东乡野生稻渐渗系基因组结构变化及多样性分析(英文)

[目的]研究东乡野生稻渐渗系基因组结构变化及对其进行多样性分析。[方法]以东乡野生稻(O.rufipogon Griff.,供体)和栽培稻协青早B(O.sativa sp.indica Kato.,受体)构建的BC1F6渐渗群体为研究材料,利用筛选出的分布在12对染色体上的25对SSR引物对群体中的239个株系进行多态性分析。[结果]适25个微卫星位点都有东乡野生稻DNA片段不同程度的渗入,群体均大部分保留亲本协青早B或东乡野生稻的DNA序列。渐渗系基因组中平均受体纯合条带百分率为78.13%,最高达94.98%(引物为RM131),最低为60.25%(引物为RM171)。平均供体纯合条带百分率为13.37%,最高达32.64%(引物为RM171),最低为2.93%(RM1095)。群体中还存在一些杂合位点,平均杂合率为5.62%,最高达10.04%(引物为RM401)。此外,BC1F6渐渗群体中发现有双亲条带的丢失和一些双亲所没有的新条带出现,其中平均亲本条带丢失率为2.88%,最高达13.39%(引物为RM311),最低为0(引物为RM401),平均新带率为1%。高世代渐渗群体平均Nei’s基因多样性(He)和平均Shannon’s信息多样性指数(I)分别为0.276和0.457。[结论]通过远缘杂交渐渗系发生了广泛遗传与表观遗传变异,扩大了遗传变异基础,为水稻品种改良与种质创新奠定了重要基础。     

利用SSR标记定位东乡野生稻苗期耐冷性基因

应用由 2 13个株系组成的协青早B/东乡野生稻的BC1F1群体 ,分析了东乡野生稻的耐冷基因与DNA标记的连锁关系。以苗期的死苗率为指标 ,对亲本和BC1F1群体各株系进行耐冷鉴定。结果表明 :死苗率在BC1F1群体中呈连续分布 ,表明耐冷性是由多基因控制的数量性状。应用单因素方差分析法 ,分别在第 4、第8染色体上发现有与耐冷性连锁的SSR标记RM 2 80、RM 337。     

东乡野生稻苗期耐冷性的QTL定位

【目的】研究东乡野生稻苗期耐冷性的QTL和连锁标记,为水稻耐冷种质资源的利用及分子标记辅助选择育种提供理论和实践依据。【方法】以东乡野生稻作为非轮回亲本,南京11号为轮回亲本,构建144株BC2F1分离群体。通过SSR标记以根电导率作为耐冷性指标,以复合区间定位法对东乡野生稻苗期耐冷性进行QTL定位。【结果】检测到2个QTL qRC10-1和qRC10-2均位于第10染色体,对表型的贡献率分别为34.13%和37.02%,是两个主效的QTL。在与2个QTL的连锁标记RM171周围发展分子标记进一步定位,检测到3个QTL位于标记RM171附近。【结论】东乡野生稻第10染色体上的2个QTLqRC10-1,qRC10-2与苗期耐冷性有关,并位于SSR标记RM304-RM1108区间,可用于水稻耐冷性分子标记辅助选择育种。     

东乡野生稻等位酶位点遗传多样性的初步研究

采用聚丙烯酰胺凝胶电泳技术 ,对东乡野生稻 9个天然小群体进行了 15个等位酶位点的遗传多样性分析。结果表明 ,东乡野生稻具有较高的遗传变异水平 ,9个小群体的多态位点百分率 (P)为 2 8.2 % ,等位基因平均数 (A)为 1.30 6 ,预期杂合度 (He)为 0 .2 34;小群体间的分化程度不高 ,基因分化系数 (Gst)为 0 .0 12 ,等位酶测定的总变异中 ,1.2 %来自小群体间 ,其余的变异 (98.8% )来自小群体内。这些结果可以为东乡野生稻的保护提供依据     

