Overview of the Study and Application of Cytoplasmic Male Sterility in Cotton

Cotton has significant heterosis. Hybrid cotton usually can increase production in lint yield by about 15% compared with conventional self-pollinated cultivars, and also can get obvious improvement in fiber quality, disease resistance, insect resistance, adversity resistance and photosynthetic efficiency. Among some links of cotton heterosis use, the most important link is the castration in the production of hybrid seeds. At present, there are four ways for the castration, such as hand emasculation, chemical male gametocide, nuclear male sterility and cytoplasmic male sterility (CMS). The production practice showed that use of cotton male sterility could not only simplify the hybrid seed production but also save the production cost on a commercial economic scale. In particular, the way of hybrid seed production by use of cotton CMS line, maintainer line and restorer line were the most effective way since it could overcome some disadvantages in the other ways. Therefore, in this paper, the study and application of the cotton CMS system in hybrid seed production were overviewed and some of problems currently limiting application were also addressed. At first, the genetic, cytological and biochemical characteristics of the cotton CMS were reviewed. Secondly, the positive/negative effects of sterile cytoplasm in hybrid F₁ were analyzed, and how to overcome these negative effects, such as pollen temperature sensitive and F₁ not expressing complete fertility, by developing strong restorer lines with a stronger ability for F₁ fertility restoration, was discussed in detail. For an example, transgenic strong restorer line could be developed by introducing the exogenous GST gene, which was assumed to have the function of enhancing pollen vitality, into some conventional restorer lines, and so that hybrids with higher heterosis could be produced by crossing this strong restorer with sterile lines. According to the characteristics of cotton as an often cross-pollination crop, this paper recommended in detail the key techniques of hybrid cotton seed production, such as rules of parent (sterile line and restorer line) selection, location selection and environment optimization for enriching native pollinators to produce more hybrid seeds. Then, the paper pointed out that compared with other crops, cotton hybrid seed production based on CMS system has four advantages in the cotton heterosis use: (1) The purity of hybrid seeds can be guaranteed because there is no pollen in anthers of cotton CMS line and its sterility is very stable and not affected by the climate and other environments; (2) The high yield of hybrid seed can be obtained since the long flowering period (about 3 months) of cotton does not result in the flowering asynchronism between sterile line and restorer line; (3) The wide ecological adaptability of cotton and the possibility of large-scale hybrid seed production will be benefited to popularize hybrid cotton; and (4) Interspecific heterosis between upland cotton (Gossypium hirsutum L.) and sea-island cotton (G. barbadense L.) can be used. It is predicted that the hybrid cotton production based on CMS system will be the main approach to utilize heterosis of cotton. Finally, the future works in study and application of CMS in cotton heterosis, especially in development of new sterile lines and restorer lines by use of modern biotechnology, was also discussed.     

棉花细胞质雄性不育的研究与利用

棉花具有十分明显的杂种优势。杂交棉通常比常规棉增产15%左右,而且在纤维品质、抗病、抗虫、抗逆境和光合效率等性状上也有明显改良。在棉花杂种优势的利用中,最重要的环节之一是杂交种子的生产（制种）。目前,杂交棉制种常有四条途径,人工去雄授粉法制种、化学杀雄法制种、利用核雄性不育的“两系法”制种和利用细胞质雄性不育的“三系法”制种。生产实践表明,利用棉花雄性不育既可简化制种又可节省成本,特别是利用棉花细胞质雄性不育系、保持系和恢复系的“三系法”制种,可较有效克服其他制种方法的一些缺点,是最有效的途径。为此,文章在阐述棉花杂种优势利用途径的基础上,重点综述棉花细胞质雄性不育的遗传学、细胞学和生理生化的特点;深入阐述不育细胞质对杂种F₁的正/负效应,并就如何培育强恢复系的问题,以培育转GST的强恢复系为例,探讨克服不育细胞质对杂种F₁负效应的可能机制;根据棉花为常异花授粉作物和花器具有虫媒花特征的特点,详细介绍三系杂交棉制种的亲本（不育系和恢复系）选配、地点选择和环境优化等条件,以及如何综合优化这些条件提高制种产量的关键技术。利用棉花细胞质雄性不育的“三系法”制种,与其他作物比较,在杂种优势利用中具有4个突出的优点：（1）不育系为无花粉不育类型,育性不受气候等环境的影响,可保证杂种的纯度;（2）棉花开花期长达3个月,不存在制种时花期不遇的现象,制种产量有保证;（3）棉花生态适应性广,育成的组合可在各地种植,种子产业化效益明显;（4）可利用种间（海岛棉与陆地棉间）杂种优势。可以预言,基于细胞质雄性不育的三系杂交棉是大有前途的,将是棉花杂种优势利用的主要途径。最后,就本领域的发展趋势,特别是在利用现代生物技术培育新的不育系和恢复系方面进行了初步探讨。     

