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古尔班通古特沙漠荒漠肉苁蓉及其对应寄主梭梭遗传多样性和营养成分的研究

Study on Genetic Diversity and Nutritional Components of Cistanche Deserticola and Its Corresponding Host Haloxylon in Gurbantonggut Desert

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【Author in Chinese】 张向前

【Supervisor】 祝建波李予霞

【Author's Information】 石河子大学, 生物化学与分子生物学, 2018, 硕士

【Abstract in Chinese】 肉苁蓉为列当科(Orobanchaceae)肉苁蓉属(Cistanche)多年生寄生性草本植物,是分布广泛的名贵中药材。肉苁蓉的多种功效导致其一直享有“沙漠人参”的美誉。梭梭是荒漠肉苁蓉的专一性寄主,为肉苁蓉生长发育提供所有的营养物质,其对维持荒漠生态平衡起着极其重要的作用。近几十年来,不合理的开发利用,造成了肉苁蓉和梭梭种质资源的破坏。研究梭梭和荒漠肉苁蓉遗传多样性对其资源评价和利用有深远意义。古尔班通古特沙漠分布有广泛的梭梭林,也是荒漠肉苁蓉的主要产地之一,为了全面了解古尔班通古特沙漠荒漠肉苁蓉居群分布的遗传多样性特点,本研究通过ISSR分子标记技术,利用Nei和Shannon等多样性指数对古尔班通古特沙漠中5个居群166个个体的荒漠肉苁蓉及其寄主梭梭进行遗传多样性分析,同时通过电感耦合等离子体发射光谱法(ICP-OES)对梭梭和肉苁蓉的铜、铁、镁、锰、锌元素进行测定,并进行相关性分析。通过高效液相色谱法测定荒漠肉苁蓉有效成分松果菊苷和毛蕊花糖苷的含量,并与微量元素含量进行相关性分析。1、肉苁蓉结论:在供试材料中,8个引物共扩增出144个多态位点,多态位点百分率达100%,5个居群的多态位点百分率差异在46.53%~77.78%之间。在物种水平上,Nei基因多样度(h)为0.2604,Shannon多样性指数(I)是0.4110。遗传变异分析表明,物种水平的居群间遗传分化系数Gst为0.2222,居群间的基因流Nm为1.7507。研究显示古尔班通古特沙漠中荒漠肉苁蓉多态位点比例高,各居群基因交流较多,不同居群间遗传变异并不明显,这些对肉苁蓉资源有效地保护和利用具有重要意义。2、梭梭结论:在供试材料中,10个引物共扩增出191个多态位点,多态位点百分率达100%,5个居群的多态位点百分率差异在57.07%~77.49%之间。在物种水平上,Nei基因多样度(h)为0.2451,Shannon多样性指数(I)是0.3878。遗传变异分析表明,物种水平的居群间遗传分化系数Gst为0.1774,居群间的基因流Nm为2.3177。塔城居群和博乐居群的遗传距离最小(0.0499),阿勒泰居群和呼图壁居群遗传距离较大(0.0986)。研究显示古尔班通古特沙漠中梭梭多态位点比例高,各居群基因交流较多,不同居群间遗传变异并不明显。3、将两种植物的遗传距离分别与地理距离、经度差和纬度差进行相关性分析发现,除了梭梭遗传距离与纬度差呈极显著相关外,其余没有显著相关性。4、微量元素含量测定结果显示:肉苁蓉和梭梭都含有铜、铁、镁、锰、锌元素,而且五种元素含量多少的变化趋势在两种植物中是相同的,都是铁元素和镁元素含量较多,铜元素含量最少;且肉苁蓉的铜、铁、锰元素含量与梭梭的铜、铁、镁、锰元素含量都存在显著或者极显著的相关性。5、肉苁蓉有效成分松果菊苷和毛蕊花糖苷含量测定结果显示呼图壁和奇台两地的荒漠肉苁蓉体内含量较其他地区多。相关性分析显示:荒漠肉苁蓉的松果菊苷和毛蕊花糖苷含量都与其本身所含镁和锌元素呈现极显著负相关;肉苁蓉的松果菊苷含量与寄主梭梭的铜元素含量呈极显著相关,而毛蕊花糖苷与寄主梭梭的镁元素呈显著负相关。同时肉苁蓉的松果菊苷和毛蕊花糖苷二者之间也呈现极显著相关。

