苦荞SSR分子遗传图谱的构建及分析

构建苦荞遗传连锁图谱,为今后有关苦荞基因组结构、重要农艺性状QTL定位、分子标记辅助育种和基因克隆等研究工作奠定基础。以栽培苦荞‘滇宁一号’和苦荞野生近缘种杂交产生的119份F4代分离材料为作图群体,利用SSR分子标记来构建苦荞的分子遗传连锁图谱。本研究构建的连锁图谱包含15个连锁群,由89个标记组成,其中偏分离的标记有22个,占24.7%,每条连锁群上的标记在2~16之间。连锁群长度在6.9~165.8 cM的范围,覆盖基因组860.2 cM,总平均长度9.7 cM。本研究构建了首张苦荞SSR遗传连锁图谱,为苦荞QTL定位、基因克隆、遗传选育等研究奠定了基础。     

Genetic analyses of agronomic traits in Tartary buckwheat (Fagopyrum tataricum (L.) Gaertn.)

The consumption of products made from Tartary buckwheat (Fagopyrum tataricum (L.) Gaertn.) has increased in recent years in Japan. Increased consumer demand has led to recognition of the need for early varieties of this crop with high and stable yields. In order to accomplish this, more information is needed on the genetic mechanisms affecting earliness and yield. We conducted genetic analysis of 3 agronomic traits (days to flowering, plant height and total seed weight per plant) to segregate F₂ and F₃ populations derived from a cross between Tartary buckwheat cultivars ‘Hokuriku No. 4’ and ‘Ishisoba’. Broad-sense heritability estimates for days to flowering, plant height and total seed weight were 0.70, 0.62 and 0.75, respectively, in F₃ population. Narrow-sense heritability for total seed weight (0.51) was highest, followed by heritability for days to flowering (0.37), with heritability for plant height (0.26) lowest. Later flowering was associated with increased plant height and higher yields. From the F₄ generation, we identified twelve candidate plants with earlier maturity and reduced plant height compared to ‘Hokuriku No. 4’, but almost the same total seed weight. These results suggest that hybridization breeding using the single seed descent (SSD) method is an effective approach for improving agronomic characteristics of Tartary buckwheat.     

Discovery and genetic analysis of non-bitter Tartary buckwheat (Fagopyrum tataricum Gaertn.) with trace-rutinosidase activity

In a screening of about 500 lines of Tartary buckwheat, we identified lines that contained no detectable rutinosidase isozymes using an in-gel detection assay. We confirmed that seeds of these individuals had only a trace level of in-vitro rutinosidase activity. To investigate the heritability of the trace-rutinosidase characteristic, we analyzed the progeny of crosses between rutinosidase trace-lines, ‘f3g-162’, and the ‘Hokkai T8’. The F₂ progeny clearly divided into two groups: those with rutinosidase activity under 1.5 nkat/g seed (trace-rutinosidase) and those with activity over 400 nkat/g seed (normal rutinosidase). The segregation pattern of this trait in F₂ progeny exhibited 1 : 3 ratio (trace-rutinosidase : normal rutinosidase), suggesting that the trace-rutinosidase trait is conferred by a single recessive gene; rutinosidase-trace A (rutA). In addition, sensory panelists evaluated the bitterness of flour from trace-rutinosidase individuals and did not detect bitterness, whereas flour from normal rutinosidase individuals was found to have strong bitterness. Although at least three bitter compounds have been reported in Tartary buckwheat seeds, our present findings indicate that rutin hydrolysis is the major contributing factor to bitterness. In addition, the trace-rutinosidase line identified here, ‘f3g-162’, is a promising material for generating a non-bitter Tartary buckwheat variety.     

