荞麦及其野生种遗传多样性分析

荞麦起源于我国西南地区,该地区分布有大量荞麦地方品种和野生近缘种。本研究利用SSR分子标记,对从我国西南地区收集的81份荞麦及其野生资源进行遗传多样性分析。结果显示,利用19对SSR引物共检测出84个等位基因,每对SSR引物平均扩增出4.421个等位基因,19对SSR引物的平均Shannon's信息指数为0.985,平均PIC为0.478。81份荞麦及其野生资源的相似系数范围为0.500~1.000,分析发现大粒组荞麦种(甜荞及其近缘种、苦荞及小米荞、金荞)与小粒组硬枝万年荞亲缘关系比较近。通过聚类分析在遗传相似系数为0.732时,可将81份荞麦种质分为4个类群,第Ⅰ类由小粒组齿翅野荞、细柄野荞、小野荞麦、疏穗小野荞麦和硬枝万年荞组成;第Ⅱ类由甜荞和甜荞近缘种组成;第Ⅲ类都是由金荞组成;第Ⅳ类是由苦荞和小米荞组成。在遗传系数为0.920时,小粒组荞麦种可以明显区分出疏穗小野荞麦、硬枝万年荞、齿翅野荞及其细柄野荞。研究表明,19对SSR引物多态性较高,能较好反应荞麦及其野生种质的遗传多样性,本研究的结果为荞麦属种之间的亲缘关系分析和荞麦起源进化研究提供科学依据。     

荞麦属2个野生荞麦种的染色体核型研究

对荞麦属皱叶野荞麦(Fagopyrum crispatifolium J.L.Liu)及其近缘种细柄野荞麦[F.gracilipes (Hemsl.) Dam.et Diels]的染色体核型进行了分析.结果表明,2个野生荞麦的染色体数目均为2n=4x=32,但二者在随体数目、染色体长度等方面差异明显,染色体核型不同,皱叶野荞麦的核型公式为:30m(4SAT)+2sm,细柄野荞麦的核型公式为:32m,属首次报道.     

荞麦属种间13S球蛋白基因序列比较研究

荞麦13S球蛋白是荞麦种子中的一类主要贮藏蛋白。本研究选用荞麦属植物甜荞栽培种及其野生类型、苦荞栽培种及其野生类型、毛野荞、左贡野荞、细柄野荞和硬枝万年荞6个种共44份材料,进行PCR特异性扩增、测序得到荞麦13S球蛋白基因的保守片段序列。对序列进行差异分析,结果发现44份供试材料13S球蛋白基因片段的285个排列位点中不变位点为24个,多态性位点S为261个(含简约信息位点数198个和单型可变位点63个),序列总突变位点Eta为503个。野生甜荞种内13S球蛋白基因序列差异明显高于栽培甜荞,但野生苦荞种内13S球蛋白基因序列差异仅稍高于栽培苦荞。推测其一方面可能与繁殖方式有关,另一方面可能与荞麦驯化过程中通常只有少数野生型群体被驯化有关。系统聚类分析发现栽培甜荞与野甜荞亲缘关系近,与左贡野荞亲缘关系次之;栽培苦荞与野苦荞亲缘关系近,与毛野荞亲缘关系次之;细柄野荞和硬枝万年荞的13S蛋白基因片段序列差异较小,说明其亲缘关系较近。上述结果为荞麦属种间遗传多样性与进化关系研究提供了理论依据。     

