秋稻品种‘Dular’广亲和基因效应的RFLP分析

利用RFLP标记对秋稻品种Dular的三交（南京11／／Dular/2533）的F2群体与灿粳品种的测交F1进行了单株带鉴定,并对广亲和性的表达进行分析。结果表明：与第6染色体上标记RG213连锁的广亲和基因S5^n效应较大;不同位点广亲和基因具有明显的累加效应;位点内的互作导致了粳型配子的部分败育,同时还存在位点间的互作。     

水稻广亲和基因（WCG）近等基因系的随机引物PCR分析

应用随机引物ＰＣＲ（ＲａｎｄｏｍＰｒｉｍｅｒＰＣＲ）技术分别在水稻广亲和基因（ＷＣＧ）的近等基因系和色素原基因（Ｃ）的分离群体库中寻找与ＷＣＧ和Ｃ基因连锁的分子标记。对于ＷＣＧ近等基因系,在２２６个随机引物中初筛到２２个显示多态性片段的引物。根据理论值计算,在２２个多态性片段中预期有２０个与ＷＣＧ连锁。在这些连锁标记中距ＷＣＧ最近的可达０．５ｃＭ。同样在分离群体库的筛选中有１０个扩增产物与Ｃ基因连锁。     

紫米基因与RFLP标记的连锁分析

选用种皮呈紫黑色的水稻体细胞无性系变异体黑珍米和其种皮呈无色的原始亲本Ｂａｓｍａｔｉ３７０配制组合,同时应用１２１个ＤＮＡ探针检测了黑珍米与Ｂａｓｍａｔｉ３７０之间的ＲＦＬＰ。应用Ｆ２和Ｆ３群体研究了紫色种皮的遗传控制。结果表明,有一个显性主效基因控制着黑珍米和Ｂａｓｍａｔｉ３７０在种皮颜色上的差异。通过多态性ＤＮＡ探针与种皮颜色的共分离分析,发现该基因与水稻第四染色体上的ＤＮＡ标记ＲＧ３２９和ＲＧ２１４连锁,与ＲＧ３２９和ＲＧ２１４的遗传图距分别为１８．９ｃＭ和２６．３ｃＭ。     

水稻广亲和性和胞质雄性不育恢复性的遗传分析

对经花培育成的广亲和恢复系ＴＧ７、ＴＧ８的遗传分析表明：ＴＧ７、ＴＧ８均带有１对广亲和基因,与ＣＰＳＬＯ１７、０２４２８的广亲和基因等位。广亲和基因与雄性不育恢复基因表现为独立遗传,野败型（籼）和滇－Ｉ型（粳）不育胞质的育性恢复基因非等位。ＴＧ７和ＴＧ８分别带有２对野败不育胞质和１对滇－Ｉ型不育胞质的恢复基因,分别来自亲本明恢６３和ＣＰＳＬＯ１７。     

水稻广亲和品种核DNA cosm id文库的构建和鉴定

用改进的方法成功构建了一个高质量的水稻广亲和品种（Cpslo17)的核基因组cosmid文库, 文库以SuperCos 1为载体,克隆总数约为12万以上,克隆平均插入片段约40kb,空载率约为5％,文库容量覆盖水稻单倍基因组约9倍以上,用RFLP标记23D12R作为探针,从文库中分离到一个位于水稻广亲和基因座S5附近的克隆R2I19。     

部分苔藓植物rbcL基因PCR—RFLP分析

运用ＲＦＬＰ方法对苔藓类６个科９种植物叶绿体ｒｂｃＬ基因的ＰＣＲ产物进行酶切分析,根据样本我态性片段,采用单联法对它们的遗传距离进行聚类分析,构建树状分支图,以此对苔藓植物的系统发育及其系统位置进行了初步探讨。     

