【作者】 王春明；

【导师】 翟虎渠；

【作者基本信息】 南京农业大学 ， 作物遗传育种， 2002， 博士

【摘要】 本研究以水稻重组近交家系为材料构建了饱和的RFLP遗传图谱，对二点黑尾叶蝉（Nephotettix virexcens Distant）抗生性性状进行了数量性状基因座（QTL）定位和遗传分析，并根据其结果，对抗黑尾叶蝉（Nephotettix cincticeps Uhler）的近等基因系接种二点黑尾叶蝉，研究了水稻对二点黑尾叶蝉抗生性的遗传规律；将抗虫基因两侧的RFLP标记成功地转换成CAPS标记，通过标记辅助选择进行了抗叶蝉基因Grh2的回交转育。此外，还对水稻杂种不育、F₂不育及抽穗期性状进行了QTL作图和遗传分析。结果如下： 1 水稻分子遗传图谱的构建 用籼稻品种ARC10313（父本）与粳稻品种台中65（母本）杂交组合衍生的125个重组自交家系F₁₀世代构建了RFLP连锁图谱。因采用了参照图谱（Harushima 1998，Tsnematsu 1996）中靠近顶端的标记，作成的图谱覆盖全基因组。全图总长1462`4cM，含113个分布匀称的标记，图中标记位置与两个参照图谱基本符合，可以用于各种农艺性状的遗传研究。在染色体3，6，9，10和11上，探测到部分标记的遗传存在偏分离。利用该图谱进行了抗虫和不育性状的QTL定位研究。 2 二点黑尾叶蝉抗生性QTL定位和遗传分析 二点黑尾叶蝉，简称GLH（Green leafhopper），分布几乎遍及所有稻区，是我国长江流域以南及温带亚洲的主要水稻害虫。以台中65（粳稻）／ARC10313（籼稻）的重组近交家系为材料，对CLH抗生性进行全基因组QTL分析，检测出的4个叮L座位中，有 3个来自于 ARC1们13，还有 1个来自于粳稻品种台中 65。 另一万面，黑尾叶蝉简称GRH（Green rice leafhopper），hi－6k4是4个抗 GRH的基因。因为本研究在第 3和 11染色体上发现的两个抗 GLH的主效 QTL分别与已报道的抗肌H基因r肋4和0厂力2位置接近，所以对0了力丫和Grh二的近等基因系进行GLH接种鉴定和杭生性遗传分析，结果表明同时拥有两个抗GRH基因的近等基困系（基因型 CfhZ／ffoZ CkhErbhFk4），对矾H表现高抗。已报道的抗从H的基因有9个（0hi－g肋尺 0功，其中G力、Wb3和W力6分别位于第4、10和 5染色体上，而其它抗队H基困的住点不详。本研究在第 3和 11染色体上检测到的这两个新的抗肌H基因，其位置和效应与2和a竹4基本一致。3抗GRH基困的回交转育和分子标记辅助选择 水稻品种DV85抗黑尾叶蝉基困6rkZ位于第11染色体上，台中65为一个综合性状好但对黑尾叶蝉敏感的品种。OSP和W40为0厂力2两侧的RFLP标记，本研究将这两个标卡己成功地转换为在亲本间具有多态的CAPS标卡己。并以台中65作为轮回亲本与抗性品种DV85连续回交得到回交高代BC人群体，在进行抗叶蝉性状的表型选择的同时，利用M2基因两侧的CAPS标记对枕J；进行了标记辅助选择，这样所选出的个体具有台中 6 5的遗传背景且携带纯合 CrhZ基因，可作为聚合抗叶蝉基因培育新品种的重要中间材料。利用所获得的分子数据和抗性数据计算了两个CAPS标记与2的连锁距离，分析了该选择方法的效果。所采用的 DNA简易提取法、CAPS引物设计和CAPS标记辅助选择等配套方法具有快速、易行、高效的特点，因而可以广泛应用于水稻各重要农艺性状的选育改良中。一水稻F；不育性状的遗传研究 以台中 6 5（粳牙）／ARC 313（粗稻）的重组自交家系 F；。代材料与亲本台中65回交得到回交F；代群体（即BF；家系），观察其小穗不育和花粉不育性状进 2行叮L分析，检测出3个。J、穗不育和1个花粉不育叮L，而且有1个。1、穗不育位点和花粉不育位点重叠于第 7条染色体的 RFLP标iC R1789处，该座位与已报道的恢复基因Rf4对应。5水稻 F。不育蛐穗期性状遗传研究 以台中65（粳稻）侣hadua（4i稻）杂交F；代群体构建的RFLP连锁图谱，含 9 4个分布较为均匀的标i乙 对 F厂小穗不育性状进行单点分析和区间分析的结果基本一致：有两个巳小穗不育叮L座位分别位于染色体1的砌N厂－伽”帖之间和染色体8的C187－－opb3P厂之间，而且该两个OTL均为新检测出的座位；检测出5个抽穗期OTL，其中3个座位在单点分析和区间分析中的结果一致，分另位于染色体1 的砌bll 3 工地夕46，染色体4的ogj－＊3工 O“－O121染色体8的o6个0121，另外，染色体6的砌bZ厂为单点分析结果，染色体10的R71dC40）为区间分析结果。由于染色体1上的F；不育叮L和抽穗期叮L重叠，该OTL效应是属于遗传效应还是属于环境效应（迟抽穗）所致有待于进一步研究。位于染色体 1和 10上的抽穗期 OTL座位为新检测的座位。更多还原

