云南苦荞种质资源主要性状的遗传多样性分析

为了从云南苦荞种质资源中挖掘优异种质资源,拓宽苦荞遗传基础,以48份苦荞种质资源为材料研究了6个主要农艺性状和5个品质性状的遗传多样性。结果表明,云南的苦荞资源存在着丰富的遗传多样性,6个农艺性状中株粒重的变异系数为34.4%最大,品质性状中总黄酮含量的变异系数为51.72%最大。聚类结果表明,将48份材料聚为3大类,可区分为低产型、矮秆高产型和中秆高产型。6个主要农艺性状和5个品质性状的主成分分析结果表明,前3个累计贡献率分别达84.105%和80.332%,各主成分性状载荷值反映了主要数量性状的育种选择潜力。综合分析种质资源的主要农艺性状,可为云南苦荞种质资源的利用提供有效的科学依据。     

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。     

苦荞地方种质资源的遗传多样性分析

利用SSR分子标记技术对中国苦荞主产区陕西、云南、四川、西藏等地的82份苦荞地方种质的遗传多样性进行了分析,以揭示中国特有的作物种质--苦荞地方种质资源的遗传多样性,促进苦荞优良品种的选育.结果显示:(1)所用25个SSR引物中有13个引物在苦荞地方品种中具有多态性,且扩增条带的稳定性较好,共扩增出208条条带,其中多态性条带200条,占总数的96.2%;(2)聚类分析结果显示,82份苦荞材料的遗传相似系数(GS)分布于0.52～0.85之间,平均值为0.69,在GS值为0.722的水平上,82份材料被聚为10大类群.研究表明82份苦荞品种间遗传多样性明显,具有丰富的遗传基础.     

In situ conservation of crop genetic resources through maintenance of traditional farming systems

A strategy is suggested for in situ conservation of crop genetic resources whereby conservation efforts are linked to rural development projects in Third World countries. We describe development projects that emphasize preservation of traditional farming systems and succeed in sustaining production by relying on the maintenance of biological and genetic diversity in these systems. Basing agricultural development efforts on indigenous knowledge, technology, and social organization can provide important guidelines for the design of cropping systems that allow lowincome farmers to produce subsistence and cash crops without dependence on external inputs and seed supplies. By incorporating landraces and wild relatives of crops into these cropping systems, major achievements in the conservation of crop genetic resources can be obtained.     

Genetic Diversity of Buckwheat Cultivars (<Emphasis Type="Italic">Fagopyrum tartaricum</Emphasis> Gaertn.) Assessed with SSR Markers Developed from Genome Survey Sequences

Tartary buckwheat is an important edible crop as well as medicinal plant in China. More and more research is being focused on this minor grain crop because of its medicinal functions, but there is a paucity of molecular markers for tartary buckwheat due to the lack of genomics. In this study, a genome survey was carried out in tartary buckwheat, from which SSR markers were developed for analysis of genetic diversity. The survey generated 21.9 Gb raw sequence reads which were assembled into 348.34 Mb genome sequences included 204,340 contigs. The genome size was estimated to be about 497 Mb based on K-mer analysis. In total, 24,505 SSR motifs were identified and characterised from this genomic survey sequence. Most of the SSR motifs were di-nucleotide (67.14 %) and tri-nucleotide (26.05 %) repeats. AT/AT repeat motifs were the most abundant, accounting for 78.60 % of di-nucleotide repeat motifs. SSR fingerprinting of 64 accessions yielded 49.71 effective allele loci from a total of 63 with the 23 polymorphic SSR primer combinations. Analyses of the population genetic structure using SSR data strongly suggested that the 64 accessions of tartary buckwheat clustered into two separate subgroups. One group was mainly distributed in Nepal, Bhutan and the Yunnan-Guizhou Plateau regions of China; the other group was mainly derived from the Loess Plateau regions, Hunan and Hubei of China and USA. The cluster analysis of these accession’s genetic similarity coefficient by UPMGA methods strongly supported the two subgroup interpretation. However accessions from Qinghai of China could be grouped into either of the two subgroups depending on which classification method was used. This region is at the intersection of the two geographical regions associated with the two subgroups. These results and information could be used to identify and utilize germplasm resources for improving tartary buckwheat breeding.     

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.     

