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Article

Foliar Spraying of ZnO Nanoparticals on Curcuma longa Had Increased Growth, Yield, Expression of Curcuminoid Synthesis Genes, and Curcuminoid Accumulation

by
Salah Khattab
1,2,
Mayyadah Abdullah Alkuwayti
1,
Yun-Kiam Yap
1,
Ahmed M. A. Meligy
3,4,
Mohammad Bani Ismail
5 and
Fadia El Sherif
1,2,*
1
Department of Biological Sciences, College of Science, King Faisal University, Al Ahsa 31982, Saudi Arabia
2
Department of Horticulture, Faculty of Agriculture, Suez Canal University, Ismalia 41522, Egypt
3
Department of Clinical Science, Central Lab, College of Veterinary Medicine, King Faisal University, Al Ahsa 31982, Saudi Arabia
4
Department of Physiology, Agricultural Research Center (ARC), Giza 13611, Egypt
5
Department of Basic Medical Sciences, School of Medicine, Aqaba Medical Sciences University, Aqaba 77110, Jordan
*
Author to whom correspondence should be addressed.
Horticulturae 2023, 9(3), 355; https://doi.org/10.3390/horticulturae9030355
Submission received: 19 January 2023 / Revised: 22 February 2023 / Accepted: 6 March 2023 / Published: 8 March 2023
(This article belongs to the Section Plant Nutrition)
Browse Figures
Versions Notes

Abstract

:
ZnO nanoparticles (NPs) can be considered a highly efficient Zn source that has been widely used in agriculture to promote crop development and productivity. The turmeric (Curcuma longa) plant has several medical properties, and its rhizome is utilized as a spice in the food sector. In this work, C. longa leaves were sprayed with various concentrations of ZnO NPs to inspect their effect on growth, yield, and bioactive compound compositions. ZnO NPs considerably increased tumeric productivity, yield, and curcuminoid content versus the control treatment. The ZnO NPs concentration of 10 mgL−1 was found to be the optimum concentration for producing the highest C. longa yields, while the concentration of 40 mgL−1 promoted positive effects on photosynthetic pigments, bisdemethoxycurcumin, demethoxycurcumin, and curcumin contents. This demonstrates that ZnO nano-fertilizer promotes plant growth, yield characteristics, and curcuminoid component synthesis, and its application is therefore notably beneficial for progressive sustainable C. longa agriculture.

1. Introduction

Assessing the influence of nanotechnology on agriculture requires studying the interactions between nanoparticles (NPs), plant growth, and yield [1,2]. Zinc (Zn) is an essential micronutrient that helps plants endure disease, maintain cell membrane integrity, protein synthesis, and pollen development, and boosts the production of a variety of antioxidant enzymes in plant tissues as well as auxin, a vital growth hormone. Zn affects carbohydrate metabolism by influencing photosynthesis and sugar conversions. Zn is a component of several photosynthesis enzymes, notably ribulose 1, 5-biphosphate carboxylase, which has been discovered to catalyze the first stage in photosynthesis’s carbon dioxide fixation [3]. Zn is essential for plant growth, and insufficient levels of zinc can slow growth, diminish reproductive sites, and reduce yields in all crops due to stunted root and tissue growth [4,5]. ZnO NPs are a highly efficient Zn source with nanoparticle features that enable them to cross the plant cell membrane, transport substances into cells, and incorporate them into metabolic processes [6,7]. They were employed to boost plant productivity and yield in a variety of species [8,9,10]. Curcuma longa L. (turmeric) is a Zingiberaceae-family herbaceous perennial herb [11]. Its rhizome contains a variety of bioactive components such as curcuminoids (non-volatile oils) and mono- and sesquiterpenoids (volatile oils) and is regarded as the most important portion of C. longa plants [12]. C. longa has long been used as a spice and cuisine ingredient due to its characteristic yellow color, flavor, and potent antioxidant property [13]. Curcuminoids (curcumin, dimethoxy- and bisdemethoxycurcumin) are the most important bioactive elements of C. longa rhizome [14]. Curcumin has remarkable biological features such as antioxidant, anticancer, and neuroprotective capabilities [15,16]. Four type III polyketide synthase genes have been identified as being engaged in the curcuminoid production pathway: diketide-CoA synthase (DCS), curcumin synthase 1 (CURS1), curcumin synthase 2 (CURS2), and curcumin synthase 3 (CURS3) [17,18,19]
The objective of this study was to see how foliar spraying of ZnO NPs affected the physiological progress and active constituents of C. longa. Curcuminoid gene expression patterns in rhizomes were examined utilizing quantitative reverse transcriptase polymerase chain reaction to investigate the molecular mechanisms in response to ZnO NPs treatments. The importance of ZnO nano-fertilizer in the advancement of sustainable C. longa agriculture will be determined.

