摘要 | 利用顶空固相微萃取-气相色谱-质谱联用(HS-SPME-GC-MS)法对枸骨(Ilex cornuta Lindl. et Paxton)雌、雄花在大蕾期、初开期、盛花期、末花期的挥发性化合物进行分离、鉴定,探究其变化对传粉的影响。结果显示:枸骨雌花挥发性化合物总释放量在末花期最大,雄花挥发性化合物总释放量在盛花期最大。在不同花期枸骨雌、雄花中共检出30种挥发性化合物,其中,雌、雄花共有成分有15种,雌花特有成分有4种,雄花特有成分11种,雌、雄花中成分均以萜烯类和醚类为主。随着花发育,雌、雄花中挥发性化合物的Jaccard相似性系数先增加后减小,在盛花期达到极相似水平,在末花期为极不相似。根据变量投影重要性(VIP)从雌花中筛选出萜烯类6种、醇类1种、酰胺类1种和醚类1种共9种差异标志物(VIP≥1),从雄花中筛选出萜烯类4种、酯类1种和醚类1种共6种差异标志物。通过偏最小二乘法判别分析(PLS-DA)可有效区分枸骨雌花的4个花期;雄花的大蕾期、初开期和盛花期、末花期也可有效区分。对苯二甲醚、α-蒎烯、长叶烯、丙氨酰丙氨酰丙氨酸、罗勒烯、蒎烯、金合欢醇和萜品油烯可用于区分雌花发育进程;α-蒎烯、肌醇六乙酸酯和芳樟醇可用于区分雄花发育进程。雌、雄花的主要访花昆虫均以蜂类昆虫为主,且访花频率的最大值均在盛花期;雄花访花昆虫的访花频率多于雌花。综合研究结果表明:不同花期枸骨雌、雄花中挥发性化合物的种类和含量存在差异;α-蒎烯、长叶烯和β-蒎烯等雌花中的差异标志物以及长叶烯、肌醇六乙酸酯和α-蒎烯等雄花中的差异标志物可能影响了蜂类等访花昆虫的访花频率;枸骨雌花在末花期会生成大量蒎烯,可能起到减少访花昆虫访问、保护子房的目的。
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Abstract | The volatile compounds in female and male flowers of Ilex cornuta Lindl. et Paxton at the big bud stage, initial opening stage, full flowering stage, and final flowering stage were separated and identified by using headspace solid-phase microextraction-gas chromatography-mass spectrometry (HS-SPME-GC-MS) method, and the effect of their changes on pollination was explored. The results show that the total release amount of volatile compounds in female flowers of I. cornuta is the largest at the final flowering stage, while that in male flowers is the largest at the full flowering stage. A total of 30 volatile compounds are detected in female and male flowers of I. cornuta at different flowering stages, among which, there are 15 common components in both female and male flowers, 4 components specific to female flowers, 11 components specific to male flowers, and terpenes and ethers are the main components in both female and male flowers. With the development of flowers, the Jaccard similarity coefficient of volatile compounds in female and male flowers first increases and then decreases, it reaches a very similar level at the full flowering stage, and is extremely dissimilar at the final flowering stage. Based on the variable importance in the projection (VIP), a total of 9 differential markers (VIP≥1) are screened from female flowers, including 6 terpenes, 1 alcohol, 1 amide, and 1 ether, and 6 differential markers are screened from male flowers, including 4 terpenes, 1 ester, and 1 ether. The four developmental stages of female flowers of I. cornuta can be effectively distinguished through partial least squares discriminant analysis (PLS-DA); the big bud stage, initial opening stage and full flowering stage, and final flowering stage of male flowers can also be effectively distinguished. The flowering process of female flowers of I. cornuta can be judged by 1,4-dimethoxybenzene, α-pinene, longifolene, trialanine, ocimene, pinene, (E,E)-farnesol, and terpinolene; the flowering process of male flowers of I. cornuta can be judged by α-pinene, myo-inositol hexaacetate, and linalool. The main flower-visiting insects for both female and male flowers are bees, and the maximum visiting frequency of flowers all occurs at the full flowering stage; the visiting frequency of flower-visiting insects for male flowers is larger than that for female flowers. The comprehensive research results show that there are differences in types and contents of volatile compounds in female and male flowers of I. cornuta at different flowering stages; differential markers in female flowers such as α-pinene, longifolene, and βpinene, and those in male flowers such as longifolene, myo-inositol hexaacetate, and α-pinene, may affect the flower-visiting frequency of flower-visiting insects like bees; a large amount of pinene are generated at the final flowering stage of female flowers of I. cornuta, which may serve the purpose of reducing the visits of flowervisiting insects and protecting the ovary.
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关键词 | 枸骨; 花发育; 挥发性化合物; HS-SPME-GC-MS; 传粉 |
Key words | Ilex cornuta Lindl. et Paxton; flower development; volatile compound; HS-SPME-GC-MS; pollination |
作者 | 崔淦莎1a,1b, 郝明灼1c, 姬晓悦1d, 邹义萍2, 彭冶1a,1b |
所在单位 | 1. 南京林业大学: a. 南方现代林业协同创新中心, b. 生命科学学院, c. 林草学院、水土保持学院, d. 现代分析测试中心, 江苏 南京210037;2. 南京青珠果园艺有限公司, 江苏 南京 210043 |
点击量 | 24 |
下载次数 | 29 |
基金项目 | 江苏省重点研发计划(现代农业)项目(BE2021307); 南京市“一区一中心”科技发展计划项目(202306003) |