属间体细胞杂交创建甘蓝型油菜细胞质雄性不育系及其鉴定

 利用原生质体融合技,获得了甘蓝型油菜品种中双4号与新疆野生油菜野油18的对称性体细胞杂种。杂种当代建成的6株植株中有2株雄性不育株。用中双4号做轮回亲本与不育株回交,不育株率随回交代数增加而升高,至BC3代大部分株系的不育株率接近100%,到BC4代不育性稳定,群体中无育性分离现象。用波里马CMS的恢复系和保持系与其测交,6个甘蓝型油菜品系均可保持该不育系的不育性,表明该雄性不育胞质与Pol不育胞质不同。     

甘蓝型油菜细胞质雄性不育研究进展

对甘蓝型油菜细胞质雄性不育的胞质类型及各自的研究进展,造成甘蓝型油菜细胞质雄性不育的分子机理,环境对甘蓝型油菜细胞质雄性不育的影响,以及甘蓝型油菜细胞质雄性不育系的微量花粉问题及解决方法的研究进展进行了综合评述.并对甘蓝型油菜细胞质雄性不育系选育的进展情况进行了扼要评论.指出只有培育出不育性稳定的甘蓝型油菜细胞质雄性不育系,才可能从根本上解决甘蓝型油菜细胞质雄性不育的微粉问题     

Cytoplasmic Male Sterility in Maize

14 isoplasmic and allonuclear cytoplasmic male sterile lines were used as female parents, 8 tester lines as male parents, 101 F1 progenies were obtained. Fertility restoration response of 101 F1 progenies were investigated through field observation and pollen stainability examination under microscope. 14 isoplasmic and allonuclear cytoplasmic male sterile lines were developed by repeated backcross with recurrent male parent lines for more than 8 generations. The result shows: tester line Zifeng1 not only restored the isoplasmic and allonuclear sterile lines of group C backcrossed with Mo17, Yu30 and Heer, but also completely restored the isoplasmic and allonuclear cytoplasm male sterile lines of group T backcrossed with Mo17, HZS , 1792 ,292 and Yu30. Therefore, nuclear background limits the use of Zifeng1 as a tester for identification of cytoplasmic male sterility. Furthermore RFLPs of mitochondrial DNA of 6 isonuclear and alloplasmic cytoplasmic male sterile lines were analyzed with Bam H Ⅰ and Hind Ⅲ restriction endonuclease and mitochondrial DNA probes pBcmH3 and Cox Ⅱ. The same RFLPs were found within sterile cytoplasm of group C, including C,Chuan G, Lei 2 and Lei 3, but a different RFLP pattern was observed among sterile cytoplasm of group S, C,T and the normal cytoplasm. This result suggested that the RFLP markers tightly linked to sterile mitochondrial genes of different groups could be applied in the identifcation of cytoplasmic male sterility.     

Development of Japonica Male Sterile Lines Integrating Cytoplasmic Male Sterility and Photosensitive Genic Male Sterility

Acknowledgement It has been previously established that the BT type of cytoplasmic male sterility （CMS） is induced by high temperatures, while photosensitive genic male sterility （PGMS） seed sets by low temperatures induce. In the current study, we have bred photosensitive cytoplasmic male sterility （PCMS） lines （2308SA and 2310SA） by crossing the CMS line with the PGMS japonica line with maintainer genes. The sterility of PCMS japonica was consequently controlled by two groups of male sterile genes resulting from the integration of PGMS and CMS genes. The results on plant fertility, at different sowing times, were as follows： （a） Under conditions of natural long-day photoperiod and at temperatures above 35~C, the PGMS gene regulated PCMS japonica sterility - the higher the temperature, the lower the pollen fertility. However, bagged seed sets of PCMS japonica, not exposed to high temperatures, induced the CMS seed set. （b） Exposure to long-day photoperiod and temperature conditions between 35℃ and the critical sterility inducing temperature of PGMS resulted in both PGMS and CMS gene controlled sterility of PCMS japonica, which exhibited stable characteristics. （c） When exposed to critical sterility inducing temperatures or short-day photoperiod and daily high temperatures below 32℃, the BT type of the CMS gene regulated PCMS sterility. Under these conditions, the PGMS gene rendered male sterility insusceptible to occasional cool summer days when this PCMS line, adopted for hybrid seed production, develops into panicle differentiation stage. The present study also investigated the fertility restoration, seed production and combining ability of PCMS japonica so as to optimize its use.     

