光照强度对温光敏细胞核雄性不育小麦育性的影响

以具有低温不育、高温可育特性的温光敏细胞核雄性不育小麦C412S和C404S为材料,以其回交转育亲本、育性正常的C412和C404为对照,用人工气候箱研究了光照强度对温光敏细胞核雄性不育小麦育性表达的影响。结果表明,在低温条件下(日温8℃/夜温6℃),C412S和C404S在不同光照强度(160μmol/m2.s和300μmol/m.2s)下自交结实率都为0,表现为完全不育。在较高温度条件下(日温18℃/夜温14℃),从花粉母细胞形成期到开花期的光照处理,C412S在160μmol/m.2s弱光照下的自交结实率仅为5.4%,表现为高不育,在300μmol/m2.s较强光照下的自交结实率高达65.0%,表现为高度可育;而另一不育系C404S在2种光照强度下的自交结实率分别为69.9%和73.2%,都达到了高度可育水平。表明光照强度对温光敏细胞核雄性不育小麦的雄性育性表达具有重要影响,但不同材料对光照强度的响应程度有所差异。     

无花瓣油菜雄蕊心皮化突变体细胞学观察及基因表达分析

油菜是我国重要的油料作物,油菜花器官具有典型的十字花科特点,无花瓣油菜在花期不存在花冠层,这种特点有助于提高油菜产量,预防茵核病的传播。雄蕊心皮化是指花器官的雄蕊结构被具有类似于雌蕊结构的器官代替,这不仅造成了花器官结构的变化也导致了雄性不育。本文通过对无花瓣油菜雄蕊心皮化突变不育分离群体中的雄性可育株和不育株的比较研究,发现心皮化现象是由遗传因素引起的。细胞学观察发现,雄蕊心皮化在花器官发育的早期就已经产生,心皮化的雄蕊中着生类似于胚珠的结构,其顶端细胞的形态和排列方式也与雌蕊柱头相似。花发育相关基因的表达分析表明,B组基因彳丹在不育株3轮花器中的表达都比可育株低,特别是在第二轮花器官中这种差异最为突出。而A组基因AP1在不育株第二轮花器官中的表达量较可育株高。c组基因AGL8、SHPI、SHP2、NAP在不育株心皮化的第二轮花器官中表达都较可育株中高。     

具野稻细胞质的胞质型雄性不育系的体细胞克隆变异II.体细胞克隆温敏核不育突变

在胞质型水稻雄性不育系IR66707A和IR69700A经离体培养而获得的136个体细胞克隆中,发现了温敏核不育突变5例。这类突变体在广州地区的自然气候下,早季至晚季前期表现为不育,晚季后期表现为可育。盛夏,在幼穗发育至花粉形成阶段对部分突变材料进行短日照处理,发现对短日照敏感的不育系农垦58S转换为可育,而供试的5例突变及另一对照培矮64S却与未经处理的材料一样仍表现为不育,表明它们的育性与日照长度的变化无关。在同一发育时期进行低温处理的结果显示,低温处理10d及10d以上者可发生育性转换,自交结实率在17.23%-42.19%之间,而未经处理的材料仍然表现不育,表明它们的育性转换与温度有关。以正常品种为父本与突变体杂交,F1全部为可育;F2可育与不育个体的分离比为3:1;以F1为父本与之测交,TF1代中可育与不育个体的分离比为1:1。遗传分析表明,这种温敏核不育突变为一对隐性核基因所控制。获得了由胞质型雄性不育变为胞核型雄性不育的突变体,这在体细胞克隆变异领域中是一种典型的突变。     

具野稻细胞质的胞质型雄性不育系的体细胞克隆变异Ⅱ.体细胞克隆温敏核不育突变

在胞质型水稻雄性不育系IR66707A和IR69700A经离体培养而获得的136个体细胞克隆中,发现了温敏核不育突变5例。这类突变体在广州地区的自然气候下,早季至晚季前期表现为不育,晚季后期表现为可育。盛夏,在幼穗发育至花粉形成阶段对部分突变材料进行短日照处理。发现对短日照敏感的不育系农垦58S转换为可育,而供试的5例突变及另一对照培矮64S却与未经处理的材料一样仍表现为不育,表明它们的育性与日照长度的变化无关,在同一发育时期进行低温处理的结果显示,低温处理10d及10d以上者可发生育性转换。自交结实率在17.23%-42.19%之间,而未经处理的材料仍然表现不育,表明它们的育性转换与温度有关,以正常品种为父本与突变体杂交,F1全部为可育;F2可育与不育个体的分离比为3：1,以F1为父本与之测交,TF1代中可育与不育具体的分离比为1：1。遗传分析表明,这种温敏核不育突变为一对隐性核基因所控制,获得了由胞质型雄性不育变为胞核型雄性不育的突变体。这在体细胞克隆变异领域中是一种典型的突变。     

