甘蓝型油菜高/低含油量近等基因系构建及鉴定

为了探讨油菜种子中脂肪酸代谢调控的机制,发现有价值的高油基因,为高油育种研究提供基础材料,构建了甘蓝型油菜高/低含油量近等基因系。以高油品系NJ9为基础试材,连续自交2代后,选择农艺性状基本一致、含油量不同的姐妹系30份,进行连续自交和含油量的选择,最终获得9份拟近等基因系材料。以来源于芸薹属DB公共数据库的156对SSR引物对9份拟近等基因系株系材料进行扩增,筛选出32对引物用于近等性分析。基于32对SSR引物PCR扩增结果,经过遗传相似性系数估算以及聚类分析,结果显示,9份拟近等基因系试材遗传相似性系数均为0.85~0.94,其中,YC13-554和YC13-559的遗传相似性系数最大,为0.94,确定其为近等基因系。成熟期农艺性状调查结果,进一步确定YC13-554和YC13-559为高/低含油量近等基因系。结果表明,连续自交和定向选择的方法可用于构建近等基因系,丰富了近等基因系的来源。     

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。     

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。     

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。     

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.     

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。     

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.     

携有不同主效白叶枯病抗性单基因的3个粳稻近等基因系的选育

自１９８６至１９９３年，我们选育了第一批携有抗水稻白叶枯病主效基因，Ｘａ－３、Ｘａ－４和Ｘａ－１２的３个粳稻近等基因系：ＣＢＢ３、ＣＢＢ４和ＣＢＢ１２（暂名）。以１个感病并具有改良株型的粳稻品种沈农１０３３为轮回亲本，通过回交、自交直至ＢＣ５Ｆ５代。各世代经抗性测定和株型选择，直至各系的抗性稳定和农艺性状趋于一致。进行了３个近等基因系与其供体基因的抗性反应型比较、农艺性状考查及抗性遗传分析。通过育     

小麦芒长抑制基因B1近等基因系的鉴定及遗传分析

芒是小麦穗部重要的光合器官之一。本研究以一对芒长度存在差异的近等基因系SN051-1(长芒)与SN051-2(短芒)为材料,对其形态性状、穗和旗叶光合能力进行了调查,对芒长性状进行了遗传分析和分子标记分析。结果表明,SN051-1与SN051-2在抽穗期、开花期、株高、穗长、每株穗数、小穗数和穗粒数等性状方面均表现一致,仅芒的长度差异明显;SN051-1穗部的光合效率高于SN051-2穗部的光合效率,其千粒重也高于SN051-2;遗传分析证明二者芒性的遗传由单基因控制,短芒为显性。此外,我们筛选获得一个与芒长性状紧密连锁的、位于5A染色体长臂的SSR标记Xgwm291,与该基因间的遗传距离为4.99cM。因此,我们初步推断SN051-1与SN051-2可能是为位于5A染色体长臂的芒抑制基因B1的近等基因系。     

SSCP Marker Development and Analysis of Candidate Restorer Gene Rf1 for Cotton Cytoplasmic Male Sterility

[Objective] Locating the cotton cytoplasmic male sterility (CMS) restorer gene Rf₁ is important for investigating restorer gene mechanisms and improving restorer lines. In our previous study, a gene cluster, with nine Pentatricopeptide repeat(PPR) genes and nine other genes, was found within the 160-kb Rf₁ target region in Scaffold 333. The objective here was to improve the density of Rf1-linked markers in the target region and determine the expression profiles of candidate genes. [Method] Using the sequences of the 18 genes, we designed 155 single-strand conformation polymorphism (SSCP) primers covering all of the gene sequences to identify the polymorphic SSCP markers between the fertile and sterile pools. Additionally, real-time polymerase chain reaction(PCR) was performed to analyze the expression profiles of eight candidate genes in the four developmental stages of buds of sterile, maintainer and restoring lines, respectively. [Result] In total, 15 polymorphic primers were identified. A genotype analysis of the F₂ population was conducted using the 15 primers and 3 other polymorphic simple sequence repeats (SSR) markers. The markers were distributed in a 4.8 cM range. In addition, owing to the influence of sterile cytoplasm or restorer genes, most of the genes showed different expression patterns in the four developmental stages of the three lines' buds. [Conclusion] SSCP markers tightly linked to Rf₁ were identified and the expression profiles of candidate genes were determined. This study provides a basis for the further fine mapping of restorer genes and for candidate gene screening.     

