利用SRAP分子标记分析47份杂草稻样品遗传多样性

 利用SRAP分子标记分析了采集于杂草稻多发地区的47份样本的遗传多样性,旨在从基因的开放式阅读框区域（ORFs）深入研究杂草稻的遗传背景,为进一步揭示其来源提供依据。结果表明,参试的我国杂草稻材料在110对引物组合中共扩增出845种类型的谱带,多态性谱带590种,多态率为6982%,平均每对引物可以检测到54条多态性谱带。说明SRAP分子标记对于杂草稻基因组开放式阅读框区域序列的多态性具有很高的检测效率,适用于遗传多样性研究。我国杂草稻材料Shannon Weaver多样性指数（I）为063,且南北方杂草稻多样性指数（I）相当,分别为063和062。聚类分析结果表明,南方收集的杂草稻与北方收集杂草稻群体间遗传分化较大,遗传差异明显。程氏指数判定结果也表明杂草稻呈南籼北粳的分布状态。     

利用SSR分子标记研究花生属种间亲缘关系

以5份花生栽培种资源和花生属六个区组的19份野生种资源为研究材料，通过SSR分子标记技术分析其DNA多态性并进行聚类分析。大多数从栽培种基因组分离出的SSR引物能在野生种中扩增出DNA片段，共筛选出21对多态性SSR引物；每对引物在花生基因组中扩增出的条带数为1～13条，在供试材料中扩增出的总条带数为5～40条，平均18.1条，其中多态性条带为4～40条，平均18.0条；SSR引物的多态性指数为0.92～9.04；供试材料间的遗传距离为0.33～0.91，平均0.76。结果表明，大多数花生SSR引物为多位点引物，花生属种间种质存在丰富的DNA多态性， A. duranensis是花生栽培种（A. hypogaea L.）的野生种亲本之一，与花生区组野生种亲缘关系最近的是异形花区组，最远的是大根区组。     

Transgene Flow from Glufosinate-Resistant Rice to Improved and Weedy Rice in China

The development of transgenic rice with novel traits in China can increase rice productivity, but transgene flow to improved or weedy rice has become a major concern. We aimed to evaluate the potential maximum frequencies of transgene flow from glufosinate-resistant rice to improved rice cultivars and weedy rice. Treatments were arranged in randomized complete blocks with three replicates. Experiments were conducted between 2009 and 2010 at the Center for Environmental Safety Supervision and Inspection for Genetically Modified Plants, China National Rice Research Institute, Hangzhou, China. Glufosinate-resistant japonica rice 99-1 was the pollen donor. The pollen recipients were two inbred japonica rice （Chunjiang 016 and Xiushui 09）, two inbred indica rice （Zhongzu 14 and Zhongzao 22）, two indica hybrid rice （Zhongzheyou 1 and Guodao 1）, and one weedy indica rice （Taizhou weedy rice）. The offspring of recipients were planted in the field and sprayed with a commercial dose of glufosinate. Leaf tissues of survivors were analyzed by polymerase chain reaction to detect the presence of the transgene. The frequency of gene flow ranged from 0 to 0.488%. In 2009, the order of gene flow frequency was as follows： weedy rice 〉 Chunjiang 016 〉 Xiushui 09 and Zhongzu 14 〉 Guodao 1, Zhongzheyou 1 and Zhongzao 22. Gene flow frequencies were generally higher in 2009 than in 2010, but did not differ significantly among rice materials. Gene flow frequency was the highest in weedy rice followed by the inbred japonica rice. The risk of gene flow differed significantly between years and year-to-year variance could mask risk differences among pollen recipients. Gene flow was generally lesser in taller pollen recipients than in shorter ones, but plant height only accounted for about 30% of variation in gene flow. When flowering synchrony was maximized, as in this study, low frequencies of gene flow occurred from herbicide-resistant japonica rice to other cultivars and weedy rice. Averaged across years, the risk     

Diversity Analysis in Selected Non-basmati Scented Rice Collection

Diversity analysis among 23 rice varieties including 16 non-basmati scented accessions, 5 basmati accessions and 2 non-scented accessions was performed by random amplified polymorphic DNA (RAPD) and inter-simple sequence repeat (ISSR) marker systems. The varieties analyzed by 11 RAPD and 8 ISSR primers yielded an average of 65% and 80% polymorphism, respectively. The average number of polymorphic bands generated per RAPD primer was 6 and per ISSR primer was 5.87. RAPD and ISSR data analysis individually could not segregate basmati and non-basmati scented rice accessions. However, the analysis using a combined data could group basmati and non-basmati scented rice accessions separately. The bands present specifically among three accessions of non-basmati scented rice were also identified. The study revealed a high genetic diversity among non-basmati scented rice accessions.     