小麦矮秆基因Rht3的RAPD和RFLP标记分析

利用PCR和RFLP技术分析了小麦赤霉酸反应不敏感的矮秆基因Rht3的近等基因系及其分离群体,RAPD分析从310条随机引物中,筛选出3个引物可以在高矮亲本中稳定地扩增出多态带,连锁分析表明仅S10601900和S10602000扩增片段与Rht3;连锁,遗传距离分别为7．1cM和9．2cM.RFLP分析选用了主要位于第4部分同源群短臂上的53个探针,其中Xper584,XksuF8和Xcdo38 3个探针在高矮亲本中揭示出多态性,;连锁分析表明仅Xper584与Rht3基因连锁,遗传距离为8．0cM。     

粳稻与几个广亲和品种杂交F1的花药培养效果

对粳稻品种秋光与几个广亲和品种及对照品种秋光的杂种F1进行花药培养,并对其中的一个组合所产生的白化苗做了PCR分析。结果表明：（1）秋光是一个培养力很高的品种,以秋光作为母本的粳／籼交组合的花药培养可以得到较高的培养力（绿苗率与愈伤组织诱导率的乘积）;（2）在供试的杂交组合中,培养力高低的一般趋势是：粳（籼）广亲和／籼（粳）广亲和＞粳／广亲和＞粳＞粳＞粳＞粳／籼;（3）与普通籼粳交相比,由于广亲和品种的参与提高了杂种F1的绿苗分化率, 白苗率;出愈率、分化率及培养力与出愈速度并无直接关系,选择适合的培养基并及时将愈伤组织转移到分化培养基上进行分化是提高花药培养的分化率和培养力的重要保证;（4）混生（或嵌合体）白化苗的形成是在分化初期愈伤组织发生遗传变异的结果,这种变异一经发生,无法用再分化的方法诱导绿苗的形成。     

水稻广亲和性遗传的主基因一多基因混合模型分析

籼、粳亚种间的F1一般表现为半不育,这限制了籼、粳杂种优势的利用。广亲和基因的发现及其遗传研究有助于揭示这种半不育现象的遗传本质,使克服籼、粳亚种间F1的半不育成为可能。本研究采用主基因－多基因混合遗传模型,分析了籼、粳杂交组合3037/02428的P1、P2、F1、B1、B2和F2六世代材料。研究结果显示：广亲和性的遗传除受单个主基因控制外还受多基因的影响。在利用广亲和基因克服亚种间的半不育性时不仅要考虑主基因对育性的作用,也不能忽视多基因对育性的影响。     

粳稻特殊广亲和系GC13的遗传分析及利用研究

粳稻品种GCl3对亚种内亲和力弱,而对亚种间亲和力强,因此特称之为特殊广亲和系(Special wide compatibility variety,SWCV)。广亲和基因等住性和遗传规律研究结果表明,GCl3的广亲和性主效基因效应明显,同时受微效基因修饰,与已知的S7、S9、S15三个育性基因住点之一等位。GCl3可与培矮64S等籼型光敏核不育系配组育成亚种间杂交稻;GCl3是粳型恢复基因源,从其杂交后代中选育出的偏粳(K’)型或偏籼(H’)型通用恢复系GR209、GR220和GR238等,对“野败”、“矮败”等多种不育细胞质和培矮64S等光敏核不育系具有强恢复性,配制出的“三系”或“二系”杂交稻具有高产潜力和利用前景。     

水稻广亲和性遗传的再研究

水稻广亲和基因的利用是克服亚种间杂种不育性的重要途径。但在广亲和性的遗传上不同研究者的结论不尽一致。以3种有广亲和品种参加的三交组合为研究材料,研究了品种Ketan Nangka的广亲和性遗传。结果表明水稻亚种杂交F1同时存在着雄性不育和雌性不育,但雄性不育对小穗育性的作用大小因组合而异;无论是在雄性不育位点还是雄性不育位点上,Ketan Nangka均具有相对应的中性基因（广亲和基因）;广亲和性的遗传特点与所用的籼粳测验品种间的杂种不育性密切相关;S－5位点的广亲和基因遗传符合单位点孢子体－配子体互作模型。     