青花菜两类雄性不育系花器官形态结构的比较

青花菜是一种重要的十字花科蔬菜作物,具有良好的抗癌功效。随着青花菜栽培面积的不断扩大和抗癌药物的开发,对青花菜种子的需求量也不断增加。利用雄性不育系生产青花菜F1杂交种子是其杂种优势利用的重要途径,但常用的细胞质雄性不育系在生产中存在前期死蕾较重、花蜜量少和种子产量不高等问题。本课题组用优良的青花菜自交系为父本,以甘蓝显性细胞核雄性不育材料79-399-3和细胞质不育材料OguraCMSR3629为不育源,通过回交转育的方法获得了多个青花菜显性细胞核雄性不育系和细胞质不育系,为了进一步有效利用其青花菜雄性不育系,寻求更好的不育源。本研究以青花菜高代自交系(保持系)8554、8590、93219及由其经多代回交转育而成的两类雄性不育系为试材,对其死蕾数、花器官形态和蜜蜂访花情况进行了比较研究。结果表明,在花蕾长、花蕾直径、单枝死蕾数、花冠直径、雄蕊长、花瓣长、花药长、蜜蜂访花次数、来访蜜蜂数及访花时间等方面,经相同自交系转育而成的显性细胞核雄性不育系(DGMS)与细胞质雄性不育系(CMS)间的差异有统计学意义,总体表现为DGMS优于CMS。     

甘蓝型油菜隐性细胞核雄性不育的研究及利用

甘蓝型油菜隐性细胞核雄性不育系育性稳定、无负效应、易转育且恢复源广,在油菜杂种优势利用中被广泛利用.目前我国主要利用的隐性细胞核雄性不育系有2类:双基因隐性细胞核雄性不育以及由2对基因互作控制的核不育类型.随着这2类不育系不育基因相继被克隆,人们对甘蓝型油菜细胞核雄性不育的分子作用机理已经有一定了解.本文综述了在甘蓝型油菜核不育分子机理的研究方面的进展,在此基础上提出了繁殖甘蓝型油菜核不育系全不育系群体可行的途径,展现了油菜细胞核雄性不育杂种优势利用广阔的前景.     

两份太空诱变玉米雄性不育突变体的遗传研究

从搭载神舟4号飞船的4份玉米自交系后代中选育出两份雄性不育突变体, 对其进行育性鉴定, 并分析不育性状的稳定性及遗传特点。以不育材料的不育株为母本, 同群体的可育株和其他自交系为父本进行杂交, 结合自交、回交分析其后代的育性表现; 同时, 以具有正常细胞质的自交系为母本, 育性完全恢复的测交F1植株为父本进行反交, 对其反交的F1及F2进行育性观察分析。结果表明：这两份不育材料不育株的花药内无花粉或含少量畸形花粉, 败育彻底, 花粉败育表现为典败型。不育性状在不同年份、不同季节、不同地点下稳定遗传, 属可遗传的单基因控制的隐性核不育类型。     

温敏雄性不育水稻培矮64S花药发育过程中钙的变化

采用焦锑酸钾沉淀法研究了温敏雄性不育水稻(Oryza sativa L.)培矮64S在高温引起雄性不育与正常可育花药发育过种中Ca2+的分布变化.结果表明,当培矮64S生长在较高温度条件下引起雄性不育,与可育花药相比,不育花粉母细胞中有较多的液泡、较多的Ca2+沉积和较少的线粒体,并且有较多的Ca2+沉积在不育花药的中间层、表皮层和绒毡层中.到四分体与单细胞花粉时期,不育花药的木质部细胞的次生加厚壁上有较多的Ca2+沉淀,连接组织中的Ca2+沉淀也大大增加,所有不育花粉外壁较厚而发育都不正常.在单核细胞早期,不育花粉的四分体细胞中有较明显的大液泡出现.不育花药中的Ca2+在花药发育的各时期均比可育花药要多.这些结果说明在高温生长条件下,花粉母细胞发育的异常、花药中Ca2+沉积的增加、绒毡层与花粉外壁发育的异常可能与培矮64S花粉败育相关.     

低温敏核不育水稻go543S育性对温、光的反应

以新型光温敏核不育（PTGMS,Photo-thermo sensitive genic male sterile)水稻go543S为材料,通过自然生态条件下和人工温、光处理条件下的育性观察,对其育性转换与温度和光周期的关系进行了研究。结果表明：go543S育性主要受温度控制,表现为低温条件下不育,高温条件下可育,育性转换的不育临界温度值为29．5℃,对温度传感的部位是幼穗,敏感时期为花粉母细胞形成致减数分裂,对应的剑叶叶枕距变化范围为－12．2－＋0．7cm;育性敏感期在人工恒温28．0℃条件下,无论长日（14－16h）或短日（10－12h）处理均表现不育,其不育性不受光周期影响,在人工恒温31.5℃条件下,无论长日还是短日处理均表现可育,但短日可明显提高其可育性。     