小麦K型不育系恢复基因近等基因系农艺性状的遗传差异分析

摘 要：对近等基因系进行遗传背景检测,有利于选育高质量的近等基因系材料。试验以17个小麦K型不育系恢复基因的近等基因系（N1-N17）和轮回亲本不育系豫麦3号为材料,通过调查这些材料的株高、成穗数、旗叶长、旗叶宽、倒二叶长、脖长、倒一节长、倒二节长、穗长、小穗数等10个农艺性状,对恢复基因恢复基因近等基因系的农艺性状进行了差异比较和聚类分析。结果表明,连续回交5代后,回交后代与轮回亲本在形态性状上表现出较高的一致性,其中N14、 N1 、N7 、N16 和N15与不育系豫麦3号的性状差异较小,在欧氏距离为5.2处把它们聚在同一类,但从各性状的变异系数看,N7和N15自身的变异较大,所以N14、 N1和N16 与不育系豫麦3号具有较好的近等性。     

Breeding of transgenic rice restorer line for multiple resistance against bacterial blight, striped stem borer and herbicide

In this paper, we described the breeding of transgenic rice restorer line for multiple resistance against bacterial blight, striped stem borer (SSB) and herbicide by conventional crossing of two transgenic parental lines transformed independently with different genes. Two stable transgenic rice lines used as donor parents were developed, one was Zhongguo91 which contained cry1Ab gene (for insect resistance) and bar gene (for tolerance of herbicide), and the other was Yujing6 which contained Xa21 gene (resistance to bacterial blight). The elite restorer line Hui773 was used as recipient and crossed with the two stable transgenic rice lines. Then five successive backcrosses were made using Hui773 as recurrent parent. Two rice elite restorers, T773-1 expressing cry1Ab and bar genes and T773-2 expressing Xa21 gene, were obtained, which were confirmed by PCR analysis and testing selectable marker genes in the hybrid progenies. The cross was made between T773-1 and T773-2 to select stable restorer line carrying Xa21, cry1Ab and bar genes. Finally, we obtained transgenic restorer line T773 with good agronomic traits and obvious multiple resistance to Xanthomonas oryzae pv. oryzae, striped stem borer (Chilo suppressalis) and herbicide. The hybrid F₁ generation produced from the cross between transgenic restorer line T773 and a corresponding male sterile line Zaohua2A maintained obvious resistance to rice bacterial blight, rice leaffolder and striped stem borer, and showed significant heterosis. Our results indicate that it is feasible to develop transgenic hybrid rice cultivar through breeding transgenic restorer lines.     

油菜Nsa CMS候选恢复基因的来源及表达

根据同源序列法克隆的Nsa CMS候选恢复基因PPR618的序列,采用PCR方法在Nsa CMS不育系、恢复系、原始体细胞杂交亲本以及其他甘蓝型油菜品种中共克隆出22个同源序列。序列分析表明,野芥亲本野油18、甘蓝型油菜亲本中双4号各含有2个同源序列,而Nsa CMS 4个恢复系则分别含有3、1、1、1个同源序列。恢复系中候选恢复基因的序列与野芥亲本野油18的同源序列的一致性在93%以上,其中恢1、恢3和恢4中至少有一个同源序列与野油18的第一个同源序列完全相同,恢2中的同源序列与野油18的第2个同源序列完全相同;但与甘蓝型油菜的同源序列一致性均低于80%,说明候选恢复基因来源于新疆野芥,而不是甘蓝型油菜。除了原来的PPR618外,获得了3个来源于恢复系的新候选恢复基因。候选恢复基因在序列上与萝卜和矮牵牛的恢复基因一致性较高。半定量RT-PCR分析表明,候选恢复基因在恢复系中多个组织都有表达,但在根、茎中表达量特别少。随着营养器官到生殖器官的发育逐渐升高,候选恢复基因在恢复系中表达量最高的是雄性败育关键时期, 即1.5~2.5 mm的花蕾中,但不育系中的同源基因在茎中的表达量则相对高于其他组织。     

The degree of phenotypic resemblance of the near-isogenic lines of the wheat cultivar thatcher with their recurrent parent