基于ITS和ndhF-rpl32序列的荞麦种间亲缘关系分析

荞麦起源于我国, 演化形成了丰富的物种和遗传多样性。为了有效研究和利用荞麦及其野生种资源, 以从四川、甘肃、贵州等地采集的荞麦属(Fagopyrum)10个种(含变种、亚种和复合体种)共71份材料为对象, 通过ITS和叶绿体ndhF-rpl32序列分析, 利用MEGA5.0构建系统进化树, 探讨了荞麦种内及种间亲缘关系。结果表明, 在ITS序列矩阵中, 序列长度为725 bp, 信息位点为150个, 占序列总长度的20.7%; 在ndhF-rpl32序列矩阵中, 序列长度为940 bp, 信息位点为158个, 占序列总长度的16.8%。由ITS序列和ndhF-rpl32序列构建的两个进化树都可以将71份材料分为大粒荞麦种组和小粒荞麦种组; 其中, 大粒荞麦种组包括栽培苦荞和米苦荞(F. tataricum)、金荞复合体(F. cymosum complex)、栽培甜荞(F. esculentum)和野生甜荞(F. esculentum ssp. ancestralis); 小粒荞麦种组包括齿翅野荞(F. gracilipes var. odontopterum)、疏穗小野荞(F. leptopodum var. grossii)、小野荞(F. leptopodum)、密毛野荞(F. densovillosum)、细柄野荞(F. gracilipes)和硬枝万年荞(F. urophyllum)。而ndhF-rpl32序列构建的系统发育树还能区分栽培甜荞和野生甜荞, 具有更好的聚类效果。另外, 与栽培甜荞相比, 金荞复合体与栽培苦荞的亲缘关系更近。该研究为荞麦属种的分类和条形码研究提供了一定的科学依据。     

六个不同类型荞麦花花粉粒形态的电镜观察比较研究

对二倍体甜荞长花柱类型 (ES2 s)、二倍体甜荞短花柱类型 (ES2 S)、四倍体甜荞长花柱类型 (ES4s)、四倍体甜荞短花柱类型 (ES4S)、四倍体有翅细野荞 (GR4HI)、四倍体无翅细野荞 (GR4HO)等 6个不同荞麦类型花的花粉形态学进行了电镜观察。结果表明 :这些荞麦类型的花粉粒都是椭圆形 ,都有网状纹饰、3孔沟等特征。二倍体甜荞染色体加倍后形成的四倍体甜荞 ,相对于二倍体甜荞而言 ,花粉粒显著增大和明显变圆。无论二倍体还是四倍体甜荞 ,其长花柱型花的花粉粒都比其对应短花柱型花的花粉粒要小。尽管四倍体甜荞和四倍体细野荞在染色体数目和倍性上一样 ,但是它们在花粉粒的大小上有显著差异。四倍体细野荞有翅类型和无翅类型的花粉粒大小和形态差异不显著     

甘薯栽培种及其近缘野生种的DAPI核型及rDNA FISH分析

利用DAPI显带和rDNA-FISH技术对栽培种甘薯(‘徐薯18’)(Ipomoea batatas cv.Xushu No.18)及2种不同产地近缘野生种(Ipomoea hederacea Jacq.)进行了细胞遗传学研究。DAPI核型分析表明,‘徐薯18’核型公式为2n=6x=90=72m+18sm(18SAT),随体位于第1、3、6染色体上;美国近缘野生种核型公式为2n=2x=30=30m(4SAT),香港近缘野生种核型公式为2n=2x=30=20m+10sm(4SAT),随体均位于第6、12染色体上。rD-NA-FISH结果显示,栽培种甘薯基因组中含有3对5SrDNA位点,分别位于着丝粒区、亚着丝粒区和染色体端部;美国近缘野生种基因组中含有2对5SrDNA位点,香港近缘野生种基因组中含有1对5SrDNA位点,均位于随体部位;两种不同地域来源的近缘野生种基因组中均含有2对45SrDNA位点,分别位于第6和第12染色体上。     

五个中国荞麦(Fagopyrum)种的核型分析

用去壁低渗法对甜荞 ( Fagopyrum esculentum)、苦荞 ( F.tataricum)、左贡野荞 ( F.zuogongense Q.F.Chen) ,大野荞 ( F.megaspartanium Q.F.Chen)及毛野荞 ( F.pilus Q.F.Chen)等大粒组荞麦种的根尖和茎尖有丝分裂染色体进行了观察 ,并对其茎尖有丝分裂染色体的核型进行了比较分析。结果表明 :这 5种荞麦在核型上类似 ,都有 2对随体染色体 ,而且都为对称核型。但它们彼此有一定的差异。甜荞、苦荞、大野荞、毛野荞及左贡野荞的核型公式分别为 1 2 m+4m( SAT)、1 2 m+4sm( SAT)、8m+4sm+4m( SAT)、1 2 m+2 m( SAT) +2 sm( SAT)及 2 4 m+4sm+4m( SAT)。     