发展中国的苦荞生产

荞麦在许多国家都有栽培,但主要集中在北半球。我国栽培荞麦的历史悠久,荞麦栽培种有甜荞 (Fagoｐyrumsagittatum )和苦荞(Fagoｐyrumtataricum)两个品种,甜荞主要分布在长江以北地区,苦荞主要分布在长江以南地区。我国是世界苦荞的集中产区,苦荞种植面积和产量均居世界第一,常年种植面积在100万hm² 以上,在四川、云南、贵州和西藏等西部省区的一些干旱高寒山区、少数民族地区、边远山区具有明显的生产优势。1　苦荞的营养价值苦荞具有很高的营养食用价值,用苦荞麦制作的荞米和面粉为我国西部一些地区人民的主食,在许多地方用苦荞制作风味小吃,苦荞是一种既无……     

Random amplified polymorphic DNA (RAPD) variation in Fagopyrum tataricum Gaertn. accessions from China and the Himalayan region

The diversity among 52 landraces and cultivars of tartary buckwheat (Fagopyrum tataricum Gaertn.) and one accession of its wild ancestor, F. tataricum ssp. potanini Batalin, from diverse geographic origins was examined using random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR) markers. Eighteen primers produced a total of 240 fragments, of which 153 (63.75%) were monomorphic and 87 (36.25%) polymorphic bands. UPGMA-based pairwise Jaccard’s coefficient of similarity was used to deduce the relationships among 53 genetically diverse accessions. The similarity between cultivated tartary buckwheat accessions ranged from 0.61 to 1.00. Four distinct clusters were formed which corresponded well with the geographic distribution of the tartary buckwheat. Nepalese accessions showed maximum diversity followed by Chinese accessions. Tartary buckwheat accessions from the Himalayan region of northwestern India revealed a narrow gene pool. The wild buckwheat accession did not group with any of the three cultivated tartary buckwheat groups, and formed its own single-entry group. Genetic similarity (0.59) of Chinese buckwheat accessions with the wild ancestor reaffirmed that cultivated tartary buckwheat originated in the Yunnan province of northwestern China. Consistent with some earlier reports, our study demonstrated the usefulness of the RAPD technique for the characterization of plant genetic resources and assessment of diversity between species. This revised version was published online in August 2006 with corrections to the Cover Date.     

ITS序列在苦荞麦种质资源鉴定中的应用

为了更精确有效地鉴定苦荞麦种质资源,根据ITS序列通用引物,以苦荞麦基因组DNA为模板通过PCR方法扩增出ITS(internal translation spacer)序列,在克隆测序的基础上对荞麦属5个栽培苦荞麦中进行分子鉴定与亲缘关系分析。结果表明：（1）5个供试材料ITS序列的变异小,序列长度在584~590 bp之间,G+C含量高,为67.5%~68.5%。（2）通过聚类方法分析发现‘建德苦荞麦’和‘红花苦荞麦’聚为一支,‘西荞2号’和‘西荞3号’聚为一支。     

In vitro germination and viability of buckwheat (Fagopyrum esculentum Moench) pollen

An in vitro method for the germination of common buckwheat pollen was developed. Pollen grains were successfully germinated in an artificial medium consisting of 0.2 g each of MnSO₄, Ca(NO₃)2.4H₂O and KNO₃, 0.04 g H₃BO₃, 15 g sucrose and 30 g polyethylene glycol (molecular weight approximately 20,000) dissolved in 100 ml of double distilled water. The viability of pollen was assessed by in vivo and in vitro germination tests at 20 °C and 25 °C over a 38 h time period. Pollen grains were collected and germinated at 4 h intervals from freshly harvested flowers grown under 16 h day length and a constant temperature. Maximum pollen viability was found 2 h and 6 h after first light when plants were maintained at 25 °C and 20 °C, respectively. Viability, as measured by germination percentage, was similar at both temperature regimes. Some pollen remained viable for approximately 34 to 38 h in intact flowers, but all pollen lost viability in less than an hour when stored at room temperature without humidity control. This revised version was published online in July 2006 with corrections to the Cover Date.     