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。     

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.     

棉花质核互作雄性不育与育性恢复的研究及利用现状

利用质核互作雄性不育系实现强优势组合三系配套是解决目前棉花杂交制种瓶颈,促进棉花杂种优势利用的有效途径之一。本文综述了棉花质核互作雄性不育与育性恢复的研究现状,介绍了棉花质核互作雄性不育系和恢复系的类型;在细胞学和分子生物学水平上探讨了质核互作雄性不育产生的机理,阐述了育性恢复的遗传方式、恢复基因的分子标记和遗传定位、恢复基因的图位克隆及三系杂交棉的研究进展;对三系杂交棉今后的研究方向进行了展望。     

油菜CMS育性不稳定的研究进展

受遗传因素和温度、光照、土壤肥力等环境因素的影响,油菜CMS的不育性状表现不稳定,不育度降低,出现少量或微量花粉,微粉在形态和功能上与正常花粉都有区别;油菜CMS育性的不稳定不但加大了不育系选育难度,而且会降低杂交种的纯度,影响杂种优势的利用;可通过大量筛选保持系,选择第二保持系,选育细胞核加细胞质不育系,利用化学杀雄剂,控制制种环境措施等来减少和消除不育系微粉的影响。关健词:油菜CMS;育性不稳定;进展     

黄绿苗辣椒生态型不育系的温敏性分析及杂交种子纯度鉴定

[目的]分析黄绿苗辣椒生态型不育系YBM-EAB12的温敏特性,为黄绿苗辣椒温敏特异资源的开发利用提供参考依据.[方法]将辣椒材料YBM-A2、YBM-EAB12、YBM-A2×C和YBM-EAB12×C分别进行日温/夜温为20 ℃/15 ℃、25 ℃/20 ℃和30 ℃/25 ℃诱导后置于温室种植管理,观测各材料花粉的数量及萌发率,并进行各亲本育性和形态学鉴定及种子纯度鉴定.[结果]YBM-EAB12具有温敏特性,育性转化临界温度为22.5 ℃,在低温(日均温度低于22.5℃)条件下表现可育,可自交结实,日均温度越低其可育程度越高,在高温(日温/夜温为30℃/25℃)条件下表现完全不育.YBM-EAB12黄叶色性状受1对隐性核基因控制.YBM-EAB12×C F2代群体分离结果表明,胞质雄性不育与不育花器官特征紧密连锁,YBM-EAB12×C F1代育性由单一核显性基因恢复,不受低温影响.[结论]黄绿苗辣椒生态型不育系具有低温可育特性,利用其进行不育系繁种,可减少不育系与保持系杂交的步骤及辣椒不育系三系制种步骤,简化繁制种子程序.     