虫媒传粉植物荞麦的生物学特性与研究进展

荞麦是禾本科之外的谷物类作物, 具有较高的营养和药用价值。栽培荞麦有甜荞(Fagopyrum esculentum)和苦荞(F. tartaricum), 这两种一年生草本分别为自交不亲和的二型花柱、自交亲和的同型花柱植物; 前者结实依赖昆虫传粉。根据国内外调查研究, 前人对蓼科荞麦属(Fagopyrum)记录了30个物种名, 已有形态学和遗传多样性的调查表明该属的物种多样性中心位于我国西南地区, 特别是长江上游的三江并流区域; 甜荞和苦荞的起源地和祖先物种也认为在该区域。本文在论述前人研究的基础上, 指出对荞麦属的分类修订、野生种质资源的分布、种间关系的调查、优良品种的选育亟待研究。孢粉学和考古学的证据显示在我国长江流域, 人们在4,500年前就开始种植荞麦。荞麦可能曾经是山区人民的主粮, 为孕育长江流域文明提供了食物资源。加强对荞麦基础生物学特性的研究, 运用现代基因组学的方法有望澄清栽培荞麦的起源并探究产量不高的原因, 挖掘和利用其经济和药用价值的性状, 为荞麦成为一类优良的粮食作物提供参考依据。     

二倍体和四倍体栽培苦荞的细胞学比较研究

观察和统计了二倍体和四倍体苦荞 (Fagopyrum tataricum)花粉母细胞减数分裂过程 TI、T 时期的异常程度及花粉粒败育性等多个指标 ,并比较分析了二倍体和四倍体苦荞在这些指标上的差异显著性。结果显示 :四倍体苦荞的异常 TI细胞频率、异常 T 细胞频率、TI细胞平均微核数、T 细胞平均微核数及花粉败育率均显著高于二倍体苦荞 ,这表明四倍体苦荞的遗传稳定性和花粉可育性都比二倍体苦荞差。     

基于PAL基因序列的地方苦荞品种遗传多样性分析

苦荞在中国具有广泛的栽培种植地区,长时间演化形成了丰富的遗传多样性。为了研究和利用苦荞资源,以国内北方苦荞产区(内蒙古、青海、陕西、山西、甘肃)、西南苦荞产区(西藏、四川、贵州、云南)、国内其他地方品种(江西、安徽、湖北、湖南、广西)及国外品种(尼泊尔)共计67份苦荞材料为研究对象,PCR扩增其PAL基因并测序。在此基础上分析苦荞的遗传多样性,并采用NJ法(neighbor-joining)对67份苦荞材料构建系统进化树。结果表明,供试的67份苦荞材料的PAL基因序列长度为2011 bp,其中,变异位点为160个,占序列总长度的7.9%,简约信息位点为33个,占序列总长度的1.64%,突变的类型主要是碱基的转换与颠换,高变异位点均集中在外显子2的N端。不同来源的苦荞材料间的遗传距离分布于0.002~0.016之间,来源于中国四川组的苦荞材料种内平均遗传距离最大(0.016),中国内蒙古组的最小(0.002)。中国四川地区的材料与其他地区来源的材料间的遗传距离位于0.010~0.016之间,而其他地区间的遗传距离为0.004~0.013。67份苦荞材料的平均π值和θ值分别为0.0034和0.0143。其中,中国四川材料的π值为0.0148。基于PAL基因序列构建的NJ进化树中,67份苦荞材料分为7个类群,分类与地理来源无关。仅中国西藏来源的5份材料聚集为一类,说明PAL基因序列较为稳定,多数材料间变化差异较小。中国四川地区的苦荞材料具有丰富的遗传多样性,中国西藏地区的某一材料中有较多的SNP位点,推测中国西藏的部分材料可能存在突变的热点区,预示着PAL基因新的突变位点区域。     

农作物遗传多样性农家保护的现状及前景

农作物地方品种的有效保护是农业生物多样性的可持续利用的基础。由于现代农业的集约化生产方式使大量农作物地方品种被少数高产改良品种所取代,造成农作物基因库的严重“基因流失”（genetic erosion)。农家保护是在农业生态系统中进行的动态保护,被保护的生物多亲性可在其生境中继续进化而产生新的遗传变异,在而是农业生物多样性就地保护的重要途径。然而,尽管人们对作物品种资源农家保护的兴趣不断增长,也有大量有关农家的保护的研究和案例分析,但目前为止还没有比较成功的农家保护实例报道。因此,对农家保护的机制及科学问题进行深入的研究,并寻求一条新的途径来充分发挥农家保护应有的作用,显得格外重要。利用生物多样性布局的水稻混合间栽的生产模式,不仅解决了病害控制的问题,而且也保护了水稻地方品种的多样性。这种混合间栽的生物多样性布局和生产方式可能成为农保护的一条新途径。     

