与甜椒隐性细胞核雄性不育基因紧密连锁的SRAP分子标记的研究

以甜椒隐性细胞核雄性不育两用系AB91为材料,采用混合集群分析法(BSA)构建了不育池与可育池.利用SRAP分子标记技术,筛选了225对SRAP引物组合及1393对EcoR Ⅰ和Mse Ⅰ引物组合,获得了与隐性核不育基因连锁的2个SRAP标记:E37 M39、E44M93,片段长度约为200 bp和500 bp,与育性...     

辣椒CMS恢复基因的遗传分析及基因定位

本研究以辣椒胞质雄性不育系131BC5A与恢复系139D-21-3为亲本,构建了包含210个单株的F2群体,采用CAPS、SSR及SCAR等分子标记技术,对辣椒胞质雄性不育恢复基因进行遗传分析和基因定位研究。田间调查和遗传分析结果表明,F2群体的表现型中可育与不育的分离比为3:1。采用分离群体分组分析法(BSA),从F2可育群体和不育群体中各随机选取10株,构建可育和不育基因池。研究选用295对引物在亲本间进行多态性筛选,其中43对引物在亲本间表现出多态。通过进一步分析43对引物在基因池间的多态性,筛选出Rf基因连锁标记14个。然后对F2群体的210个单株进行连锁分析,最后将恢复基因定位在SSR标记pep43和pep20之间,约3.9 c M(或498.6 kb)的区间内,与两个标记的遗传距离分别为1.3 c M与2.6 c M。本研究为Rf基因的精细定位和克隆奠定了良好基础,也为辣椒雄性不育恢复系的分子标记辅助选择育种提供了理论参考。     

甜高粱含糖量与出汁率的SRAP分子标记

甜高粱(Sorghum bicolor L.Beauv)作为重要的能源作物已经为大家所认可。本研究利用集群分离分析(BSA)法结合SRAP分子标记技术相结合,以141个甜高粱"W455"和粒用高粱"忻粱52"杂交形成的重组自交系F2:3群体为材料,寻找与甜高粱含糖量性状相关的SRAP分子标记。结果表明:含糖量遗传属于单基因加显或单基因加性效应模式,出汁率遗传是由多基因共同控制的数量性状;通过SRAP分子标记发现,在7号染色体上找到1个与含糖量基因连锁的SRAP标记M3E7-S248,通过测序比对发现其与高粱腺苷二磷酸葡萄糖焦磷酸化酶亚基SH2有79%的相似度;在6号染色体上找到了3个与出汁率基因连锁的SRAP标记M8E2-J727(M8E2-J712)、M8R12-J241和F13E9-J150,其中M8E2-J712与M8E2-J727为共显性标记;通过测序比对发现标记M8E2-J712与多种植物叶绿体上的氧捕获增强蛋白高度一致;标记M8E2-J727与双色高粱未知功能蛋白m RNA中的一段序列具有99%的一致性;标记M8R12-J241和F13E9-J150均与6号染色体上的未知基因序列具有高度一致性。     

利用SRAP与SSR标记分析不同类型甜菜的遗传多样性

为选育优质甜菜新品种, 指导种质资源引进和利用, 为进行分子标记辅助选择育种提供科学依据, 采用SRAP和SSR两种分子标记方法相结合, 对甜菜单胚雄性不育系及保持系等49份材料进行遗传多样性分析。利用4个表型差异显著的甜菜品系对SRAP的64对引物组合及SSR的11对引物组合进行扩增, 分别筛选出有效引物组合11对和9对。SRAP的11对引物组合共产生199条扩增带, 其中有86条多态性带, 多态性带的比率平均为43.7%。SSR的9对引物共产生35条扩增带, 多态性比率为100%。全部材料的平均遗传距离为0.3860, 平均遗传相似系数为0.6795, 大约30%的材料遗传距离或遗传相似系数具显著或极显著差异。遗传相似系数平均值比较, 多胚四倍体品系0.7264＞单胚杂交组合0.7243＞国外品种0.7060＞多胚二倍体品系0.6908＞单胚品系0.6837。在遗传距离0.20处, 将49个甜菜材料划分为A、B、C、D 4个类群, D类群又分为4个亚类, 较好地显示了甜菜材料丰富的遗传多样性。表明不同甜菜品种具有相当高的异质性, 国外与国内材料的遗传基础存在一定差异, 但生产应用的甜菜品种间存在亲缘关系较近、遗传基础较窄的倾向。     