抗稻瘟病优质籼型水稻不育系青A选育及应用

[目的]培育抗稻瘟病优质水稻不育系并组配选育抗稻瘟病优质杂交稻品种,为广西水稻安全生产提供品种更新换代保障.[方法]以抗稻瘟病优质水稻材料桂2为亲本,与汕B×博B后代杂交并复交,制保后再与博A回交选育成籼型水稻不育系青A,并与恢复系配组选育抗稻瘟病优质杂交稻品种.[结果]青A稻瘟病抗性1~3级,整精米率58.5%,粒长5.4 mm,长宽比3.0,垩白粒率4%,垩白度0.5%,透明度2级,碱消值6.3,胶稠度68 mm,直链淀粉25.1%,花粉不育度100.000%.采用青A组配的杂交稻组合具有高产、优质和抗性好等特点.[结论]青A为稻瘟病抗性强、稻米品质优、花粉败育彻底、育性稳定、开花习性好、农艺性状优良和配合力好的不育系,与不同熟期恢复系配组的杂种呈感温性,其杂交组合在应用推广中表现适应性广、高产性强、米质优和田间稻瘟病抗性强.     

小麦T型细胞质雄性不育系T9023A的选育及育性恢复研究

为了丰富T型杂交小麦的不育系和恢复系资源,以优质多抗小麦品种郑麦9023为轮回亲本,对T型小麦细胞质雄性不育系T504A连续回交选育不育系,研究其雄性败育特点,并进行恢复系鉴定和育性恢复的遗传分析。结果表明:回交转育获得不育系T9023A,其不育性稳定,雄性败育彻底,花药干瘪细长、不开裂,I_2-KI染色显示,花粉败育类型为圆败和典败。以具有野燕麦(Q型)细胞质的小麦品种川农26和川农27与T9023A测交,其测交F_1自交结实率(国内法)分别为80.3%和62.6%,川农26对T9023A的恢复能力更强一些。T9023A/川农26的F_2群体可育株与不育株分离比符合15∶1,表明川农26携带有T型细胞质雄性不育的2对主效恢复基因。     

棉花细胞质雄性不育研究进展

棉花具有十分明显的杂种优势,棉花细胞质雄性不育在棉花杂种优势的利用上具有重要作用。利用棉花细胞质雄性不育和育性恢复系统突破传统人工去雄杂交育种的瓶颈,为棉花杂交种制种的商业化推广展现了光明的前景。综述了近年来棉花细胞质雄性不育在细胞学、生理生化、分子生物学和育性恢复4个方面的研究进展。详细阐明了棉花细胞质雄性不育小孢子败育时期及细胞学形态、生理生化指标研究、胞内基因组和核内恢复基因的分子生物学研究和获得理想恢复系的方法,并提出了研究中存在的主要问题,展望了今后工作的研究方向。     

几个水稻同核异质雄性不育系育性遗传的比较研究

本文分别以花粉可育率和田间结实率为指标，采用保持系（ｐ１）×恢复系（ｐ２）杂交组合的六个群体及Ｐ１、Ｐ２、Ｆ１、Ｆ２、Ｂ１（Ｆ１×ｐ１）和Ｂ２（Ｆ１×ｐ２），分别与不育系测交的方法，对三个同核异质雄性不育系的育性遗传进行了分析和比较，其结果表明：（１）恢复系为古２２３时，以花粉可育率和田间结实率为指标，Ｄ汕Ａ、Ｇ汕Ａ和Ｗ汕Ａ三个不育系的雄性不育及其育性恢复均受两对独立主效基因的控制；（２）恢复系为ＩＲ２４时，以花粉可育率为指标，Ｄ汕Ａ和Ｗ汕Ａ雄性不育及其育性恢复均受三对主效基因的控制，其中一对独立，两对连锁，交换值为２５％；而Ｇ汕Ａ的雄性不育及其育性恢复仍受控于两对独立主效基因。以田间结实率为指标，该三个不育系的雄性不育及其育性恢复均受两对独立主效基因的控制。     