应用微卫星标记分析东乡野生稻遗传多样性初探

 利用SSLP标记对东乡野生稻进行多样性分析。结果表明 ,在 14对SSLP标记引物中 ,12对引物扩增的产物呈现多态 ,多态性探针百分率为 85 .71% ,共扩增出 70条多态性带 ,平均每对引物扩增出多态性带 5 .83条 ,表明东乡野生稻具有较丰富的遗传多样性。聚类分析结果表明 ,东乡野生稻 9个居群内及居群间存在一些差异较大的材料 ,同时也存在一些相同或遗传距离很小的材料 ;居群内平均遗传距离为 0 .2 3～ 0 .4 7,居群间平均遗传距离为0 .4 0～ 0 .5 5 ,居群间的遗传距离略大于居群内的遗传距离 ,为此 ,必须对东乡野生稻现有居群进行严格的保护。     

青蛤不同地理群体遗传结构差异的AFLP分析

[目的]探讨青蛤种群遗传多样性水平和遗传分化特征。[方法]采用AFLP技术对我国山东潍坊(WF)、江苏南通(NT)、浙江宁波(NB)、浙江温州(WZ)沿海共4个青蛤地理群体的遗传结构差异进行了分析。[结果]5对引物共得到261个位点,4个群体的多态位点比例平均为82.95%,其中WZ群体最高(86.97%),NT群体最低(78.93%);4个群体Nei’s基因多样性指数分别为0.258 1、0.252 6、0.271 3和0.277 6,香农多样性指数分别为0.394 7、0.383 0、0.412 0和0.421 4;群体遗传变异主要来自于群体内,占96.39%;WF和NT群体、NB和WZ群体遗传关系较近,聚类分析分别先聚在一起。[结论]研究的青蛤4个群体遗传基础较好,且尚未有明显的遗传分化。     

长白山区北五味子种质资源的遗传多样性分析

在长白山区选择具有代表性的56个野生北五味子自然群体,采用RAPD方法对其进行不同群体间遗传多样性分析。结果表明,在20个随机引物中共扩增出125条带(其中62个为多态性条带),平均每个引物扩增出6.25条带,多态率为49.6%;根据RAPD结果对其进行聚类分析的结果,群体间的遗传距离为0～0.56,显示出丰富的遗传变异。了解长白山区北五味子种质资源的遗传多样性将在保护和利用好种质资源、培育新品种等诸方面起到积极的作用。     

广西种源红锥栽培群体遗传多样性评价

[目的]揭示红锥栽培群体遗传多样性水平和结构。[方法]采用ISSR分子标记技术对人工选育的广西3个种源的红锥居群进行遗传多样性分析。[结果]9条引物共扩增出113条带,其中62条具有多态性,多态位点比例(PPF)为54.87%,单个居群的多态带百分率PPF为46.90%～47.79%。在物种水平上,期望杂合度(Hpop)和Shannon信息指数(I)分别为0.1366和0.2198。遗传分化系数GST为0.1275,表明经人工选育后的红锥遗传变异发生在居群内的个体占87.25%,12.75%的遗传变异发生在居群间。各群体之间的遗传一致度I为0.9593～0.9765,红锥群体水平的基因流Nm为3.423。[结论]ISSR-PCR方法可以有效地揭示红锥栽培群体遗传多样性水平和结构。     

野生大麻居群遗传多样性ISSR分析的取样策略

[目的]分析野生大麻居群的合理取样样本量,为全面了解我国大麻资源的遗传多样性及制定野生大麻保护策略提供参考依据.[方法]随机抽取96个辽宁科尔沁沙地野生大麻自然居群个体,采用计算机模拟方法从中随机抽取不同样本量(分别为5、10、15、20、25、30、35、40、50、60、70、80、90和96个个体)的抽样群体各20次,利用ISSR分子标记对其遗传多样性进行分析,确定合理的取样样本量.[结果]利用从60条ISSR引物中筛选出的9条引物对随机抽取的96个野生大麻个体进行PCR扩增,共获得76个位点,其中多态位点48个,多态率达63.16%,观测等位基因数(Na)为1.632,有效等位基因数(Ne)为1.250,Nei's基因多样性指数(H)为0.156,Shannon's信息指数(I)为0.245.由各遗传参数拟合曲线变化趋势分析结果可知,初始的4种抽样群体(分别为5、10、15和20个个体)遗传多样性水平呈急速增加趋势,之后随着样本量的加大,遗传多样性水平增加趋势明显变缓.当抽样群体样本量超过25个个体时,Na、Ne、H和I均包含了野生大麻居群90%以上的遗传变异.[结论]采用ISSR分子标记评估野生大麻群体遗传多样性时,群体取样样本量不宜小于25个个体才能较好地反映该居群总体的遗传多样性水平.     

