【作者】 唐丽；

【导师】 刘友全；

【作者基本信息】 中南林业科技大学 ， 森林培育， 2007， 博士

【摘要】 南天竹(Nandina domestica Thunb.),属小檗科南天竹属植物,是一种具有观赏、生态、药用等多种价值的树种。针对目前国内外对于南天竹的种质资源、遗传多样性、生长发育规律和抗性选育研究较少以及园林资源开发和利用混乱等问题,本论文对湖南地区南天竹种源进行调查,研究南天竹在湖南的分布、资源类型及生态适应性。通过对南天竹形态特征、生长发育规律、繁殖特性及园林利用特性、遗传多样性的系统研究,分析南天竹在园林中的观赏特性和观赏效果,并对不同种源南天竹进行种源遗传结构分析及分子鉴别,建立一套南天竹种质资源开发利用价值评价标准和体系。主要结论如下：南天竹在湖南地区的物候变化规律：春季萌芽早,能在土壤解冻后、春季大风来临之前很快萌芽,约在2月中旬至3月初树芽开始萌动,3月上旬至3月下旬底为芽子膨大期,4月初抽生出尚未展开的花序和嫩叶,4月上旬至4月下旬,为花蕾期,从5月上旬至7月中旬,为花期,近3个月,种子10月上中旬成熟,成熟期晚,且红叶和果实观赏期长。南天竹主芽具有较强的萌芽力,主芽受损后,副芽和隐芽往往同时萌发,抽生成枝条；其新梢的年生长、叶片生长规律遵循Logistic方程的拟合曲线。南天竹属于当年花芽分化,翌年开花结实类型；从开花到果实成熟大致需要180-210 d左右。表型性状研究表明,南天竹地理种源遗传变异丰富,25个形态特征平均变异系数达到了29.956%。13个种源间的欧氏遗传距离为0.001～1.000。平均遗传距离为0.5005。25个表型性状中,叶片形状、果实形状、花器官特征和嫩梢(叶)秋冬季颜色等反映了南天竹70.986%的表型信息,利用这些特征可基本对不同地方来源的南天竹进行分类。根据聚类及主成分分析结果,并结合不同地方种源性状特征,可将13个湖南南天竹种源分为5大类型：观赏效果较差的南天竹主要分布于第一类,有张家界南天竹、宜章南天竹、新化南天竹；而树体高大、果实圆形或椭圆形,颜色红色或棕红色,观赏效果较好的南天竹主要分布于第二类,有岳阳南天竹、韶山南天竹、衡山南天竹、祁阳南天竹；秋冬季节叶片颜色为紫红色的V02(柏加南天竹)和V10分别单独为一类,属于第四类和第五类；其余种源属于第三类,其表型性状特征不明显。根据对南天竹过氧化物酶、超氧化物岐化酶、细胞色素酶、酯酶等同工酶的研究,探求湖南南天竹不同种源之间的亲源关系及遗传多样性。同工酶电泳染色后,绘制图谱,测Rf值。结果表明南天竹13个种源不同同工酶的酶带数目、位置差异较大,表明湖南南天竹种源变异十分丰富、亲缘关系复杂。根据13个种源不同同工酶的酶带电泳图谱,采用Dice相似系数计算方法,计算各种源之间的相似系数。结果表明,不同种源之间相似系数差异较大。其中POD相似系数为0.500-1.000,EST相似系数为0.500-1.000,SOD相似系数为0.364-1.000,COD相似系数为0.556～1.000。采用SPSS13.0软件进行种源聚类,根据各种源之间的距离绘制的分类树状图,13个种源在不同的相似距离处分开,表明13个种源大致可分为不同的三大类群。利用NTSYSpc2.1分析软件,进行主坐标分析。将Dice遗传相似性系数矩阵进行Dcenter数据转化,求其特征值和特征向量,生成主坐标二维和三维图。从二维和三维图上可以看出,同一种同工酶的主坐标聚类与系统聚类结果是一致的。通过对ISSR反应程序中的一些重要参数进行摸索和优化试验,建立了稳定的、适用于南天竹13个种源遗传多样性和遗传分化研究的ISSR-PCR分析反应体系和反应程序,对湖南13个南天竹种源进行遗传多样性分析。从100条引物中筛选出10条重复性好、条带特异、多态性明显的引物。该10条引物扩增出56条清晰可重复的条带,其中48条是多态带,平均每条引物为4.8条多态带,总多态带百分率为85.7%,扩增片段长度范围为300～1600 bpo Nei遗传距离和遗传一致度的结果表明：V05和V12之间的遗传距离最小(0.1133),相应的遗传一致度最大(0.9286)；而V09与V10之间的遗传距离最大(0.6581),相应的遗传一致度最小(0.5179)。这说明V05和V12两个种源的生长环境相类似,而V09和V10之间的差异可能较大。通过NTSYS-pc软件和MVSP32软件对南天竹的13个种源的Nei的遗传距离分析发现,V05、V07、V08、V12在遗传上较相似,聚为.一类；V1l、V13很相似,聚为一类；V02、V03、V04的相似性较近,可以聚为一类；V01、V06、V09、V10则同其它三类的相似性较远,且彼此单独为一类。形态标记与分子标记之间的相关性系数为,0.213,两者相关性不高。南天竹观赏特性较高,其观赏特性主要表现为直接观赏性和间接观赏性；观赏方式有绿化观赏、室内点缀观赏和盆栽观赏；绿化手法多种多样,常见的有片植、从植、绿篱、地被植物、庭院绿化、盆景盆栽以及插花等。采用灰色关联分析方法综合评价南天竹的观赏价值。从最终评价结果来,13个南天竹种源,按从优到劣排序,依次为浏阳柏加南天竹、岳阳县南天竹、.黄花机场南天竹、衡山南天竹、宜章南天竹、韶山南天竹、祁阳南天竹、洪江南天竹、泪罗南天竹、株洲芦淞区南天竹、娄底新化南天竹、张家界南天竹、益阳桃江南天竹。浏阳柏加的南天竹在13个测试种源排名第一,观赏特性极好,景观美观最优。试验分析结果与园林实际应用效果的一致性较好。利用模糊数学的多级综合评判法(AHP法)进行南天竹开发利用价值评价。从最终评价结果来看：柏加南天竹、岳阳县南天竹、黄花机场南天竹、祁阳南天竹、衡山南天竹和宜章南天竹列入第Ⅰ或第Ⅱ级,其观赏价值、资源潜力、生物学特性各方面分值都相对很高；桃江南天竹、韶山南天竹等观赏价值很高却被列入第Ⅲ级,主要原因是资源数量、抗逆性或生物学特性等方面的限制,造成综合评价值偏低。通过对观赏植物南天竹种源开发价值评价体系的建立,使评价方法,不再局限于对观赏植物南天竹观赏价值的定性描述和评价,能比较全面准确地反映特定区域特定生态环境下观赏植物南天竹种质资源开发利用的特点,可以避免开发过程中对生态环境的严重破坏和资源浪费等问题,是一种比较理想的评价方法。此评价方法应用在湖南地区观赏植物南天竹资源的评价结果,将为湖南观赏植物南天竹资源的开发利用起到一定的指导作用。综合试验结果可得：从园林利用的角度上,湖南地区南天竹种质资源以浏阳柏加南天竹、岳阳县南天竹这两个地方种源为最佳种源,可以在园林中广泛开发利用,且利用价值高。而张家界南天竹、邵东南天竹,应结合其特性进行其它方面的开发利用,其余各种源可以根据开发利用途径和目的,进行如盆景和水土保持等方面的开发利用。AHP模型评价湖南南天竹的结果,将为今后开发利用湖南南天竹资源提供理论依据。更多还原

