甜瓜雌雄异花同株近等基因系的构建及近等性评价

雌雄异花同株类型在甜瓜杂交制种时可部分免除人工去雄,确保杂交种纯度,所以,对杂种优势利用中有着重要价值.为了研究和利用控制甜瓜雌雄异花同株基因A,用雌雄异花同株材料Mr作母本P1,雄花两性花材料Mm作父本P2,用P1做轮回亲本与F1杂交,经连续回交并最终自交纯化,获得雌雄异花同株近等基因系(BC6F4).用形态相似法和AFLP标记对获得的近等基因系和轮回亲本进行分析,结果表明:(1)Mr xMm的F2与BC6S1分离群体中,雌雄异花同株株数与雄花两性花株数符合3:1比例,回交群体中雌雄异花同株株数和雄花两性花株数符合1:1的比例;在发育期、果皮色、网纹等农艺性状上没有明显差异;(2)用448对AFLP引物检测轮回亲本和近等基因系之间多态性,有417对引物没有多态性,31对有多态性,统计得其遗传相似系数为99.73%;(3)平均每个AFLP引物对可扩增出46.8条带,说明AFLP揭示的遗传信息量很高,是一种评价近等性较理想的分子标记,这与以往结果一致.     

3个粳稻抗白叶枯病近等基因系的构建

自１９８９－１９９６年,选用分别个有抗水稻白叶枯病主效基因Ｘａ２,Ｘａ７和Ｘａ１４的３个品种为供体亲本,以一个不携有抗病基因的改良型粳稻品种沈农１０３３为轮回亲本,通过６次回交,自交直至ＢＣ６Ｆ４代,经各世代抗性和农艺性状筛选,转育成３个粳型抗白叶枯病近等基因系,命名为ＣＢＢ２,ＣＢＢ７和ＣＢＢ１４。这些ＮＩＬ除所带不同的抗病基因外,其生育期,株高,分蘖力,穗型大小等农艺性状与轮回亲本基本相似。     

大白菜抗根肿病近等基因系的选育及其评价

本研究以具有抗根肿病基因CRb的大白菜‘CR Shinki DH’系为供体亲本,大白菜自交系‘91-12B’为轮回亲本,通过回交和4个阶段的分子标记辅助选择,选育出7个携带CRb基因的抗根肿病近等基因系.通过CRb基因所在大白菜A3染色体的分子标记检测和抗病性鉴定,筛选出抗根肿病、并插入有供体亲本最短染色体片段的3个近等基因系B1S1、B2S2和B3S1.表型分析表明这3个近等基因系的结球相关性状与轮回亲本无显著差异,而且保持了‘91-12B’的配合力.所选育的近等基因系不仅能应用于育种实践,也为通过转录组分析研究根肿病的抗病机理提供了材料.     

水稻小穗簇生性近等基因系的构建及其近等性评价

为了研究育种中间材料Z1820突变产生的小穗簇生基因对农艺性状的影响和对该基因进行定位克隆，我们用恢复系泸恢17(LH17)和N45作母本，Z1820作父本杂交，并用上述恢复系作轮回亲本，连续回交并自交，获得了簇生性近等基因系Cl-LH17和Cl-N45。用形态相似法和SSR标记对获得的两对近等基因系进行多态性分析，结果表明：（1）LH17和Cl-LH17在株高、穗长上有显著差异，在所考查的其他性状上差异不显著；N45和Cl-N45仅在穗长上有显著差异。（2）在所选用的120对SSR引物中，对LH17和Cl-LH17，只有第6染色体的两个引物（RM7434、RM5957）揭示了多态性，而对N45和Cl-N45则有位于3条染色体上的4个引物能揭示多态性。综合形态和SSR标记分析说明，LH17和Cl-LH17是一对近等性理想的簇生性近等基因系，有利于该簇生基因的进一步研究；Z1820的簇生基因能使穗长、株高负增长，对有效穗、每穗总粒数、结实率等重要性状无显著影响。（3）簇生性在N45的遗传背景中的表现强于在LH17的遗传背景中的表现。     

