Microsatellite Marker Based Assessment of Genetic Diversity among Cultivars, Landraces and Wild Relatives in Rice

India being the primary center of origin for rice had a very large treasure of local land races, most of which are out of cultivation today. The exact genetic potential and their differences from commercial varieties and the magnitude of heterogeneity still present in them are not well catalogued. Hence the need to characterize the available land races has become imminent in the modem day concept of crop improvement （Rezai and Frey, 1990）. The present study addresses the utility of SSR markers in divulging the genetic relationships at molecular level among the major component factions office gennplasm viz., local cultivars, land races and wild species collected from a wide range of agro-geographical regions of Indian subcontinent.     

高通量分子标记检测方法的研究进展

分子标记直接反映基因组序列水平上的遗传多样性,被广泛地应用于基因定位、多样性分析、作物遗传育种及医疗等领域。近几年,伴随着芯片杂交技术和测序技术的快速发展,高通量、高自动化的分子标记检测技术有许多新发展。本文针对目前常用的分子标记检测新技术,包括dCAPS、KASPar、基因芯片、简化基因组测序和靶向测序基因型检测等技术的原理、方法、优缺点和其应用前景进行综述,为分子标记的检测和遗传相关研究提供信息支持。     

水土流失防治措施对马尾松林土壤微生物群落分子生态网络的影响

水土流失防治措施可以提高侵蚀退化林地的土壤质量。为了探究典型水土流失防治措施对土壤微生物分子生态网络的影响,在南方红壤区马尾松林下建立了水土流失防治措施小区,设置3种不同处理,分别为挖设鱼鳞坑加种植草本,挖设鱼鳞坑加种植草本与灌木,以及对坡面土壤不设置任何措施的对照。利用16S rRNA和18S rRNA基因 Illumina MiSeq 高通量测序技术,测定3种处理下的土壤微生物群落组成,基于随机矩阵的方法构建微生物网络。结果表明,措施实施后,绿弯菌门相对丰度显著降低,变形菌门与酸杆菌门的相对丰度显著升高（P < 0.05）。2种措施处理下微生物网络总节点数、总连接数、平均连通度以及模块性均较高,微生物网络的规模增大,微生物间互相作用更复杂。3个微生物网络均以负互相作用（60.59%～67.49%）为主,措施实施后物种间竞争作用进一步加强。绿弯菌门、放线菌门和变形菌门中的部分菌群在本研究区的微生物网络中起着重要的连接作用,此外,3个微生物网络的部分关键节点所属的菌群相对丰度较低（< 1%）,对于构建微生物网络也具有关键作用。2种措施处理下微生物网络平均路径距离较长,微生物作用的响应速度慢,群落结构稳定性提高,其中挖设鱼鳞坑并种植草本与灌木的措施较挖设鱼鳞坑并只种植草本的措施效果更优。冗余分析表明,土壤容重（R2 = 0.465,P < 0.05）、pH（R2 = 0.377,P < 0.05）、有机质（R2 = 0.383,P < 0.05）、全氮（R2 = 0.545,P < 0.01）、对细菌群落结构有显著影响,土壤含水量（R2 = 0.485,P < 0.05）对真菌群落结构有显著影响。上述研究结果表明,水土流失防治措施实施后土壤微生物群落结构发生明显变化,网络规模增大、物种间互作强度以及微生物群落结构的稳定性提高。     

苹果杂交后代植株果实着色性状的遗传分析和预测

苹果着色问题是影响我国许多产区苹果质量的重要因素,本研究利用苹果着色相关基因MdMYB1（GenBank登录号：DQ886414）设计引物进行PCR扩增,开发出一个酶切位点为HaeⅢ的CAPS标记Mb2,该标记可用于区分杂交亲本‘富士’和‘嘎拉’苹果及分析其杂交后代植株。进一步利用Mb2标记对该杂交组合的后代植株进行遗传分析,结果表明,可将77个后代植株分为2组,一组42个后代植株含有来自‘嘎拉’亲本的非红色性状等位基因以及来自‘富士’亲本的非红色性状等位基因或者红色性状等位基因,其果实颜色预测为非红色与红色或者非红色与桔红色;另一组35个后代植株含有来自‘嘎拉’亲本的红色性状等位基因以及来自‘富士’亲本的非红色性状等位基因或者红色性状等位基因,其果实颜色预测为桔红色与红色或者只有红色。本研究结果将为选育优质苹果新品种提供依据和方法参考。     