【Abstract】 Common Nandina(Nandina domestica Thunb)belonges to Berberidaceae. It is a plant that has many values in Ornamental,ecology,medicine and so on.In order to solve the problems of researches on its species resource,genetic diversity,growing regularities,stress breeding study,and less exploitation and utilization in landscaping,the article inquired into the different endemic species and studied it’s distribution,resource type and ecological adaptability of Common Nandina in Hunan. The systemic researches of morphology,growth regularities and of the reproductive characteristics and exploitation and utilization in garden,and genetic diversity have been done in this article.The ornamental characteristics and impression were analyzed,and the genetic structure analysis and molecular identification among different species have been done,result in the standard system for exploitation and utilization were established in Hunan.Primary conclusion as follows:In Hunan area,the change rules of Common Nandina Phonology:The bud in early spring sprout after the soil thaw and before gale coming,about mid-February to early March tree buds begin to germinate,The time from early March to late March to the end is buds bulked period,the leaves and inflorescence,which are not starting expanding, take out in early April.The bud period is the time from in early April to late April.And the florescene period is as long as sustained about three months from early May to mid-July,seeds mature in the mid-October,which belong to late mature is later,the ornamental force,after the strong injury,auxiliary shoots and bud often sprout soon,and the branches germinates.The annual growth regularity of new shoots and leaves follows the fitted curve of the Logistic equation.Common Nandina completes bud differentiation in the first year,and blessoms next year.belongs bud differentiation 180-210 days is needed from blossoms to fruit.The results of Morphological research showed that genetic diversity was much abundant in Common Nandina in Hunan. The average variation coefficient(CV)was 29.956% among the 25 morphological characters. Genetic distance(Nei’s)ranged from 0.001 to 1.000,and the average genetic distance was 0.5005.In 25 morphological characters,the shape of leaf and the fruit,characters of flower,color of soft tip(leaf)were the decisive factors limit classification of Common Nandina,of which revealed about 70.986% morphological information.Common Nandina from different place can classify by these characters.According to UPGMA clustering results and principal components analysis(PCA)and their geographical origins’characters,13 sources from Hunan were classified in 5 clusters:Having not good ornamental effect were the first type,such as zhangjiajie,Yizhang,Xinhua; And the second type has big body,circular or ellipse fruit,which were red or brown red,has better ornamental effect such as Yueyang,Shaoshan,Hengshan Mount,Qiyang; the V02(Baijia)of purple-red leaf in fall and winter and the V10 were different type alone,belong to the fourth type and the fifth typerespectively;Other sources belong to the third type,their Morphological characters were not obvious.According to these researches on Peroxidase(POD),Superoxid dismutase Isoenzyme(SOD),Enzyme(EST)Cytochrome(COD),Esterase isozymes,the, connection and genetic diversity were selected in different source of Hunan.After isoenzyme electricity swimming and