4.3. Inducing Mutations Using Acute Gamma Irradiation

Radiation causes ionization of atoms within cells, generating reactive chemical radicals that induce chemical changes. These changes can damage cellular functions, leading to cell destruction or abnormal cell division. Radiation can damage meristematic tissues [38], and severe exposure may lead to plant death [39]. In addition to DNA-level changes, radiation can also alter chromosome number or structure [10]. Chromosomal breakage and subsequent rejoining may result in deletions, additions, inversions, or translocations, potentially causing mutations. The extent of these chromosomal changes can significantly impact gene expression and regulation, depending on the types and numbers of genes involved, as well as their functional roles within the genome [40]. In this study, the number of surviving shoots declined as the gamma radiation dose increased. A high survival rate was noted at a dose of 20 Gray (Figure 3 and Table 3). Our study agreed with El-Khateeb et al. [8] who reported that gamma radiation-induced mutations in Philodendron scandens at doses of 0, 0.5, 2, 4, and 8 Krad resulted in decreased survival rates with increasing radiation doses. Similarly, Li et al. [16] found that tulip (Tulipa gesneriana L.) bulbs irradiated with gamma rays at 10, 20, 40, 60, and 100 Gray showed decreased survival rates with increasing doses. Cytological studies have shown that radiation slows down mitotic cell division, causes chromosomal aberrations, and reduces differentiation capacity, ultimately leading to plant death in gamma-irradiated specimens [41].
As the radiation dose increased, new shoot formation, leaf width, and leaf length decreased (Table 4). Radiation affects plant growth and development [42]. Low doses of radiation can stimulate plant growth and development, while higher doses can cause damage. Yamaguchi et al. [43] found that chrysanthemum ‘Taihei’ callus exposed to gamma radiation doses of 15–60 Gray showed decreased shoot formation with increasing radiation doses. Leaf variegation occurs due to uneven chlorophyll distribution caused by radiation exposure, resulting in chimeras and alterations in leaf coloration [12]. These findings align with those of El-Khateeb et al. [8], who reported that increasing gamma radiation levels led to decreased plant height, stem diameter, and leaf count. The most significant morphological changes were observed at 8 Krad. Similarly, Pallavi et al. [7] found that Zinnia elegans var. (Dreamland) exposed to high radiation doses (75, 100, and 125 Gray) exhibited reduced germination, growth, and alterations in morphological characteristics such as plant height, flower count, and flower diameter compared to control plants. Plants exposed to radiation levels between 20 and 40 Gray produced one to two new plants with small, pale-yellow leaves, particularly yellow-green leaves, which are favored by customers (Figure 5). Previous studies reported that gamma irradiation mutagenesis using 60Co in Philodendron billietiae induced low mutation rates, resulting in new pale-yellow leaves without any yellow-green vegetative leaves [6]. However, utilizing cesium-137 as the primary source of gamma radiation for inducing mutations may achieve the characteristics desired by consumers. At elevated gamma radiation doses from cesium-137, certain irradiated ginger plants displayed yellowish-green leaves and malformed leaflets. These phenotypic alterations reflect qualitative genetic changes that arise within major or oligogenic regions of the genome [44]. Cesium-137 emits gamma rays with a relatively lower average energy of 0.662 MeV, resulting in reduced penetration depth [10]. This shallower penetration may be advantageous for experiments focusing on specific tissue layers or smaller samples, as it allows for more localized mutation induction, and may also be preferable for generating subtle or targeted mutations, particularly in surface tissues. In contrast, Cobalt-60 (60Co), with its higher gamma-ray energy, is suited for applications requiring greater penetration and broad mutational effects [10].
In previous report, African violets subjected to acute gamma radiation showed changes in leaf color, flower color, leaf size, and flower size compared to the control group [45]. Abdullah et al. [44] observed that Zingiber officinale Roscoe exposed to varying levels of gamma radiation exhibited distinct leaf color changes compared to non-irradiated controls. The effects included stunted growth, altered plant stature, leaf deformation, and chlorophyll mutations. Notable leaf characteristics included incomplete formation, variegation, twisted midribs, and detachment from the midrib. Some leaflets were rudimentary, displaying rough surfaces and uneven lamina and margins. At high doses, certain irradiated ginger plants showed yellowish-green coloration and irregular leaflet forms. Phenotypic mutations can be classified as macro or micro mutations; the observed leaf characteristics were macro mutations, easily detectable in individual plants and morphologically distinct. These qualitatively inherited genetic changes likely occurred in major genes or oligogenes [44]. The leaf characteristics may result from actual mutations, such as chromosomal alterations, or from genes with pleiotropic effects, where a single gene influences multiple unrelated phenotypic traits [44].