多花黑麦草杂交种SSR分子标记鉴定及表型比较分析

多花黑麦草为异花受粉植物,杂交过程中易出现杂交种生物性混杂现象,有必要对杂交后代进行真假鉴定分析.本研究旨在筛选出多花黑麦草优势杂交组合和优异杂交后代材料,为多花黑麦草杂交育种工作提供基础资料.实验利用SSR分子标记辅助鉴定5个多花黑麦草(Lolium multiflorum)骨干品种为亲本创制的杂种后代真实性,并对亲本与杂种后代主要表型进行比较.结果表明:(1)用20对引物组合共得到多态性条带426条,多态性位点百分率为82.41％,说明SSR分子标记能够很好的揭示多花黑麦草材料间的差异;(2)经SSR分子鉴定,将后代分为有父本特征谱带的杂种后代、无父本特征带的后代2类,在150个杂交后代单株中有父本特征谱带的杂种后代有108个为真杂种,其中杂交组合GC、CG真杂种最多均为24个,WG组合真杂种最少为19个;(3)杂交组合CG的F1代部分形态特征显著优于亲本,5个杂交组合内(间)在营养器官与生殖器官性状均存在显著差异,其中杂交组合MG的变异系数最高,WG的变异系数最低,杂交组合CG形态特征显著高于其他杂交组合.由研究结果可以初步判断长江2号×赣选一号多花黑麦草为优势杂交组合,其杂交种为优异杂交后代材料,可为后续杂交选育新品系提供后备材料.     

ITS鉴定木材种类：一例司法鉴定研究实例

本研究选取一例非法收购国家重点保护植物油丹的司法鉴定案作为研究实例,在确认鉴定技术可行的前提下,应用ITS序列分析对待检测的原木进行了分子鉴定。本研究采用CTAB法提取原木DNA,对其提纯后进行ITSrDNA的PCR扩增,再对PCR阳性产物进行克隆、测序,将测序结果进行Blast分析和构建系统发育树。结果表明,送检的47根原木中有9根被鉴定为油丹。本研究采用ITS序列系统发育分析方法,成功鉴定了原木种类,为今后类似司法鉴定案件的审理提供了可靠的鉴定意见。     

大白菜InDels标记开发及其在剩余杂合体鉴定中的应用

大白菜(Brassica rapa ssp.pekinensis)参考基因组序列的公布及测序成本的降低为大规模开发插入/缺失(Insertion/Deletion,InDel)标记提供了可能.本研究以性状差异明显的大白菜自交系He102与06-247为亲本,开展了全基因组重测序、标记开发和验证工作.二者测序深度分别为10×和8×,经筛选共获得330 218个InDels差异位点,出现频率为1.2个/kb,其中有11 238个差异位点位于编码区,包括5 184个差异基因,每个基因平均包含2.2个差异位点.分别以He102与06-247测序数据中筛选出的多态性InDels位点为参照,从10条染色体上随机选择933个位点进行了PCR扩增及电泳检测验证.结果表明,测序深度对假阳性率有直接影响,以测序较深的材料中筛选到的差异位点为参照,其验证结果的假阳性率亦较高,反之则较低.本研究中以He102与06-247为参照选择InDels位点验证的平均假阳性率分别是51.9％和22.5％.从933个位点中共筛选出593个在亲本之间实际表现为多态性的位点,这些差异位点中有375个位于编码区,是潜在的功能性标记.利用上述差异位点,检测了He102与06-247衍生的F7代重组自交系群体,明确了F7代各株系在593个位点的纯/杂合状态,为利用剩余杂合株系精细解析和精准定位大白菜耐抽薹、抗病毒、抗干烧心等重要农艺性状奠定了基础.     