78份杧果种质资源AFLP分析

利用AFLP标记技术对78份杧果种质进行遗传多样性分析,结果显示,9对AFLP引物共产生908个等位位点,平均每对引物产生100.89个等位位点,多态性带为794条,共产生42个特异性位点,能区分21份种质,单态带25条,缺失带17条,特异性条带比例为4.63％,多态性比例为87.44％。不同种质间的相似系数范围是0.603~0.913,平均相似系数为0.735。聚类分析结果显示,以遗传相似性系数0.726为阈值,供试的78份种质可分成5个类群。主成分分析结果将供试的78份种质可分成4个类群,通过空间距离更能直观地表现种质间关系远近。     

用EST-SSR标记分析巴西橡胶树的遗传多样性

利用19对EST-SSRs引物对41份巴西橡胶树材料(6个野生材料和35个栽培种)进行遗传多样性分析.结果表明:19对引物均获得了预期的扩增结果,得到92个多态性位点,每对引物检测到的等位基因数为2～7个,平均为4.84个.在相似系数为0.64的水平上,野生材料和栽培种区分开来,在相似系数0.75的水平上,又可将栽培种材料分为四个类群;栽培种和野生材料间的遗传距离在0.11～1.16之间,栽培种间遗传距离较小,野生材料间遗传距离相对较大.在栽培种和野生材料之间,遗传距离最大的是AC/T/15/114与PR261(1.16),最小的是保亭155与热研7-20-59(0.11).     

利用SRAP标记分析花生属花生区组种质亲缘关系

利用54对SRAP引物分析花生属花生区组11个物种28份材料间的遗传变异,结果表明, 28份材料共扩增出327条多态性DNA带,平均每对引物扩增出5.95条多态性DNA带,扩增变幅为2～25条。聚类分析表明,28份花生材料间的遗传距离变幅为0.12～0.75,平均为0.42,A基因组物种A. duranensis与栽培种花生的亲缘关系最近,可能是栽培种花生的A基因组的供体。28份种质分为两大类,第一大类包含四倍体种质及A基因组的A. villosa和A. duranensis,第二大类包含B基因组、D基因组及其他A基因组。主成分分析结果与聚类分析结果相似,仅将第二大类中的B基因组种质A.batizocoi独立作为第三大类;第一和第二个主成分可以解释81%（57.5%和23.5%）的总变异。     

Diversity of weedy red rice (Oryza sativa L.) in Arkansas,U.S.A. in relation to weed management

Weedy rice, specifically red rice (Oryza sativa L.), is a major weed in rice which causes up to 80% yield loss and reduction of grain quality. Red rice accessions from Arkansas, U.S.A., were characterized to classify red rice accessions into certain phenotypic groups relevant to weedy rice management. The red rice accessions were 70% strawhull, 22% blackhull, 7% brownhull and <1% goldhull. Generally, blackhull red rice was the tallest (139 cm) and strawhull the shortest (133 cm) among all accessions. Blackhull red rice had more tillers (102/plant), smaller flag leaves (13 cm wide, 34 cm long), and flowered later (1225 heat units) than strawhull red rice which had 85 tillers/plant, 15 cm-wide and 34 cm-long flag leaves, and flowered after accumulating 1195 heat units. Morphological differences between accessions within each hull type were highly significant, showing great diversity within a hull color group as indicated by large ranges in traits. For example, blackhulls were 75–190 cm tall with 18–69 cm long flag leaves, 21–188 tillers and produced 40–949 g seed. Strawhulls were 46–189 cm tall with 18–66 cm flag leaf length, 16–172 tillers and produced 100–608 g seed. Some traits, such as seed production, differed widely between accessions within each hull color group such that the average seed production/accession for blackhull did not differ from that of strawhull weedy rice (196 vs. 192 g/plant). The onset of flowering among all accessions ranged from 56 to 126 d after planting. Red rice accessions formed six phenotypic clusters generally segregated by plant size or flowering time. Each morphotype would have different competitive abilities; thus, weedy rice management could be geared toward plant types. Highly competitive plant types would require intensive control measures to minimize yield losses and reduce the soil seed bank. Flowering dates impact stewardship strategies for herbicide-resistant or any genetically modified rice.     

