【作者】 雒新艳；

【导师】 戴思兰；

【作者基本信息】 北京林业大学 ， 园林植物与观赏园艺， 2009， 博士

【摘要】 菊花(Chrysanthemum×morifolium Ramat.)是菊科(Compositae)菊属(Chrysanthemum)植物,在我国有一千六百多年的栽培历史,是深受人民群众喜爱的传统名花。菊科植物是被子植物中进化程度最高的类群,而由于长期的栽培、自然选择和人工杂交,菊属中的菊花则位于进化的顶点。我国菊花品种数量众多,形态变异丰富,适应性强,分布广泛,遗传背景非常复杂,这些都为菊花品种资源调查、品种分类与鉴定、品种多样性分析等带来了困难,严重限制了菊花的遗传改良进程和园林实践应用。本文对北京地区的大菊品种资源进行了详细的调查,并在控制的栽培条件下,分别在表型和分子水平上对其进行了遗传多样性评价,并对数量性状进行概率分级,筛选出变异幅度及对分类作用最大的性状,期望为菊花品种资源的收集保存及合理利用提供依据,为菊花的品种分类与鉴定、品种保护及育种工作提供一定的参考资料。主要研究结果如下:1.品种资源调查经调查统计鉴定出北京地区的常见大菊品种1001个,5类瓣型中,平瓣类和管瓣类品种数量最多,分别占总品种数的42%和33%,桂瓣类(2%)较为稀少;在30个花型中,叠球型(205个)、翻卷型(97个)和卷散型(86个)品种数量较多,而管球型和全桂型仅各有一个品种,极为珍贵;在9个色系类型中,品种数量以白色和黄色居多(226个,213个),占44%;红色(29个)、绿色(29个)和橙色(15个)等花色在菊花品种中较为稀少;花色在不同花型中分布不均,在翻卷型、卷散型、钩环型和管盘型4种花型中具有齐全的9类花色,其余26个花型中均存在不同程度的花色缺失现象。调查中还发现:目前生产中仍然存在大量同名异物、同物异名的混杂现象。2.数量性状分级选择代表性强的400个大菊品种,参照《菊花DUS测试指南》的形态性状测试方法,检测其18个主要的数量性状,并对性状的分布范围、平均值、变异系数进行统计计算,在此基础上,绘制了分布频次图,结合非参数检验判断性状的分布类型。研究结果表明:叶柄长、叶片长、花径、花梗粗等13个数量性状均符合正态分布,而花瓣宽、节间长、花梗长、舌状小花数和筒状小花数5个数量性状呈现非中心的偏态分布,其中筒状小花数变异情况最为特殊,是严重的偏态分布。在此基瓷?对符合正态分布的13个数量性状进行了概率分级,确定分点值;对其余5个不符合正态分布的性状按照实际测试值分级,并在小于1/10的标准差范围内适度调整,方便其在实际中应用。3.质量性状分布参照《菊花DUS测试指南》的形态性状测试方法,观察记录大菊39个主要质量性状的表现,计算各性状的表现式样在品种群内的分布比例,结果显示舌状花的重瓣性、花瓣曲反状况、叶片基部性状、托叶大小等32个性状的表现式样在品种群内分布极不均匀,接近或超过70%的品种集中在性状的1种或2种表现式样中;而花瓣形状、外花瓣表面色、外花瓣背面色、花中心部颜色、叶裂刻基部有无突起、茎的曲直性和茎色等7个性状的表现式样在品种群内的分布则相对均匀。4.性状的稳定性及变异计算性状的变异系数,结果表明菊花表型性状均具有较高的多样性,花部性状相对于叶部性状变异系数更高。对22个稳定性状进行R聚类分析发现菊花性状之间相关性不大,大都独立演化。主成分分析结果显示:影响品种分类的第1主成分主要包含舌状小花数、花瓣长、总苞片基部到花顶部的高度、花瓣形状等花部性状;第2主成分主要为两个花色性状(花瓣表面色彩、花瓣背面色彩),第3主成分主要是筒状花部直径、筒状花长度,第4主成分主要是托叶相关性状。5.ISSR分子标记研究从50个ISSR引物中筛选出10个条带清晰、重复性良好的引物,对具有典型代表性的150个大菊品种的遗传多样性进行分析。结果共检测清晰可重复的229个位点,其中220个位点是多态性的;多态性位点百分率(PPB)是96.07%,Nei’s基因多样性指数(He)和Shannon信息指数(I)分别是0.3485和0.5154;而五个瓣型内的遗传多样性水平都比较高,其中平瓣类的遗传多样性水平最高(PPB=95.20%),而桂瓣类(PPB=86.90%)最低。利用瓣型间的遗传相似系数进行UPGMA聚类分析,结果将瓣型首先分为两大类:平瓣类、管瓣类、匙瓣类、畸瓣类聚在一类中,桂瓣类单独作为一类,在第一类中,平瓣类与管瓣类是最先聚在一起的两种类型,然后是匙瓣类,最后是畸瓣类。综合以上的研究结果表明:(1)北京地区现有大量大菊品种,且花型与花色较为丰富;(2)珍稀花型、花色品种亟需保护;(3)大部分数量性状是符合正态分布的,少数重点观赏性状迫于选择压力呈现偏态分布,概率分级法能够辅助大菊的数量性状建立新的分级标准;(4)大部分质量性状在品种群内的分布是不均匀的,可能是由菊花性状遗传分化的不均衡性或受到选育重点性状的连锁作用而造成的;(5)大菊花部性状相对稳定,对品种分类影响较大,其托叶部分也具有一定程度的稳定性,可以作为品种分类的合理补充,而叶部性状稳定性较差,在品种分类中应作为更次一级的分类依据;(6)平瓣类、管瓣类和匙瓣类的亲缘关系最近,畸瓣类品种花瓣先端特化受到人们的喜爱,长期的选择压力使其区别于3种基本瓣型,而这四种瓣型与桂瓣类在亲缘关系上较远;(7)ISSR分子标记检测表明:大菊品种多态性位点百分率较高,平瓣类品种的遗传分化最为丰富。更多还原

【Abstract】 Chrysanthemum×morifolium Ramat.,which belongs to Chrysanthemum and Compositae,is much loved by Chinese people as a traditional flower with a history of more than one thousand and six hundred years..Since the Compositae is the most advanced group in angiosperm,the cultivars of C. morifolium were deemed to be situated on the peak of the evolution owing to long periods of cultivation, natural selection and artificial crossing.As a result of numerous cultivars,abundant morphological variation,powerful suitability,wide distribution and extremely complicated genetic background,the investigation,classification,identification and diversity analysis of the cultivars became to be very difficulty,which restricted the breeding and landscape application furtherly.On the basis of investigation and identification of the cultivars of large-flowered chrysanthemum in Beijing area,the genetic diversity of phenotypic character and genic differences were analyzed under the condition of controlled cultivation,the system of probability grading for quantitative characters was built and some characters with bigger variation