亚麻显性雄性核不育基因的RAPD标记

应用252条10-mer随机引物对遗传背景相似的可育株和不育株亚麻进行了RAPD分子标记,在不育株与可育株之间寻找DNA的多态性差异带,结果发现,在252条随机引物中有2条引物(即S62和S135)可分别得到1个与显性核不育的雄性基因有关的RAPD分子标记为S62-500和S135-350。     

小麦新种质YW243抗条锈病新基因的AFLP标记

小麦新种质YW243抗条锈病新基因对条锈菌具有较宽抗谱，对26个不同毒性谱的条锈菌菌系免疫到近免疫，与已知基因系抗病谱不同，可能为新的基因或基因组合。遗传分析表明，YW243对条锈菌条中31号小种的抗性由显性单基因YrX控制。通过BSA法对1056对MseI/EcoRI AFLP引物进行筛选，结果表明7对引物可扩增出多态性片段，通过对F2群体单株遗传连锁分析和作图软件分析，结果表明，2个AFLP 标记M54E63-700、M54E64-699与YrX连锁较紧密、遗传距离为9.27 cM，为该基因的精细作图和分子标记辅助育种打下了一定基础。     

青花菜胞质不育相关基因的SRAP标记筛选

以青花菜Ogura胞质不育系XY及其保持系XYF基因组DNA为材料进行SRAP分析,共筛选了146对SRAP引物,其中8对引物在两系之间表现差异,经单株验证只有引物对Me6＋Em7没有交换单株,找到不育系与保持系间的特异扩增片段M6E7-700,该片段仅在不育系中稳定扩增。测序结果表明该片段全长为689bp,经Blast分析比对,该片段与已报道的所有育性相关基因序列均不同源,而其部分序列与大白菜BAC克隆KBrB042N05和KBrB041L12的部分序列高度同源,表明该片段是一段新发现的与胞质不育育性相关片段,且该片段可能来自核DNA。根据测序结果设计了1对特异引物,将M6E7-700标记转化为更稳定的SCAR标记。本研究首次发现芸薹属作物Ogura-CMS不育系和保持系的核DNA也存在差异,该结果为从质核互作角度解释细胞质雄性不育的分子机制提供了新线索。     

谷子显性雄性不育基因Msch的AFLP标记

利用雄性不育是实现谷子杂种优势利用最经济、有效的途径之一.为了寻找与不育基因Msch紧密连锁的分子标记,提高不育系的选育效率,本研究构建了Msch不育/可育近等基因系(NILs),通过对400对AFLP引物组合进行筛选,找到了与不育基因紧密连锁的两个AFLP标记(P17/M37224和P35/M52208),与不育基因的遗传距离分别是2.1 cM和1.4 cM,而且位于不育基因的同一侧,标记间相距0.7 cM.这两个AFLP标记可有效用于分子标记辅助选择育种.     

鲜食番茄茄红素基因的AFLP-SCAR分子标记

番茄红素(Lycopene)是功能性天然色素,因其高抗氧化功能而备受关注.为筛选高番茄红素品种,加速鲜食番茄分子标记辅助育种进程,本试验以高茄红素番茄骨干系F-516F8和低茄红素骨干系Nor作为试验材料,研究了鲜食番茄高茄红素基因的AFLP-SCAR分子标记.利用杂交F1代、自交F2代遗传群体,通过64对E/M引物组...     