SSR标记遗传距离与粳稻杂种优势的相关性分析

 利用SSR分子标记对30个粳稻品种进行遗传多样性分析,继而研究分子标记遗传距离与按照NCⅡ设计获得的200个杂交组合主要产量性状杂种优势的相关性,探讨分子标记遗传距离预测杂种优势的可行性。结果表明,64对SSR引物共检测到185条多态性片段,平均每对引物2.9条,每个SSR位点的多态性信息含量指数（PIC值）变化范围为0.064~0.844,平均为0.380。以SSR标记遗传相似系数为原始数据,按UPGMA聚类方法将30个亲本材料划分为7大类群,分类结果与系谱关系基本相符。分子标记遗传距离与杂种性状平均值的相关除每穗总粒数外均达到显著或极显著水平,与杂种优势的相关均达到极显著水平,相关系数介于-0.361~0.359,说明分子标记可用于水稻杂种优势群的划分和遗传多样性分析,但相关程度还不足以预测产量杂种优势。     

杂草稻和栽培稻叶片下表皮结构特征的观察及聚类分析

 利用光学显微镜对33份杂草稻及6份栽培稻材料的叶片下表皮中央长细胞及其各级乳突、气孔器及其乳突的结构特征进行了观察、统计后作主成分分析及聚类分析。主成分分析后得到3个在各个性状因子中贡献率最大的主成分,分别是中央长细胞大乳突、中乳突及小乳突的数目、大小,气孔密度。叶表皮结构特征在各供试材料之间有一定的变异和特异性,在杂草稻分类鉴定及亲缘关系的研究中具有一定的价值。     

江淮流域杂草稻叶绿体DNA的籼粳分化

 为了探明江淮流域杂草稻的细胞质来源,根据水稻叶绿体DNA ORF100（Open Reading Frame 100）序列在籼粳间存在69 bp差异的特征,设计了InDel标记cpDNA69。利用该标记对2个分别以籼稻和粳稻为母本的F2群体、22份栽培稻（Oryza sativa L.）、3份普通野生稻（O. rufipogon Griff.）进行了分析验证。PCR结果可以获得缺失和非缺失两种带型,缺失带型与以籼稻为母本的F2群体、10份籼稻材料完全对应;非缺失带型与以粳稻为母本的F2群体、11份粳稻材料完全对应,3份广西普通野生稻为非缺失带型,属粳型,1份以粳稻为母本的籼粳杂交材料为粳型。因此,cpDNA69可以用作叶绿体籼粳鉴定标记。利用该标记对22份杂草稻的叶绿体DNA鉴定的结果表明,7份早年发现的杂草稻,即江苏省连云港穭稻和安徽省怀远、来安、全椒、肥东的塘稻的叶绿体DNA为非缺失带型,属粳型,而近年来在江苏省扬中、高邮、灌云、洪泽、盐都、兴化、如皋等地直播稻田发现的15份红米杂草稻的叶绿体DNA为缺失带型,属籼型。这为进一步研究江淮流域杂草稻的来源提供了依据。     

中国龙血树属SRAP遗传多样性分析

利用SRAP标记技术对来自我国龙血树属（Dracaena）的24份材料进行遗传多样性和亲缘关系分析。结果显示,从80对SRAP引物组合中筛选出的15对引物组合共扩出155条条带,其中多态性条带为135条,占87.1％,平均每对10条,最好的引物组合为F18Em6。利用NTSYS软件分析数据得出24份材料间的Jaccard相似系数变化范围为0.27～0.97,平均值为0.56,UPGMA聚类显示,在相似系数为0.34处将一未定名种与其它材料分开,其他23份材料在相似系数0.46处又可分成2类,其中勐腊龙血树     