width and greatest impact in classification were screened out in order to provide the basis for collection,preservation and reasonably application and reference for classification, identification,protection and breeding of the cultivars.The main results were as follows:1.The investigation of cultivars resource Through the author’s survey and record in 6 year,we got 1001 cultivars of large-flowered Chrysanthemum in Beijing.The results of statistic analysis showed: the Flat-petaled Group(42%) and Quilled-petal Group(33%) took the most part of cultivars in the 5 groups,whereas the Anemone-petal group(2%) had even less cultivars.The Globular Types(205), Reflexed Types(97) and Mobile-petaled Types(86) had the most cultivars in 30 flower types,but the Tubular-globular Types and the Whole Anemone Types only had one rare cultivar.The white(226,23%) and the yellow cultivars(213,21%) were the two greatest teams in the 9 groups of flower colors, whereas the red(29),green(29) and orange(15) cultivars were much fewer.The colors distributed in different flower types were unever.Only the Reflexed Types,Mobile-petaled Types,Open bead Types and Tubular-pan Types had all nine colors;all of the other 26 flower types existed deficiency of flower color in various degrees.Additionally,a lot of mistake with homonym and synonym phenomenon still existed in the production..2.The probability grading for quantitative characters 400 large-flowered cultivars were chosed and their eighteen chief quantitative characters were tested and recorded according to the test method of morphological characters in the <Guidelines for the Conduct of Tests for Distinctness,Homogeneity and Stability in China>.The empirical distributions were determined by nonparametric test after the distribution range,mean value and variation coefficients of these characters were calculated and frequency diagrams were drawn.The results indicated that 13 quantitative characters(petiole length, leaf length,inflorescence diameter,peduncle diameter and so on) obeyed to the normal distribution, whereas the other 5 characters(floret width,internodes length,peduncle length,number of ray flowers and number of disc flowers) agreed with skewed distribution,in which the number of disc flowers was the most special character with extreme skewness.The grading criterials of the 13 quantitative characters obeyed to the normal distribution were determined with the method of probability grading, the grading criterials of another 5 characters were calculated on the basis of their actual test values. these criterials were moderately adjusted in less than 1/10 of standard deviation for practical application.3.The distribution of the qualitative characters The performance of the 39 chief qualitative characters were observed and recorded,according to the test method of morphological characters in the <Guidelines for the Conduct of Tests for Distinctness,Homogeneity and Stability in China>.The results of distribution proportion of each character in cultivars group showed that the distribution of 32 characters(number of ray flower rings,floret curliness,shape of leaf base,size of stipule and so on) were very asymmetrical with near or above 70%cultivars concentrated upon one or two performance, however,the distribution of the other 7 characters(floret type,surface color of outer floret,back color of outer floret,color of disc floret,claw in base of sinus between lateral lobes,stem erectness and stem color) in the cultivars group were relatively symmetrical. 