2. Materials and Methods

2.1. Plant Materials and Growth Conditions

Plant Resources and Growth Environmental Turmeric daughter rhizomes with one or two buds (40 g) [20] from Agriculture and Veterinary Research and Training Center, King Faisal University, Saudi Arabia (25.266223133116906, 49.69556316077168) were planted into sandy soil (Supplementary Table S1) on 1 April 2021 and cultured 40 × 40 cm apart in a greenhouse located at the same center mentioned above. The experiments were set up in a completely randomized block design with four (5, 10, 20, and 40 mgL−1) concentrations of ZnO NPs (catalogue no. 677450, with a particle size of 50 nm, Sigma-Aldrich (St. Louis, MO, USA)) as treatment groups and distilled water as a control. After soil-cultured the rhizomes for two months, the entire foliage of each plant were sprayed with around 50 mL of ZnO NPs solutions in the morning at two-month intervals. The control plants were given the identical quantity of deionized water as the experimental plants. All plants were irrigated with groundwater as needed. The soil and irrigation water component (Supplementary Tables S1 and S2) was determined according to [21]. After 240 days of growth, the entire plant was collected, and the plant height (cm), leaves, roots, and rhizomes number per plant (n), leaves, roots, and rhizomes dry weight per plant (g), and rhizome diameter (mm) were measured using ten random plants from each treatment.

2.2. Chemical Evaluation

2.2.1. Photosynthetic Pigment Measure

Four 240-day-old turmeric plants were chosen at random and the third-top fresh leaf of each plant was used to measure photosynthetic pigment composition. The quantities of chlorophyll a and b, as well as carotenoids, were separated with 80 percent acetone and measured according to [22].

2.2.2. Mineral Composition

On day 240 after planting, plant leaves from various treatments were dried for 48 h at 60 °C and degraded with Sulfuric acid [23]. The nitrogen content was determined using the modified micro-Kjeldahl technique [24]. Calorimetry was employed to assess phosphorus (P) levels [25], and atomic absorption flame photometry was utilized to evaluate potassium (K) as well as zinc (Zn) levels [26]. At the completion of the experiment, soil samples were gathered and analyzed. Soil and water studies were carried out in accordance with [27].

2.2.3. Analysis of C. longa Rhizomes Powder Compositions

According to [28,29], the near-infrared spectroscopy (NIRS) approach was used to evaluate fat, fiber, moisture, protein, starch, and ash% in C. longa rhizomes from three independently picked plants for each treatment (control, 5,10, 20, and 40 mgL−1 ZnO NPs). The average from three duplicates of near-infrared spectroscopy was calculated. Using fecal near-infrared spectroscopy in the 1105 to 2450 range, the model was calibrated.

2.3. GC/MS Analysis of Dried C. longa Rhizomes Ethanolic Extract

According to [18], the GC 1310-ISQ mass spectrometer (Austin, TX, USA) with a TG-35MS direct capillary column (30 m 0.25 mm 0.25 m film thickness) was used to analyze ethanolic extracts of air-dried C. longa rhizomes from three randomized picked plants from each treated group (control and 5, 10, 20 and 40 mgL−1 ZnO NP).

2.4. Determination of Curcumin, Bisdemethoxycurcumin, and Demethoxycurcumin Contents in Ethanolic Extracts of Dried C. longa Rhizome by High-Performance Liquid Chromatography (HPLC)

The contents of curcumin, bisdemethoxycurcumin, and demethoxycurcumin in air-dried C. longa rhizomes powder from three plants randomly chosen from each treatment (control and 5, 10, 20, and 40 mgL−1 ZnO NPs) were determined using a Waters 2690 Alliance HPLC system (Waters, Milford, MA, USA) equipped with a Waters 996 photodiode array detector (Waters, Milford, MA, USA) and a C18 Inertsil (4.6 mm 250 mm, and 5 m) column according to the methods described by [18].

2.5. Determined CURS1, -2, -3, and DCS Gene Expression Using Real-Time Reverse Transcriptase Polymerase Chain Reaction (Real-Time RT–PCR)

The transcript levels of curcuminoid genes (Supplementary Table S3) were determined using Real-Time RT-PCR in C. longa rhizomes from four 240-day-old plants chosen at random from each of the experimental groups according to the methods described by [18].

2.6. Statistical Analysis

With ten repetitions, the experiment used a completely randomized block design. ANOVA was employed to test the variance homogeneity and statistically examine the data using StatSoft’s [30] Statistica 6 software. At p = 0.05, Duncan’s test was employed to establish the significance of mean differences.

