Startseite Composition and diurnal variation of floral scent emission in Rosa rugosa Thunb. and Tulipa gesneriana L.
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Composition and diurnal variation of floral scent emission in Rosa rugosa Thunb. and Tulipa gesneriana L.

  • Lu Yang EMAIL logo , Xiang Liao , Ping Cheng , Zhi-Gang Zhang und Hong Li EMAIL logo
Veröffentlicht/Copyright: 14. August 2020

Abstract

This study was aimed to explore the composition and diurnal variation analyses of floral scent emission from Rosa rugosa Thunb. and Tulipa gesneriana L. The floral scent from the fresh flower were collected at different time points (9:00, 12:00, 15:00, 18:00, and 21:00) using dynamic headspace collection and were analyzed using autothermal desorber-gas chromatography/mass spectrometry (ATD-GC/MS). The results showed that a total of 62 volatile flavor compounds were detected from Rosa rugosa Thunb and a total of 70 volatile flavor compounds were detected from Tulipa gesneriana L. They were identified with eight functional categories: alcohols, fatty hydrocarbons, terpenes, aldehydes, ketones, esters, and other substances. The total release amount first decreased, and then increased with time, and arrived at the lowest at 15:00. The release amounts of different categories present distinct change patterns. Among the components, phenylethyl alcohol, citronellol, methylene chloride, hexane, and acetone showed relatively higher release amounts and were thought as the main components in floral scent of Rosa rugosa Thunb. Alpha-Farnesene, ethanol, pentadecane, beta-ocimene, longifolene, caryophyllene, and acetone showed relatively higher release amounts and were thought as the main components in floral scent of Tulipa gesneriana L. Research of roses and tulips in aromatic in the garden provides a theoretical basis and research and improvement of the aroma components of aroma.

1 Introduction

The release of flower fragrance is an important characteristic of flowering plants, and it is also the main ornamental character. It is an important factor to form and influence the ornamental value of flowers. Floral scent is widely used for perfumes, cosmetics, flavorings, and therapeutic applications [1,2]. Floral scent is a composite character determined by a complex mixture of low-molecular-weight volatile molecules [3,4,5,6] and is one of the main factors that make up and influence the ornamental value of flowers [7]. As an important part of plant volatile compounds, the floral scent plays a key role in plant ecophysiology and represents a decisive communication channel between plants and animals [8,9]. To achieve this, the floral scent is often developmentally and rhythmically regulated to be associated with the activity of corresponding pollinators. As insect pollinator exhibit rhythmic activities, the flowers emit scent on specific timing in a day. Light directly affects floral scent emission. Diurnal variations of floral scent emission have been observed in numerous plant species [10]. The release of flower scent of Prunus davidiana [11], Gardenia jasminoides from grandiflora [12], Chrysanthemum, and Siberia kirilowii [13] showed obvious diurnal variations.

With the development of society, aromatherapy has been gradually moving to people’s lives, and the establishment of the aromatic botanical garden, night garden, and blind botanical garden and so on, with the characteristics of plant aroma, has the health care effect of green space. Rose and tulip with high ornamental value have attracted more and more attention because of their unique fragrance and curative effect widely used in landscaping. Experiments proved that aromatic plant volatiles can alleviate depression, anti-inflammatory and analgesic activities, and regulate the human nervous system, and it has a health function on the human body [14,15]. Roses are the most important crop in the floriculture industry and used as cut flowers, pot plants, and garden ornamentals. They are also a source of natural fragrances. Rose is used in producing rose water, attar of rose, and essential oils in the perfume industry. Tulip is world-famous flowers with bright colors, unique shape, and delightful scent. It is called “the queen of flowers”. Tulip bulbs and roots can be used as a medicine, have a sedative effect, and can be used in the treatment of dirty mania.