【Abstract】 Cistanche is a perennial parasitic herb of the genus Cistanche of Orobanchaceae.It is a rare and precious Chinese herbal medicine widely distributed.Cistanche has a variety of effects,Therefore,Cistanche has always enjoyed the reputation of “desert ginseng”.Haloxylon is a special host of desert Cistanche,providing all nutrients for the growth and development of Cistanche,which plays an extremely important role in maintaining the ecological balance of desert.In recent decades,unreasonable development and utilization have caused the destruction of the germplasm resources of Cistanche and Haloxylon.Studying the genetic diversity of Haloxylon and Cistanche in desert have far-reaching significant meanings to evaluate their resources and utilization.The Gurbantunggut desert has a wide range of Haloxylon ammodendrons and is also one of the main producing areas of Desert Morsels.In order to understand the genetic diversity of Desert Cistanche in the Gurbantunggut Desert comprehensively,the ISSR molecular markers were used,by means of Nei and Shannon diversity index,to detect diversity of 166 individuals in five populations of Cistanche and Haloxylon in Gurbantunggut Desert.At the same time,copper,iron,magnesium,manganese,and zinc in Haloxylon and Cistanche were measured by Inductively Coupled Plasma Optical Emission Spectrometry(ICP-OES),and correlation analysis was performed.The content of echinacoside and verbascoside in the effective components of Desert Cistanche was determined by HPLC,and the correlation analysis was carried out with the contents of trace elements.1.Conclusion of Cistanche: 144 polymorphic bands were induced by amplification primers.Percentage of polymorphic bands is 100% and the difference among five populations ranged from 46.53% to 77.78%.Shannon’s indices of diversity(I)is 0.411 0 and Nei’s gene diversity(h)is 0.2604 respectively.Genetic variance analysis showed that the coefficient of gene differentiation in the species is0.222 and the gene flow is 1.7507 among populations.Studies have shown that Cistanche deserticola in the Gurbantunggut Desert is high proportion of polymorphic loci,and there was more gene communication among different populations.The genetic variation among different populations was not obvious,these are of great significance to the effective protection and utilization of Cistanche deserticola.2.Conclusion of Haloxyne: 191 polymorphic bands were induced by amplification primers.Percentage of polymorphic bands is 100% and the difference among five populations ranged from 57.07% to 77.49%.Shannon’s indices ofdiversity(I)is 0.3878 and Nei’s gene diversity(h)is 0.2451 respectively.Genetic variance analysis showed that the coefficient of gene differentiation in the species is0.1774 and the gene flow is 2.3177 among populations.The genetic distance between Tacheng population and Bole population was the closest(0.0499),and the genetic distance between Altay population and Hutubi population was the farthest(0.0986).Studies have shown that Haloxyne in the Gurbantunggut Desert is high proportion of polymorphic loci,and there was more gene communication among different populations.The genetic variation among different populations was not obvious.3.When compared with geographic distance,longitude and latitude difference respectively,correlation analysis found that apart from haloxyne ammodendron whose genetic distance was significantly correlated to the latitude,the rest have no significant correlation.4.The content of trace element showed that both Cistanche and Haloxylon contained copper,iron,magnesium,manganese and zinc elements,and the variation tendency of the five elements was the same in both plants,which was the content of iron and magnesium were higher,while the content of copper was the least.The content of copper,iron and manganese in Cistanche deserticola had significant or extremely significant correlation with Haloxylon ammodendron.5.The results of the determination of echinacoside and verbascoside in the effective components of Cistanche showed that the content of desert cistanche in Hutubi and Qitai was higher than that in other regions.The correlation analysis showed that the content of echinacoside and verbascoside in desert cistanche showed extremely significant correlation with magnesium and zinc contained therein.The content of echinacoside in Cistanche was significantly correlated with the content of copper in Haloxylon ammodendron,and the content of verbascoside was significantly negatively correlated with magnesium in Haloxylon ammodendron.At the same time,there are also extremely significant correlations between echinacoside and verbascoside.

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