4.The stability and variation of the characters The results of every character’s variation coefficient indicated that they all had higher diversity,the variation coefficient of floral characters is higher than that of foliar.The characters evolved almost independently according to the R cluster analysis of 22 stable characters.The principal component analysis showed that the number of ray flowers, the floret length,height from point of attachment of involucral bracts to top of flower head,floret type and height should be the most important factors.The second principal component contained the surface and back color of outer floret,which followed by the disk flower part diameter and the length of corolla tube. The fourth principal component included some factors about the stipule.5.Research of ISSR molecular marker The Genetic Diversity of 150 cultivars with typical representation in large-flowered chrysanthemum were investigated using 10 primers with clear bands and steady repetitiveness chosen from 50 ISSR primers.A total number of 229 bands were amplified,of which 150 were polymorphic loci.The main index were:PPB=96.07%,He=0.3485,I=0.5154(at species level).The genetic variation in each floral types were quite high,in which the Flat-petaled group is highest(PPB=95.20%),whereas the Anemone-petal group(PPB=86.90%) and the Spoon-petaled group(PPB=%9.52%) were the lowest.UPGMA cluster analysis which according to genetic similarity among floral types indicated that 5 groups were clustered into two groups,the Anemone-petal group was one group solely;the other four groups grouped together,in which the Flat-petaled group and the Quilled-petal group clustered firstly,followed by the Spoon-petaled group and the Filiform-petal group.The results above showed:(1) The cultivars of large-flowered chrysanthemum are widely distributed in Beijing area with plentiful floral types and floral colors;(2) The cultivars with the rare types and colors are urgent for protection;(3) Most of the quantitative characters obeys to the normal distribution,some important traits agreed with skewed distribution.The probability grading can assist quantitative character in establishing new grading standards;(4) Most of qualitative characters is asymmetrical in cultivars,which might be caused by unbalanced differentiation of characters or linkages coming from emphasis characters selection;(5) The floral characters are relatively stable and have comparably large influence on classification,the characters of stipule also have a certain degree of stability and can act as an reasonable supplement for cultivars classification,but the foliar characters have worse stability and can only be recognized as lower level criterion.(6) The genetic relationship among the Flat-petaled group,the Quilled-petal group and the Spoon-petaled group are closest,which followed by the Filiform-petal group because of long-term selection pressure for the specialization of the petal tip loved by most people.The Anemone-petal group has further genetic relationship with these four groups.(7) ISSR molecular marker detection indicates that the proportion of polymorphic loci of lardge-flowered chrysanthemum is higher,in which the Flat-petaled group has the highest genetic diversity level.更多还原