平欧杂种榛主栽品种（系）遗传关系的ISSR分析

采用ISSR分子标记的方法,对达维、辽榛3号等17个平欧杂种榛主栽品种(系)进行了遗传关系研究。试验从60条ISSR引物中筛选出7条进行电泳分析,使用NTSYS pc 2.11F和Popgene 1.32软件进行数据统计分析并作图。结果显示:7条ISSR引物共获得58条谱带,平均每条引物8.3条谱带,引物平均多态性比率84.6%;7条ISSR引物可将所有样品完全分开,当遗传相似性阈值为0.687时可将样品分成3个类群:第Ⅰ类群14个品种(系)可分成4个亚类,第Ⅱ类群1个品种,第Ⅲ类群2个品系。样品间的遗传相似性系数为0.448~0.879(平均0.678)。群体的有效等位基因数、基因多样度、Shannon信息指数分别为1.6751、0.3701和0.5308。上述结果表明,平欧杂种榛是一个具有高度遗传多样性的群体,主栽品种(系)间存在复杂的亲缘关系。该研究结果对于平欧杂种榛的遗传关系研究具有重要意义,也可为其他榛属植物的相关研究提供参考。     

山西省榛属植物居群的SSR遗传多样性研究

利用9对SSR引物对山西省平榛(Corylus heterophylla Fisch)和毛榛(C.mandshurica Maxim.et Rupr.)野生居群、欧榛(C.avellana L.)和平欧杂种榛(C.heterophylla Fisch.×C.avellana L.)的人工栽培居群,共205个样本进行PCR扩增,共扩增出172个等位基因。每个位点的等位基因数为5~18个,平均等位基因数为12.5个。居群观测杂合度(Ho)和预期杂合度(He)的变化范围分别为0.395~0.665和0.778~0.906,表明榛属植物遗传多样性较高,其中平欧杂种榛的遗传多样性最高(He=0.867,I=2.271),毛榛遗传多样性最低(He=0.825,I=2.006)。不同物种居群间遗传分化系数FST=0.106,平均基因流Nm=2.609,表明居群间的遗传分化水平较低。各居群在大多数位点上偏离Hardy-Weinberg平衡,主要原因是人工选择或近交所致。分子方差分析(AMOVA)表明,遗传变异主要发生在物种居群内。NJ聚类结果显示毛榛和平榛多数个体聚在各自居群内,平欧杂种榛和欧榛个体交互混合组成一小支后再与平榛聚在一起,表明平欧杂种榛与欧榛、平榛的亲缘关系较近,而毛榛与其它3种榛属植物的亲缘关系较远。本研究还分析讨论了山西省榛属植物居群具有较高遗传多样性的原因,并提出了野生榛子的保护利用策略。     

不同品种(系)榛子坚果表型性状差异性分析及其综合评价

该研究以目前主栽的平榛、毛榛及25个平欧杂种榛品种(系)的坚果为研究对象,测定了不同品种(系)榛子坚果的17个表型性状指标,用方差分析、相关性分析、聚类分析和主成分分析方法对其进行差异性分析和综合评价,为国内榛树品种(系)的选优、开发及加工利用提供科学依据。结果表明：(1)不同品种(系)榛子坚果的17个表型性状指标存在显著差异,其中果壳质量、坚果体积和坚果质量差异最为明显,变异系数分别为31.49%、30.44%、27.50%,横纵径比、果形指数、伸长率、圆球度的变异系数均小于10%。(2)平欧杂种榛坚果的纵径、横径、侧径及坚果质量、果仁质量、出仁率等指标显著优于毛榛和平榛,其中杂种榛的平均果仁质量(0.70～1.46 g)和出仁率(31.48%～56.12%)显著高于平榛(0.51 g、35.13%)和毛榛(0.37 g、38.12%)。(3)榛子坚果表型性状指标间的相关性分析表明,榛子坚果体积越大,其质量和果仁质量也越大,果壳越薄则出仁率更高。(4)主成分分析结果发现,影响榛子坚果综合得分的主要指标为坚果的纵径、横径、侧径、质量和果仁质量,其次是果形指数、横纵径比、出仁率、果仁饱满...     