The collection and adsorption of plant volatiles or flower fragrance are one of the key links in the identification of flower fragrance components. Automated thermal desorption (ATD), also known as thermal desorption, is based on the design principle of dynamic headspace. It completes the whole process of sampling, enrichment, concentration, and injection by thermal desorption instruments. It is a more comprehensive sample of pretreatment technology. Because the thermal desorption method is more direct and close to nature in the collection of volatile components, the biggest advantage of the thermal desorption method is that it can directly collect the volatiles of living plants and fully retain the original state of the samples. The sensitivity of this technology is high, and the detection limit can reach 10−9 levels [16,17,18]. ATD-gas chromatography/mass spectrometry (GC/MS) is a kind of experimental techniques that can be used to collect, absorb, and analyze the volatile components of living plants [19]. It can effectively eliminate the interference of external volatile components and reflect the components and their release amount of flower fragrance more truly. It is suitable for the qualitative and quantitative analyses of plant volatile components in the near-natural state [20]. Therefore, the study takes the living rose and tulip flowers as materials, collects, and analyzes the chemical components of the volatiles, summarizes the main aromatic characteristics of the rose and tulip, explores the significance of landscape value, and provides a clear reference for rational utilization.

Therefore, in this study, rose and tulip are used as experimental materials. These were collected using dynamic headspace collection method and ATD-GC/MS analysis technique combines the research and analysis of the diurnal variation of the aroma for further development and application research of roses and tulips in aromatic in the garden and provides a theoretical basis and research and improvement of the aroma components of aroma.

2 Experimental section

2.1 Materials

Selecting healthy, growing white tulip and red rose plants, used in this experiment was verified at the flowering stage as experimental materials cultivated in Xinjiang Academy of Forestry Shuixigou planting base in May 2018. The temperature range ranged from 19–31°C and relative humidity of 20–35%. The soil physical and chemical properties were weak alkaline (pH = 8.1) with organic matter of 36.06 g kg−1, and contained total N 2.7 g kg−1, CEC 15.49 cmol kg−1, and conductivity 2.24 s mol−1, soil mechanical composition: clay 15.24%, sand 21.34, and silt 63.42%. Sampling was done at different time points (9:00, 12:00, 15:00, 18:00, and 21:00), and each time point was done 3 times for the test.

2.2 Floral scent collection

A dynamic headspace method was used to collect the volatiles released from roses and tulip. An individual flower was put in a sampling bag (355 mm × 355 mm; Reynolds, USA), which releases and absorbs a few volatiles. A stainless-steel tube (0.25 × 3.5; USA) containing Tenax-TA (60–80 mesh) was used as the volatile trap, which avoided touching the flower. A Portable air sampler (QC 1; Beijing Municipal Institute of Labor Protection, China) served as the pump, and air filtered through a drying column filled with charcoal was pumped into the bag. In a short time, extract the air in the sampling bag, connect the adsorption tube, and then fill the bag with the air filtered by activated carbon. When the air in the bag reaches 3/4 full, stop filling and start to enter the headspace circulation sampling. The collected volatile substances are absorbed into the stainless steel adsorption tube containing Tenax-TA adsorption material. The volatiles were collected for 15 min at a flow rate of 300 mL to min−1 for each flower. Each variety was sampled 3 times during the same test session. Empty bags served as control samples. Afterward, the stainless steel tubes were sealed and placed in a refrigerator.

2.3 Floral scent analysis

2.3.1 Thermal desorption conditions

The ATD-GC/MS technique was used to analyze the floral scent. Install the adsorption tube with volatile components on the thermal desorption device. The floral scent collected in the stainless steel tube was desorbed by heating in an ATD (TurboMatrix 350; PerkinElmer Corp. Norwalk, CT, USA). Set the first desorption temperature as 250°C and the first desorption time as 10 min on the thermal desorption device, so that the volatile components absorbed in the adsorption tube can be resolved and brought into the cold well by carrier gas for condensation and concentration. The temperature of cold hydrazine is −30°C, the second desorption temperature is 300°C, and the second desorption time is 5 min, vaporize the concentrated volatiles in the cold trap and directly enter the gas chromatographic column head of GC-MS, the injection time is 1 min. Carrier gas (helium) pressure: 20 kPa, sample inlet temperature: 260°C.