YM型小麦温敏雄性不育系不育基因的QTL定位

YM型小麦温敏雄性不育系的不育基因被定位在1Bs染色体片段上, 但已发现的相邻分子标记与该基因的遗传距离较大, 达10 cM以上。为寻找与该基因连锁更紧密的分子标记, 以YM型温敏雄性不育系ATM3314与恢复系中国春杂交的F₂代200株为作图群体, 从1Bs的22个SSR引物中筛选出5个在亲本和F₂代中分离的SSR引物, 构建了1个包含5个标记的1Bs局部遗传连锁图谱。结合F₂代个体的育性调查, 采用复合区间作图法在YM型温敏雄性不育系的1Bs染色体上检测到不育基因的1个主效QTLrfv1-1和1个微效QTLrfv1-2。rfv1-1位于SSR标记Xgwm18和Xwmc406之间, 与两标记的遗传距离分别为6.0 cM和4.6 cM, LOD值为8.80, 加性效应23.87, 显性效应10.44, 可解释表型变异的23.91%; rfv1-2位于Xwmc406和Xbarc8之间, 与两标记的遗传距离分别为4.0 cM和3.4 cM, LOD值为3.10, 加性效应17.59, 显性效应5.99, 可解释表型变异的7.78%。本研究初步定位了YM型小麦温敏雄性不育系1Bs染色体片段上不育基因的QTL, 为进一步准确定位该基因奠定了基础。     

蓝标型小麦核雄性不育、保持系的选育研究

将蓝粒小麦中携带有蓝色胚乳基因和育性恢复基因的4E 染色体,附加到小麦核型雄性不育系上,选育出蓝标型小麦雄性不育、保持系。该体系浅蓝粒种子长出植株自交结实,粒色分离为深蓝、浅蓝和白粒。其中,白粒种子长出植株,全部为雄性不育,为普通小麦品种所恢复,杂种优势显著;浅蓝粒种子植株则自交结实,粒色仍分离为深蓝、浅蓝和白     

棉花核不育系豫98-8A育性遗传分析

为了阐明1999年从转基因后代遗传群体中发现的1株雄性不育植株不育基因的遗传规律及其与现有不育基因的等位性,采用表型观察测量,以及经典的自交和测交手段,研究了该不育材料败育性状的遗传规律。花器官形态特征调查表明：不育株花柱长和花柱外露长度均明显高于同质系的正常可育株,而每朵花的子房直径及花药数量没有明显差异。遗传分析表明：杂合体可育株自交,后代不育株与可育株呈3:1分离,不育株与杂合姊妹可育株测交,不育株与可育株呈1:1分离,表明该核不育材料受隐性单位点控制;与阆A(msc1)、洞A(msc3)等育性位点杂合可育株分别杂交,其F1代单株育性均得到恢复。由其F1代产生的F1:2家系中均出现不育株与可育株呈1:3和7:9两个育性分离群体,表明该材料败育基因为不同于阆A、洞A的不育基因位点。     

Chromosomal location of fertility restoring genes for ‘wild abortive’ cytoplasmic male sterility using primary trisomics in rice

Summary Identification and location of fertility restoring genes facilitates their deployment in a hybrid breeding program involving cytoplasmic male sterility (CMS) system. The study aimed to locate fertility restorer genes of CMSWA system on specific chromosomes of rice using primary trisomics of IR36 (restorer), CMS (IR58025A) and maintainer (IR58025B) lines. Primary trisomic series (Triplo 1 to 12) was crossed as maternal parent with the maintainer line IR58025B. The selected trisomic and disomic F₁ plants were testcrossed as male parents with the CMS line IR58025A. Plants in testcross families derived from disomic F₁ plants (Group I crosses) were all diploid; however, in the testcross families derived from trisomic F₁ plants (Group II crosses), some trisomic plants were observed. Diploid plants in all testcross families were analyzed for pollen fertility using 1% IKI stain. All testeross families from Group I crosses segregated in the ratio of 2 fertile: 1 partially fertile+partially sterile: 1 sterile plants indicating that fertility restoration was controlled by two independent dominant genes: one of the genes was stronger than the other. Testcross families from Group II crosses segregated in 2 fertile: 1 partially fertile+ partially sterile: 1 sterile plants in crosses involving Triplo 1, 4, 5, 6, 8, 9, 11 and 12, but families involving triplo 7 and triplo 10 showed significantly higher X² values, indicating that the two fertility restorer genes were located on chromosome 7 and 10. Stronger restorer gene (Rf-WA-1) was located on chromosome 7 and weaker restorer gene (Rf-WA-2) was located on chromosome 10. These findings should facilitate tagging of these genes with molecular markers with the ultimate aim to practice marker-aided selection for fertility restoration ability.     