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。     

Purification and characterization of a 24 kDa protein from tartary buckwheat seeds

A 24 kDa protein was isolated from tartary buckwheat seeds by using chromatography of Superdex 75 gel filtration and Resource Q ion-exchange column. SDS-PAGE and Sephacryl S-200 gel filtration were used to provide information about the molecular mass of the protein purified from tartary buckwheat. The protein was composed of 215 amino acid residues and showed strong IgE binding activity in an ELISA test to the sera colleted from two patients allergic to buckwheat. These results suggested that the purified 24 kDa protein from tartary buckwheat seeds was an important functional protein and was relatively specific for buckwheat-allergic patients. It should be a very useful tool in the diagnosis of buckwheat allergy in the future.     

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。     

Mycorrhizal status and diversity of fungal endophytes in roots of common buckwheat (Fagopyrum esculentum) and tartary buckwheat (F. tataricum)

To determine the mycorrhizal status and to identify the fungi colonising the roots of the plants, common buckwheat (Fagopyrum esculentum) and tartary buckwheat (F. tataricum) were inoculated with an indigenous fungal mixture from a buckwheat field. Root colonisation was characterised by the hyphae and distinct microsclerotia of dark septate endophytes, with occasional arbuscules and vesicles of arbuscular mycorrhizal fungi. Sequences of arbuscular mycorrhizal fungi colonising tartary buckwheat clustered close to the Glomus species group A. Sequences with similarity to the Ceratobasidium/Rhizoctonia complex, a putative dark septate endophyte fungus, were amplified from the roots of both common and tartary buckwheat. To the best of our knowledge, this is the first report of arbuscular mycorrhizal colonisation in tartary buckwheat and the first molecular characterisation of these fungi that can colonise both of these economically important plant species.     

Combined effects of elevated UV-B radiation and the addition of selenium on common (<Emphasis Type="Italic">Fagopyrum esculentum</Emphasis> Moench) and tartary [<Emphasis Type="Italic">Fagopyrum tataricum</Emphasis> (L.) Gaertn.] buckwheat

The combined effects of UV-B irradiation and foliar treatment with selenium on two buckwheat species, common (Fagopyrum esculentum Moench) and tartary [Fagopyrum tataricum (L.) Gaertn.] buckwheat, that underwent different intensity of breeding, were examined. Plants grown outdoors under three levels of UV-B radiation were studied for 9 weeks, from sowing to ripening. At week 7 they were sprayed with solution containing 1 g(Se) m⁻³ that presumably mitigates UV-B stress. Morphological, physiological, and biochemical parameters of the plants were monitored. Elevated UV-B radiation, corresponding to a 17 % reduction of the ozone layer, induced synthesis of UV absorbing compounds. In both buckwheat species it also caused a reduction in amounts of chlorophyll a during the time of intensive growth, an effect, which was increased in tartary buckwheat in the presence of selenium. The respiratory potential, measured as terminal electron transport system activity, was lower in plants subjected to enhanced UV-B radiation during the time of intensive growth. The effective quantum yield of photosystem 2 was also reduced due to UV-B radiation in both buckwheat species and was mitigated by the addition of Se. Se treatment also mitigated the stunting effect of UV-B radiation and the lowering of biomass in common buckwheat.     

Integrating gene flow, crop biology, and farm management in on-farm conservation of avocado (Persea americana, Lauraceae)

Maintaining crop diversity on farms where cultivars can evolve is a conservation goal, but few tools are available to assess the long-term maintenance of genetic diversity on farms. One important issue for on-farm conservation is gene flow from crops with a narrow genetic base into related populations that are genetically diverse. In a case study of avocado (Persea americana var. americana) in one of its centers of diversity (San Jerónimo, Costa Rica), we used 10 DNA microsatellite markers in a parentage analysis to estimate gene flow from commercialized varieties into a traditional crop population. Five commercialized genotypes comprised nearly 40% of orchard trees, but they contributed only about 14.5% of the gametes to the youngest cohort of trees. Although commercialized varieties and the diverse population were often planted on the same farm, planting patterns appeared to keep the two types of trees separated on small scales, possibly explaining the limited gene flow. In a simulation that combined gene flow estimates, crop biology, and graft tree management, loss of allelic diversity was less than 10% over 150 yr, and selection was effective in retaining desirable alleles in the diverse subpopulation. Simulations also showed that, in addition to gene flow, managing the genetic makeup and life history traits of the invasive commercialized varieties could have a significant impact on genetic diversity in the target population. The results support the feasibility of on-farm crop conservation, but simulations also showed that higher levels of gene flow could lead to severe losses of genetic diversity even if farmers continue to plant diverse varieties.     