dying,drawing up the patterns,measuring the Rf value,the results showed that 13 sources different isoenzyme’belt number,position had bigger difference and 13 sources diversity were much abundant,the blood relationship were complex in Hunan. According to 13 sources of different isoenzyme electrophoresis patterns,We use the method of the Dice coefficient to calculate their coefficient. The results showed that the difference of coefficient in sources was bigger. The POD Dice coefficient was 0.500-1.000, the EST Dice coefficient was 0.500-1.000,the SOD Dice coefficient was 0.364-1.000, the COD Dice coefficient was 0.556-1.000. By using Statistical Package for the Social Sciences (SPSS13.0)software,to carry out the source UPGMA clustering,13 sources were detached in different distance and were placed into three different clusters.By using NTSYSpc2.1 analysis software,the principal components analysis was carried out. the Dice coefficient matrix were transformed by the Dcenter data means,eigenvalue and eigenvector were calculated. two-dimensional chart and three-dimensional chart were drew.The 2D and 3D charts showed that the cluster from the same isozyme by the Principal Coordinates Analysis(PCOD)and the cluster of UPGMA clustering method were consistent.The exploration and optimization tests have been carried on through some important parameters in ISSR response procedure,which established the reacting system and the response procedure which have been stable and suitable to Common Nandina 13 sources genetic diversity and genetic differentiation research by ISSR-PCR analysis. The genetic diversity was analyzed to 13 Common Nandina sources of Hunan. From 100 ISSR primer combinations,10 were selected for the analysis based on numeber and quality of polymorphic fragments,which repetition were better,the strip belt was special,polymorphric were obvious. A total of 56 ISSR markers were identified,of which 48(85.7%)were polymorphic,there were 4.8 polymorphic in every primer combinations.The fragments of amplified bands ranging were from 300 to 1600 bp.The results of genetic distance(Nei’s)and genetic similar coefficient showed:the genetic distance was the least between the V05 and the V12 (0.1133),the corresponding genetic similar coefficient was the biggest (0.9286). But the Nei’s genetic distance was the farthest between V09 and the V10(0.6581),the corresponding genetic similar coefficient was the smallest(0.5179).This indicated the V05 and the V12 sources habitats were similar,but the difference of the V09 and the V10 may be bigger.Through NTSYS-pc software and MVSP32 software to analyze 13 sources Common Nandina Nei’s distance,we found that V05,V07,V08,V12 comparatively were similar in the heredity,and they were clustered as one type kind;V11,V13 were more similar,and they were clustered as another type; the similarity of V02,V03,V04 was nearer,its may be clustered as one type; V01,V06,V09,V10 were farther with other three kinds of similarities, and each was clustered as one type respectively. The relevant coefficient between the morphological mark and the molecular mark was 0.213,both correlation were not higher.The