2.3.2 Chromatographic conditions

The GC (7890 A Network GC System; Agilent Technologies, Santa Clara, CA, USA) was equipped with a capillary TR-5MS column (60 m × 0.25 mm, with a 0.25 µm film thickness). Carrier gas: high purity helium (purity ≥ 99.999%); programmed temperature rise: initial temperature 40°C, hold for 2 min, rise to 280°C at the rate of 6°C/min, hold for 3 min; inlet temperature: 250°C; split ratio: 1:1.

2.3.3 Mass spectrometry conditions

The MS (5975C Network MSD; Agilent Technologies, Santa Clara, CA, USA) was operated in EI source energy: 70 eV; MS interface temperature: 250°C; ion source temperature: 250°C; quadruple temperature: 150°C; sampling mode: full scan; EM voltage: 1576 V; mass scanning range: M/Z 40–500.

Table 1

Analysis result of floral composition and relative content of rose

No.Retention timeMain componentRelative content/%
9:0012:0015:0018:0021:00
Alcohol
13.92Ethanol, 2-(methylamino)-0.2941.4341.237
24.104Ethanol0.2810.7861.482
310.5933-Hexen-1-ol, (Z)-0.7461.111
415.4211-Hexanol, 2-ethyl-1.12
515.952Benzyl alcohol0.4211.5230.999
618.139Phenylethyl alcohol29.43259.47449.4412.8311.623
720.699Citronellol17.9761318.3010.7250.623
821.245Geraniol16.1475.3212.808
923.502Propyl mercaptan0.909
Fatty hydrocarbon
104.564Methylene chloride7.3071.4821.19114.27728.56
114.721Pentane, 2-methyl-1.1970.5671.3631.891
124.856Pentane, 3-methyl-1.0040.9341.065
135.018n-Hexane7.823.8931.0478.95616.69
145.489Trichloromethane1.9232.1045.607
155.922Hexane, 2-methyl-0.5190.547
166.003Cyclohexane0.499
176.074Hexane, 3-methyl-0.706
189.127Hexanal1.230.634
1917.132Undecane0.290.7711.295
2019.714Dodecane0.693
2122.144Tridecane0.4990.9050.663
2223.833Dodecane, 2,6,10-trimethyl-0.563
2324.428Tetradecane0.9620.5972.7541.737
2425.678Tetradecane, 4,11-dimethyl-0.4111.0791.1471.067
2525.781Heneicosane0.7510.582
2626.577Pentadecane1.3015.1583.406
2728.612Hexadecane0.8920.6393.6691.03
2828.607Octadecane3.669
2929.451Hentriacontane0.731
3030.528Heptadecane1.536
Terpenes
3112.775Alpha-pinene0.561
3215.546d-Limonene0.5410.502
3315.849Beta-Ocimene0.328
3423.4272,6-Octadiene-2,6-dimethyl-0.6733.218
3525.381Longifolene2.3821.4312.330.785
3626.847Alpha-Farnesene1.8420.625
Aldehydes
376.653Acetaldehyde1.465
3814.804Octanal0.882
3917.554Nonanal0.5272.519
4020.168Decanal1.363
4121.0882,6-Octadienal, 3,7-dimethyl-, (Z)-0.433
4221.792Citral1.018
Ketones
434.277Acetone2.7910.7570.677.50710.642
4414.2475-Hepten-2-one, 6-methyl-0.8610.5140.5012.5660.64
4516.953Acetophenone1.6850.436
Esters
464.472Acetic acid, methyl ester0.594
475.278Ethyl acetate1.3480.9172.7443.314
4821.576Acetic acid, 2-phenylethyl ester0.7161.2812.274
4924.119Geranyl acetate0.3040.6050.709
Arenes
506.058Benzene1.5221.822
518.298Toluene0.5671.1711.577
5210.821Ethylbenzene0.7581.1461.284
5311.081p-Xylene1.1050.6461.752.174
5411.768Benzene, 1,3-dimethyl-0.5451.1321.561
Others
556.62Mercaptamine0.761
5614.047Benzaldehyde2.5370.742
5719.205Benzoic acid0.4972.12
5820.228Naphthalene0.954
5921.576
6022.956Naphthalene, 2-methyl0.55
6123.946Eugenol1.749.696

2.4 Data analysis

Data processing and mass spectrometry retrieval: using the NIST 11 and WIEIY Library in the TurboMass Version 5.4.2 software of GC-MS, we can automatically retrieve and analyze the mass spectrum data of the components, and search all the results and refer to the relevant standard maps for checking and supplemental retrieval. The composition of each volatile matter is qualitative, and the relative percentage content of the components in the total volatiles is calculated by the method of the total ion flow peak area normalization.