稻属AA染色体组8个种间SSR多样性与亲缘关系

选用平均分布于水稻基因组的30对SSR 引物,对AA染色体组8个野生稻种共42份材料的遗传多样性及遗传关系进行了研究。结果显示,本试验选取的30个SSR标记均具有多态性,多态性位点百分率为100%。30个多态性位点共扩增出的等位基因数为224, 每个位点可扩增出3～10个等位基因,平均7.47个;等位基因有效数(Ae)变幅为1.25～891,平均5.45。多样性指数中,Shannon多样性指数(I)为0.454～2.386,平均1.826;而Nei基因多样性指数变幅为0.199～0.888,平均0.774。系统聚类和带型分析结果表明,亚洲栽培稻(Oryza sativa) 与普通野生稻(O. rufipogon) 的亲缘关系最近,非洲栽培稻(O. glaberrima) 则与巴蒂野生稻(O. barthii) 关系最为密切,杂草稻(O. spontanea) 与普通野生稻 (O. rufipogon)、亚洲栽培稻(O. sativa) 之间有较近的亲缘关系,而展颖野生稻(O. glumaepatula)、长雄蕊野生稻(O. longistaminata)与AA组其他稻种之间的亲缘关系较远。     

温室条件下抗除草剂转基因水稻与杂草稻杂交和回交后代的适合度分析

在温室条件下,将抗除草剂转基因水稻Y0003和99-1与马来西亚杂草稻和国内安徽塘稻人工授粉获得的携带抗性基因的F1、F2与相应杂草稻回交,统计回交结实率和携带抗性基因的比例,并对携带抗性基因的杂交/回交代的适合度进行了测定,综合判断抗性基因漂移的风险。研究表明,在人工授粉的条件下,携带抗性基因的杂交代都能与相应的杂草稻回交并结实,结实率为15%～60%。F1的种子绝大多数都对草丁膦表现出了良好的抗性,F2和回交代分别表现为3∶1和1∶1的抗性分离比例,符合孟德尔遗传规律。适合度研究表明,杂交代、回交代与相应的杂草稻相比没有明显差异,大多数的杂交种的适合度和回交代相差不大,个别杂交代的适合度没有回交代高。以上研究结果表明在无除草剂选择压下,携带转基因水稻Y0003和99-1的抗性基因的2种杂草稻的杂交和回交后代有在适宜环境条件下生存定植的可能性。     

24份西番莲种质SSR分子标记分析

采用SSR标记对24份西番莲材料的遗传多样性进行研究,从45对引物组合中筛选出16对扩增带清晰、重复性好的引物组合。结果表明：16对SSR引物共产生68条扩增带,其中58条为多态性条带,多态性条带比率平均为85.3 %,24份材料的遗传相似系数(GS)的范围为0.09-1.0;UPGMA法聚类分析结果表明,24份西番莲种质以0.45为阈值可分为7大类,SSR标记丰富的多态性可在西番莲分子生物学领域中广泛应用。     

Rice landrace diversity in Nepal: Variability of agro-morphological traits and SSR markers in landraces from a high-altitude site

Nepal has a wide diversity of rice landraces in all rice-growing areas from low to high altitude. A baseline survey in Jumla, a high-altitude site (2200–3000 m), identified rice landraces with different names that differed in phenotype and were adapted to the extreme high-altitude environment. Jumli Marshi was the most common traditional rice variety, accounting for 85% of samples collected. The genetic diversity of these landraces was assessed by agro-morphological variability and microsatellite marker polymorphism. Forty-two qualitative and quantitative traits and 39 microsatellite (simple sequence repeats, SSR) markers were assessed among accessions with 10 different names. The accessions showed low morphological diversity with an average Shannon Weaver diversity index of 0.23. Only 16 morphological traits showed significant variation among the accessions. Discriminant function analysis showed that only 36% of accessions could be clustered according to name by morphological traits. Only one SSR locus was polymorphic, distinguishing only one accession. The names used by farmers to describe the phenotypic characteristics of panicle and grains were inconsistent indicators of genetic identity. We conclude that the Jumla landrace population has a narrow genetic base. The diversity detected was sufficiently low for it to be possible that there was a single origin for all of the Jumla landraces.     

