荞麦属种质资源发芽种子过氧化物酶同工酶研究

用聚丙烯酰胺凝胶电泳对荞麦属9个种(含大粒组8个和小粒组1个)32个收集系栽培及野生荞麦种子的过氧化物酶同工酶进行了研究。结果表明:过氧化物酶同工酶酶带23条,不同物种的酶带数4到8条。其中,甜荞有7条带,而苦荞为4条。酶带分析及聚类分析表明:大粒组荞麦种的谱带与F.gracilipes等小粒组荞麦种间差异极大,甜荞和苦荞酶带分别与F.megaspartanium和F.pilus相似,并分别与F.megas-partanium和F.pilus聚类最近,支持F.megaspartanium和F.pilus可能分别是甜荞和苦荞祖先种的假说。     

荞麦属种质资源发芽种子酯酶同工酶研究

用聚丙烯酰胺凝胶电泳对荞麦属8个种(含大粒组7个和小粒组1个)33份材料发芽种子的酯酶同工酶进行了研究。结果表明:酯酶同工酶不同酶带合计22条,不同物种的酶带数4到8条。其中,甜荞有8条带,苦荞为7条。酶带及聚类分析表明,大粒组荞麦种的谱带与细野荞等小粒组荞麦种间差异极大,甜荞和苦荞酶带分别与大野荞和毛野荞相似,并分别与F.megaspartanium和F.pilus聚类最近,支持F.megaspartanium和F.pilus可能分别是甜荞和苦荞祖先种的假说。     

荞麦属植物三叶期幼叶酯酶同工酶研究

用聚丙烯酰胺凝胶电泳技术对荞麦属(Fagopyrum Mill)8个种(含大粒组7个种和小粒组1个种)29份栽培及野生荞麦植株三叶期幼叶的酯酶同工酶进行了研究.结果发现:酯酶同工酶酶带共31条,不同物种的酶带数4～9条,其中甜荞有8条带,而苦荞为9条.酶带分析及聚类分析表明:大粒组荞麦种的谱带与细野荞(F.gracilipes)等小粒组荞麦种间差异极大,甜荞(F.esculentum)和苦荞(F.tataricum)酶带分别与大野荞(F.megaspartanium)和毛野荞(F.pilus)相似,并分别与大野荞和毛野荞聚类最近, 提示大野荞和毛野荞可能分别是甜荞和苦荞的祖先种.     

荞麦属植物三叶期幼叶过氧化物酶同工酶研究

用聚丙烯酰胺凝胶电泳技术对荞麦属(Fagopyrum Mill)8个种(含大粒组7个种和小粒组1个种)28份栽培及野生荞麦植株三叶期幼叶的过氧化物酶同工酶进行了研究.结果发现:过氧化物酶同工酶酶带共23条,不同物种的酶带数4～9条,其中甜荞有6条带,而苦荞为9条.酶带及聚类分析表明:大粒组荞麦种的谱带与细野荞(F.gracilipes)等小粒组荞麦种间差异极大,甜荞(F.esculentum)和苦荞(F.tataricum)酶带分别与大野荞(F.megaspartanium)和毛野荞(F.pilus)相似,并分别与大野荞和毛野荞聚类最近, 支持Chen(1999)提出的大野荞和毛野荞可能分别是甜荞和苦荞的祖先种的假说.     

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。     

自交可育甜荞、金苦荞、米苦荞不同品系总黄酮、粗蛋白及其蛋白组分含量研究北大核心CSCD

为探究荞麦新品种的营养保健价值,该文对自交可育甜荞、金苦荞、米苦荞共56个不同品系荞麦种子的粗蛋白、总黄酮、蛋白组分含量及其果实性状的变异进行了研究。结果表明:(1)甜荞、金苦荞、米苦荞种子中粗蛋白含量平均值分别为13.19%、15.44%、11.75%,总黄酮含量平均值分别为0.14%、2.50%、2.09%,清蛋白含量的平均值分别为5.22%、6.13%、4.56%,球蛋白含量的平均值分别为1.29%、1.15%、0.91%,醇溶蛋白含量的平均值分别为0.42%、0.58%、0.55%,谷蛋白含量的平均值分别为2.66%、3.36%、2.80%,三种荞麦的蛋白组分均符合清蛋白>谷蛋白>球蛋白>醇溶蛋白。(2)果实性状中,甜荞果实千粒重、果实面积、果实直径的变异系数最大,米苦荞果实周长、果实长宽比、果实长、果实宽和50 mL容重的变异系数最大。(3)相关分析表明甜荞种子粗蛋白含量与果实长宽比、果实长,金苦荞种子粗蛋白含量与果实周长、果实长,米苦荞种子粗蛋白含量与果实宽、总黄酮含量与果实面积、果实宽、果实直径、50 mL容重的相关性均达到了显著或极显著水平。(4)该研究筛选出甜荞(1808-166贵甜2号优系)、金苦荞(多苦74、多苦78)、米苦荞(1906-136黑米荞麦、43-2)等高蛋白含量或高黄酮含量的荞麦品系。该研究结果对荞麦优良品种的选育和荞麦新产品的开发具有一定指导意义。     