皖北地区平欧杂种榛优势品种坚果性状评价

【目的】对皖北地区栽培的平欧杂种榛优势品种进行性状评价,明确品种加工适宜性。【方法】2011年起在皖北地区引进平欧杂种榛品种（系）22个,依据适应性和丰产性等指标筛选出9个优势品种,选择与坚果加工利用直接相关的经济性状指标、榛仁营养成分、矿质元素、油脂理化性质及脂肪酸组成进行研究,从烤制、仁用、油用、鲜食等不同加工利用角度进行分析评价,以期为平欧杂种榛选择与推广提供科学依据。【结果】结果表明,皖北地区9个平欧杂种榛优势品种坚果质量分布范围为2.54～4.40 g,三径均值为18.99～21.49 mm,形状指数为1.04～1.31,榛仁质量分布范围为1.00～1.66 g,果壳厚度为1.53～2.41 mm,出仁率为34.36%～47.01%。榛仁脂肪含量为50.50～58.00 g/（100 g）,蛋白质含量为21.10～25.10 g/（100 g）,可溶性糖含量为4.08%～4.94%。油脂中鉴定出棕榈酸、棕榈烯酸、硬脂酸、油酸、亚油酸、亚麻酸、花生酸和花生烯酸8种脂肪酸成分,其中油酸含量最高,为81.21%～88.53%,不饱和脂肪酸含量为92.19%～94.61%。在矿质营养方面,榛仁中钾、磷、钙和镁含量较为丰富。对加工产能进行折算,9个平欧杂种榛优势品种单株坚果产量分布范围为1.52～2.20 kg,单株脂肪产量为330.50～503.47 g,单株蛋白质产量为141.68～224.40 g。【结论】根据榛果消费方式的不同,研究认为皖北地区平欧杂种榛品种中,辽榛3号最适作为带壳烤制品种,辽榛3号、达维和辽榛4号最适作为仁用加工品种,其中辽榛4号可作为油用加工品种,辽榛1号可作为功能性蛋白饮料加工品种,达维和辽榛9号可作为高钙类加工品种,达维、辽榛3号、平欧545号是皖北地区较为适宜的鲜食品种。     

盐胁迫对3种平欧杂种榛幼苗叶片解剖结构及离子吸收、运输与分配的影响

研究盐胁迫下3个品种平欧杂种榛幼苗叶片解剖结构和离子代谢特征,以揭示盐胁迫响应与适应机制及不同品种的耐盐性差异。以‘达维’、‘辽榛7号’、‘玉坠’2年生压条苗为材料,在盆栽条件下经轻度、中度、重度(分别为50、100、200 mmol/L NaCl)盐胁迫处理,设对照为0,研究幼苗叶片显微解剖结构参数和Na~+、K~+、Cl~-、Ca²⁺含量的变化及其在根、茎、叶中的吸收、运输和分配特征。不同品种平欧杂种榛叶片厚度、上表皮厚度、下表皮厚度、栅栏组织和海绵组织厚度随着盐胁迫程度的增强呈现出先增加后降低的特点,轻度和中度胁迫下各参数显著高于对照。中度盐胁迫显著提高了各品种叶片结构紧密度。盐胁迫导致平欧杂种榛根、茎、叶Na~+和Cl~-含量明显高于对照。盐胁迫下,Na~+和Cl~-在叶中的绝对含量明显高于茎和根,但二者的增幅以根中最大,叶中最小,表明平欧杂种榛根系首先会吸收并截留一定数量的Na~+和Cl~-,然后将其运输至茎和叶中。与对照相比,轻度和中度盐胁迫下根、茎对K~+和Ca²⁺的吸收保持稳定或减少,叶对K~+和Ca²⁺...     

三倍体白杨杂种无性系指纹图谱的构建

目的:构建三倍体白杨杂种无性系指纹图谱,鉴定三倍体白杨杂种无性系。方法:分离纯化三倍体白杨杂种DNA模板,采用扩增片段长度多态性(AFLP)分子标记技术构建三倍体白杨杂种无性系指纹图谱。结果:从64对引物组合中筛选出M-CTA/E-CAG、M-CAC/E-CCA、M-ACT/E-CTC和M-CTT/E-CTG等4对多态性较高的引物组合,并应用该引物组合对21个三倍体白杨杂种无性系进行了AFLP分析,构建了21个三倍体白杨杂种无性系指纹图谱。结论:构建无性系指纹图谱是鉴别三倍体白杨杂种无性系的有效方法,能够有效鉴别21个三倍体白杨杂种无性系。本研究为品种鉴定及新品种权保护奠定了基础。     