核质互作不育和光敏核不育聚合的粳稻不育系选育

 针对粳型核质互作不育系高温自交结实和粳型光敏核不育系低温自交结实的现状,选择BT型核质互作不育系与含有BT型保持基因的粳型光敏核不育系,通过杂交和回交,使雄性核不育基因与核质互作不育基因聚合,育成了粳型光敏核质互作不育系2308SA和2310SA。该不育系的不育性是由两套相互独立的基因系统控制的,分期播种育性观察结果表明,在长日高温下,雄性光敏核不育隐性纯合基因控制水稻不育性,避免了BT型不育基因因穗期受高温影响而导致不育系自交结实;在长日适温下,水稻不育性由核不育基因和核质互作不育基因共同控制而表现稳定;在低温（低于光敏不育系不育临界温度）或短日适温（日最高气温<32℃）下,BT型核质互作不育基因控制水稻不育性,消除了育性敏感期低温导致核不育系"打摆子"造成杂交种子不纯的风险。还对粳型光敏感核质互作不育系的可恢性、可繁性和配合力进行了研究。     

粳稻细胞质雄性不育系育性稳定性研究概述

粳稻细胞质雄性不育系的育性稳定性是其杂种优势利用中不可忽视的问题之一.较为详细地对影响粳稻细胞质雄性不育系育性稳定性的因素,育性变化的敏感阶段,育性稳定性研究及鉴定方法,不育系育性不稳定解决途径4个方面进行了综述.     

甘蓝型油菜双低显性细胞核+细胞质雄性不育系DGCMS-3A的构建及遗传模式研究

 【目的】验证显性核不育两用系与波里马三系的基因型,明确Gd1AB和D3AB显性核不育的遗传模式,并构建显性细胞核+细胞质不育系统。【方法】通过显性核三系与波里马三系相互杂交验证其基因型,分析Mf和Rfc基因的等位性,采用临保系与显性纯合两用系×显性核不育恢复系F2代核不育株测交的方法,推论显性纯合两用系Gd1AB和D3AB的遗传控制模式。C3B为轮回父本与不育胞质的显性核不育株定向回交,以创建显性核不育+细胞质不育。【结果】鉴别了Gd1AB和D3AB的遗传控制模式,确定甘蓝型油菜显性纯合两用系Gd1AB由一对复等位遗传模式控制,而显性纯合两用系D3AB由两对基因互作控制。初步构建出双低显性杂合核不育+细胞质不育系统DGCMS-3A,并证实了选育显性纯合核不育+细胞质不育系的可行性。【结论】显性抑制基因Mf与波里马恢复基因Rfc不等位,Gd1AB的基因型为N（MsMsrfcrfc）×N（MsMfrfcrfc）),D3AB的基因型为N（MsMsmfmfrfcrfc）×N（MsMsMfmfrfcrfc）。显性核不育+细胞质不育可细分为四种遗传模式：双位点互作显性杂合核不育+细胞质不育[S（Msmsmfmfrfcrfc）+S（msmsmfmfrfcrfc）]、单位点复等位显性杂合核不育+细胞质不育[S（Msmfrfcrfc）+S（mfmfrfcrfc）]、双位点互作显性纯合核不育+细胞质不育[S（MsMsmfmfrfcrfc）+S（MsMsMfmfrfcrfc）]、单位点复等位显性纯合核不育+细胞质不育[S（MsMsrfcrfc）+S（MsMfrfcrfc）]。     

水稻细胞质雄性不育育性恢复研究进展

论述了水稻细胞雄性不育及育性恢复基因的主要类型、恢复基因的遗传、定位和育种的研究进展,并提出其存在的问题和展望。     

细胞质雄性不育棉花的转基因恢复系的选育

 用根癌农杆菌介导法 ,将谷胱甘肽S 转移酶基因 (gst)导入待改良的棉花细胞质雄性不育的恢复系中。从转化植株的后代中 ,筛选到一个对雄性不育系具有强恢复力的恢复系 ,暂记为“浙大强恢”。“浙大强恢”与受体恢复系“DES HAF2 77”比较 ,对不育系的恢复力提高了 2 5 .8% ,从而使杂种F1单株结铃数多 3.6个 ,不孕籽率降低了10 .1% ,皮棉产量提高了 10 .6 %。以基因 gst作为探针进行的Southern和Northern核酸杂交分析表明 ,“浙大强恢”中含有外源基因 gst,并有较高水平的表达。     

Development of Transgenic Restorer of Cytoplasmic Male Sterility in Upland Cotton

A glutathione S-transferase gene (gst) has been introduced into restorers of cytoplasmic male sterility in upland cotton ( Gossypium hirsutum L. ) using Agrobacterium-mediated transformation. A transviable pollens of hybrid (sterile line×restorer) F1. The hybrid gave 3.6 more bolls per plant, 10.1% less aborted seeds and 10.6% more lint yield when ‘Zheda strong restorer’ was used as male parent than when ‘DES-HAF277’ was. Southern and Northern bloting analysis showed that the foreign gst gene was detectable and highly expressed in ‘Zheda strong restorer’.     