  1. Ethical approval: The conducted research is not related to either human or animal use.

3 Results and discussion

3.1 Analysis of the composition of floral scent

Figure 1 showed the chromatographic profiles of the floral scent emitted from rose and tulip at different time points, and a significant difference was found.

According to Table 2, 70 volatile compounds were identified in five time points. They were divided into eight kinds, including alcohols, fatty hydrocarbon, terpenes, aldehydes, ketones, esters, and other substances. The variety and release of volatile substances of flower fragrance at different time periods were different. Of all the ingredients, the volatilization of alpha-Farnesene was the largest, and at 21:00 it accounted for 30.7% of the total flower fragrance release. Followed by ethanol, pentadecane, beta-ocimene, longifolene, caryophyllene, and acetone, these may be the main sources of the smell of tulip. The composition of volatile substances in the diurnal variation showed a law that decreased first and then increased. At 15:00, the least volatile substances were detected (26 species), and the most volatile substances released at 9:00 were 44 species. Among the aroma components of the tulip, the number of fatty hydrocarbons (20 species) is the largest, and the number of terpenes is the least, only three kinds of compounds. The proportion of terpenes, arenes, and fatty hydrocarbons accounts for most of the results.

Table 2

Analysis result of floral composition and relative content of tulip

No.Retention timeMain componentRelative content/%
9:0012:0015:0018:0021:00
Alcohol
13.909Glycidol0.3870.9251.672.16
24.077Ethanol2.3353.6614.07511.3566.46
34.711-Propanol, 3-ethoxy-2.0110.7341.4450.829
46.415Silanediol, dimethyl-0.9041.284
510.4423-Hexen-1-ol, (E)-0.6591.054
610.613-Hexen-1-ol0.84713.54822.2174.2881.138
715.4321-Hexanol, 2-ethyl-0.388
819.2481-Nonanol0.696
Fattyhydrocarbon
94.542Methylene chloride11.8472.2392.4379.4874.193
105.013n-Hexane10.4251.4850.8585.985
115.468Trichloromethane3.544
125.895Hexane, 2-methyl-0.599
136.599Propane1.17
148.6341-Octene0.716
158.818Octane0.5151.375
1611.568Nonane0.3480.8240.747
1717.127Undecane0.352
1824.423Tetradecane0.3490.6541.17
1925.651Hentriacontane0.8
2025.765Decane, 3,8-dimethyl-0.649
2126.572Pentadecane1.0631.6561.3373.0561.058
2228.612Hexadecane0.5560.6230.771.9170.629
2329.435Heneicosane1.6230.585
2430.4632-Hexene, 2,5,5-trimethyl-2.5287.1017.2873.343
2530.512Heptadecane1.332
2633.511Eicosane1.187
2733.992Heptadecane0.933
2836.12Nonadecane1.534
Terpenes
2915.546Alpha-pinene0.643
3015.86Beta-ocimene0.7687.7555.0651.0843.069
3125.37Longifolene1.031.6933.0754.4871.033
3225.473Caryophyllene1.2791.5941.9731.2650.872
3326.853Alpha-Farnesene12.00820.05623.75212.98530.734
Aldehydes
344.84Methacrolein1.5480.796
356.675Acetaldehyde1.2362.621.48
366.702Pentanal1.36
379.116Hexanal0.651.1031.029
3814.025Benzaldehyde1.3220.651.4244.114
3914.804Octanal0.4460.70.661
4016.293Benzeneacetaldehyde0.385
4120.168Decanal1.1180.8981.0421.716
Ketones
424.255Acetone7.6745.1352.3635.4115.298
435.0892-Butanone1.2010.90.8541.292
4411.8763-Nonanone0.291
4514.2745-Hepten-2-one, 6-methyl-1.4361.8381.4080.637
4616.921Acetophenone0.7790.8591.439
4733.592Megestrol acetate0.572
Esters
485.257Ethyl acetate1.3160.777
4917.581Benzoic acid, methyl ester2.4696.1162.6922.9442.289
5020.212Methyl salicylate0.984
5135.508Dibutyl phthalate0.971
5238.7822-Propenoic acid, 3-(4-methoxyphenyl)-, 2-ethylhexyl ester0.831.45
5341.1052-Propenoic acid, 3-(4-methoxyphenyl)-, 2-ethylhexyl ester0.962
Arenes
546.042Benzene0.833
558.288Toluene14.2622.5981.0650.803
5610.815Ethylbenzene0.59
5711.07p-Xylene1.202
5811.757Benzene, 1,3-dimethyl-0.48
5919.362Benzene, 1,4-dimethoxy-1.9043.233.4722.6521.839
6021.8953,5-Dimethoxytoluene7.386.8335.8888.9126.159
Others
614.932Acetic acid0.8580.6320.885
624.9972-Nonenoic acid0.85
6314.22Phenol1.263
6414.7343-Hexen-1-ol, acetate, (E)-1.5112.126
6516.6295H-Naphtho[2,3-c]carbazole0.488
6619.178Benzoic acid0.7332.039
6723.492Phthalic anhydride0.5821.259
6831.271-Methyl-1,3,3-triphenylindane0.51
6934.3662,6-Diphenylpyridine0.933
7040.358[1,1′:3′,1″-Terphenyl]-2′-ol0.941
Figure 1 Total ion chromatography profiles of floral scent emitted from rose and tulip during the daytime ((a) Rose, (b) Tulip, (A) 9:00, (B) 12:00, (C) 15:00, (D) 18:00, (E) 21:00).
Figure 1