Summary Thatcher, 16 of its near-isogenic lines (NIL) and 8 donors were investigated for the degree in which the NILs phenotypically resembled their recurrent parent. In general the NILs have a good phenotypic resemblance. In a few NILs characters occur that are not seen in Thatcher. In some cases these are found in the donor indicating the presence of donor genes other than the marker gene in the NIL. In other cases a character possessed by a NIL is not present in Thatcher or in its donor. In such cases either inhibitor genes may play a part, or the donor accession that accompanies the set of NILs may not be the donor used to breed the NIL. Two causes are possible. One is that the donor was a mixture of genotypes, the other is contamination.The presence of donor genes other than the marker Lr gene(s) in a NIL may lead to unexpected results or, what is worse, to wrong conclusions.     

小麦抗白粉病近等基因系遗传背景的分子标记检测

利用AFLP分子标记技术,对近年来培育的几套近等基因系的遗传背景进行了检测,发现所检测到的遗传多样性主要集中于供体亲本之间,而近等基因系内部已达到很高的遗传一致性;包含Pm4b的近等基因系、包含Pm2/Pm6的近等基因系与轮回亲本遗传上比较接近,其内部选系之间也达到很高的一致性,是理想的近等基因系;发现Pm21中簇毛     

Altered tissue-specific expression at the Wx gene of the opaque mutants in rice

Amylose content is a major determinant of the eating quality in rice. To elucidate the allelic diversity at the Waxy (Wx) gene which controls the amylose synthesis, two cultivated strains having opaque endosperms were studied. The gene responsible for opaque endosperms was introduced into the genetic background of the Japonica type of rice by successive backcrosses, and the two near-isogenic lines (NILs) were selected from the B5 generation. The genetic experiments revealed that an allele, Wx^op, controls opaque endosperms which show chalky as wx endosperms in spite of the production of amylose. Immunoblotting analysis was carried out to compare the gene expression by using the NILs with 4 different alleles (Wx^a, Wx^b, Wx^op and wx). The level of the gene product bound to starch granules was slightly lower in the NILs with Wx^op than that with Wx^b, showing a positive correlation with amylose content in the endosperm. Extracts from mature anthers indicated that the gene product was markedly reduced in the NILs with Wx^op as well as that with wx, showing an altered expression in the tissue specificity in the Wxop lines. Sequence analysis suggested that the Wx^op had been derived from Wx^a, independently of the origin of Wx^b. The importance of the gene regulation was discussed in relation to diversified phenotypes established during the domestication process.     

T型细胞质雄性不育小麦T763A的败育特点及育性恢复

为了明确T型细胞质雄性不育小麦T763A败育的形态特征和细胞学特点及对T763A恢复系的选用提供依据,以不育系T763A,保持系763B,恢复系Tm3315B、Tm504B和TP731B为供试材料,进行外部形态特征观察和花粉粒制片(醋酸洋红、I2-KI和DAPI);并以中国春和黑麦为对照试材,对所有供试材料进行核型鉴定。结果表明:T763A败育类型为典败和圆败,成熟花粉粒皱缩无规则,内含物少,花粉败育,败育主要发生在单核晚期到二核期;所有供试材料均为非1B/1R类型;3个恢复系(Tm3315B、Tm504B和TP731B)恢复能力均较强,其中以Tm504B对T763A的恢复能力相对最好,这可能与T763A的胞质类型及与恢复系所含的恢复基因数量有关。     

云南地方老品种水稻中恢复基因位点遗传差异分析

目前生产上使用的水稻恢复系之间的遗传基础过于狭窄,应积极开发水稻新恢复系的选育工作。云南地理气候条件特殊,地方老品种资源丰富,是筛选新恢复系的理想材料。以61份云南地方老品种水稻资源为父本,通过测交筛选到6份可以恢复野败型不育系的品种。再利用5对特异引物,对这6份恢复材料和7份恢复系的恢复基因位点遗传差异进行分析。结果表明：5对特异引物对6份恢复材料的扩增结果各不一样,而与对照恢复系仅有2.8 kb的一条扩增带型一致。说明云南地方老品种水稻中蕴藏的恢复基因与目前生产上应用的恢复基因的亲缘关系较远。从云南地方老品种水稻中发掘利用新型恢复基因和资源,是突破水稻恢复基因遗传基础狭窄、筛选新质源恢复系的一种有效策略。