19个枇杷杂交新品种(系)的SSR鉴定和指纹图谱构建

为探究枇杷(Eriobotryajaponica)杂交品种真实性和DNA指纹图谱,对新育成的19个枇杷杂交品种(系)进行SSR标记鉴定分析。结果表明,从已发表的89对SSR引物中筛选出扩增条带清晰稳定的多态性引物19对,在24份枇杷材料中共扩增到83条带,每对引物平均扩增4.37条,PIC值为0.234～0.983,平均为0.764。经12对具有父本特征带多态性引物鉴定, 19个杂交新品种(系)全部为真杂种,真杂种率为100%。UPGMA聚类分析表明,19个杂交新品种(系)的遗传相似系数为0.728～0.969,与杂交亲本‘新白2号’和‘贵妃’聚为同一个大类,并可细分为4个亚类,红肉与白肉的枇杷品种(系)间无明显划分。同时利用8对多态性SSR引物组合,构建了19个枇杷杂交新品种(系)的分子指纹图谱。这为枇杷品种鉴定、新品种权保护和杂交育种提供重要参考依据。     

盐胁迫对平欧杂种榛幼苗生长、光合荧光特性及根系构型的影响

以平欧杂种榛3个品种(新榛1号、新榛2号、新榛3号)幼苗为材料,研究不同盐胁迫程度(对照、轻度、中度、重度)对幼苗生长性状、光合荧光特性以及根系构型的影响.结果表明: 3个品种幼苗的新稍长度、基径、叶面积,以及根、茎、叶、总生物量随着盐胁迫的增强而降低,根冠比则增加.中度、重度盐胁迫下3个品种幼苗的净光合速率较对照分别显著降低20.5%和43.2%.轻度、中度、重度盐胁迫下的蒸腾速率和气孔导度较对照分别降低2.0%、16.3%、32.0%和10.2%、35.7%、60.1%.随着盐胁迫的增强,胞间CO₂浓度缓慢上升,水分利用效率呈先上升后下降的趋势,并在轻度盐胁迫下达到最大值.3个品种幼苗的初始荧光随着盐胁迫的增强而升高.最大荧光、最大光化学效率、潜在光化学活性、实际光化学效率、电子传递速率、光化学淬灭系数随着盐胁迫的增强而下降,非光化学淬灭系数则随着盐胁迫的增强先升高后降低.根系构型分析表明,盐胁迫导致3个品种幼苗的根系生物量、长度、表面积和体积下降.同一盐胁迫下,新榛2号各径级根系构型参数的下降幅度低于其他2个品种.平欧杂种榛幼苗生长性状、光合荧光特性以及根系构型参数受品种和盐胁迫程度的双重影响.盐胁迫下新榛2号表现出较强的生长与光合生理适应性,其耐盐性优于其他2个品种.     

佳木斯地区榛子引种试验初报

2000～2003年间先后引进平欧杂交榛(Corylus heterophylla Fisch.×C.avellana L.)品系33个、土耳其榛(C.colurna L.)栽培种1个、欧洲大果榛子(C.maxima Mill.)栽培种21个材料的树苗,栽培于佳木斯大学植物园内进行以抗寒性鉴定为主要内容的引种试验.通过3～6年的观察结果证明:从辽宁省经济林研究所引入33个平欧杂交榛81.8%可在这个地区存活并生长良好,55.6%品系可以正常结果,已从中筛选出单株结果率高,坚果大,果形好,出仁率高达40%左右的繁殖推广类型4个;2001年之后相继从乌克兰中部森林草地区引入土耳其榛和欧洲大果榛子的22个品种表现只有根存活,每年地上枝条均因抽干而枯死,但每年蘖发新枝条平均生长量可达1.3 m高,因此只能在保护地条件下作为杂交的原始材料.     