3. Results

3.1. The Impact of ZnO NPs on the Growth and Yield of Plants

Table 1 shows the means of plant development metrics. The result demonstrated that there were varied significantly in plant height, leaf and root counts, and leaf dry weight between ZnO NPs treatment groups and the control group. Among all treatments, 10 mgL−1 ZnO NPs produced the most root number, root length, and root and leaf dry weight, and this increase was significant when compared to the control and other ZnO NPs treatments. On the contrary, the optimum plant height and number of leaves were achieved with 40 mgL−1 ZnO NPs (Table 1).
In regards to yield, the ZnO NPS groups had more rhizomes number, rhizome dry weights, and rhizome diameters than the control, with a statistically significant difference. The 10 mgL−1 ZnO NPS treatment resulted in the optimum rhizomes number (26.14), rhizomes dry weight (27.1 g), and rhizomes diameter (22.7 mm). The control treatment, on the other hand, had the lowest number of rhizomes (16.67), rhizome dry weight (9.7 g), and rhizome diameter (8.0 mm) (Table 2).

3.2. Photosynthetic Pigments, Mineral Contents, and Rhizomes Powder Compositions

Table 3 shows that ZnO NPs treatments had a positive and significant effect on chlorophyll a and b and carotenoid concentration in contrast to the control group. The treatment with 40 mgL−1 ZnO NPs produced the highest values for all measured parameters.
Table 4 shows that ZnO NPS treatments reduced the nitrogen (N), phosphorus (P), and potassium (K) content of C. longa leaves as compared to control plants. ZnO NPs foliar application promotes zinc (Zn) accumulation in the leaves. Among different ZnO NPs concentrations and control treatments, the 40 mgL−1 ZnO NPs treatment resulted in the highest Zn levels in the C. longa leaves.
Data in Table 5 revealed that the rhizomes produced in the 5 mgL−1 ZnO NPs treatment had the highest fat percentage (3.9%), followed by the 40 mgL−1 ZnO NPs treatment (3.88%), and the control (3.74%), but the control treatment had the highest fiber content (10.02%). The moisture and starch percentages were higher in 5 mgL−1 ZnO NPs, while the ash percentage was determined to be between 3.19% (40 mgL−1 ZnO NPs) and 2.02% (control), and the protein content was higher in 20 mgL−1 ZnO NPs (11.08%).

3.3. GC-MS Analysis

GC-MS was utilized to evaluate ethanolic extracts of C. longa rhizomes from treatments with various ZnO NPs concentrations, as well as the control treatments. The composition (area %) of common compounds found in plants from all experimental groups are listed in Table 6. The composition (area %) of unique compounds found in each experimental group are listed in Table 7. Lastly, the composition (area %) of compounds differentially produced in two or more treatment groups but not in all treatments were listed in Table 8. The full list of all detected compounds by GC-MS in all experimental groups were provided in Supplementary Table S4.
Seven common compounds were found in varying levels in the extracts of all five experimental groups (control, 5-, 10-, 20-, and 40 mgL−1 ZnO NPs) (Table 6). They were Coumaran, 4-Hydroxy-3-methylacetophenone, alpha.-curcumene, ar-tumerone, beta-sesquiphellandrene, curlone, and vanillin. The comparison of the composition (area %) of these compounds indicated that foliar spraying with ZnO NPs at various concentrations significantly increased the levels of coumaran, 4-Hydroxy-3-methylacetophenone, and vanillin when compared to control treatments. Low level (5 mgL−1) of ZnO NPs had the greatest effect as it resulted in a 6.06-, 4.69-, and 7.52-fold increase in coumaran, 4-Hydroxy-3-methylacetophenone, and vanillin, respectively, as compared with the control. In addition, foliar spraying with 5 mgL−1 ZnO NPs also significantly enhanced the alpha.-curcumene and beta-sesquiphellandrene by 1.59- and 1.41-fold, respectively, as compared with control, even though the negative impact on the levels of these two compounds were observed when higher concentration (10-, 20-, and 40 mgL−1) of ZnO were applied. Ar-tumerone was the only common compound that was down-regulated in all the ZnO NPs (5-, 10-, 20-, and 40 mgL−1) treatment groups.
A few distinct chemicals were discovered in a specific treatment group (Table 7). Among them were the germacron (control), 4-propylguaiacol and benzene, (1,1-dimethylnonyl) (10 mgL−1 ZnO), benzene, 1,4-dimethyl-2-(2-methylpropyl)-, camphor, (+)-.alpha.-bisabolol, and palmitic acid ethyl ester (20 mgL−1 ZnO), and dicumene, palmitic acid methyl ester and palmitic acid (40 mgL−1 ZnO).
Table 8 indicated that eight compounds were detected only in the plants of ZnO NPs treatment groups but not in the control. Among the eight compounds, three (P-hydroxybenzaldehyde, cis, cis-linoleic acid, and humulane-1,6-dien-3-ol) were found in all the ZnO NPs treatments and the 5 mgL−1 ZnO NPs treatment had produced the highest quantities for all these three compounds. Three other compounds were found in only three out of the four ZnO NPs treatment groups. They were Cis oleic acid and Oleic acid, ethyl ester, which were found in 5, 10, and 20 mgL−1 ZnO NPs treatments, while pentadecanoic acid was only detected in 10-, 20-, and 40 mgL−1 ZnO NPs treatment. The remaining two compounds were differentially elicited in only two of the ZnO treatment groups. They were (-)-Zingiberene (5- and 10 mgL−1 ZnO NPs) and thymol (20- and 40 mgL−1 ZnO NPs). Two compounds, namely, caryophyllene and tumerone were found in the control and some of the ZnO NPs treatment groups.