Screening of RAPD Markers Linked to the Photoperiod-Sensitivity Gene in Rice Chromosome 6 Using Bulked Segregant Analysis

Bulked segregant analysis was used to determine randomly amplifiedpolymorphic DNA (RAPD) markers in a specific interval in themiddle of chromosome 6 of rice for tagging the photoperiod sensitivitygene.Two pools of F₂ individuals (japonica cv. Nipponbare and indicacv. Kasalath) were constructed according to the genotypes ofthree restriction fragment length polymorphism (RFLP) markerslocated at both ends and the middle of the targeted interval.Then another pair of pools were constructed based on the "graphicalgenotype," which was made with our high density linkage map.RAPD analysis was performed using these DNA pools as templates,and polymorphic fragments were detected and mapped. Using 80primers, either singlyor pairwise, we tested 2,404 primer pairsand established 14 markers tightly linked to the photoperiodsensitivitygene. The obtained RAPD markers were converted intosequence-tagged sites bycloning and sequencing of the polymorphicfragments and they can be used directlyfor construction of physicalmaps. This bulked segregant method can be applied for any speciesand any region of interest in which detailed linkage maps orphysical maps are needed.     

Identification of Two Blast Resistance Genes in a Rice Variety, Digu

Blast, caused by Magnaporthe grisea is one of most serious diseases of rice worldwide. A Chinese local rice variety, Digu, with durable blast resistance, is one of the important resources for rice breeding for resistance to blast (M. grisea) in China. The objectives of the current study were to assess the identity of the resistance genes in Digu and to determine the chromosomal location by molecular marker tagging. Two susceptible varieties to blast, Lijiangxintuanheigu (LTH) and Jiangnanxiangnuo (JNXN), a number of different varieties, each containing one blast resistance gene, Pik^s, Pia, Pik, Pi‐b, Pi‐k^p, Pi‐ta², Pi‐ta, Pi‐z, Pi‐i, Pi‐k^m, Pi‐z^t, Pi‐t and Pi‐11, and the progeny populations from the crosses between Digu and each of these varieties were analysed with Chinese blast isolates. We found that the resistance of Digu to each of the two Chinese blast isolates, ZB13 and ZB15, were controlled by two single dominant genes, separately. The two genes are different from the known blast resistance genes and, therefore, designated as Pi‐d(t)1 and Pi‐d(t)2. By using bulked segregation method and molecular marker analysis in corresponding F₂ populations, Pi‐d(t)1 was located on chromosome 2 with a distance of 1.2 and 10.6 cM to restriction fragment length polymorphism (RFLP) markers G1314A and G45, respectively. And Pi‐d(t)2 was located on chromosome 6 with a distance of 3.2 and 3.4 cM to simple sequence repeat markers RM527 and RM3, respectively. We also developed a novel strategy of resistance gene analogue (RGA) assay with uneven polymerase chain reaction (PCR) to further tag the two genes and successfully identified two RGA markers, SPO01 and SPO03, which were co‐segregated toPi‐d(t)1 and Pi‐d(t)2, respectively, in their corresponding F₂ populations. These results provide essential information for further utilization of the Digu's blast resistance genes in rice disease resistance breeding and positional cloning of these genes.     

矮泰引-3中半矮秆基因的分子定位

矮泰引-3的矮生性状受两对独立遗传的半矮秆基因控制,利用SSR标记将这两个矮秆基因分别定位到第1和第4染色体上。等位性测交的结果表明,位于第1染色体上的矮秆基因与sd1是等位的,所以仍然称其为sd1;而位于第4染色体上的矮秆基因是一个新基因,暂命名为sdt2。利用SSR标记将sd1定位于RM297、RM302和RM212的同一侧,而与OSR3共分离,它们之间的位置关系可能是RM297-RM302-RM212-OSR3-sd1,遗传距离分别为4．7cM、0cM、0．8cM和0cM,这与sd1在第1染色体长臂上的确切位置是基本一致的。利用已有的SSR标记和拓展的SSR标记将sdt2定位于SSR332、RM1305和RM5633、RM307、RM401之间,它们的排列位置可能是SSR332-RM1305-sdt2-RM5633-RM307-RM401,它们之间的遗传距离分别为11．6cM、3．8cM、0．4cM、0cM和0．4cM。     