莲品种DNA指纹图谱的构建

为了克服单纯依据形态特性鉴定品种的局限性, 我们开展了莲品种DNA指纹图谱构建研究, 旨在对其品种的快速准确鉴定及专利权保护等起一定作用。本研究以圆明园保存的72个莲品种为实验材料, 用来自不同地点的1,409份野生莲(Nelumbo nucifera)和58份美洲黄莲(N. lutea)群体样本作遗传背景参照。从104对核微卫星引物(nSSR)中筛选出15对, 从17对叶绿体微卫星(cpSSR)引物中筛选出2对, 共17对引物作为72个莲品种DNA指纹鉴定的条码。15对nSSR引物共检测到94个等位基因(平均6.27个), 其中11个属于美洲黄莲, 65个属于野生莲, 18个不能区分; 多态信息含量(PIC)介于0.3899-0.8023之间 (平均0.5748)。2对cpSSR引物共检测到13个单倍型, 其中9个属于野生莲, 4个属于美洲黄莲。全部17对引物标记结果显示, 共有19个品种含有美洲黄莲遗传组分, 其中8个母系来源于美洲黄莲; 有36个品种(涉及12对引物)具有至少1个特有基因型; 最少8对引物组合可完全区分开68个品种。有2组共4个品种组内全部17对引物均不能区分。本研究通过核心引物组合法使68个莲品种获得特异性DNA指纹。推荐13对nSSR和2对cpSSR共15对引物作为莲品种鉴定的核心条码, 并建议将形态特征与DNA指纹相结合作为莲品种的鉴定标准。     

HPLC screening of natural vitamin E from mediterranean plant biofactories--a basic tool for pilot-scale bioreactors production of alpha-tocopherol

The study was performed in order to investigate a simple, efficient, reliable and rapid method of extracting and quantifying natural vitamin E for pressurized liquid extraction (PLE) as well as high-performance liquid chromatography (HPLC) analysis. Lyophilized Corylus avellana L. nut samples were powdered by high-speed milling with Waring blender for 40 s. alpha-Tocopherol was extracted from the nut tissue powder using dehydrated hexane fortified with 0.01% butylated hydroxytoluene (BHT, co-antioxidant). The rate of alpha-tocopherol accumulation showed differences among nut samples collected in different areas of Italy. Sarda Piccola nut biofactory contained higher amount (81.17 microg/g d.w) of alpha-tocopherol than other-local eleven Italian cultivar nuts. These results provide insight into the biofactory basis for alpha-tocopherol accumulation in hazelnut and give the suitable cultivar tissues to establish pilot-scale bioreactors production of natural bioactive vitamin E.     

De novo sequencing and development of EST-SSR for Gevuina avellana (Proteaceae), a nut tree from South America

?Premise of the study: The aim of this study was to develop molecular tools to investigate the genetic structure and diversity of natural populations of Gevuina avellana (Molina, Proteaceae), the Chilean hazelnut. Simple sequence repeats (SSRs) derived from expressed sequence tags (EST-SSRs) were developed, optimized, and characterized. ?Methods and Results: The microsatellite-containing sequences were selected from a cDNA library developed from the nut. The eight marker loci showed two to seven alleles in 60 unrelated trees, from two different natural populations. The observed heterozygosity (H(o)) was 0.44, ranging from 0.07 to 0.92 for different loci. When the multilocus genotype of the eight EST-SSRs was considered, all trees could be differentiated. ?Conclusions: The newly identified EST-SSRs in G. avellana and the multi-pooling strategy involving eight markers will facilitate future studies of clone identification, genetic structure, and diversity.     

Molecular genetic diversity of the Turkish national hazelnut collection and selection of a core set

European hazelnut (Corylus avellana L.) is an economically and nutritionally important nut crop with wild and cultivated populations found throughout Europe and in parts of Asia. This study examined the molecular genetic diversity and population structure of 402 genotypes including 143 wild individuals, 239 landraces, and 20 cultivars from the Turkish national hazelnut collection using simple sequence repeat (SSR) markers. A total of 30 SSR markers yielded 407 polymorphic fragments. Diversity analysis of the Turkish hazelnut genotypes indicated that they fell into three subpopulations according to ad hoc statistics and neighbor-joining algorithm. Although all cultivars clustered together, they overlapped with the wild accessions and landraces. Thus, the dendrogram, principal coordinate, and population structure analyses suggest that they share the same gene pool. A total of 78 accessions were selected as a core set to encompass the molecular genetic and morphological diversity present in the national collection. This core set should have priority in preservation efforts and in trait characterization.     