甜荞品种内与品种间的遗传多样性研究

以9个甜荞品种为材料,1个苦荞品种为对照,每个品种随机取10粒种子发芽,提取基因组DNA,采用长随机引物和内含子切接点引物的PCR分子标记技术,对甜荞品种内和品种间的遗传多样性进行了分析.结果表明:(1)所使用的引物均能在 9份甜荞品种和1份苦荞品种中稳定扩增出条带,共获得了45条条带,其中多态性条带42条,占总数的93.3%;(2)甜荞同一品种内存在一定的遗传差异,但遗传距离较小,大部分不同甜荞品种间的遗传差异大于其品种内的遗传差异.因而,在甜荞品种鉴定和遗传多样性研究中,通过品种内适当混合取样可以较好地代表和反映甜荞品种特性.     

几种荞麦的抗氧化酶活性研究

测定了荞麦属六种十个居群的SOD、CAT、POD和ASP活性。结果表明在栽培荞麦中,苦荞的抗氧化酶活性高于甜荞;野生荞麦中差异很大,抗氧化酶活性高的有金荞和贵州云台山大野荞。     

苦荞和甜荞查尔酮合成酶基因的克隆及序列比较

以苦荞品种‘西农9920’和甜荞品种‘西农9976’为材料,根据其它植物查尔酮合成酶(chalcone synthase,CHS)基因DNA序列的保守区域设计的一对简并引物,进行PCR扩增,从2种荞麦基因组中克隆出了长度均为860 bp的CHS基因片段,对其进行回收、克隆,挑选阳性克隆测序;序列分析表明这2个片段含有CHS基因的N端和C端的结构域,分别为苦荞和甜荞的CHS基因片段,命名为FtCHS和FeCHS。对获得的2种荞麦CHS基因的DNA序列进行比较分析,发现两者间存在多达43处单碱基多态性,这些单碱基多态性可能是苦荞和甜荞种子中类黄酮含量差异的重要原因之一。苦荞和甜荞CHS与其它植物CHS的氨基酸序列的进化分析表明,其与同为蓼科的掌叶大黄和石竹科的满天星的同源性较近。     

普通荞麦资源的耐铝性研究

利用小容器溶液培养法对耐铝性鉴定条件和52份普通荞麦栽培品种资源的耐铝性进行了研究。结果发现普通荞麦耐铝性鉴定的适宜条件为发芽种子于500μmol/LAlCl3溶液(pH4.5)处理3d,以发芽种子在这三天内的根伸长量衡量耐铝性程度。在该处理条件下,普通荞麦不同品种间的耐铝性有显著差异。其中,陕西大红花甜荞品种、日本大粒荞、织金红花甜荞的耐铝毒胁迫能力最强,值得在荞麦耐铝性育种和耐铝机制研究中利用。     

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.     

Hybridization between Fagopyrum (Polygonaceae) species native to China

Thirty-eight crosses between seven Fagopyrum species native to China were studied. Following normal pollination, there were very low inter-group crossabilities (only 0 3%) between five species in the large-achene group and two species (E gracilipes and E pleioramosum) in the small-achene group, and high (15 50%) within the large-achene group, i.e. E esculentum, E tataricum, E zuogongense, E pilus and E megaspartanium. Crosses between various types of (lowers (homostyle, heterostyle etc.) suggested that the intraspecific incompatibility also occurs in interspecific crosses. The hybrid seeds obtained from interspecific crosses all withered or shrivelled and lacked activity. Only one hybrid plant was obtained, tetraploid E esculentum x tetraploid E zuogongense. The hybrid displayed anomalous behaviour at meiosis, pollen grain sterility and withering of achenes, confirming the genetic differences between E esculentum and E zuogongense. The results support the hypothesis that E megaspartanium and E pilus are the possible ancestors of cultivated buckwheat     

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。     

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.     

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)。     

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.