中国四川蓼科荞麦属一新种——皱叶野荞麦

描述了蓼科Polygonaceae一新种--皮叶野荞麦Fagopyrum crispatifolium J.L.Liu.本种近似于细柄野荞麦F. gracilipes(Hemsl.)Damm.& Diels,但以其叶片表面泡状突起,叶缘皱波状,具不规则波状圆齿、圆齿或小圆齿,聚伞花序密集与之相区别.此外,还报道了该物种的染色体数目,发现它是一个四倍体,染色体数目为2n=4x=32.     

中国荞麦属果实形态特征

应用解剖镜和扫描电镜对中国产荞麦属Fagopyrum Mill．8种1变种的果实形态和微形态特征进行了观察。结果表明这些种类的果实可分为3类：(1)果实三棱锥状,表面不光滑,无光泽,具皱纹网状纹饰,此种类型的植物有苦荞麦、金荞麦。(2)果实卵圆三棱锥状,表面光滑,有光泽,具条纹纹饰,此种类型的植物有荞麦、长柄野荞麦、线叶野荞麦。(3)果实卵圆三棱锥状,表面光滑,有光泽,具大量的瘤状颗粒和少数模糊的细条纹纹饰,此种类型的植物有硬枝野荞麦、细柄野荞麦、小野荞麦、疏穗小野荞麦。研究结果认为金荞麦与栽培种苦荞麦关系较近。     

两个荞麦品种的核型分析

本文对南方两个荞麦品种高荞3号和溪荞5号的根尖细胞有丝分裂染色体进行核型分析。结果表明,高荞3号和溪荞5号的核型都为2A,高荞3号和溪荞5号的核型公式分别为2n=2x=16=10m(2SAT)+4sm+2st和2n=2x=16=8m+6sm(2SAT)+2st。     

Morphological characteristics of fruits of Fagopyrum (Polygonaceae) from China

Macro- and micro-morphological characteristics of fruits in eight species and one variety of the genus Fagopyrum Mill. (Polygonaceae) from China were observed under stereoscope and scanning electron microscope(SEM). Based on the results, the fruits of the species studied are divided into type Ⅰ , Ⅱ and Ⅲ. The fruits of type Ⅰ are triangular-pyramidal; their surface are rugosely reticulate, neither smooth nor shiny. Two species, F. tataricum and F. dibotrys have this fruit type. Those of type Ⅱ are ovoid-triangular-pyramidal; their surface are smooth and shiny, and striately reticulate. Three species, F. esculentum, F. statice and F. lineare, have this fruit type. In type Ⅲ, the fruits are ovoid-triangular-pyramidal; their surface are smooth and shiny, and covered with many warty grains and sparsely finely striate. F. urophyllum, F. gracilipes, F. leptopodum var. leptopodum, and F. leptopodum var. grossii have this fruit type. Judging from the morphological characteristics of fruits, F. dibotrys might be more closely related to F. tataricumthan to F. esculentum.     

Search for the wild ancestor of buckwheat III. The wild ancestor of cultivated common buckwheat, and of tatary buckwheat

By surveying wild Fagopyrum species and their distribution in southern China and the Himalayan hills, I arrived at the conclusion that the newly discovered subspecies F. esculentum ssp. ancestralis Ohnishi is the wild ancestor of cultivated common buckwheat, while previously known wild tatary buckwheat,F. tataricum ssp. potanini Batalin is the wild ancestor of tatary buckwheat. Their original birthplace is revealed to be northwestern corner of Yunnan province for common buckwheat judging from the distribution of wild ancestor, and to be the northwest part of Sichuan province for tatary buckwheat judging from allozyme variability in wild tatary buckwheat. F. cymosum is not the ancestor of cultivated buckwheat; it is only distantly related to cultivated buckwheat, in morphology, isozymes and cpDNA. Several genetic, ecological and taxonomic categories which should be taken into consideration in examining the origin of buckwheat were discussed. Key Words: Fagopyrum esculentum ssp. ancestralis; Fagopyrum tataricum ssp. potanini; southern China theory of origin of buckwheat. Contribution from Plant Germ-Plasm Institute, Faculty of Agriculture, Kyoto University. No. 78.     