Determination of Anomeric Configuration of Furanoside Derivatives by Carbon-13 NMR

A 24 kDa protein was isolated from tartary buckwheat seeds by using chromatography of Superdex 75 gel filtration and Resource Q ion-exchange column. SDS-PAGE and Sephacryl S-200 gel filtration were used to provide information about the molecular mass of the protein purified from tartary buckwheat. The protein was composed of 215 amino acid residues and showed strong IgE binding activity in an ELISA test to the sera colleted from two patients allergic to buckwheat. These results suggested that the purified 24 kDa protein from tartary buckwheat seeds was an important functional protein and was relatively specific for buckwheat-allergic patients. It should be a very useful tool in the diagnosis of buckwheat allergy in the future.     

Crop Genetic Resources Diversity and Sustainable Agriculture in Yunnan Province,China

Yunnan is well known as a "Kingdom of Plant". Yunnan is also rich in natural resources of crop. Lots of studies on collection, conservation, protection, evaluation and utilization of crop genetic resources have been done in recent years. But there are some unfavourable factors, too. With the agricultural intensity, some new problem such as decreasing diversity, simplifying varieties and narrowing genetic basis of crop have arisen to restrict the development of agriculture in Yunnan. The only way to achieve a sustainable agriculture in Yunnan is doing our utmost to protect the crop diversity, develop new crop or new varieties and widen the genetic basis of crop varieties.     

Proximity to wildflower strips did not boost crop pollination on small,diversified farms harboring diverse wild bees

The yield of many agricultural crops depends on pollination services provided by wild and managed bees, many of which are experiencing declines due to factors such as reductions in floral resources. Thus, improving pollinator habitat on farmlands using management strategies like planting wildflower strips is vital for wild bee conservation and sustainable crop pollination. Yet, few studies have examined whether and at what spatial scales wildflower strips enhance crop pollination and yields, and most research has been conducted in large-scale commercial agriculture. Therefore, we investigated the effects of wildflower strips on crop pollination on small, diversified farms (i.e., those growing a variety of crop species) where wild bee diversity and abundance is predicted to be comparatively high. Over three years, on four diversified farms in Montana USA, we tested the hypothesis that distance (20, 60, and 180 m) of crops from native perennial wildflower strips planted alongside crop fields affected wild bee visitation, pollination, and yields of squash and sunflower crop plants. We found that distance to wildflower strips did not affect bee visitation or pollination in crops. Squash yield was pollen-limited in the growing season prior to wildflower strip establishment, and in one of the two years after wildflower strip establishment, but proximity to wildflower strips did not influence the magnitude of pollen limitation. Sunflower seed production was not pollen-limited in any year. Our findings demonstrate that even on diverse farms with wildflower strips and a demonstrated high diversity of bees, some crops do not necessarily receive maximum pollination, regardless of distance from the wildflower strips. However, the value of wildflower strips for supporting wild bee diversity, and other ecological or economic benefits, needs consideration for a full understanding of this pollinator habitat management strategy.     

Nutritional consequences of the transformation from subsistence to commercial agriculture in Tabasco,Mexico

Recent changes in the area of the Plan Chontalpa in Tabasco have greatly reduced the production of subsistence crops by rural families, resulting in decreased crop diversity and a concomitant increase in the degree of dependence on outside sources of food. Results from a nutrition survey of 149 families demonstrate that dietary diversity, dietary quality, and nutritional status of preschool children are negatively associated with lower crop diversity and increased dependence on purchased foods. Dietary deterioration is illustrated by the negative relationship found between nutritional status and increased sugar consumption. The assumption that a rise in income accompanying the adoption of commercial production will automatically lead to improved nutrition is challenged: income levels were not found to be consistently related to nutritional status. Children of families that have converted to cattle production, despite greater land availability and family incomes, do not have improved nutritional status. In the study area, where wages are low and food prices are very high, the value of a higher degree of self-sufficiency in food is recognized, yet families continue to switch to cash crops due to the environmental, economic, and time constraints imposed by the system of commercial agriculture in which they participate. The solution is not to return to traditional subsistence farming, however, but to determine under what conditions a more progressive form of agricultural change can occur.