3.4. HPLC Results

Using high-performance liquid chromatography (HPLC), the impact of various concentrations of ZnO NPs on the accumulation of bisdemethoxycurcumin, demethoxycurcumin, and curcumin in ethanolic extracts of C. longa rhizomes were measured. Table 9 findings revealed that all concentrations of ZnO NPs treatments enhanced significantly bisdemethoxycurcumin, demethoxycurcumin, and curcumin levels when compared to the control treatment. Foliar spraying with (5 and 40 mgL−1) ZnO NPs increased bisdemethoxycurcumin, demethoxycurcumin, and curcumin levels by approximately (2.79- and 2.85-), (2.65- and 2.94-), and (2.78- and 3.17-) fold, respectively, compared to the control treatment (Table 9 and Figure 1).

3.5. The Influence of ZnO Nanoparticles on the Expression of Curcuminoid Biosynthesis Genes

Curcuminoid gene expression levels in C. longa rhizomes treated with ZnO NPs and control treatments were determined using real-time PCR. The results demonstrated that foliar application of ZnO NPs increased the expression of curcuminoid genes as compared to the control treatment (Figure 2). The CURS1, -2, -3, and DCS genes were shown to be differently elevated by ZnO NPs treatments in this investigation. The results showed that when ZnO NPs were applied foliarly, the expression levels of the CURS1, -2, -3, and DCS genes were higher than the control treatment. The CURS1, -3, and DCS gene expression levels were greatest in the 40 mgL−1 ZnO NPs treatment as compared with the control and other (5, 10, and 20 mgL−1) ZnO NPs treatments. The highest increase in CHS2 expression was observed in the 20 mgL−1 ZnO NPs group.

4. Discussion

Nanotechnology has a definite role to play in revolutionizing agriculture and food production, with the possibility to adapt conventional agricultural operations, save fertilizer, and reduce environmental pollution [1,2]. The impact of foliar treating C. longa plants with varying concentrations of ZnO NPs on plant development, production, phytochemical composition, and patterns of curcuminoid gene expression were studied to improve the quality and quantity of C. longa plant growth and yield.
In comparison to the control treatment, foliar spraying with ZnO NPs boosted plant growth metrics, rhizome dry weight, rhizome yield, and photosynthetic pigments in C. longa plants. This could be due to the increased nutrient uptake efficiency linked with nanostructured formulated zinc oxide fertilizers, which promote nutrient uptake to plants while conserving nutrient resources [11,31,32]. This result was consistent with [9,33,34]. In our investigation, foliar spraying of ZnO NPs reduced N, P, and K accumulation on C. longa leaves while Zn deposited in the leaves [35] ZnO NPs may function as an efficient slow-release Zn source for plant metabolic reactions [36,37]. Foliar spraying of ZnO NPs improved the nutritional properties of C. longa rhizomes by increasing the fat, starch, and protein percentages. Similarly, employing ZnO NPs at low and high concentrations increased moisture and ash%. The same results were reported by [34,38,39]. Zinc is required for carbohydrate and protein metabolism due to its function in influencing plant sugar transfer through starch metabolism and as an important component of RNA polymerase, which is required for protein synthesis [40]. High levels of ZnO (1000 mgL−1–4000 mgL−1) nanoparticles have been implicated in the reduction of seed germination, seedling growth, or root growth in numerous plant species, such as corn, radish, cucumber, and ryegrass [41]. Due to the low concentration range (5–40 mgL−1), we did not observe any adverse effects on the plant growth and yield.
Due to the plethora of chemically varied metabolites found in higher plants, no single analysis method has yet been capable of detecting the entire molecular basis of these plants, particularly medicinal and aromatic species [20,42]. GC-MS and HPLC were utilized to detect unique chemical components in C. longa plants treated with varying doses of ZnO NPs, as well as control plants. The findings indicate that the ZnO NPs treatments boosted the bioactive components of C. longa’s ethanolic rhizome extract which, were identified using GC-MS. Coumaran, 4-Hydroxy-3-methylacetophenone, alpha.curcumene, beta.-sesquiphellandrene, curlone, vanillin, (-)-zingiberene, p-hydroxybenzaldehyde, caryophyllene, cis oleic acid, cis,cis-Linoleic acid, and humulane-1,6-dien-3-ol were enhanced by foliar spraying ZnO NPs. HPLC analysis of polyphenolic curcuminoids (bisdemethoxycurcumin, demethoxycurcumin, and curcumin) indicated that ZnO NPs treatments considerably increased these molecules as compared to the control, ZnO NPs have the highest concentrations of the three compounds, at 40 mgL−1, followed by 5 mgL−1. These compounds were previously discovered by GC-MS and HPLC analysis in the rhizome of C. longa and have potent therapeutic properties [13,14,19,43]. The effects of ZnO NPs on plants are caused by changes in the physical, chemical, and biological properties of the materials utilized as nano-fertilizers, as well as their catalytic properties. These changes have an effect on chemical and biological processes in plants that can produce oxidative stress and toxicity, while also boosting antioxidant mechanisms [44]. ZnO NPs have a higher transport potential and thus improved bioavailability and absorption, allowing them to interact with intracellular structures that promote ROS formation and rise in non-enzymatic antioxidant molecules such as phenolic compounds and flavonoids as a result of ROS buildup [45,46,47]. The use of ZnO NPS has been linked to higher concentrations of effective (active) chemicals in a number of plants [34,48,49]. RT-PCR amplification and HPLC analysis demonstrated a relationship between curcuminoid gene expression and curcumin synthesis in C. longa. The accumulation of essential secondary metabolic products has been linked to the production of coordinate genes in response to adequate elicitors in plants [19,50,51]. The bioactivities, such as antimicrobials, antioxidants, or anticancer activities of rhizome extracts from plants treated with various concentrations of ZnO nanoparticles, will be examined in the future.

