摘要
【目的】探讨中国野生葡萄果皮和叶片中白藜芦醇(Resveratrol,Res)含量的特点,为进一步开发利用中国野生葡萄资源提供理论依据。【方法】以51个野生葡萄株系为材料,利用高效液相色谱法(HPLC)连续2 a测定成熟果皮和叶片中白藜芦醇的含量,并结合气候条件进行对比分析。【结果】在51个野生葡萄株系中,约60%的葡萄果皮中白藜芦醇含量高于叶片,株系间果皮、叶片Res含量(ω)存在很大差异,其中果皮中含量最高的‘双溪腺枝葡萄03’(2013年与2014年分别为67.82μg·g^(-1)和68.44μg·g^(-1))比含量最低的‘洪江刺葡萄04’(2013年0.08μg·g^(-1),2014年0.09μg·g^(-1))高855.5倍,叶片中含量最高的‘高山二号’(2013与2014年分别为10.27μg·g^(-1)和11.69μg·g^(-1))比含量最低的‘冯举沟桑叶葡萄02’高256.8倍。结合采样地2 a的气候条件分析,果皮中的Res含量几乎未受气候条件影响,而叶片中Res含量在采样的2 a间变化幅度较大。【结论】腺枝葡萄中的‘双溪腺枝葡萄03’‘芷江水腺枝葡萄’‘双溪腺枝葡萄01’及山葡萄中的‘山葡萄N43-3’‘长白9号’‘双优’等果皮富含白藜芦醇。研究为有效利用和开发野生葡萄资源中的白藜芦醇及促进功能性食品开发提供理论依据。
【Objective】Grapevine is one of the most important fruit trees in the world. China is the important origin center of grapevine and possesses many species and large diversity of germplasm resources.Resveratrol, as a phytoalexin, can not only enhance plant resistance to various abiotic and biotic stresses,but also be highly beneficial to human health. So far resveratrol content had been detected only in 72 plant species. Moreover, table grapevines and wines have become one of main resveratrol sources as food. We have known in general that the genotypes with high content of resveratrol are mostly found in wild grapevines. In order to explore and ultilize more grapevine resources from Chinese native species evaluation of extractable amounts of resveratrol in berry skins and leaves in 51 genotypes from Chinese wild grapevines were carried out for two consecutive years.【Methods】The wild grapevines were grown under natural field conditions in the National Grape Germplasm Resources Repository of Zhengzhou Fruit Research Institute,Chinese Academy of Agricultural Sciences. Grape berries were collected from June to September andleaves were picked up in the end of June every year. And the fruits were peeled and the juice was soaked up using filter paper. Trans-resveratrol in berry skins and leaves was investigated in 2013 and 2014, respectively. Trans-resveratrol extraction was performed using HPLC method described by Li et al. Three gram samples were ground to powder using a porcelain mortar and pestle in liquid nitrogen, extracted by 15 m L ethyl acetate in the dark at 25 ℃ for 48 h, and centrifuged at 10 000 r·min-1for 10 min. The supernatants were transferred into a tube containing 5 m L ethyl acetate, followed by centrifugation at 10 000 r·min-1for10 min. All supernatants were evaporated to dryness by rotary vacuum evaporator at 40 ℃. Dried samples were then dissolved in 2 m L of methanol and stored at-80 ℃. The samples were filtered through a 0.22μm PTFE membrane filter before resveratrol analysis. Extractable amounts of resveratrol were analyzed using a Waters 2695 HPLC system. Elution was carried out with a mobile phase delivered using a Waters C18 HPLC pump at a flow rate 0.8 m L·min-1. A Waters 2996 detector was used for analysis of UV detection at 306 nm. HPLC chromatograms of resveratrol was performed with a standard solution. Resveratrol contents in berry skins and leaves were compared in 51 grapevine genotypes. In addition, according to the climate data in the two years（including mean temperature, relative humidity, rainfall and sunshine hours）,comparative analysis of resveratrol amounts were conducted between skins and leaves.