影响小麦提型不育系种子质量的遗传因素

根据Ｔ型不育系和恢复系,不育系和保持系的两套双列杂交试验以及川７Ａ和其它保持系的４套回交试验。对影响Ｔ型不育系种饱满度和发芽率的遗传因素进行了分析,发现不育系异交子粒的发育进程和充实程度主要取决于母体的基因型;而不育系种子的发芽率则受母体基因型和花粉供体基因型的共同影响。用川７Ｂ给其它不育系授粉得到的杂交种子,其发芽率明显地高于这些不育系与同型保持系杂交产生的种子;不育系种子的饱满度也因有了川７Ｂ     

玉米细胞质雄性不育与恢复基因研究进展

玉米作为我国重要的粮、饲两用作物在保障粮食安全,推动社会经济发展和提供工业能源等方面扮演着关键的角色。细胞质雄性不育是高等植物中普遍存在的生物学现象,玉米细胞质雄性不育可以分为T型、C型与S型3种类型。不育基因来源于线粒体基因重排形成嵌合基因,新形成的嵌合基因对花药中小孢子的发育产生危害导致败育的发生。恢复基因的存在可以消除不育基因的危害,使小孢子正常生长。由于不育基因为线粒体基因,恢复基因为细胞核基因,对细胞质不育与恢复机理的研究同样是探究质-核互作关系的桥梁。同时对雄性不育系的利用是玉米利用杂种优势的一个重要技术手段,玉米生产上利用细胞质雄性不育对于作物杂种优势的利用、杂交种的制种都有重要意义,不但能够解放劳动力降低制种成本,而且提高了制种纯度增加产量。对玉米细胞质雄性不育的分类、特征及近年来发现的玉米细胞质雄性不育基因与育性恢复基因进行概述,并探讨了玉米细胞质雄性不育应用过程中的问题与发展前景,以其为细胞质雄性不育在生产上的推广利用提供参考。     

Inheritance of Fertility Restoration for Cytoplasmic Male Sterility in a New Gossypium barbadense Restorer

In order to clarify inheritance mechanism of fertility restoration for cytoplasmic male sterility （CMS） in a new Gossypium barbadense restorer line Hai R which was found in the fertility test crossing of G. hirsutum CMS lines with G. barbadense germplasms. 23 fertility segregation populations of F2 and backcross were used to analyze the inheritance of fertility restoring gene（s） of Hai R. The result showed that Hai R had one major dominant gene （RfB） to control the CMS fertility restoration and this fertility restoration gene functioned at the sporophytic level. The sterile cytoplasm background might not only influence the transmission rate of male gamete but also that of female gamete when the restorer gene was recessive. It could be deduced that this fertility restoration gene might come from G. harknessii cotton, Hai R is of value in the application of cotton interspecific hybrid breeding.     

Establishment and Identification of Cytoplasmic Male Sterility in Brassica napus by Intergeneric Somatic Hybridization

Exploitation of novel cytoplasmic male sterility(CMS)is a main approach for widening the cytoplasmic genetic background of hybrid oilseed rape and avoiding epidemic risk in oilseed rape production.In this study,symmetric somatic hybrids between Brassicanapus var.Zhongshuang4 and Sinapis arvensis(Yeyou18)were produced by protoplast fusion.Two of the six established hybrids were male sterile showing trace or no pollen release upon flowering with non-or slightly extended stamens.Using Zhongshuang4 as a recurrent parent to pollinate the male sterile plants,the ratio of male sterile plants increased with the number of backcrosses.As early as in BC3 generation,most of the sterile families had nearly 100%sterile plants.Up to BC4 generation,the male sterility became stable and no fertility segregation was observed.All F1 progenies from tested crosses using restorer and maintainer lines of Polima CMS were 100%sterile,indicating that the established CMS by somatic hybridization is different from Polima CMS.The origin of the cytoplasm and potential use of this hovel CMS in oilseed rape breeding were discussed.Key wotds:Oilseed rape,Protoplast fusion,Cytoplasmic male sterility,Sinapis arvensis     