5. Conclusions

This is the initial investigation of the influence of ZnO NPs on C. longa growth, yield, and chemical compounds in a greenhouse condition. In conclusion, ZnO-NPs is an effective elicitor for increasing C. longa growth, yield, nutritional quality, and biochemical continues. The result suggested that the effect of ZnO NPs is proportional to the concentration used. According to the findings of this study, 10 mgL−1 of ZnO NPs is deemed to be the optimum Zn concentration for producing higher C. longa yields, while 40 mgL−1 was best in promoting the increase of photosynthetic pigments, bisdemethoxycurcumin, demethoxycurcumin, and curcumin contents, which were associated with highest fold increase in the CURS1, -3, and DCS gene expression levels.

Supplementary Materials

The following supporting information can be downloaded at: https://www.mdpi.com/article/10.3390/horticulturae9030355/s1, Table S1: Physical and chemical properties of the experimental soil; Table S2: Chemical properties and compositions of the irrigation water; Table S3: Sequences of forward and reverse primers for real-time RT-PCR; Table S4: Components identified by GC-MS analysis in the C. longa ethanolic rhizome extracts from different zinc oxide nanoparticles (ZnO NPs) treatments.

Author Contributions

Conceptualization, F.E.S., M.B.I., M.A.A. and S.K.; methodology, A.M.A.M. GC MS analysis, F.E.S., M.A.A. and Y.-K.Y.; formal analysis, F.E.S. and S.K.; investigation, F.E.S., Y.-K.Y. and S.K.; visualization, F.E.S., S.K. and Y.-K.Y.; writing—original draft preparation, F.E.S. and S.K.; writing—review and editing, F.E.S. and M.B.I.; project administration, S.K.; funding acquisition, F.E.S. All authors have read and agreed to the published version of the manuscript.

Funding

This research was funded by Deanship of Scientific Research, King Faisal University, grant number grant 2653.

Data Availability Statement

Not applicable.

Acknowledgments

We would like to express our gratitude to the greenhouse staff at King Faisal University’s Agriculture and Veterinary Research and Training Centre for their invaluable assistance.

Conflicts of Interest

The authors declare there are no conflict of interest.