Total ion chromatography profiles of floral scent emitted from rose and tulip during the daytime ((a) Rose, (b) Tulip, (A) 9:00, (B) 12:00, (C) 15:00, (D) 18:00, (E) 21:00).

3.2 Analysis of diurnal variation of floral scent

There is a significant difference in the release of the aroma at different periods of the day in a rose (as shown in Table 1 and Figure 2). Among all volatile species, alcohols exhibited the highest release amount in volatile substances at 9:00, 12:00, and 15:00, and the highest release number of fatty hydrocarbons in volatile substances in 18:00 and 21:00 for two periods. At 15:00, the light is the strongest, but the aroma of the aroma is the least, which may be affected by transpiration. Terpenes, aldehydes, aromatic hydrocarbons, and other classes showed significant differences at 15:00, and aldehydes and aromatic hydrocarbons were released at 15:00 at zero. Fatty hydrocarbons, aldehydes, ketones, and aromatic hydrocarbons, and the total release rate of volatile substances is V type at 9:00, 12:00, 15:00, 18:00, and 21:00, that is to say, descend first and then rise.

Figure 2 The diurnal variation of total release amount of floral scent from rose ((a) alcohol, (b) fatty hydrocarbon, (c) terpenes, (d) aldehydes, (e) ketones, (f) esters, (g) arenes, (h) others).
Figure 2

The diurnal variation of total release amount of floral scent from rose ((a) alcohol, (b) fatty hydrocarbon, (c) terpenes, (d) aldehydes, (e) ketones, (f) esters, (g) arenes, (h) others).

Figure 3 shows the diurnal variation of the total release amount of aroma components of tulip. At different time points, the relative content of aroma components is distinct. The terpenes were at 12:00, 15:00, and 21:00 three time points, and the volatile substances showed the highest release amount. At 15:00, the light is the strongest, but the aroma of the aroma is the least, which may be affected by transpiration. The release of aldehydes at 15:00 is zero. At five time points, 9:00, 12:00, 15:00, 18:00, and 21:00, the release of alcohol substances first increased and then decreased, whereas in aldehydes, the amount of alcohol released first decreased and then increased.