粳稻糙米钙含量QTL分析

以'十和田'为轮回亲本,'丽粳2号'为供体,培育出糙米钙含量近等基因系下的重组近交系261个BC_5F_6株系,采用BSA法从遍布水稻12条染色体上的600对SSR引物中筛选到1个与糙米钙含量有关联的SSR标记RM5536.进一步据其在水稻染色体上的位置,结合PCR找到了与糙米钙含量有关的3个SSR标记(RM5794、RM5362和RM12178).用MAPMAKER/EXP3.0软件做出了这4个标记的连锁群,最后采用混合线性模型找到了糙米钙含量QTL位点.QTL分析结果显示:该位点位于1号染色体引物RM12178和RM5362之间,贡献率为9.62%,为新发现的糙米钙含量QTL位点,暂命名为qBRCA-1.并发现与糙米锶含量有关的QTL位点,其位于标记RM5362和RM5794之间,贡献率为3.93%.同时本试验首次从分子水平上验证了偏相关比简单相关更准确解释元素间的相关性.     

Identification of peanut (Arachis hypogaea L.) RAPD markers diagnostic of root-knot nematode (Meloidogyne arenaria (Neal) Chitwood) resistance

DNA markers linked to a root-knot nematode resistance gene derived from wild peanut species have been identified. The wild diploid peanut accessions K9484 (Arachis batizocoi Krapov. & W. C. Gregory), GKP10017, (A. cardenasii Krapov & W. C. Gregory), and GKP10602 (A. diogoi Hoehne) possess genes for ressitance to Meloidogyne arenaria. These three accessions and A. hypogaea cv. Florunner were crossed to generate the hybrid resistant breeding line TxAg-7. This line was used as donor parent to develop a BC₄F₂ population segregating for resistance. Three RAPD markers associated with nematode resistance were identified in this population by bulked segregant analysis. Linkage was confirmed by screening 21 segregatingh BC₄F₂ and 63 BC₅F₂ single plants. Recombination between marker RKN410 and resistance, and between marker RKN440 and resistance, was estimated to be 5.4±1.9% and 5.8±2.1%, respectively, on a per-generation basis. These two markers identified a resistance gene derived from either A. cardenasii or A. diogoi, and were closely linked to each other. Recombination between a third marker, RKN229, inherited from A. cardenasii or A. diogoi, and resistance was 9.0±3.2% per generation. Markers RKN410 and RKN229 appeared to be linked genetically and flank the same resistance gene. All markers were confirmed by hybridization of cloned or gel-purified marker DNA to blots of PCR-amplified DNA. Pooled data on the segregation of BC₅F₂ plants was consistent with the presence of one resistance gene in the advanced breeding lines. Different distributions of resistance in the BC₅F₂ progeny and TxAG-7 suggest the presence of additional resistance genes in TxAG-7.     

Genetic Analysis and Mapping of the Dominant Dwarfing Gene D-53 in Rice

The dwarfing gene D-53 Is one of a few dominant genes for dwarfing In rice （Oryza satlva L.）. In the present study, our genetic analysis confirmed that mutant characteristics including dwarfing, profuse tlllerlng, thin stems and small panicles are all controlled by the dominant D-53 gene. We measured the length of each Internode of KL908, a D-53-carrylng line, and classified the dwarfism of KL908 Into the dn-type. In addition, we measured elongation of the second sheath and a-amylase activity In the endosperm, and we characterized KL908 as a dwarf mutant that was neither glbberelllc acid-deficient nor glbberelllc acid-Insensitive. Using a large F2 population obtained by crossing KL908 with a wild-type variety, NJ6, the D-53 gene was mapped to the terminal region of the short arm of chromosome 11, with one simple sequence repeat marker, Ds3, co-segregating, and the other, K81114, located 0.6 cM away.