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Figure 1. HPLC chromatogram of organic extract of C. longa exposed to 40 mgL−1 ZnO NPs. The peaks for the different curcuminoid compounds [bisdemethoxycurcumin (Bi), dimethoxycurcumin (De), and curcumin (Cu) were shown.
Figure 1. HPLC chromatogram of organic extract of C. longa exposed to 40 mgL−1 ZnO NPs. The peaks for the different curcuminoid compounds [bisdemethoxycurcumin (Bi), dimethoxycurcumin (De), and curcumin (Cu) were shown.
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Horticulturae 09 00355 g002 550
Figure 2. Expression patterns of curcuminoid synthase genes, CURS1, CURS2, CURS3, and DCS, in C. longa rhizome from control and zinc oxide nanoparticles (ZnO NPs) (5, 10, 20, and 40 mgL−1) treated plant. The Actin gene was used as an internal reference gene.
Figure 2. Expression patterns of curcuminoid synthase genes, CURS1, CURS2, CURS3, and DCS, in C. longa rhizome from control and zinc oxide nanoparticles (ZnO NPs) (5, 10, 20, and 40 mgL−1) treated plant. The Actin gene was used as an internal reference gene.
Horticulturae 09 00355 g002
Table
Table 1. Zinc oxide nanoparticles (ZnO NPs) impact the plant height (cm), number (n) of leaves and roots, and dried weight (g) of leaves and roots of C. longa. Mean values followed by the same letter within a column are not significantly different at p = 0.05, as measured by Duncan’s test.
Table 1. Zinc oxide nanoparticles (ZnO NPs) impact the plant height (cm), number (n) of leaves and roots, and dried weight (g) of leaves and roots of C. longa. Mean values followed by the same letter within a column are not significantly different at p = 0.05, as measured by Duncan’s test.
ZnO NPs (mgL−1)Plant Height
(cm)		Number of Leaves (n)Number of Roots (n)Root Length (cm)Weight of the Dried Root (g)Weight of Dried Leaves (g)
Control120.0 d ±
0.605551		10.0 d ±
0.0516611		20.25 c ±
0.429101		12.25 b ±
0.00331		2.90 b ±
0.22073		23.53 d ±
0.208167
5159.7 b ±
0.50287		15.5 b ±
0.081666		36.7 b ±
0.507571		13.3 b ±
1.00525		2.33 b ±
0.52758		49.7 bc ±
0.568624
10177 ab ±
0.888194		13.3 c ±
0.886751		46.3 a ±
1.527525		22.7 a ±
1.527525		3.6 a ±
0.688878		88.73 a ±
1.113553
20148.7 c ±
0.74223		14.17 bc ±
0.57735		33.7 b ±
0.767453		11.7 b ±
0.763763		2.4 b ±
0.616276		34.2 c ±
0.556776
40187.3 a ±
00098		16 a ±
0.033223		39 b ±
0557439		16.7 ab ±
0.732051		2.93 b ±
0.960109		62.77 b ±
0.85049
Table
Table 2. Zinc oxide nanoparticles (ZnO NPs) impact the number (n) of rhizomes, rhizomes dried weight, and rhizome diameter (mm) of C. longa. Mean values followed by the same letter within a column are not significantly different at p = 0.05, as measured by Duncan’s test.
Table 2. Zinc oxide nanoparticles (ZnO NPs) impact the number (n) of rhizomes, rhizomes dried weight, and rhizome diameter (mm) of C. longa. Mean values followed by the same letter within a column are not significantly different at p = 0.05, as measured by Duncan’s test.
ZnO NPs (mgL−1)Number. of Rhizomes (n)Weight of Dried Rhizome (g) Diameters of Rhizomes (mm)
Control16.67 c ± 0.509259.7 c ± 1.931328.0 d ± 1.113553
519.7 b ± 0.13160116 b ± 1.25539814.3 b ± 1.123892
1026.14 a ± 0.61880227.1 a ± 1.0518322.7 a ± 0.923398
2018.37 b ± 0.64575111.07 b ± 1.874609.0 c ± 1.042874
4022.25 a ± 1.52752515.27 b ± 1.4110913.7 b ± 1.562861
Table
Table 3. Zinc oxide nanoparticles (ZnO NPs) impact the chlorophyll a (chl a), b (chl b), and carotenoid concentrations of C. longa leaves. Mean values followed by the same letter within a column are not significantly different at p = 0.05, as measured by Duncan’s test.
Table 3. Zinc oxide nanoparticles (ZnO NPs) impact the chlorophyll a (chl a), b (chl b), and carotenoid concentrations of C. longa leaves. Mean values followed by the same letter within a column are not significantly different at p = 0.05, as measured by Duncan’s test.
ZnO NPs
(mgL−1)		Chl a
(mg/100 g F.W.)		Chl b
(mg/100 g F.W.)		Carotenoids
(mg/100 g F.W.)
Control64.126 c ± 0.422089 29.003 c ± 0.0236970.0825 c ± 0.94639
584.483 b ± 1.1550538.860 b ± 0.1201872.8609 d ± 0.05155
1077.577 b ± 0.1212530.287 c ± 0.465488.733 c ± 0.28711