利用InDel标记解析中国花生地方品种的遗传多样性与群体结构

为揭示InDel标记对我国地方花生品种遗传多样性和遗传结构的解析能力,本研究利用13个多态性InDel 标记检测4个典型生物学类型的90份中国花生地方品种材料。共扩增出42的等位基因,平均3个;Nei’s多样性指数 变幅为0.011~0.705,平均0.443;Shannon’s信息指数变幅为0.034~1.497,平均0.758。龙生型、普通型、珍珠豆型与多 粒型花生的Nei’s遗传多样性指数分别为0.3198、0.4407、0.4435和0.4372,结果表明龙生型的遗传多样性明显小于其 它生物学类型。种质间的遗传相似系数范围为0.077 ~ 1.000,平均为0.636;且普通型与龙生型花生之间遗传相似系 数最高,为0.636。群体分析结果表明密枝亚种与疏枝亚种、不同植物学类型之间遗传分化较小,且普通型与珍珠豆 型遗传结构相似,龙生型与多粒型花生遗传结构不同。利用非加权平均法（UMPGA）聚类分析将90份花生材料划分 为两类（G1、G2）,其中G1包括全部的龙生型、61.90%的普通型以及20.00%的珍珠豆型。本研究结果表明InDel标记 能够有效地揭示栽培种花生的遗传变异,可为花生杂交亲本的选择以及优异基因的挖掘提供重要参考。     

北方杂交粳稻亲本遗传背景分析

 为了揭示北方杂交粳稻亲本的遗传背景,利用微卫星(SSR)分子标记对北方杂交粳稻亲本及其高代后续材料进行了遗传差异鉴定和籼性程度分析。26对SSR引物共扩增出71个等位基因,每对引物平均等位基因数为2.73个。用非加权类平均法(UPGMA)进行聚类分析,供试材料在相似系数0.66处可分成5组,各组间材料的遗传差异相对较大;64个恢复系材料的籼性程度参数值(ADi)的变幅为0.056～0.684;24个不育系材料ADi的变幅为0.087～0.381。分析表明,强优势杂交粳稻组合双亲之间遗传距离应在0.30～0.60,恢复系的ADi应在0.26～0.50,不育系的ADi不应高于0.20,杂交组合的ADi应在0.20～0.40,同时杂交粳稻组合优势受母本的遗传背景的影响较大,母本自身应具有较高的产量潜力。     

广东地区野生狗牙根遗传多样性的ISSR分析

应用ISSR标记,对广东地区30份野生狗牙根进行遗传多样性研究。从50个ISSR引物中筛选出15个多态性明显、反应稳定的引物,共扩增出151条谱带,平均每个引物能扩增出10.07条带,其中多态性条带总数为142条,多态性条带比率达93.7%。材料间遗传相似系数GS=0.58278～0.92715。基于遗传相似系数,利用UPGMA聚类分析表明,供试材料可聚为4类。POPGENE分析软件结果表明,平均Shannon信息指数I=0.5689,平均Nei's基因多样性h=0.3813,每位点平均有效等位基因数ne=1.6196。     

Polymorphisms in the ALS gene of weedy rice (Oryza sativa L.) accessions with differential tolerance to imazethapyr

Herbicide-resistant Clearfield™ rice technology allows the use of ALS inhibitors to control weedy rice. Weedy rice plants have differential tolerance to imazethapyr, perhaps due to ALS polymorphisms. We aimed to assess ALS polymorphisms in weedy rice accessions from Arkansas, USA, with differential tolerance to imazethapyr in seedling growth bioassays. Six base changes were identified in the ALS of 14 weedy rice accessions. Three of these nucleotide changes resulted in amino acid substitutions — Pro⁹³Thr, Glu⁶³⁰Asp, and Val⁶⁶⁹Met — in four accessions: Ark-4, Ark-9, Poi-1 and Poi-6. The GR₅₀ values and inhibition of root and shoot growth (%) of these accessions differed. The Glu⁶³⁰Asp substitution occurs in the herbicide binding domain B and Val⁶⁶⁹Met occurs at the C-terminal tail where the co-factor binds. Variability in weedy rice ALS exists, but polymorphism patterns did not relate to tolerance levels. The observed mutations presented the possibility that sustained selection pressure will eventually lead to selection of a herbicide-tolerant individual that will be the progenitor of a resistant population. Concomitantly, pollen-mediated gene flow from Clearfield™ rice to weedy rice will lead to the evolution of ALS-resistant weedy rice populations.