基因差异表达技术cDNA-SCoT在植物上的研究应用

目标起始密码子多态性(start codon targeted polymorphism,SCoT)分子标记是一种源于起始密码子ATG翻译起始位点保守区域来设计其侧翼单引物的标记技术.自从首次应用SCoT标记进行甘蔗(Saccharum officinarum)基因差异表达分析以来,该技术己被广泛应用于花生(Arachis hypogaea)、玉米(Zea mays)、铁皮石斛(Dendrobium officinale)、芒果(Mangifera indica)、西瓜(Citrullus lanatus)等植物的研究.本文介绍了cDNA-SCoT技术的原理、引物设计方法及特征,着重总结cDNA-SCoT技术体系筛选及其在抗病、抗寒、抗旱、遗传进化等方面的应用研究,以期为科研工作者在选择差异表达分析技术提供参考.     

Agronomical,quality, and molecular characterization of twenty Italian emmer wheat (<Emphasis Type="Italic">Triticum dicoccon</Emphasis>) accessions

Emmer wheat (Triticum dicoccon Schrank, 2n = 4x = 28) consists in a hulled wheat; its cultivation has been drastically reduced during the last century as a consequence of its low yield. Recently, its agronomic and nutritive values, as well as the increase of popularity of organic agriculture, have led to a renewed interest making its cultivation economically viable in the marginal lands with an increase of the cultivated areas. In Italy, it mainly survives in few marginal lands of central and southern Italy, where local varieties, adapted to the natural environment from where they originate, are used; moreover, some selected lines have also been developed. In the present work, agro-morphological and qualitative traits, together with molecular analyses of 20 emmer accessions consisting of Italian landraces, breeding lines, and cultivars, were performed. The field experiments were conducted for two consecutive years (2001/2002–2002/2003) in two locations: Viterbo in central Italy, and Foggia in south Italy. The analyzed emmer wheat accessions showed a good amount of genetic variability for both evaluated agro-morphological and molecular traits. This study illustrates an increase in earliness, GY, TW, TKW, and YI going from landraces, breeding lines to cultivars, while the variability does not change proportionally.     

Nomenclature and genetic relationships of apples and pears from Terceira Island

Heritage apple (Malus domestica Borkh. hybrids) and pear (Pyrus communis L. hybrid) trees grow in villages throughout Terceira Island, Azores, Portugal. Some of these pears have different names but similar morphology. The objective of this study was to determine synonymy, homology, and phylogeny of apples and pears from Terceira and to examine potential relationships of the island pears with standard apples and pears of Portuguese or American descent. Nine apple microsatellite markers were used to determine genetic relationships. Distance- and parsimony-based cluster analysis grouped these genotypes into separate apple and pear clades. The Terceira apples were divided into two clades: the maçā and the reineta-reinette. Among the 17 heritage apple genotypes, seven unique accessions were identified and four groups of synonyms, or possibly clones, were detected including: ‘Reineta Agosto’ and ‘Reineta Verde’ from Altares; ‘Reineta Castanha’ and ‘Reineta Verde Miuda’; ‘Maçā Pêra,’ ‘Maçā Calhau’, ‘Pêro Branco’ from Salga and from Terra-Chā and ‘Maçā Marmelo’; and the five genotypes ‘Maçā Sao Joao’, ‘Malápio Rosa’, ‘Maçā Gaspar’, ‘Maçā Branca’ and ‘Maçā Pato’. In addition, two homonyms were detected. ‘Pêro Vermelho’ from Terra Chā was a separate genotype from a tree from Doze Ribeiras of the same name, but Pêro Branco from Terra Chā appears to be a clone that can be distinguished by an additional allele at CH1F07a from a tree with that name from Salga. One pair of apple clones, ‘Reineta Agosto’ and ‘Reineta Verde’ from Altares appear to be derived from an unreduced gamete of ‘Golden Delicious.’ Another apple genotype ‘Maçā Acida’ could be a sibling of the ‘Maçā Pêra’ clonal group. Other tested standard apples from the US genebank were unrelated to Terceira genotypes. Of the seven heritage pears, five unique genotypes and one pair of synonyms were detected. ‘Pêra Papo Pintassilgo’ from Raminho and ‘Pêra Vermelha’ from the nursery of Serviço de Desenvolvimento Agario da Terceira (SDAT) were synonyms. ‘Passans du Portugal’ was related to ‘Pêra Cabaca’ but other standard pears from the US genebank were unrelated to Terceira genotypes. Future studies will include additional apple and pear cultivars from other Islands of the Azores and continental Portugal, and wild Asian species to further explore genetic relationships.