Figure 3 The diurnal variation of total release amount of floral scent from tulip ((a) alcohol, (b) fattyhydrocarbon, (c) terpenes, (d) aldehydes, (e) ketones, (f) esters, (g) arenes, (h) others).
Figure 3

The diurnal variation of total release amount of floral scent from tulip ((a) alcohol, (b) fattyhydrocarbon, (c) terpenes, (d) aldehydes, (e) ketones, (f) esters, (g) arenes, (h) others).

3.3 Discussion

Previous reports have concluded that the aroma of rose results from major volatiles with high relative contents. However, the composition of volatile organic compounds (VOCs) obtained from oils or plant organs is significantly different from those produced and emitted by living plant systems. The living plant approach better reveals the composition of volatile compounds, resulting in more accurate and reliable results [21]. The VOCs emitted from six commercial rose cultivars were collected in vivo by HS-SPME in green-houses and analyzed by gas chromatography coupled with mass spectrometry, which have abundant terpenes, aromatic hydrocarbons, and esters [22]. Linalool, a-pinene, b-pinene, and limonene, these volatile moieties were also detected by Knudsen, which is consistent with our analysis [23,24]. Used HS-SPME detected citronellol and the 2-phenylethyl alcohol, which are commonly found in the essential oil of Rosa, were detected in our study [22,25].

Oyama-Okubo [26] found two aromatics, two terpenoids and minor volatiles, which are consistent with our research. Benzyl alcohol and linalool were produced by Oriental hybrid lily flowers with a mild pleasant aroma. Although these two minor compounds were not detected in this study, the quantities of ethanol, beta-ocimene, longifolene, caryophyllene, and other compounds identified in Tulipa gesneriana L. make the fragrance for some consumers [27]. We found longifolene and caryophyllene are so-called green leaf volatiles (GLVs), which are de novo synthesized after herbivore damage as indicators of tissue disruption to the plant [28].

Our results confirmed that the emission of volatiles fluctuates in a diurnal rhythm before anthesis. This diurnal rhythm was also reported in Rosa rugosa Thunb. and Tulipa gesneriana L., producing high level of volatiles at night from flowers after anthesis. There is a significant difference in the release of the aroma at different periods of the day in rose and tulip in this study. Floral production is of great significance in plant biology and ecology. It promotes pollination through interaction with insects and other organisms, thus playing a role in reproduction, and it also provides value for many horticultural flowers with commercial value [29].

4 Conclusion

Limonene terpene compounds, α-pinene, β-pinene, and linalool alcohols can enhance the freshness of the air and can regulate the nervous system of the human body, can make people relax, has obvious sedative effect, with health effects on the human body. Roses and tulips are all aromatic plants used for landscaping. ATD-GC/MS analysis detected 62 and 70 compounds in the floral scent of rose and tulip, respectively. Volatile compounds were categorized as alcohols, fatty hydrocarbon, terpenes, aldehydes, ketones, esters, and other substances. There is a significant difference in the release of the aroma at different periods (9:00, 12:00, 15:00, 18:00, 21:00) of day in rose and tulip. Phenylethyl alcohol, citronellol, methylene chloride, hexane, and acetone may be the main source of the smell of roses. Ethanol, alpha-Farnesene, pentadecane, beta-ocimene, longifolene, caryophyllene, and acetone may be the main source of the smell of tulip. Research of roses and tulips in aromatic in the garden provides a theoretical basis and research and improvement of the aroma components of aroma.

Acknowledgments

This work was supported by special fund for basic research of forest, fruit industry (XJLG201906), and special fund for forestry science and technology of Xin Jiang China (XLK2014-049).

  1. Conflict of interest: The authors of this paper declare that there is no conflict of interest in relation to the publication of this paper.

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Received: 2019-01-07
Revised: 2020-02-24
Accepted: 2020-02-25
Published Online: 2020-08-14

© 2020 Lu Yang et al., published by De Gruyter

This work is licensed under the Creative Commons Attribution 4.0 International License.

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