2083.129 b ± 0.8641936.4314 b ± 0.35553102.918 b ± 1.7836
40105.934 a ± 1.0502749.509 a ± 0.63638133.499 a ± 0.48326
Table
Table 4. Zinc oxide nanoparticles (ZnO NPs) impact the composition of Nitrogen (N) (g kg−1), Phosphorus (P) (mg kg−1), Potassium (K) (mg kg−1), and Zinc (Zn) (mg kg−1) in C. longa leaves. Mean values followed by the same letter within a column are not significantly different at p = 0.05, as measured by Duncan’s test.
Table 4. Zinc oxide nanoparticles (ZnO NPs) impact the composition of Nitrogen (N) (g kg−1), Phosphorus (P) (mg kg−1), Potassium (K) (mg kg−1), and Zinc (Zn) (mg kg−1) in C. longa leaves. Mean values followed by the same letter within a column are not significantly different at p = 0.05, as measured by Duncan’s test.
ZnO NPs (mgL−1)N (g kg−1)P (mg kg−1)K (mg kg−1)Zn (mg kg−1)
Control61.525 a ± 0.002645750.0375 a ± 0.00264611.01538 a ± 0.1891801520.03215 b ± 0.004709
539.75 a ± 0.0028284270.021 b ± 0.0028289.32245 b ± 1.5066324190.0417 b ± 0.009051
1040.35 a ± 0.0007071070.0235 b ± 0.0007079.9004 ab ± 0.2975505340.033375 b ± 0.003323
2051.3 a ± 0.02345070.0245 b ± 0.00070710.2673 ab ± 0.1374615580.0502 b ± 0.006223
4050.75 a ± 0.002121320.0255 b ± 0.0021219.45515 b ± 0.3049751550.0773 a ± 0.024466
Table
Table 5. Zinc oxide nanoparticles (ZnO NPs) impact fat, fiber, moisture, protein, starch, and ash percentage (%) in the C. longa rhizome. Mean values followed by the same letter within a column are not significantly different at p = 0.05, as measured by Duncan’s test.
Table 5. Zinc oxide nanoparticles (ZnO NPs) impact fat, fiber, moisture, protein, starch, and ash percentage (%) in the C. longa rhizome. Mean values followed by the same letter within a column are not significantly different at p = 0.05, as measured by Duncan’s test.
ZnO NPs (mgL−1)Fat (%)Fiber (%)Moisture (%)Protein (%)Starch (%)Ash (%)
Control3.74 b ± 0.11313710.02 a ± 1.56977714.43 c ± 0.2899144.02 e ± 0.68589424.21 cd ± 2.0011122.02 ab ± 1.19501
53.9 a ± 0.0212135.41 c ± 0.00707116.81 a ± 0.0070715.95 c ± 0.07071129.95 a ± 0.0707110.29 c ± 0.014142
103.48 b ± 0.0282847.32 b ± 0.02121314.88 b ± 0.0282845.05 d ± 0.07071126.39 b ± 0.0070711.61 bc ± 0.007071
203.05 c ± 0.0707118.42 ab ± 0.02828414.02 d ± 0.02828411.08 a ± 0.03535527.29 b ± 0.0212131.96 ab ± 0.007071
403.88 a ± 0.0282849.52 a ± 0.02828414.08 d ± 0.0353558.42 b ± 0.02828422.81 d ± 0.0070713.19 a ± 0.021213
Table
Table 6. Comparison of the compositions of common compounds in the ethanolic rhizome extracts from C. longa plants of various experimental groups. Ethanolic extracts from three plants were measured to obtain the average value as presented. Mean values followed by the same letter within a column are not significantly different at p = 0.05, as measured by Duncan’s test.
Table 6. Comparison of the compositions of common compounds in the ethanolic rhizome extracts from C. longa plants of various experimental groups. Ethanolic extracts from three plants were measured to obtain the average value as presented. Mean values followed by the same letter within a column are not significantly different at p = 0.05, as measured by Duncan’s test.
PhytochemicalMolecular FormulaComposition (Area %)
ControlZnO NPs
(5 mgL−1)		ZnO NPs
(10 mgL−1)		ZnO NPs
(20 mgL−1)		ZnO NPs
(40 mgL−1)
CoumaranC9H6O20.77 d4.67 a2.35 b1.76 c1.26 c
4-Hydroxy-3-methylacetophenoneC9H10O21.31 d6.15 a3.44 b3.24 b2.33 c
alpha.-CurcumeneC15H222.84 b4.53 a1.41 c0.8 d1.08 c
Ar-tumeroneC15H20O56.45 a36.11 c53.75 a51.36 ab46.65 b
beta.-SesquiphellandreneC15H243.02 b4.25 a1.33 c1.12 c1.19 c
CurloneC15H22O19.85 b9.28 c21.83 a22.60 a20.59 a
VanillinC8H8O30.25 c1.88 a1.15 a0.66 b0.81 b
Table
Table 7. Comparison of the compositions of unique compounds in the ethanolic rhizome extracts from C. longa plants of various experimental groups. Ethanolic extracts from three plants were measured to obtain the average value as presented. ND: not detected. Mean values followed by the same letter within a column are not significantly different at p = 0.05, as measured by Duncan’s test.