油菜MICMS恢复基因近等基因系的构建与近等性分析

以宁R7为恢复基因源,利用连续回交法构建以T78、T84和ZS9三个优异种质为遗传背景供体的近等基因系候选株系。于成熟期考察并比较候选株系农艺性状,同时采用SSR标记对候选株系进行PCR扩增估算遗传相似系数并进行聚类分析。农艺性状分析(t测验)表明,不同候选株系与其供体的农艺性状具有不同程度的相似性,株高和分支点高度的差异程度较其他性状高。采用75对SSR引物对21个候选株系PCR扩增共得到157个目的片段,发现三个遗传背景候选株系的平均遗传相似系数分别为0.86、0.92和0.98。农艺性状与SSR标记综合分析后,认为wh13、wh59和wh118分别为三个种质的最优近等基因系。综上表明,采用农艺性状比较与分子标记相结合分析近等性是一种有效的近等基因系的检测方法。中选近等基因系可用于恢复基因的定位及克隆。     

利用分子标记辅助选择优质蛋白玉米近等基因系

为了扩增优质蛋白玉米育种新种质,用普通玉米种质转育创制优质蛋白玉米(QPM)近等基因系;利用SSR标记phi057进行辅助选择,以优质蛋白玉米种质为o2基因供体,转导、构建来自不同遗传背景普通玉米种质为受体的QPM近等基因系。结果表明：利用共显性SSR标记引物phi057在o2基因供体自交系与普通玉米受体自交系间表现多态性,能区分O2O2、O2o2和o2o2等3种基因型,来自不同遗传背景种质所构建获得的QPM近等基因系赖氨酸含量有不同程度提高,赖氨酸含量达到0.36%~0.42%,不同遗传背景受体自交系赖氨酸增加幅度9.1%~64.0%。普通玉米种质导入供体o2基因后能够提高种质赖氨酸含量。     

糯玉米opaque2基因近等基因系的创制

opaque2基因能够提高糯玉米赖氨酸、色氨酸等必需氨基酸的含量。用2种优质蛋白玉米(QPM)CA339和鲁2548自交系作为opaque2基因供体,25个品质较好的糯玉米自交系作为受体,利用回交技术和SSR分子标记辅助选择育种,创制糯玉米opaque2近等基因系。分子标记结果表明,不同受体以及两种供体间都存在多态性。5套创制成功的opaque2近等基因系的赖氨酸含量比其轮回亲本分别提高了59.0%、52.7%、48.5%、46.3%和61.9%,分别由0.308%、0.313%、0.309%、0.341%、0.323%提高到0.489%、0.478%、0.458%、0.498%、0.522%。本研究表明,可以利用此方法通过向不同遗传背景的多种受体导入opaque2,选取赖氨酸含量提高较大、透明表型的近等基因系,提高糯玉米的营养价值和经济价值。     

转Bt基因抗虫棉近等基因系的研究

培育了 7个转 Bt基因抗虫棉近等基因系。Bt基因转入陆地棉基因组后 ,对棉花的衣分、铃重、单株铃数无明显影响 ,对皮棉产量有一定影响 ,部分组合的抗虫植株产量高于感虫植株 ,对绒长、整齐度、比强度、伸长率、麦克隆值等品质指标亦无明显影响。通过回交可将 Bt基因转育到优良品种的遗传背景上 ,实现抗虫性状与高产、优质等性状的结合。     

甘蓝型油菜近等基因系构建及应用研究进展

【摘要】 近等基因系只有一两个基因差异,可以减少很多不必要的工作,因此在作物育种、分子标记和基因定位等方面研究有着广泛的应用。综述了甘蓝型油菜近等基因系材料的构建及近等基因系材料在抗逆、分子标记和蛋白质组学等研究中的应用情况。认为随着构建技术的推广,新型近等基因系材料将会发挥更大的作用。     

利用近等基因系定量评价穗稻瘟病田间抗性基因的预防效果

为了从数量上评价衍生于籼稻品种Modan并授予水稻穗瘟病田问抗性的Pb1基因的预防效果，用两年时间在两个试验点的水田中进行了稻瘟病抗性试验；试验所用的3种近等基因系是从大约2000个重组自交系中选出的，而这些重组自交系则又是通过日本水稻品种黄金晴（对稻瘟病敏感，不含Pb1基因）与月之光（抗稻瘟病，含有Pb1基因）杂交而育成的。     

Uniqueness of the Gossypium mustelinum Genome Revealed by GISH and 45S rDNA FISH

Gossypium mustelinum [-(AD)4"] is one of five tetraploid species in Gossypium.Three pairs of nucleolar organizer regions (NOR) in (AD)4 were detected by FISH with 45S rDNA as a probe,they also were observed with genomic DNA (gDNA) from Gossypium D genome species as probes.Of the three NORs or GISH-NORs,one was super-major and other two were minor,which was distinctly different from other tetraploid cottons.     