硼、锌对平欧杂交榛光合生理、结实和果仁品质的影响

通过对黑龙江省东宁地区的4年生平欧杂交榛（Corylus heterophylla×Corylus avellana）联合施用不同浓度的硼（0.2%、0.3%、0.4%）、锌（0.3%、0.4%、0.5%）肥处理,研究两种微量元素对平欧杂交榛光合生理、结实、果仁和叶片营养状态的影响。研究结果表明：（1）硼、锌对净光合速率影响显著,其交互作用对净光合速率影响极显著。（2）硼对出仁率影响极显著,对果仁重影响显著,对单果重和座果率影响不显著。锌对单果重和果仁重影响极显著,对出仁率影响显著,对座果率影响不显著。硼、锌交互作用对出仁率影响显著,对单果重、果仁重和座果率影响不显著。（3）硼、锌和其交互作用对果仁脂肪酸含量、果仁蛋白质含量影响不显著。硼、锌和其交互作用对4年生平欧杂交榛的生理和结实影响明显,提高了平欧杂交榛叶片的净光合速率,同时对果仁重、出仁率和单果重有不同程度的促进作用。本地区4年生平欧杂交榛硼、锌叶面施肥的浓度分别以0.3%和0.4%为宜。     

CaMADS1, a MADS box gene expressed in the carpel of hazelnut

Hazelnut (Corylus avellana L.) is a species of economic interest that shows a peculiar floral biology. Unlike most of the angiosperms, which produce ovules during floral development such that they are ready for pollen at anthesis, hazelnut ovary development is delayed and triggered by compatible pollination. In order to elucidate the mechanisms regulating this unusual process and the role of the MADS box genes in ovary development, a cDNA library from pollinated styles of hazelnut was screened with a mixture of MADS box genes from different plant species. CaMADS1 (Corylus avellana MADS box), a floral-specific MADS box gene, was isolated, and characterized as belonging to the sub-family of the AGAMOUS genes. Northern blot, RT-PCR analyses and in situ hybridization experiments show a precise correlation between ovary development and CaMADS1 expression, indicating a role of this MADS box gene in the processes of floral organogenesis.     

DNA typing and genetic relations among European hazelnut (Corylus avellana L.) cultivars using microsatellite markers.

In this work, 78 hazelnut (Corylus avellana L.) cultivars from various germplasm repositories were studied at 16 simple sequence repeat (SSR) loci in order to identify the genotypes and investigate their genetic relations. Polymorphism at SSR loci was evaluated on the basis of number of alleles (mean: 9.4), expected heterozygosity (mean: 0.78), and power of discrimination (mean: 0.91). Several synonyms reported in the literature were confirmed, and new cases of synonymy were identified. The parentage of North American cultivars 'Butler', 'Ennis', and 'Royal', the French selection 'Fercoril-Corabel', and 'Impératrice Eugenie' was investigated on the basis of the alleles present at 16 loci and analysis at 8 additional loci. A dendrogram generated from cluster analysis using the unweighted pair group method with arithmetic mean grouped cultivars according to their pedigrees or geographical origins. There was an evident differentiation of the northern European cultivars from the southern European ones and from the Turkish cultivars. The latter clustered close to but separate from the Italian and Spanish clusters. It is very likely that exchanges of cultivars occurred between the central and western Mediterranean basin as a result of human migration and trade. A database containing the SSR profiles of the most important hazelnut cultivars will be useful for identification of cultivars and synonyms, legal protection, and parentage analysis.     

A bacterial artificial chromosome library for 'Jefferson' hazelnut and identification of clones associated with eastern filbert blight resistance and pollen-stigma incompatibility

European hazelnut (Corylus avellana L.) is the only economically important nut crop in the family Betulaceae. Because of its small genome size (~385 Mb / 1C), relatively short life cycle, availability of a dense linkage map, and amenability to transformation by Agrobacterium, the European hazelnut could serve as a model plant for the Betulaceae. Here we report the construction of a bacterial artificial chromosome (BAC) library for 'Jefferson' hazelnut using the cloning enzyme MboI and the vector pECBAC1 (BamHI site). The library consists of 39,936 clones arrayed in 104,384-well microtitre plates with a mean insert size of 117 kb. The genomic coverage of the library is estimated to be about 12 genome equivalents. This library provides a valuable resource for the map-based cloning of two important genes, the resistance gene from 'Gasaway' that confers resistance to eastern filbert blight caused by the fungus Anisogramma anomala (Peck) E. Müller and the S locus that controls pollen-stigma incompatibility. Fine-resolution mapping near the two loci was carried out using random amplified polymorphic DNA (RAPD) and simple sequence repeat (SSR) markers. Fine mapping at the disease resistance locus showed that markers W07-375 and X01-825 flanked the resistance locus at distances of 0.06 and 0.05 cM, respectively. The S locus is flanked by markers 204-950 and KG819-200 at distances of 0.14 and 0.24 cM, respectively. Assuming that 1 cM corresponds to a physical distance of 430 kb, it will take approximately two to three chromosome walks to assemble BAC contigs that span both loci.