苦荞漆酶基因的克隆与生物信息学分析

利用RT-PCR技术从苦荞中克隆出2条漆酶（laccase）基因,运用生物信息学技术对2个基因序列进行分析,预测结构域、二级和三级蛋白结构,进行蛋白同源比对及进化树分析。结果表明,FtLAC-1基因序列开放阅读框为1695bp,编码564个氨基酸,FtLAC-2基因序列开放阅读框为1707bp,编码568个氨基酸。FtLAC-1和FtLAC-2蛋白预测分子量分别为61.41和62.47kDa,理论等电点分别为9.45和9.41,为亲水性蛋白,2个基因序列相似性较低,氨基酸序列同源性不高。qRT-PCR分析出其在苦荞不同组织器官存在差异性表达。结论为进一步探索FtLAC-1和FtLAC-2在苦荞薄果壳形成中的功能提供理论基础。     

Seed protein variation for identification of common buckwheat (Fagopyrum esculentum Moench) cultivars

Summary Total seed proteins of 24 common buckwheat cultivars and cultivated populations within a molecular weight range of 30 to 54 kDa were analysed by SDS-PAGE. Single seed analysis of six cultivars identified a total of 18 alternative protein bands with different mobilities. Differences of individual protein band frequencies extracted from single seeds among six buckwheat cultivars varied distinctively, indicating high intravarietal polymorphism. The relation between frequencies of protein bands revealed by single seed analysis and their appearance on the bulk seed analyses was demonstrated. Regarding to band mobility rate and relative band intensity among 24 bulk samples analysed, 14 had distinctive electrophoregrams while the other 10 were ranged into four distinct groups. Analysis of endosperm and cotyledon proteins showed that proteins stored in these main seed parts are tissue specific. The observed electrophoretic polymorphism related to proteins stored in the cotyledons while there was no apparent variability with endosperm proteins.     

四川省苦荞麦的开发利用

苦荞为我国独有的栽培种,其味略苦,营养价值和药用价值一向鲜为人知。近年来,随着科学研究的深入,苦荞麦的营养价值逐渐引起国内外人们的关注和重视。目前,利用苦荞麦加工而成的各种糕点、快餐食品、保健食品、营养食品深受人们欢迎,其原粮和成品已逐渐销往国际市场。四川省苦荞麦栽培面积大,种植历史悠久,品种资源丰富,随着开发利用的深入,这种融营养、保健于一身的绿色食品将越来越受到人们青睐。1四川省苦荞麦开发利用现状四川省荞麦栽培面积约10万公顷,其中以凉山州为最大,常年种植面积达6万公顷,其它分布在阿坝州、甘孜州及盆周山区,盆…     

Effect of sowing time on buckwheat (Fagopyrum esculentum Moench) growth and yield in central Korea

Buckwheat (Fagopyrum esculentum MOENCH) is a traditional Korean crop that gaining global attention as a health food due to its rich nutrition. Climate change effects such as high temperature, cold weather, flooding, and drought induce plant stress, reduced production yield of upland crops. In general, buckwheat is grown during the fall season in Korea. Flooding is a severe environmental stress factor, and flooding during the early growth stage of buckwheat causes large reductions in yield. Therefore, we established optimum growth parameters to maximize buckwheat yield. The optimum temperature for harvesting of buckwheat was approximately 1,083 ~ 1,515°C. The number of growing degree days (G.D.D.) from flowering to harvest was 509 ~ 673°C. Plant hight, stem diameter, and dry matter weight were reduced as planting date was delayed (p < 0.05). We investigated buckwheat sowing time during fall, and established an optimum cropping system according to the local climate conditions in central Korea. The growth period of buckwheat cultivar ‘Yangjueol’ was approximately 56 ~ 60 days during fall, and yield was higher than that of cultivar ‘Daesan’. The results indicated that buckwheat yield in central Korea was maximized by sowing cultivar on 25th August and cultivar ‘Daesan’ on 15th August. The yield of Yangjeol sown in spring was 119 kg 10a^-1, whereas the yield of Daesan sown in summer was 158kg 10a^-1. The results of this study established the effects of sowing time on growth and yield in central Korea. We propose an optimum cropping system for improving quality.     

