Abstract
1H Nuclear Magnetic Resonance (1H NMR) combined with multivariate statistics was adopted to discriminate female and male flower buds of Populus tomentosa in the study. Samples of 11 female and 16 male flower buds of P. tomentosa were collected in Beijing, China. 1H NMR spectra were acquired on a 400 MHz spectrometer. In total, 30 chemical compounds were identified with standards and literature according to chemical shifts, peak areas, and multiplicity. Principal component analysis (PCA), hierarchical clustering analysis (HCA), and supervised orthogonal partial least squares-discriminant analysis (OPLS-DA) were applied to discriminate female and male flower buds. An apparent grouping trend (R2X, 0.809; Q2, 0.903) between female and male groups was exhibited with PCA and HCA. The two groups were also well discriminated with OPLS-DA (R2X, 0.808; R2Y, 0.976; Q2, 0.960). Combined with variable importance in projection (VIP) > 1.0 and p < 0.05 of OPLS-DA, it was found that the content of daucosterol, β-sitosterol, ursolic acid, and betulonic acid in male group was higher than that in female, which should be the key differences of chemical constituents in female and male flower buds of P. tomentosa. The study demonstrated that 1H NMR combined with multivariate statistics could be used to discriminate female and male plants and clarify differences, which provided a novel method to identify the gender of dioecious plants.
Highlights
Dioecious plants refer to the plants that have female and male flowers grown in different individuals
The result of hierarchical clustering analysis (HCA) confirms the classification with Principal component analysis (PCA) model. These results demonstrate that there are differences in chemical compositions between female and male flower buds of Figure 5, 27 samples could be clearly separated into two groups, i.e., all of the 11 female samples are classified into Group 1 and 16 male samples are classified into Group 2
The 27 samples could be divided into two groups with PCA, HCA, and OPLSDA analysis successfully, which are coincident with their gender
Summary
Dioecious plants refer to the plants that have female and male flowers grown in different individuals. The incidence of dioecy in flowering plants globally is relatively low (6~7%) [1], there are several traditional Chinese medicines from dioecious plants, such as Duzhong (Eucommiae Cortex) from Eucommia ulmoids Oliv. (Fam. Eucommiaceae), Yinxingye (Ginkgo folium) from Ginkgo biloba L. (Fam. Bilobaceae), and Tianhuafen (Trichosanthis Radic) from Trichosanthes kirilowii Maxim. T. rosthornii Harms (Fam. Cucurbitaceae) [2]. Most traditional Chinese medicines derived from dioecious plants are applied in clinical and pharmaceutical practice regardless of their gender
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