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。     

柑橘黄龙病菌分泌蛋白04580基因的克隆及原核表达

为进一步了解柑橘黄龙病菌亚洲种CLas的致病机理,从Prasad预测的166种分泌蛋白中选取1个在柑橘植株中表达量较高的CLIBASIA–04580基因进行研究。提取感染柑橘黄龙病的柑橘叶片总DNA,经PCR扩增得到CLIBASIA–04580基因,切胶回收PCR产物,双酶切连接到pET–28α载体上,使用菌落PCR和限制性内切酶双酶切鉴定筛选阳性克隆;转化大肠杆菌BL21(DE3),用0.5、1.0、1.5、2.0 mmol/L的异丙基硫代半乳糖苷(IPTG)诱导重组蛋白表达融合组氨酸标签的CLIBASIA–04580重组蛋白,聚丙烯酰氨凝胶电泳(SDS–PAGE)检测重组蛋白的表达水平,再用蛋白纯化镍柱进行纯化。结果表明,携带组氨酸标签的CLIBASIA–04580重组蛋白得到表达,且异丙基硫代半乳糖苷(IPTG)浓度为1.5mmol/L时相对表达量达到峰值,通过SDS–PAGE凝胶分析,与未经过镍柱纯化的蛋白原液比较,可知纯化了CLIBASIA–04580重组蛋白。     

辣椒细胞质雄性不育突变体CMS102的败育机理及其育性遗传

以辣椒细胞质雄性不育突变体CMS102及其相应的恢复系ST–8为材料,对其造孢期、小孢子母细胞期、小孢子减数分裂期、四分体时期、单核小孢子期、双核小孢子期和花粉粒成熟期的花蕾进行细胞学形态观察,测定小孢子减数分裂期、四分体时期、单核小孢子期和双核小孢子期发育时期的花蕾中与育性相关的生理活性物质的含量。结果表明：CMS102败育发生在四分体时期,绒毡层过度膨大挤压四分体,使其不能正常发育成单核小孢子,属于孢子体不育;CMS102花蕾中可溶性蛋白和游离脯氨酸含量低于ST–8的,可溶性糖、MDA含量及POD酶活性高于ST–8的;构建F₂遗传分离群体并调查其育性,对CMS102的育性进行遗传学分析,CMS102育性恢复由1对显性基因调控。     

辣(甜)椒细胞质雄性不育胞质基因的遗传效应

以辣(甜)椒细胞质雄性不育系(CMS)21A、延A、8A及相应的保持系21B、延B、8B和恢复系湘紫、LS7为材料,首次系统研究3个CMS不育胞质基因的遗传效应。结果表明,辣(甜)椒雄性不育细胞质基因的遗传效应之一表现为单性结果在不育系间有明显差异。不育胞质基因对F1代株高、株幅、单株结果数、果实大小、产量等主要农艺性状有一定正效应和促进作用,不育胞质基因对繁种没有影响。不育系的扩繁与保持以及F1杂交种子的繁制以大棚人工辅助授粉为最佳,单株结果数和单果种子数显著高于露地人工辅助授粉和自然授粉。     

细胞质雄性不育海岛棉恢复系恢复基因的遗传分析

 【目的】在陆地棉细胞质雄性不育系与海岛棉种质材料进行测恢筛选时,选得一个对不育有恢复力的海岛棉材料“海R”,本研究的目的是了解其育性恢复的遗传方式。【方法】构建23种不同的F2群体和回交群体,分析和明确恢复基因的遗传。【结果】“海R”恢复基因属孢子体遗传,由一个主效显性基因（RfB）控制。在恢复基因呈隐性时不育胞质不但对雄配子的传递有影响,而且对雌配子的传递也有影响。【结论】推测此海岛棉恢复系的育性恢复基因来源于哈克尼西棉基因组,在陆地棉与海岛棉的种间杂种优势利用中具有应用价值。