【Results】The results showed that resveratrol contents in berry skins were higher than those in leaves in about 60% of 51 Chinese wild grapevine genotypes. The resveratrol content of berry skins varied from 0.08 to 67.82 μg·g-1,the mean value was 7.04 μg·g-1,and the coefficient of variation was 192.33% in 2013. The resveratrol content of berry skins varied from 0.09 to 68.44 μg·g-1, the mean value was 7.24 μg·g-1, and the coefficient of variation was 191.45% in 2014. There was greater difference in the resveratrol content of berry skins among the tested grapevine genotypes, of which, the resveratrol content of‘Shuangxi 03’（V. Adenoclada）with the highest resveratrol content（67.82 μg·g- 1in 2013, 68.44 μg·g- 1in 2014） was 855.5-fold higher than‘Hongjiang 04’（V. davidii） with the lowest resveratrol content（0.08 μg·g-1in 2013, 0.09 μg·g-1in2014）. The highest concentration of resveratrol was followed by V. adenoclada accession Shuangxi 03, such as V. adenoclada accession Zhijiangshui, Tangwei seeding, and V. adenoclada accession Shuangxi 01,whereas, the lowest of that was followed by V. davidii accession Hongjiang 04, such as V. davidii accession Hongjiangtongmu 07, Gaoshan No.1, and V. davidii accession Hongjiang 10. Meanwhile, the resveratrol content of leaves varied from 0.04 to 10.27 μg·g-1, the mean value was 1.61 μg·g-1, and the coefficient of variation was 127.63% in 2013. The resveratrol content of leaves varied from 0.20 to 11.69 μg·g- 1, the mean value 2.23 μg·g-1, and the coefficient of variation was 112.34% in 2014. There was not obvious difference in the resveratrol content of leaves among the tested grapevines, of which, the resveratrol content of Gaoshan No. 2（V. davidii） with the highest resveratrol content（10.27 μg·g- 1in 2013, 11.69 μg·g- 1in2014） was 256.8-fold higher than‘Fengjugou 02’V. ficifolia with the lowest resveratrol content（0.04 μg·g-1in 2013）. The highest was followed by Gaoshan No.2, such as V. ficifolia accession Fengjugou 03, V. davidii accession Wuhan, and V. betulifolia accession Songxian, whereas, the lowest was followed by V. ficifolia accession Fengjugou 02, such as V. ficifolia accession Qingling No.2, V. ficifolia accession Shibanyan 02 and V. adstricta. In addition, comparative analysis of resveratrol content based on climate data in summer of2013 and 2014 showed that there was a greater difference in leaves than that in skins in the two experimental years. The change range of resveratrol content between 2014 and 2013 in berry skins in only one genotype was over 100% while those in 29 genotypes were below 10%. However, the change range of resveratrol content between 2014 and 2013 in leaves in 14 genotypes were over 100% while those in 13 genotypes were below 10%.【Conclusion】The resveratrol contents in berry skins were higher than those in leavesabout 60% of 51 genotypes. It was noted that some genotypes from V. adenoclada Hand.-mazz（such as‘Shuangxi 03’‘Zhijiangshui’and‘Shuangxi 01’） and V. amurensis Rupr.（such as‘N43-3’,‘Yanshan’‘Changbai No.9’and‘Shuangyou’） were rich in resveratrol in berry skins. This founding would provide atheoretical basis for exploring the use of natural resveratrol in Chinese wild grapevines.
出处
《果树学报》
CAS
CSCD
北大核心
2016年第9期1092-1102,共11页
Journal of Fruit Science
基金
国家自然科学基金(31201591)
遗传工程国家重点实验室开放课题项目(20150321)
河南省重大科技专项(151100110900)
河南省高校科技创新团队支持计划(14IRTSTHN011)
关键词
中国野生葡萄
白藜芦醇
高效液相色谱
果皮
叶片
种质资源利用
Chinese wild grapevine
Resveratrol
HPLC determination
Skins
Leaves
Utilization of germplasm resources