Improving Fiber Quality Traits of Xinluzao 45 Using Gossypium barbadense Chromosome Segment Substitution Lines

Gossypium barbadense chromosome segment substitution lines could be used as an important germplasm resource for improving fiber quality traits of upland cotton. In this study, we used TM-1 background substitution lines CSSL-122, the Xinjiang upland cotton Xinluzao 45 and hybrid, backcross progeny BC₂F₁ populations comprising 120 individual plants as test materials. Nineteen SSR markers located in Gossypium barbadense chromosomes land inked with fiber length and strength were used to screen out obvious polymorphic primers between the parents. We then traced and detected chromosome segments of Gossypium barbadense in the BC₂F₁ populations. Simultaneously, we compared fiber quality traits of positive plants that contained Gossypium barbadense chromosome segments with non-positive plants in the BC₂F₁ populations. The results showed that two markers, NAU2987 and BNL3145, which were linked with fiber length, could accurately identify the positive plants. In addition, compared with non-positive plants, the increased fiber length and strength of the positive plants were very significant (P < 0.01). Our research suggested that alien Gossypium barbadense chromosome segments significantly improved the fiber quality traits of Xinluzao 45. Thus, the Gossypium barbadense chromosome segment substitution lines will provide a vital theoretical basis and practical reference for improving Upland Cotton fiber quality traits.     

棉属九个同核异质系酯酶、过氧化物酶同工酶分析

利用聚丙烯酰胺凝胶电泳分析了棉属中九个同核异质系不同器官（或同一器官不同发育时期）脂酶、过氧化物酶同工酶。结果表明，异质系在酯酶、过氧化物酶同工酶酶谱上均表现出特异酶带。由于不同特异酶带代表了特定细胞质，作者提出了对这几种胞质进行分类、鉴定的可能方案，并对核质关系展开了讨论。     

陆地棉同核异质系异源胞质的遗传效应

通过核代换培育了具有亚洲棉(G.arboreum)、草棉(C.herbaceum)、异常棉(G.anomalum)、长萼棉(G.longicalyx)、哈克尼西棉(G.harknesii)、海岛棉(G.barbadense)、和陆地棉(C.hirsutum)野生种系蓬蓬棉(G.h.purpurascens)、墨西哥野生棉(C.h.mexicanum)等8个棉属种和种系胞质的中棉所7号同核系,并培育了中棉所7号自交系。在检验同核异质系核的同质性基础上研究了胞质遗传效应,结果所有供研究的胞质对产量、产量性状及除花器官外的形态特征均没有显著影响,胞质的遗传效应主要表现对花器官及抗枯萎病性的影响;哈克尼西棉胞质导致雄性完全不育并显著增加花药数量,笔形冠长度和直径显著变小,亚洲棉和异常棉胞质引起花药数量显著减少,笔形冠形状发生显著变化,亚洲棉胞质对抗枯萎病有较显著效应。     

Evaluation of Chromosome Segment Substitution Lines Related to Fiber Yield and Quality Traits from Gossypium hirsutum×Gossypium barbadense

Three generations of chromosome segment substitution lines lines (BC₄F₃, BC₄F_3:4, and BC₄F_3:5, Anyang, Korla) were used to evaluate phenotypic traits related to fiber yield and quality. The average performance of the traits in the population was near to the control, CCRI45, in the four populations. Seventy-eight percent of the lines showed longer and stronger fibers than the recurrent parent, and 22.59%～53.61% of lines had bigger bolls and higher lint percentages than the recurrent parent. Correlation analysis showed that the yield and fiber quality traits were positively correlated among the four populations. These results indicated that most of the genetic background in the substitution lines has been restored to the recurrent parent with abundant genetic variations. In addition, most of substitution lines had good yield and fiber quality. This study provided a large amount of basic research materials for cotton quantity genetics, gene pyramiding, and gene function analysis, and provided parent materials and new strains that can directly applied for cotton breeding.     