Table 7. Comparison of the compositions of unique compounds in the ethanolic rhizome extracts from C. longa plants of various experimental groups. Ethanolic extracts from three plants were measured to obtain the average value as presented. ND: not detected. Mean values followed by the same letter within a column are not significantly different at p = 0.05, as measured by Duncan’s test.
PhytochemicalMolecular FormulaComposition (Area %)
ControlZnO NPs
(5 mgL−1)		ZnO NPs
(10 mgL−1)		ZnO NPs
(20 mgL−1)		ZnO NPs
(40 mgL−1)
(-)-ZingibereneC15H24ND0.60 a0.32 aNDND
P-hydroxybenzaldehydeC7H6O2ND0.93 a0.43 b0.16 d0.29 c
CaryophylleneC15H240.09 c0.38 a0.16 bNDND
cis oleic acidC18H34O2ND1.02 a0.52 b0.61 bND
cis,cis-Linoleic acidC17H30O2ND1.83 a0.79 c0.95 b0.90 b
Humulane-1,6-dien-3-olC15H22OND3.57 a1.67 b1.58 b1.13 b
Oleic acid, ethyl esterC20H38O2ND0.17 b0.19 b0.28 aND
Pentadecanoic acidC15H30O2NDND2.56 b3.49 a2.06 b
ThymolC10H14ONDNDND0.16 b0.35 a
TumeroneC15H22O6.2 aNDNDND5.63 a
Table
Table 8. Comparison of the compositions of compounds detected only in some but not all of the ethanolic rhizome extracts from C. longa plants of various experimental groups. Ethanolic extracts from three plants were measured to obtain the average value as presented. ND: not detectable.
Table 8. Comparison of the compositions of compounds detected only in some but not all of the ethanolic rhizome extracts from C. longa plants of various experimental groups. Ethanolic extracts from three plants were measured to obtain the average value as presented. ND: not detectable.
PhytochemicalMolecular FormulaComposition (Area %)
ControlZnO NPs
(10 mgL−1)		ZnO NPs
(20 mgL−1)		ZnO NPs
(40 mgL−1)
4-propylguaiacolC10H14O2ND0.11NDND
Benzene, (1,1-dimethylnonyl)-C17H28ND2.45NDND
Benzene, 1,4-dimethyl-2-(2-methylpropyl)-C18H22NDND1.54ND
GermacronC15H22O2.56NDNDND
Palmitic acid methyl esterC17H34O2NDNDND1.26
Camphor C10H16ONDND1.34ND
(+)-.alpha.-Bisabolol C15H26ONDND1.26ND
Dicumene C18H24CrNDNDND1.25
Palmitic acid, ethyl esterC18H36O2NDND1.42ND
Palmitic acidC16H32O2NDNDND3.57
Table
Table 9. Effects of zinc oxide nanoparticles (ZnO NPs) treatments on bisdemethoxycurcumin, demethoxycurcumin, and curcumin (µg/mL) accumulation of C. longa. Mean values followed by the same letter within a column are not significantly different at p = 0.05, as measured by Duncan’s test.
Table 9. Effects of zinc oxide nanoparticles (ZnO NPs) treatments on bisdemethoxycurcumin, demethoxycurcumin, and curcumin (µg/mL) accumulation of C. longa. Mean values followed by the same letter within a column are not significantly different at p = 0.05, as measured by Duncan’s test.
ZnO NPs
(mgL−1)		Bisdemethoxycurcumin
(µg/mL)		Demethoxycurcumin
(µg/mL)		Curcumin
(µg/mL)
Control140.02985935 e ± 0.00019972.339401155 e ± 0.227121225.70146805 e ± 0.002076
5390.9309998 b ± 0.001414191.9427214 b ± 0.010293628.0079489 b ± 0.002901
10297.7843364 c ± 0.022152155.2782524 c ± 0.313598 502.7190404 c ± 0.001357
20235.8495247 d ± 0.043099129.44198 d ± 0.011342469.839536 d ± 0.000656
40398.4860058 a ± 0.019791212.6428497 a ± 0.000213715.05218605 a ± 0.003092
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Khattab, S.; Alkuwayti, M.A.; Yap, Y.-K.; Meligy, A.M.A.; Bani Ismail, M.; El Sherif, F. Foliar Spraying of ZnO Nanoparticals on Curcuma longa Had Increased Growth, Yield, Expression of Curcuminoid Synthesis Genes, and Curcuminoid Accumulation. Horticulturae 2023, 9, 355. https://doi.org/10.3390/horticulturae9030355

AMA Style

Khattab S, Alkuwayti MA, Yap Y-K, Meligy AMA, Bani Ismail M, El Sherif F. Foliar Spraying of ZnO Nanoparticals on Curcuma longa Had Increased Growth, Yield, Expression of Curcuminoid Synthesis Genes, and Curcuminoid Accumulation. Horticulturae. 2023; 9(3):355. https://doi.org/10.3390/horticulturae9030355

Chicago/Turabian Style

Khattab, Salah, Mayyadah Abdullah Alkuwayti, Yun-Kiam Yap, Ahmed M. A. Meligy, Mohammad Bani Ismail, and Fadia El Sherif. 2023. "Foliar Spraying of ZnO Nanoparticals on Curcuma longa Had Increased Growth, Yield, Expression of Curcuminoid Synthesis Genes, and Curcuminoid Accumulation" Horticulturae 9, no. 3: 355. https://doi.org/10.3390/horticulturae9030355

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