5. Conclusions

This is the first report to identify the optimal mutation induction of Philodendron billietiae using gamma radiation from cesium-137 as the primary sources and in vitro tissue culture techniques. The optimal medium for shoot induction from Philodendron billietiae shoot tips was MS medium supplemented with 1.5 mg L−1 BA, while the most suitable medium for root induction was MS supplemented with 0.5 mg L−1 NAA. The LD50 of gamma radiation for Philodendron billietiae was determined to be 26.17 Gray. Increasing radiation doses resulted in decreased new shoot formation, leaf width, and leaf length. Using cesium-137 as the primary source of gamma radiation for inducing mutations can produce the characteristics desired by consumers. Gamma radiation doses between 20 and 40 Gray induced morphological changes in the leaves of Philodendron billietiae, resulting in yellow-green leaves, favored by customers. These findings can serve as guidelines for breeding programs, utilizing gamma radiation-induced mutations to enhance the value and diversity of this ornamental plant.

Author Contributions

Conceptualization, R.K. and N.J.; methodology, R.K.; software, R.K.; validation, R.K. and N.J.; formal analysis, R.K.; investigation, R.K.; resources, R.K.; data curation, R.K.; writing—original draft preparation, R.K. and N.J.; writing—review and editing, R.K. and N.J.; visualization, R.K. and N.J.; supervision, R.K. and N.J.; project administration, R.K.; funding acquisition, R.K. All authors have read and agreed to the published version of the manuscript.

Funding

This research was funded by the Thailand Science Research and Innovation for their financial support (Funding number: 181398).

Data Availability Statement

The data presented in this study are available within the article.

Acknowledgments

Grateful acknowledgment to the Thailand Science Research and Innovation for their financial support. The acknowledgment is extended to the Faculty of Science and Technology, Loei Rajabhat University, for providing laboratory facilities and research equipment. Assistance was also received from Chaichat Boonyanusith, at the Faculty of Science and Technology, Nakhon Ratchasima Rajabhat University, for his expertise in regression analysis of LD50.

Conflicts of Interest

The authors declare no conflicts of interest.