西林水牛群体的RAPD遗传多样性分析

【目的】了解西林水牛群体的遗传变异和遗传结构,为其辅助标记育种、遗传资源保护及利用提供参考依据。【方法】从广西西林县的8个乡（镇）采集184份西林水牛血样,采用优化后的RAPD技术检测其多态性DNA,统计多态性条带数,并应用Popgene32软件对西林水牛群体的有效等位基因数、Nei氏基因多样性指数及Shannon多样性指数进行分析。【结果】从18条RAPD引物中筛选出8条扩增产物稳定、条带清晰可辨的引物,扩增出的DNA片段分子量为100-1000 bp;从184个样本中共扩增出4968个多态性条带,平均每条引物可扩增出621个多态性条带,多态性频率达60.0%-100.0%,平均为89.7%。西林水牛群体的平均有效等位基因数为1.6745,平均Nei氏基因多样度指数为0.3583,平均Shannon多样性指数为0.5510。【结论】西林水牛群体的遗传变异、遗传分化及遗传多样性均较高,但选育程度较低,育种潜力较大,有待进一步保种和开发利用。     

不同土壤pH对普通野生稻生理特性的影响

[目的]研究不同土壤pH对普通野生稻生理特性的影响。[方法]以雁山普通野生稻为试验材料,通过盆栽试验设置不同土壤pH栽培,比较不同pH下普通野生稻的生理特性变化。[结果]在pH为6.8的中性条件下,普通野生稻叶片叶绿素相对含量和可溶性糖含量最高,脯氨酸含量和MDA含量维持最低水平,且CAT和POD活性均为最高。[结论]雁山普通野生稻对中性偏碱环境的适应性较强,适宜的环境pH范围为6.0～7.4。     

Molecular Marker Assisted Selection for Yield-Enhancing Genes in the Progeny of Minghui63 × O.rufipogon

Two yield-enhancing genes (yldl.1 and yld2.1) are located on chromosomes 1 and 2 respectively in a weedy relative of cultivated rice, Oryza rufipogon. SSR markers RM9 and RM166 are closely linked with the two loci respectively. Minghui63 (MH63) has been a widely used restoration line in hybrid rice production in China during the past two decades. The F1 of cross "MH63 × O.rufipogon" was backcrossed with MH63 generation by generation. RM9 and RM166 were used to select the plants from the progeny of the backcross populations. The results were as follows: (1) In BC2F1 population, the percentage of the individuals which have RM9 and RM166 amplified bands simultaneously was 12.2%, while in the BC3F1 population, that was 16.3%. (2) Among 400individuals of BC3F1, four yield-promising plants were obtained, with yield being 30% more than that of MH63. (3) The products amplified by primer RM166 in O. rufipogon and MH63 were sequenced. It was found that the DNA fragment sequence amplified by RM166 from MH63 was 101 bp shorter than that from O. rufipogon. The 101 bp sequence is a part of an intron of the PCNA (proliferating cell nuclear antigen) gene.     