A study of resources of Fagopyrum (Polygonaceae) native to China

Morphology, taxonomy, reproductive biology, esterase isozyme patterns and chromosome number were studied in 16 accessions of cultivated and wild buckwheat native to Tibet, Sichuan, Guizhou and Yunnan. Among these are four established species: Fagopyrum esculentum, F tataricum, F. pleioramosum and F gracilipes. The first three species are diploid (2 n = 16) and the last one is tetraploid (2 n = 32). In addition to the above, three new species are described: F. zuogongense, F. megaspartanium and E pilus. There are clear differences in esterase isozyme patterns between these species. The zymographs of esterase isozyme of diploid F. megaspartanium (native to Sichuan) and F. pilus (native to Tibet) are similar to those of F. esculentum and F tataricum respectively. F. gracilipes , in the small achene group of the genus, has a zymograph of esterase isozyme that is very different from the species in the large-achene group. The diploid accessions of F. pilus and F. megaspartanium may be ancestors of cultivated buckwheat.     

中国荞麦栽培品种的核型比较分析

通过对我国栽培的10个甜荞（Fagopyrun esculentum)品种和7个苦荞（F.tatricum)品种根尖染色体数目观察和核型比较分析,结果表明：甜荞和苦荞染色体数目都是由16条染色体构成的,2n=2x=16。甜荞有2对随体,而苦荞有1对随体,它们的核型公式分别为12m 4m(SAT)、12m 2sm 2sm(SAT);种内各品种在染色体相对长度、染色体长度比、随体染色体数目和形态等方面差异较小,但在种间的差异明显。     

Species relationships in Fagopyrum revealed by PCR-based DNA fingerprinting

Random amplified polymorphic DNA (RAPD) markers were used to distinguish between 28 different accessions belonging to 14 species and two sub-species of Fagopyrum. Of the 75 random 10-mer primers tested, only 19 generated robust, easily interpretable amplification products. A total of 364 bands were observed with an average of 19.15 bands per primer, of which 99.45% were polymorphic. Primer OPN-08 produced the maximum number of fragments and UBC-183 produced the minimum number of fragments. The data were utilized to elucidate genetic relationships among 14 species and two sub-species of Fagopyrum. Cluster analysis using the unweighted paired group method of arithmetic means (UPGMA) showed four main clusters, two each of the cymosum and urophyllum groups. The results showed that Fagopyrum tataricum is closer to its wild ancestor F. tataricum ssp. potanini Batalin, closely followed by Fagopyrum giganteum. Cultivated common buckwheat ( Fagopyrum esculentum) showed affinity with its putative wild ancestor F. esculentum ssp. ancestrale and the other closely related diploid species Fagopyrum homotropicum. In the urophyllum group, Fagopyrum macrocarpum and Fagopyrum pleioramosum formed one cluster, whereas Fagopyrum capillatum, Fagopyrum gracilipes and Fagopyrum gilessii clustered separately. Except for a few cases, our results correspond with previously reported studies on Fagopyrum using the isozyme, RFLP and RAPD methods. Species-diagnostic amplification products specific to some species in the cymosum and urophyllum groups were identified. Our results show that RAPDs can be successfully used to analyze species relationships in Fagopyrum and also for constructing linkage maps.     

川西北荞麦种间亲缘关系初步研究

采用RAPD和同工酶技术对川西北具有代表性的10份荞麦材料进行分析.结果表明,筛选出的15个RAPD引物扩增出388条带.其中352条具有多态性,多态性比率为90.72%;过氧化物同工酶分析获得15条酶带,酯酶同工酶获得10条酶带.3种方法聚类结果基本一致,10份材料可初步聚为4个类群.其中,与金荞亲缘关系相比,甜荞较近而苦荞较远;普格县野荞和齿翅野荞、细柄野荞亲缘关系较近,划为同类;阿坝野荞单独划为一类,对其在分类中的地位有待进一步研究.