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Horticulturae 10 01164 g001
Figure 1. Growth of Philodendron billietiae plantlets cultured on MS medium supplemented with varying concentrations of BA and NAA (with BA at concentrations of 0 (a), 0.5 (b), 1 (c), 1.5 (d), and 2 (e) mg L−1 alone, and NAA at concentrations of 0.5 mg L−1 in combination with, 0.5 (f), 1 (g), 1.5 (h), and 2 (i) mg L−1 BA) after 12 weeks of culture.
Figure 1. Growth of Philodendron billietiae plantlets cultured on MS medium supplemented with varying concentrations of BA and NAA (with BA at concentrations of 0 (a), 0.5 (b), 1 (c), 1.5 (d), and 2 (e) mg L−1 alone, and NAA at concentrations of 0.5 mg L−1 in combination with, 0.5 (f), 1 (g), 1.5 (h), and 2 (i) mg L−1 BA) after 12 weeks of culture.
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Figure 2. Root characteristics of Philodendron billietiae plantlets cultured on solid MS medium supplemented with varying concentrations of NAA (with NAA at concentrations of 0 (a), 0.5 (b), 1 (c), 1.5 (d), and 2 (e) mg L−1) after 8 weeks of culture.
Figure 2. Root characteristics of Philodendron billietiae plantlets cultured on solid MS medium supplemented with varying concentrations of NAA (with NAA at concentrations of 0 (a), 0.5 (b), 1 (c), 1.5 (d), and 2 (e) mg L−1) after 8 weeks of culture.
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Figure 3. Characteristics of Philodendron billietiae plants exposed to radiation doses of 0 (a), 10 (b), 20 (c), 30 (d), 40 (e), and 50 (f) Gray, cultured on MS medium supplemented with 1.5 mg L−1 BA for 6 weeks.
Figure 3. Characteristics of Philodendron billietiae plants exposed to radiation doses of 0 (a), 10 (b), 20 (c), 30 (d), 40 (e), and 50 (f) Gray, cultured on MS medium supplemented with 1.5 mg L−1 BA for 6 weeks.
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Figure 4. Relationship between gamma radiation dosage and the survival percentage of Philodendron billietiae plants after 6 weeks of exposure to gamma radiation.
Figure 4. Relationship between gamma radiation dosage and the survival percentage of Philodendron billietiae plants after 6 weeks of exposure to gamma radiation.
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Figure 5. Characteristics of Philodendron billietiae plants exposed to radiation doses of 0 (a), 10 (b), 20 (c), 30 (d), and 40 (e) Gray, cultured on MS medium supplemented with 1.5 mg L−1 BA for 22 weeks.
Figure 5. Characteristics of Philodendron billietiae plants exposed to radiation doses of 0 (a), 10 (b), 20 (c), 30 (d), and 40 (e) Gray, cultured on MS medium supplemented with 1.5 mg L−1 BA for 22 weeks.
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Table 1. Percentage of shoot regeneration, number of shoots per explant, and number of leaves per shoot of Philodendron billietiae obtained from the culture of shoot tips on MS medium supplemented with varying concentrations of BA and NAA after 12 weeks of culture.
Table 1. Percentage of shoot regeneration, number of shoots per explant, and number of leaves per shoot of Philodendron billietiae obtained from the culture of shoot tips on MS medium supplemented with varying concentrations of BA and NAA after 12 weeks of culture.
BA
(mg L−1)		NAA
(mg L−1)		Shoot Generation Percentage (%)Number of Shoots per Explant ± SENumber of Leaves per Shoot ± SE
0000.00 ± 0.00 e0.00 ± 0.00 e
0.50400.30 ± 0.15 de0.30 ± 0.15 de
1.00700.90 ± 0.17 bcd1.10 ± 0.19 cd
1.501003.00 ± 0.06 a3.37 ± 0.02 a
2.00700.70 ± 0.15 cd0.70 ± 0.15 cd
0.50.5700.90 ± 0.18 bcd1.50 ± 0.23 bc
1.00.5800.80 ± 0.13 bcd1.00 ± 0.16 cd
1.50.51002.00 ± 0.05 ab2.82 ± 0.02 ab
2.00.5801.70 ± 0.19 abc1.32 ± 0.18 bc
Same letters in the same column indicate non-significant differences at the level of α = 0.0595% confidence level, as determined by the Tukey test.
Table 2. Percentage of root regeneration, number of roots per plant, and root length of Philodendron billietiae obtained from the culture of plantlets on MS medium supplemented with varying concentrations of NAA after 8 weeks of culture.
Table 2. Percentage of root regeneration, number of roots per plant, and root length of Philodendron billietiae obtained from the culture of plantlets on MS medium supplemented with varying concentrations of NAA after 8 weeks of culture.
NAA
(mg L−1)		Percentage of Root Regeneration (%)Number of Roots per Plant ± SERoot Length (cm) ± SE
0700.80 ± 0.19 c3.39 ± 0.22 a
0.51005.70 ± 0.11 a2.53 ± 0.05 ab
1.01003.70 ± 0.16 ab2.41 ± 0.09 ab
1.5803.00 ± 0.30 bc1.60 ± 0.07 b
2.0802.50 ± 0.25 bc1.45 ± 0.06 b
Same letters in the same column indicate non-significant differences at the level of α = 0.0595% confidence level, as determined by the Tukey test.
Table 3. Survival rates of Philodendron billietiae plants following gamma radiation exposure for 6 weeks.
Table 3. Survival rates of Philodendron billietiae plants following gamma radiation exposure for 6 weeks.
Gamma Radiation (Gray)Survival Rates Percentage (%) ± SE
0100.00 ± 0.00 a
1096.67 ± 3.33 a
2086.67 ± 5.44 a
3026.67 ± 8.31 b
406.67 ± 4.44 c
500.00 ± 0.00 c
Same letters in the same column indicate non-significant differences at the level of α = 0.0595% confidence level, as determined by the Tukey test.
Table 4. Number of new shoots, leaf width, and leaf length of Philodendron billietiae plants after acute gamma radiation exposure for 22 weeks.
Table 4. Number of new shoots, leaf width, and leaf length of Philodendron billietiae plants after acute gamma radiation exposure for 22 weeks.
Gamma Radiation (Gray)New Shoot Number ± SELeaf Width (cm) ± SELeaf Length (cm) ± SE
03.11 ± 0.03 a0.85 ± 0.01 a1.17 ± 0.05 a
102.73 ± 0.05 ab0.64 ± 0.02 a0.81 ± 0.02 ab
202.05 ± 0.07 ab0.56 ± 0.02 ab0.67 ± 0.03 b
301.73 ± 0.16 b0.35 ± 0.06 bc0.54 ± 0.08 bc
400.35 ± 0.18 c0.27 ± 0.09 c0.31 ± 0.10 c
Same letters in the same column indicate non-significant differences at the level of α = 0.0595% confidence level, as determined by the Tukey test.
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Khamrit, R.; Jongrungklang, N. Determining Optimal Mutation Induction of Philodendron billietiae Using Gamma Radiation and In Vitro Tissue Culture Techniques. Horticulturae 2024, 10, 1164. https://doi.org/10.3390/horticulturae10111164

AMA Style

Khamrit R, Jongrungklang N. Determining Optimal Mutation Induction of Philodendron billietiae Using Gamma Radiation and In Vitro Tissue Culture Techniques. Horticulturae. 2024; 10(11):1164. https://doi.org/10.3390/horticulturae10111164

Chicago/Turabian Style

Khamrit, Rattana, and Nakorn Jongrungklang. 2024. "Determining Optimal Mutation Induction of Philodendron billietiae Using Gamma Radiation and In Vitro Tissue Culture Techniques" Horticulturae 10, no. 11: 1164. https://doi.org/10.3390/horticulturae10111164

APA Style

Khamrit, R., & Jongrungklang, N. (2024). Determining Optimal Mutation Induction of Philodendron billietiae Using Gamma Radiation and In Vitro Tissue Culture Techniques. Horticulturae, 10(11), 1164. https://doi.org/10.3390/horticulturae10111164

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