Influence of water availability on fertility restoration of CMS lines with the 'M35', A4 and '9E' CMS-inducing cytoplasms of sorghum

Male fertility restoration in new types of sorghum cytoplasmic male sterility‐inducing cytoplasms (A4, ‘9E’, ‘M35’), characterized by the formation of non‐dehiscent anthers, is difficult. Lines with fertility‐restorer genes for these unique cytoplasms do occur, but rarely, and when found tend to be unstable in their inheritance and expression. The aim of this research was to explore reasons for this instability. Seven lines in three unique cytoplasms, ‘9E’, A4 and ‘M35’, and six lines that restore with these cytoplasms were grown at the Agricultural Research Institute for South‐East Region in Saratov, Russia from 1993 to 2004. Levels of male fertility restoration and various environmental factors were recorded. It is reported that for sorghum hybrids in the A4, ‘9E’ and ‘M35’ male‐sterile cytoplasms, the level of plant male fertility is determined by the level of water available to plants during anther and pollen formation that which ‘switches on’ the expression of fertility‐restoring genes, and is possibly involved in an unusual type of male fertility inheritance in these cytoplasms. The creation of reliable line‐fertility restorers capable of the restoration of male fertility of F₁ hybrids in ‘M35’ cytoplasm under conditions of water stress is also reported. Current research explore mechanisms involved possible in responses to water levels at various growth stages and their influence on fertility within these cytoplasms.     

温敏型雄性不育亚麻的研究

通过在自然条件下对亚麻雄性不育材料的特征特性和不育性表现的研究表明, 该不育材料农艺性状优良, 雄性不育特征明显. 温度对不育性有重要影响, 一定温度范围内, 高温能使育性提高, 结果率和结实率增加, 低温使育性下降, 结果率和结实率下降, 同时还发现不同材料对温度的敏感程度不同, 通过对杂交后代育性分离的分析, 表明几     

陆两优996种子纯度的SRAP指纹图谱鉴定

利用270对SRAP引物对陆两优996和2个亲本的DNA指纹图谱，获得能在父、母本间表现出多态性的特异SRAP标记引物有54对，其中差异性条带136个。利用Me3Em8引物，条带在杂交种完全互补，对200株样本进行了纯度鉴定，结果鉴定纯度为97.50%，与田间种植鉴结果97.50%(海南鉴定)一致，表明SRAP标记技术适用于种子纯度鉴定。     

Inheritance of male sterility mutations induced in haploid sorghum tissue culture

Summary A high frequency of male sterile mutants regeneration was shown in callus cultures derived from leaves and panicles of haploid sorghum (Ms_c1, A1 cytoplasm) and a spontaneous autodiploid obtained from this haploid. The cultures derived from the embryos of this autodiploid yielded significantly fewer mutants. Absolutely or partially male sterile mutants appeared among the regenerants or in the progeny of fertile regenerants. In the self-fertilized progenies of partially male sterile mutants and in the hybrids of sterile mutants with autodiploid line (i.e. under one and the same nuclear genome) male sterility mutations were inherited as cytoplasmic. Non-Mendelian segregation of sterile, partially male sterile and fertile plants was observed in these progenies. Partially male sterile plants were characterized by somatic segregation of male sterility genetic factors. In test-crosses with some CMS A1 fertility restorers, mutations were manifested as nuclear recessive while with others as nuclear dominant. These differences are supposed to be the result of interaction of fertility restorer genes of these testers with the novel cytoplasm. Male sterility mutations accompanied with female sterility were inherited as nuclear recessives.Abbreviations f fertile - ps partially male sterile - s male sterile plants     

Influence of Tetracycline on the Expression of Cytoplasmic Male Sterility (cms) in Chives (Allium schoenoprasum L.)