广西邕宁普通野生稻(Oryza rufipogon Griff.)种群遗传多样性及核心种质构建研究

[目的]研究广西邕宁普通野生稻种群的遗传多样性和核心种质构建,为普通野生稻种群保护提供依据。[方法]从水稻12条染色体上均匀选取24对SSR标记对243份样本进行遗传多样性分析。[结果]结果表明,24对引物均表现出多态性,多态位点比率为100%;24个多态位点共检测出103个等位变异,平均等位基因数A为4.291 7,平均有效等位基因数E是2.511 2,平均期望杂合度He为0.573 2,平均多态信息含量PIC为0.572 0。根据聚类结果分析,从243份材料中筛选出31份核心种质,包含了24个SSR位点检测到的所有的遗传变异,其平均期望杂合度He是0.595 8,平均多态信息含量PIC是0.586 3,基本上可以代表广西邕宁普通野生稻种群的遗传多样性。[结论]广西邕宁普通野生稻资源具有丰富的遗传多样性,建议对该地区普通野生稻种群的遗传多样性进行重点保护和利用     

Identification of quantitative trait loci and candidate genes associated with ABA sensitivity in common wild rice(Oryza rufipogon Griff.)

Abscisic acid(ABA),as one of the foremost signaling molecules in plants,is an important hormone which plays versatile functions in regulating developmental process and adaptive stress process.A set of introgression lines were previously generated via a backcrossing program using an elite indica cultivar rice Teqing(O.sativa L.) as recipient and an accession of Yuanjiang common wild rice(O.rufipogon Griff.) as donor.In this study,the previously developed introgression lines were evaluated for ABA sensitivity.Here we reported that a total of 14 quantitative trait loci(QTLs) associated with ABA sensitivity were identified.An ABA sensitive introgression line,YIL53,was identified and characterized.Physiological characterization,including chlorophyll content,malondialdehyde content,soluble sugar content,and stomata movement,demonstrated that YIL53 exhibited the characteristics associated with ABA sensitivity.Genotypic analysis revealed that YIL53 harbored one QTL related to ABA sensitivity,q ASS1-2,which was located on chromosome 1 within one introgressed segment derived from the Yuanjiang common wild rice.Furthermore,the qASS1-2 was finally narrowed down to a 441-kb region between simple sequence repeats(SSR) marker RM212 and single nucleotide polymorphism(SNP) marker M3 using the segregation population derived from the cross between Teqing and YIL53,and three candidate genes associated with ABA sensitivity were identified using a strategy combined gene expression analysis with QTL mapping.Identification of the QTLs related to ABA sensitivity and characterization of the ABA sensitive line YIL53 would provide a helpful basis for isolating novel genes related to ABA sensitivity.     

Genetic Diversity and Genetic Changes in the Introgression Lines Derived from Oryza sativa L. Mating with O. rufipogon Griff.

The objectives of the present study were to estimate genetic diversity and genetic changes of introgression lines (ILs) which derived from cultivated rice (Oryza sativa L. cv. Xieqingzao B, XB) mating with common wild rice (O. rufipogon Griff., CWR). The genetic data of 239 ILs were based on a total of 131 polymorphic microsatellite (SSR) markers distributed across the 12 chromosomes of rice. On average, these ILs possessed 77.1 and 14.31% homozygous bands from XB and CWR, respectively. Most of the ILs were clustered together with XB individual, which was revealed by principal coordinate analysis (PCA) and the program STRUCTURE analysis. The result from PCA demonstrated that some intermediate genotypes between XB and CWR were also found. Moreover, there were some genomic sequence changes including parental bands elimination and novel bands emergence in the ILs. The average Nei's gene diversity (He) was 0.296, which was higher than that of cultivated rice. It suggested that interspecific hybridization and gene introgression could broaden the base of genetic variation and lay an important foundation for rice genetic improvement. These different genotypic ILs would provide a better experimental system for understanding the evolution of rice species and the mechanism of alien gene introgression.     