Development of semidwarf breeding line with shattering resistance and preharvest-sprouting resistant in buckwheat (Fagopyrum esculentum)

An important breeding issue in buckwheat is to increase its resistance to lodging, which can result in the loss of yield and quality of buckwheat foods. We developed the semidwarf buckwheat ‘Kyukei 50’, which has a short plant height and a strong lodging resistance. The internode length of ‘Kyukei 50’ was shorter than that of the summer-sown standard variety ‘HITACHIAKISOBA’. Progeny analysis revealed that the semidwarf trait was dominated by at least one recessive gene. Compared with ‘HITACHIAKISOBA’, in ‘Kyukei 50’, the time of maturation was earlier, and the pre-harvest sprouting resistance was the same, indicating that ‘Kyukei 50’ is suitable for sowing in both spring and summer. ‘Kyukei 50’ exhibits the ‘green flower type’ shattering resistance trait and, therefore, has a thick pedicel; breaking tensile strength is approximately two-fold stronger than that of ‘HITACHIAKISOBA’. The yield of ‘Kyukei 50’ is the same as that of ‘HITACHIAKISOBA’. In addition, the lowest seed height of ‘Kyukei 50’ was >17 cm. These results indicate that the ‘Kyukei 50’ is a promising breeding line for lodging resistance and loss of harvesting machinery.     

Morpho-physiological response of common buckwheat (Fagopyrum esculentum) to flooding stress at different growth stages

Journal of Crop Science and Biotechnology - Flooding is one of the major abiotic stresses which accounts for considerable damage to plant growth and development and induces multiple...     

苦荞品种(系)主要农艺性状与蛋白质含量的聚类分析

对来自5个国家的55份苦荞的9个主要农艺性状及蛋白质含量进行了主成分分析和聚类分析.结果表明:供试材料具有中、矮秆,主茎节数、有效花序数、蛋白质含量适中,一级分枝数偏多、总分枝数偏少,千粒重、单株粒重偏低和生育期较长等特点.主成分分析将9个主要农艺性状简化为5个综合指标,其累积贡献率达87.77%.入选材料应是主茎节数、一级分枝数、总分枝数适中,有效花序数和千粒重偏高,中矮秆的材料.据此标准,筛选到九江苦荞和苦刺荞2份综合农艺性状优异的材料.基于性状表现,供试材料可分为低产晚熟型、多分枝高蛋白型、早熟粒重型等3类,但聚类结果与地理来源并不完全一致.     

Emasculation of Tartary buckwheat (<Emphasis Type="Italic">Fagopyrum tataricum</Emphasis> Gaertn.) using hot water

Hot water treatment was applied to Tartary buckwheat (var. ‘Hokkai T8’) for emasculation. Apical clusters of flower buds were soaked in a constant temperature water bath (42–44°C) in the afternoon and then the flowers which opened in the next morning were used for the investigation of self-fertilization and the artificial pollination. Hand pollination was carried out by rubbing the anthers taken from the male parent (var. ‘Hokkai T10’) to the stigma of the flowers treated by hot water. The seed set by self-fertilization was almost 0% when treated at 42°C for 8 min or more, at 43°C for 5 min or more and at 44°C for 3 min or more. With artificial pollination, the best performance was obtained when treated at 44°C for 3 min, in which the seed set was 55.1% and the matured seed was obtained 37.7% of the hand-pollinated flowers. Nine out of 26 progeny seedlings emerged and the cotyledonal color of them were all reddish green, probably indicating the hybrid of ‘Hokkai T8’ and ‘Hokkai T10’. The hybridization was reconfirmed by the segregation in F₂. The hot water treatment at 44°C for 3 min was available to other seven varieties. From 24.5% to 100% of the hand-pollinated flowers set mature seeds though only one self-fertilized seed remained without artificial pollinations. This hot water treatment must be practically useful in hybridization breeding of Tartary buckwheat.     

Inter-varietal variations of rutin content in common buckwheat flour (Fagopyrum esculentum Moench.)