To influence the expression of cytoplasmic male sterility (cms), different clone members of male sterile genotypes of chives were treated with different chemicals, especially antibiotics which are known to act as protein synthesis inhibitors. A remarkable number of male sterile plants turned to partial or full male fertility after the treatment with tetracycline HCl and tetracycline base. Pollen grains produced by tetracycline on male sterile plants were viable and able to fertilize the egg cells. By using the perennial nature of Allium schoenoprasum L. it could be shown that the fertility induced on male sterile plants by tetracycline is reversible. This could be of practical importance for the multiplication of male sterile parents in hybrid breeding. None of the chemicals had any effect on the fertility of male fertile plants. Differences in reaction indicated that genetic differences exist between male sterile plants responding to tetracycline and those which do not. The results are discussed in view of the influence of tetracycline on the mitochondrial protein biosynthesis.     

水稻质核互作雄性不育系选育的反向杂交法研究

不同的部分保持系可能存在不同的微效恢复基因,通过有性杂交,产生基因重组,能够获得完全保持系类型,但只有在不育细胞质中才能观察得到微效恢复基因是否存在以及它们的作用大小.反向杂交法以不育细胞质为选择背景,在杂交后代植株中直接观察到微效恢复基因的表达,获得的完全不育株,在一定程度上排除微效恢复基因,不育株再通过高温处理转换为可育后自交,来自不育株的微效恢复基因可以进一步排除掉,从而产生出没有(或很少)微效恢复基因的"亲本",利用该"亲本"高温处理后仍可转换为可育的特性,作为父本进行杂交或回交育种,在其后代中获得没有微效恢复基因的完全保持系.该研究为Cp 26不育细胞质源创造出了完全保持系.如果在田间鉴定出优异的完全不育株,对其进行单倍体育种(诱导孤雌生殖或花培),选择到没有(或很少)微效恢复基因的纯合不育株.再对其进行花培,筛选可育的突变体;或者利用纯合不育株的原生质体与一个已破坏细胞核的可育系原生质体融合,都可能得到具有纯合不育株细胞核和可育细胞质的保持系,而能够完善和改进反向杂交法.反向杂交法不但能够为所谓不能保持的不育细胞质源创造出保持系,而且有利于加强新不育系选育的目标性和预见性,提高不育系配合力和培育不同类型优异不育系.     

食荚豌豆雄性不育突变体的遗传研究

对国内首例豌豆雄性不育突变体的不育度、遗传特点及稳定性进行研究。观察发现：在生育前期。不育株外部形态特征与正常株没有明显差异;现蕾后,剥开花蕾可看到不育株的花药呈淡黄色半透明状。而可育株的花药呈橙黄色。用I2-KI染色法镜检花粉的可染性,发现不育株的花药内没有花粉粒,败育彻底,为典型的“无花粉型”雄性不育。用不育株作母本,与同品系的正常可育株进行姊妹交,F1全部可育,F2可育株与不育株的分离比例为3：1。用不育株作母本,其他品系作父本进行测交,同时用其他品系作母本,姊妹交F1作父本进行反交,正反交后代的育性表现一致。F1全部可育。F2可育株与不育株呈3：1分离。结果表明：该雄性不育突变体的不育性是可遗传的,属单隐性基因控制的核不育类型,与细胞质遗传物质无关。在不同年份、不同季节下,不育性状表现稳定。