Cloning and Sequence Analysis of Disease Resistance Gene Analogues from Three Wild Rice Species in Yunnan

Two sets of degenerate oligonucleotide primers were designed according to amino acid conservedregions of reported plant disease resistance genes which encode proteins that contain nucleotide-binding site andleucine-rich repeats(NBS-LRR), and the plant disease resistance genes which encode serine/threonine proteinkinase(STK). By polymerase chain reaction(PCR), disease resistance gene analogues have been amplified fromthree wild rice species in Yunnan Province, China. The DNA fragments from amplification have been clonedinto the pGEM-T vector respectively. Sequencing of the DNA fragments indicated that 7 classes, 2 classes and6 classes NBS-LRR disease resistance gene analogues from Oryza rufipogon Griff. , Oryza officinalis Wall. ,and Oryza meyeriana Baill. were obtained respectively. The two representative fragments of TO12 from Ory-za officinalis Wall. and TR19 from Oryza rufipogon Griff. belong to the same class and homology of theirsequences are 100%. The result shows that the sequences of the same class disease resistance gene analogueshave no difference among different species of wild rice. 5 classes STK disease resistance gene analogues werealso obtained among which 4 classes from Oryza rufipogon Griff. , 1 class from Oryza officinalis Wall. Bycomparison analysis of amino acid sequences, we found that the obtained disease resistance gene analogues havevery iow identity(low to 25%) with the reported disease resistance gene L6, N, Bs2, Prf, Pto, Lr10 and Xa21etc. The finding suggests that the obtained disease resistance gene analogues are analogues of putative diseaseresistance genes that have not been isolated so far.     

欧洲肉鸽与卡奴鸽不同杂交组合生产性能的比较

[目的]比较欧洲肉鸽与卡奴鸽不同杂交组合的生产性能,以寻求最佳杂交配套。[方法]对欧洲肉鸡3个品系与白卡奴鸽正反交,组成6个杂交组合,对6个杂交组合的生产性能进行测定,比较6个杂交组合的总产蛋数、产蛋周期、蛋重、受精率、受精蛋孵化率、初生重、28日龄体重、28日龄成活率等指标。[结果]总产蛋数以E2BC组合最高(22.32个),显著大于BCE1、BCE2、BCE3、E1BC组合(P0.05);BCE2组合总产蛋数最少(19.70个),显著小于其他5个组合(P0.05)。E2BC组合产蛋周期最短(34.37 d),显著小于BCE2组合(38.91 d)(P0.05)。蛋重基本在23 g左右。受精率E2BC组合最高(79.85%),显著高于BCE3组合(71.57%)(P0.05)。BCE1组合受精蛋孵化率最高78.79%,显著高于E3BC组合(74.72%)(P0.05)。各杂交组合的初生重基本都在16 g左右。28日龄上市体重以E2BC组合最高(599.94 g),显著大于E3BC组合(556.13 g)(P0.05)。BCE2组合28日龄成活率为84.77%,显著低于其他5个组合(P0.05)。[结论]E2BC组合的年产蛋量、受精率和28日龄体重均最高,其繁殖性能和生长性能都比较优良,可在生产中加以利用。E3品系在杂交利用上不太理想。     

云南3种野生稻中抗病基因同源序列的克隆及序列分析

 根据已报道的NBS LRR类和STK类抗病基因结构中的氨基酸保守区域 ,设计简并引物 ,通过PCR扩增及克隆 ,从普通野生稻 (OryzarufipogonGriff.)、药用野生稻 (OryzaofficinalisWall.)、疣粒野生稻 (Oryzameye rianaBaill.)中共获得 14类NBS LRR类抗病基因同源序列 ,其中普通野生稻中的有 7类 ,药用野生稻中的有 2类 ,疣粒野生稻中的有 6类。药用野生稻中TO12代表序列与普通野生稻中TR19代表序列 ,同属一类 ,且具有 10 0 %的同源性 ,说明不同种野生稻中的同一类 (聚类 )抗病基因同源序列是完全一致的。同时 ,还获得 5类STK类抗病基因同源序列 ,其中普通野生稻中的有 4类 ;药用野生稻中的有 1类。通过氨基酸同源性比较分析 ,发现笔者克隆到的抗病基因同源序列与已克隆的抗病基因L6、N、Bs2、Prf、Pto、Lr10和Xa2 1等的氨基酸同源性都相当低 (均低于 2 5 % ) ,暗示了这些抗病基因同源序列可能是目前尚未报道的抗病基因的同源序列。