1份玉米胞质雄性不育材料的分类研究

[目的]报道1份败育彻底、育性稳定、农艺性状优良的玉米胞质雄性不育材料TFMs。[方法]采用综合鉴定、逐级分类的方法,对1份不明类型的玉米胞质雄性不育材料进行了研究。[结果]不同播期的育性表现为：5月15日播种的有3株花药较饱满,部分花药外露,但未见花药开裂现象,其他株和其他播期植株花药均瘦瘪,无花粉,完全不育;育性恢复专效性测定结果表明,获白、自330、N6测交F1,花药大小颜色、开花散粉正常,可育株率100%,镜检可染花粉率90%以上,推测其为孢子体不育。[结论]经初步判定,TFMs属于C组胞质雄性不育。     

Abortive Process of a Novel Rapeseed Cytoplasmic Male Sterility Line Derived from Somatic Hybrids Between Brassica napus and Sinapis alba

Somatic hybridization is performed to obtain significant cytoplasmic male sterility （CMS） lines, whose CMS genes are derived either from the transfer of sterile genes from the mitochondrial genome of donor parent to the counterpart of receptor or production of new sterile genes caused by mitochondrial genome recombination of the biparent during protoplast fusion. In this study, a novel male sterile line, SaNa-IA, was obtained from the somatic hybridization between Brassica napus and Sinapis alba. The normal anther development of the maintainer line, SaNa-IB, and the abortive process of SaNa-IA were described through phenotypic observations and microtome sections. The floral organ of the sterile line SaNa-IA was sterile with a shortened filament and deflated anther. No detectable pollen grains were found on the surface of the sterile anthers. Semi-thin sections indicated that SaNa-IA aborted in the pollen mother cell （PMC） stage when vacuolization of the tapetum and PMCs began. The tapetum radically elongated and became highly vacuolated, occupying the entire locule together with the vacuolated microspores. Therefore, SaNa-IA is different from other CMS lines, such as ogu CMS, pol CMS and nap CMS as shown by the abortive process of the anther.     

辣椒胞质雄性不育（CMS）不育系的转育技术研究

纯合自交系的CMS基因类型有N（msms）、N（MsMs）、S（MsMs）3种;N（Msms）、S（Msms）2种基因型后代会出现分离,根据不同CMS基因型的特点,从理论和实际上探讨了不同的基因型最快速有效的转育技术,并成功利用此技术转育了10对不育系及其相应的保持系。     

甘蓝型油菜新型细胞质雄性不育系NCa不育胞质类型的分子鉴定

 以4个限制性内切酶（Eco RI、Bam HI、HindIII、PstI）进行酶切、14个线粒体基因为探针，对NCa和pol、nap、ogura、tour等5个甘蓝型油菜细胞质雄性不育系的不育胞质线粒体DNA进行RFLP分析。在总计54个有效探针/酶组合中，除12个组合在5种不育细胞质间没有检测到差异外，其余42个探针/酶组合在5个不育系中均检测到差异。其中， atp1/EcoRI、atp1/HindIII、atp6/Eco RI、atp8/Bam HI、orf139/ Bam HI等5个探针/酶组合在5个不育系的不育胞质间检测到各不相同的条带，从而可以将它们两两完全区分开。RFLP分析结果从分子水平上进一步证明了NCa不育系确实具有与pol、nap、ogura、tour等不育系不同的的不育胞质，是一类新的不育胞质类型。这5个探针/酶组合也可以作为鉴定甘蓝型油菜不育细胞质的分子标记。根据控制pol、nap、ogura等不育系的线粒体基因序列和拟南芥线粒体基因组序列设计了22对引物，对上述5种甘蓝型油菜不育系的线粒体DNA进行了 PCR分析，没有找到可以将NCa不育胞质与其它类型不育胞质明显区分开来的引物。     

原生质体非对称融合法获得花椰菜与ogura CMS甘蓝型油菜种间杂种

 摘要：甘蓝型油菜Ogu CMS系幼叶原生质体经剂量分别为0.0250、0.0750、0.3150 、0.4680和0.9600 J·cm-2 的紫外线辐照后， 与花椰菜下胚轴原生质体通过PEG法诱导融合。其中0.0250~0.4680J·cm-2获得再生植株68株，0.9600J· cm-2辐射剂量下没有获得再生株。通过根尖染色体计数、流式细胞仪倍性鉴定、同工酶酶谱分析、RAPD及Ogu CMS线粒体基因orf138特异引物扩增等方法对再生株进行了鉴定，结果表明，其中32株为杂种植株，Ogu线粒体基因orf138已转入花椰菜中。同时也发现，紫外线辐照剂量的增加对供体染色体的丢失程度影响不大。     