Genetic Diversity for Adult Plant Resistance to Leaf Rust (Puccinia recondita) in Near-Isogenic Lines and in Indian Wheats

Variation for adult plant resistance in near-isogenic wheat lines carrying Lrl4b, Lrl4ab and Lr30 in a ‘Thatcher’ background indicated the possible presence of novel adult plant resistance genes effective against the Indian leaf rust population. Sixty-one wheats released for cultivation in India were grown in isolated nurseries. Each nursery was separately inoculated with one of four leaf rust pathotypes which had been selected to aid identification of resistance effective only in the adult plant stage. Seven distinct response groups were recognised and a minimum of six sources of adult plant resistance were postulated. In a group of 14 wheats, resistance was explained on the basis of the seedling response genes that were identified. Similar results for two years with pathotype 77-1 gave support to the reliability of field tests. Adult plant resistance (APR) sources were either race-specific or effective against all pathotypes used. Seedlings of cultivars with APR showed susceptible reactions. The possible presence of Lr34 in Indian wheats and its role in durable leaf rust resistance are discussed.     

利用黄褐棉染色体片段导入系定位产量和纤维品质性状QTL

陆地棉的遗传基础狭窄,阻碍了棉花的遗传改良进程。为有效拓宽陆地棉的遗传基础,本试验利用野生种黄褐棉(AD4)为供体亲本,以综合性状优良的B0011品系为受体亲本(国审棉华杂棉H318的亲本之一),构建了含71个株系的导入系BC5S5群体。基于SLAF-seq的基因分型和多年多点田间试验的综合分析表明,该导入系在产量和纤维性状方面具有很大的变异,共检测到48个QTL,其中包含19个产量和29个纤维构成因素相关的QTL。在At亚组检测到9个性状的32个QTL,在Dt亚组为16个。进一步对QTL加性效应方向分析显示,其中有30个QTL的加性效应为正,18个QTL的加性效应为负。本研究结果为利用黄褐棉重要农艺性状有利等位基因改良陆地棉产量和品质奠定了基础。     

半野生棉与亚洲棉对草甘膦抗性评价及比较分析

采用叶片棉球定位鉴定法,初步鉴定并评价了半野生棉和亚洲棉对草甘膦的自然抗性,结果表明:参试材料抗性水平从高抗至高感分6级,极端抗性水平相差33.3倍(0.2%对0.006%)。其中,半野生棉对草甘膦的自然抗性分级丰富,抗性水平分布自高抗至高感;但大部分半野生棉不耐草甘膦,近86%(346份)的材料抗性水平处于低耐级别以下。而亚洲棉抗性水平都处于高抗至低耐的4个级别,抗性多样性较差,但整体抗性水平较高,70.3%(71份)表现为耐或以上级别。从半野生棉和亚洲棉中分别筛选到2份和5份高抗材料,半野生棉中有1份高感材料,亚洲棉中无对草甘膦敏感材料。本研究报道的极端材料为棉花草甘膦抗性遗传机制研究、新品种的培育、内源抗性基因的挖掘和感抗机理研究,奠定了良好的材料基础。     

Mapping QTL of Fiber Yield and Quality Traits in F2 Populations of Chromosome Segment Substitution Lines from Gossypium hirsutum × Gossypium barbadense

In this study, F₂ populations from two chromosome segment substitution lines MBI7455 and MBI7358, were quantified using SSR to evaluate the parents' genotype and detect QTL related to fiber quality plus yield traits of cotton. Results show that the recurrent parent (CCRI45) hosted 96.70% and 95.60% of chromosome segment substitution in MBI7455 with 12 chromosome segments and in MBI7358 with 16 chromosome substitution segments of Gossypium barbadense, respectively. In the F₂ population, the average rate of chromosome substitution of the recurrent parent (CCRI45) was 96.44%, and the average segments of chromosome substitution of Gossypium barbadense was 13.42, with an average segments of homozygous donor chromosome value of 3.90. Analysis showed 19 fiber quality-related QTL with a phenotypic variance of between 2.52%-13.11% and seven yield traits-related QTL with a range of 2.93%-11.40% phenotypic variance, resulting in a total of 26 QTL. The CSSLs could be used to detect QTL for fiber yield and quality traits, which offer an important foundation for the cotton molecular-assisted breeding.