【Abstract】 An RFLP linkage map based on the recombinant inbred FIO lines (RILs) derived from a cross between a japonica cultivar Taichung 65 and an indica cultivar ARC 10313 was constructed. QTL mapping of antibiosis to Green Leafhopper(GLH), Nephotettix virescens Distant, was conducted and the genetic basis of the resistance was identified with near isogenic lines of Grh2 and Grh4 genes. Grh2 gene was transferred through backcrossing and CAPS markers assistant selection. In addition, QTLs controlling spikelet and pollen sterility, F2 sterility and heading date were also identified. The results are as follows:1 Construction of RFLP linkage mapTo exploit the genetic potential of rice, 125 recombinant inbred F10 lines (RILs) derived from a cross between & japonica cultivar Taichung 65 and an indica cultivar ARC 10313 were developed. As a first step, an RFLP linkage map based on the RILs was constructed. The RFLP map contained 113 well-dispersed RFLP makers. Total map length was 1462.4cM. Linkage arrangement of the RFLP markers was in good agreement with that of the previously constructed maps.2 QTL mapping of antibiosis to Green Leafhopper, N. virescens and identification of genetic basis of the resistanceGreen Leafhopper (GLH), is one of the major insect pests of rice in the temperate Asian rice region. Nine GLH resistance genes (Glhl-glh8, Glh} and 4 Green Rice Leafhopper (GRH), Nephotettix cincticeps Uhler, resistance genes (Grhl, Grh2, Grh3 and Grh4) have already been identified. In this paper, QTL for antibiosis to GLH were investigated on Chromosome 3, 5, 11 and 12. Among them, the QTLs nearXNpb292 on chromosome 12 for increasing GLH mortality was fromjaponica variety. Among the other three QTL from indica variety, two QTL on chromosome 3 and 11 were close to Grh2 , and Grh4 , respectively. Therefore, the resistance to GLH was studied with near isogenic lines of Grh2 and Grh4 derived from susceptible variety Kinmaze(japom’ca), and resistant variety DV85 (indica). The results revealed that these two GRH resistance genes interaction also expressed strong resistance to GLH. None of the 9 Glh genes reported was located on chromosome 3 or 11, but 2 QTL for resistance to GLH newly identified in ARC 10313 were detected on chromosome 3 and11 in this study, which may correspond to Grh2 and Grh4.3 Green rice leaf leafhopper, N. dncticeps resistance gene transferring through backcrossing and CAPS Markers assistant selectionGrh2, one of green rice leafhopper, N.dncticeps resistance genes was- located on chromosome 11 of resistant variety DV85 (indica). Taichung 65 is ajaponica cultivar with elite characters, but susceptible to green rice leafhopper. C189 and G1465 are two RFLP markers flanking Grh2 gene, which were transformed into CAPS markers in this study. Both phenotypic selection and CAPS marker assistant selection were conducted in BC6F2 population derived from the cross of Taichung65 and DV85 in order to pick out the important breeding materials with Taichung65 background and resistance to green rice leafhopper. The linkage distance was calculated with the molecular and phenotypic data, meanwhile the effect of the selection method was analyzed. Since the experimental methods in this paper, including rude extraction of DNA, CAPS primer design, and the method of CAPS marker assistant selection, were characterized by fast, easy, and high effective to operate, it can be applied broadly to the selection for various agronomic traits.4 Identification of QTLs controlling spikelet and pollen sterilityQuantitative trait locus (QTL) analysis has been carried out to identify genes conferring spikelet sterility and pollen sterility in rice. One hundred twenty five F10 RILs derived from the cross between ajaponica cultivar Taichung 65 and an indica cultivar ARC 10313 were developed. Sixty three recombinant inbreed F, (BF,) lines derived from backcrossing RILs to Taichung65 were used as a segregating population for QTL analysis. Three QTLs for F, spikelet sterility were detected on Chromosome 1, 7 and 11 usin更多还原