棉花细胞质雄性不育与育性恢复的研究与利用

对棉花细胞质雄性不育与育性恢复的研究现状进行了较为全面的综述。介绍了棉花细胞质雄性不育系、恢复系及“三系”杂种棉的选育进展,不育系花药的细胞学及其生理生化特征,细胞质雄性不育产生的分子机理,不育胞质对杂种一代的遗传效应。详细阐明了细胞质雄性不育恢复系的类型,育性恢复的遗传方式,育性恢复基因与育性增强基因之间的关系以及恢复基因的遗传定位。文中对“三系”杂种棉选育进展缓慢的原因进行了剖析,并从利用新的胞质不育类型选育优良不育系,聚合多个恢复基因和育性增强基因选育强恢复系,优化以蜜蜂为媒介的传粉体系等方面,探讨了“三系”杂种棉的发展对策。     

水稻细胞质雄性不育育性恢复遗传及其环境效应研究

在江西环境条件下利用"协青早A/(协青早B/密阳46)F6"、"珍汕97A/(协青早B/密阳46)F6"、"协青早A/(珍汕97B/密阳46)F6"及"珍汕97A/(珍汕97B/密阳46)F6"4套测交群体,进行了水稻细胞质雄性不育育性恢复遗传研究。结果表明:水稻CMS的育性恢复表现为质量—数量性状;不同遗传背景对水稻育性恢复的影响可能主要表现在微效QTL的不同;不同环境条件对水稻育性恢复的影响不仅表现为微效QTL的变化上,主效基因的作用亦可能因环境而异。     

辣椒胞质雄性不育分子生物学研究进展

文中对辣椒胞质雄性不育的来源、分子标记、基因分布和定位、基因克隆和表达等方面的最新进展进行了综述,并对分子生物学在辣椒胞质雄性不育方面的应用前景进行了展望。     

细胞质雄性不育玉米的根系特性和氮效率研究

【目的】探讨细胞质雄性不育玉米的根系特性和氮素吸收利用规律。【方法】在土柱栽培条件下,选择目前生产中广泛应用的骨干自交系478和齐319的细胞质雄性不育系（CMS）和其同型保持系（可育系）为试验材料,比较分析雄性不育系及其可育系灌浆期根系特性的差异及对缺氮的响应。【结果】CMS玉米的籽粒产量和千粒重均高于其同型可育植株（P＜0.05）,生物产量差异不显著（P＞0.05）,收获指数明显较高（P＜0.05）,且缺氮条件下优势更为明显。CMS玉米灌浆期具有较高的根系干重、根系体积和根系总活力,且深层根系分布相对较多,根系活力较强,有效延长了根系的功能期,促进了植株对养分和水分的吸收。CMS玉米氮素积累总量高于其同型可育系（P＜0.05）,成熟期氮素在茎秆和籽粒中分配较多;低氮水平下,CMS玉米的氮素转移率、贡献率和氮素利用率均显著高于其同型可育系（P＜0.05）;CMS玉米单株氮效率较高,氮响应度相对较低。【结论】细胞质雄性不育玉米籽粒产量高、氮效率高与其深层根量多、根系活力强密切相关。     

甜菜细胞质雄性不育与内源激素含量的关系

 【目的】研究甜菜细胞质雄性不育与内源激素含量的关系。【方法】以表现型具明显差异的两对甜菜单胚CMS系及其O型系作为试验材料,采用间接酶联免疫（ELISA）检测技术,分析营养生长期和生殖生长期的功能叶片和花蕾中生长素、细胞分裂素、脱落酸、赤霉素等4大类内源激素的含量和比值变化。【结果】在营养生长的糖分积累期,ABA、IAA、GA3含量均为O型系高于CMS系,差异显著,而DHZR、IPA、ZR含量在营养生长期间均差异不显著;在开花初期的叶片,除IPA外,IAA、ABA、DHZR、ZR、GA3含量均为O型系高于CMS系;饱满花蕾的IAA、ABA、ZR、GA3含量均为O型系高于CMS系。甜菜激素变化较有规律的时期和器官依次为开花盛期的饱满花蕾、开花初期的叶片和和营养生长阶段糖分积累期的叶片。在上述3个时期, CMS系与O型系甜菜的激素含量变化较大,CMS系的IAA/ABA、ZR/ABA和GA3/ABA值分别为O型系的0.55倍、1.43倍、1.64倍;2.66倍、1.55倍、1.49倍和1.14倍、1.60倍、1.53倍。【结论】各种激素的含量变化与甜菜CMS相关密切。影响甜菜CMS的时期、器官和激素比值分别为饱满花蕾IAA/ABA的降低、ZR/ABA和GA3/ABA的提高,开花初期和糖分积累期的叶片为IAA/ABA、ZR/ABA和GA3/ABA的提高。该研究为探讨甜菜细胞质雄性不育的生理机制和育性的调控提供了理论依据。