ornamental characters of Common Nandina was higher,of which main performance were directly or indirectly to be viewed; the manners of view had the landscape,indoor potted ornamental plants and ornamental decorations; The green technique were varied and the common such as mass planting, clump planting,hedgerow,and the grassprot planting,the garden virescence,bonsai and pot plantingand ikebana.Common Nandina ornamental Characters were analyzed by Grey correlative Degree comprehensive evaluation.From finally appraising the results,the order of 13 different Common Nandina sources were in turn Liuyang Baijia Nandina,Yueyang County Nandina,the Huanghua airport Nandina,Hengshan Nandina, Yichang Nandina, Shaoshan Nandina, Qiyang Nandina,the Hong Chiang Nandina;Miluo Nandina,the Zhuzhou Lusong area Nandina,Loudi Xinhua Nandina, Zhangjiajie Nandina,the Yiyang Taojiang from good to poor.The Liuyang BaiJia Nandina was the No.1 in 13 source,and the ornamental Characters was. extremely good,and the landscape impression was the best. The uniformity between the experimental analysis result and the garden application actual effect was better.The evaluation system for exploitation and utilization value of Common Nandina was established by using Analytic hierarchy process(AHP). Based on final evaluation result,BaiJia Nandina,Yueyang County Nandina,Huanghua airpot Nandina,Qiyang Nandina,Hengshan Nandina and Yizhang Nandina were listed in the gradeⅠorⅡ, its score of ornamental value,rescourcese potential, biology characteristic were the highest. Taojiang Nandina,Shaoshan Nandina and so on which had very high ornamental value actually,but they were ordered in the gradeⅢ, the main reasons were some restrictively factors such as resisting ability, resources quantity,nature or biology characteristic property, of which brought about that its appraisal synthesis value were the lowest.By establishment of the evaluation system for exploitation and utilization value of Common Nandina in Hunan,the evaluation method was no longer restricted to the qualitative description and appraisal of Common Nandina viewing value to ornamental plants. It can quite comprehensively accurately reflect the exploitation and utilization character of the ornamental plant resources;This was a perfect evaluation method to avoid environment havoc and resource waste in exploitation and utilization. It was one quite ideal evaluation method. The evaluation result which was applied to ornamental plants in the Hunan area,will play a guiding role in the local ornamental plants for the development and utilization of Common Nandina resources.In conclusion, the resources of Baijia,Yueyang Common Nandina were the best and the exploitation and utilization value were the highest and may be exploited and utilized to the garden widely. The other aspects value of Zhangjiajie Common Nandina,Shaodong Common Nandina were exploited for the purpose, such as minisize bonsai and keeping natural environment.This method system was proved to be satisfied in flexibility and accuracy and it was a scientific foundation for exploiting and utilizing the Nandina domestic in Hunan.更多还原