Summary To improve the quality of buckwheat flour, we investigated the effect of the cropping season on the rutin content in various cultivars of common buckwheat (Fagopyrum esculentum Moench.). Thirty cultivars of buckwheat were grown under both long day conditions (the summer cropping) and short day conditions (the late summer cropping). The inter-varietal variations and the effect of the cropping season on the rutin content were examined. The rutin content was determined by high performance liquid chromatography (HPLC). Results show that the average rutin content in the late summer cropping was about half that of the summer cropping. There were wide inter-varietal variations of rutin content in the summer cropping. The differences in rutin content in the late summer cropping were less pronounced. Rutin content was closely correlated with the flowering period in summer (r=0.735^***). The later flowering cultivars produced a higher content of rutin than the earlier flowering cultivars in the summer cropping. The differences in the cumulative solar radiation received in the two seasons seems to be a reasonable explanation for this increase in the rutin content of the buckwheat flour. The rutin content of early flowering cultivars, which are suitable for long day conditions, is positively correlated with yield in the summer cropping. However, the rutin content in the intermediate and late summer ecotype cultivars, which are suitable for short day conditions, shows no correlation with yield in the late-summer cropping. In conclusion, there are wide inter-varietal variations of rutin content in buckwheat flour and the rutin content increases under long day conditions.     

Rapid genotyping with DNA micro-arrays for high-density linkage mapping and QTL mapping in common buckwheat (Fagopyrum esculentum Moench)

For genetic studies and genomics-assisted breeding, particularly of minor crops, a genotyping system that does not require a priori genomic information is preferable. Here, we demonstrated the potential of a novel array-based genotyping system for the rapid construction of high-density linkage map and quantitative trait loci (QTL) mapping. By using the system, we successfully constructed an accurate, high-density linkage map for common buckwheat (Fagopyrum esculentum Moench); the map was composed of 756 loci and included 8,884 markers. The number of linkage groups converged to eight, which is the basic number of chromosomes in common buckwheat. The sizes of the linkage groups of the P1 and P2 maps were 773.8 and 800.4 cM, respectively. The average interval between adjacent loci was 2.13 cM. The linkage map constructed here will be useful for the analysis of other common buckwheat populations. We also performed QTL mapping for main stem length and detected four QTL. It took 37 days to process 178 samples from DNA extraction to genotyping, indicating the system enables genotyping of genome-wide markers for a few hundred buckwheat plants before the plants mature. The novel system will be useful for genomics-assisted breeding in minor crops without a priori genomic information.     

Tartary buckwheat breeding (<Emphasis Type="Italic">Fagopyrum tataricum</Emphasis> L. Gaertn.) through hybridization with its Rice-Tartary type

Tartary buckwheat (Fagopyrum tataricum) is a highly nutritious crop that is widely grown in Asia, but the flour contains a large portion of the hull if it is ground with industrial processing since the hull is very hard to remove as it adheres to the testa layer of the groat. Rice-Tartary, a particular type of Tartary buckwheat, has seeds with a loose hull and the presence of splits on the sides of the seeds that make dehulling easily. The present study reports on the first attempt of crop improvement in Tartary buckwheat breeding through hybridization. Hybrids were obtained by hybridization of crosses between Tartary buckwheat and Rice-Tartary. Additional crosses were made among selected progenies of these crosses. Based on progeny analysis of the F₁, F₂, and F₃, the character of Rice-Tartary, as exhibited as the presence of splits on the sides of the seeds, is controlled by one gene which is homozygous recessive. A Tartary buckwheat breeding program has been conducted for 6 years based on these crosses. Advanced lines with easy dehull and yield potential are at the stage of ready for yield trials. Production of easy dehulling Tartary buckwheat in the future could boost Canada’s domestic and international markets.     

Effects of cultivar and environment on phenolic acids in buckwheat

Summary Phenolic acid contents were determined in seeds from five buckwheat cultivars grown at three locations in western Canada for four years. Buckwheat contained 12–16 g/kg of total phenolic acids, about 3 g/kg of esterified phenolic acids, and 8–13 g/kg of etherified phenolic acids. The latter represented 70 to 79% of the total phenolics. Variation in phenolic acids was mainly due to cultivar, seasonal effects and their interaction, while growing location had no significant effect. Phenolic acid contents of buckwheat were independent of seed color and protein content. Esterified phenolic acids exhibited a negative significant association with etherified phenolic acids in buckwheat.