芥菜型油菜细胞质雄性不育系WJS01A的比较鉴定北大核心CSCD

芥菜型油菜细胞质雄性不育(CMS)系WJS01A是一种稳定不育系,其败育彻底,不受环境条件的影响。该研究从形态、细胞特征、遗传和分子生物学等方面对WJS01A进行鉴定,以揭示其败育机制,为该不育系在油菜育种中的应用提供理论基础。结果表明:(1)不育系WJS01A的花序结构与正常芥菜型油菜差异不大,但在花蕾饱满度、花朵张开度及花瓣长度和宽度方面略低于正常的芥菜型油菜;它的雌蕊发育正常,但花药、花丝缩短,致使雄蕊高度显著低于柱头,花药白化无花粉产生。(2)将WJS01A衍生的甘蓝型油菜背景不育系WNJ01A以及Polima(Pol)、Ogura(Ogu)和Kosena(Kos)不育系分别与其恢复系或保持系测交,结果显示来源于WJS01A的不育类型与Pol、Ogu和Kos等材料的恢保关系明显不同,仅Hui01可以恢复WNJ01A的育性。(3)不育系WJS01A属于无花粉囊型不育,败育时期为花药原基到孢原细胞时期。(4)线粒体不育基因多重PCR可以明显区分WJS01A、WNJ01A与Pol、Ogu、Kos,但是目前的引物组合不能区分WJS01A与正常的芥菜型油菜。(5)线粒体基因组的限制性片段长度多态性(RFLP)分析表明,在所检测的8个探针/酶组合中均可以将不育系WJS01A与其他4种细胞质雄性不育系区分开,说明WJS01A是一种显著不同于Pol、Ogu和Kos等的细胞质雄性不育类型。WJS01A的利用可以丰富和拓宽当前油菜杂种优势利用的遗传基础,为缓解当前油菜杂种优势利用中不育胞质单一性问题提供新的种质。     

Testing cms-P-linked AFLPs for selection of rye hybrid components

Application of AFLPs linked to pollen fertility restoration and non-performing genes evaluated in the C394-F2 hybrid was studied using a set of male sterile lines in the sterilising Pampa cytoplasm, several restorers and maintainer lines and, finally, two inbred lines backcrossed into cms-P, cms-R, cms-S and cms-C cytoplasms each. The set of male sterile lines based on the Pampa cytoplasm exhibited gradual variation in their ability to restore pollen fertility (starting from low and closing with high) in crosses with three unrelated restorers. Variations in the AFLPs between the analysed materials were observed, however, no clustering of the lines according to their sterile and fertile phenotypes was observed. The same markers, when applied to the population restorer (cv. Walet) that formed the C394-F2 cross permitted identification of plants with genotypes that could be recognized as restorers.     

Inducing male sterility of tomato using two component system

Production of hybrid seeds and pursuing heterosis breeding of many crops have been accomplished using male sterile lines. However, not all crops have valuable male sterile lines due to instability of male sterility and absence of a restorer system. In this study, male sterile lines have been induced using a two-component system. The extracellular ribonuclease Barnase was cleaves into two inactive yet complementary fragments, designated as ??Bn-5?? and ??Bn-3??. Both components were controlled by a TA29 promoter. They were transferred into the tomato inbred line ??Yellow tomato?? by Agrobacterium method. Southern blotting identified that 11 transgenic Bn-5 plants (T₀) and 10 transgenic Bn-3 plants (T₀) were obtained. The vegetative phenotypes of all T₀ plants were similar to wild-type, and they were capable of producing viable pollen grains and normal fruit with seeds, indicating that Barnase had lost its function after it being split two partial fragments. After self-pollination, homozygous progenies (T₁) of transgenic Bn-5 and Bn-3 plants were chosen to cross each other, Barnase could be reconstituted and co-expressed in the same cell, which caused the hybrid plants to produce collapsed pollen grains with no viability and thus100?% male sterile plants were obtained. Stamens of male sterile plants were shorter than those of the wild type plants. PCR detection demonstrated that all male sterile plants contained Barnase, but male fertile plants did not. The male sterile plants were crossed with the male fertile inbred lines, and the result showed that hybrid (F₁) plants were capable of producing normal fruit with seeds, and their pollen grain fertility was restored. The co-segregation ratio of Bn-5 and Bn-3 fragments showed 1:1 among hybrid plants. In conclusion, the results verified that the male sterility could be generated by two component system and be used in hybrid seed production. The F₁ between the male sterile plant and the inbred line showed heterotic comparing to both parents. This system needs not breed restoration line.     

TA29-barnase基因转化菜心

利用根癌农杆菌导入法, 以菜心带柄子叶为外植体, 对TA29-barnase基因转化菜心进行研究。获得转化植株,进行PCR、Southern blotting杂交和半定量RT-PCR检测, 表明目的基因已经整合到转化植株中, 并且目的基因在转基因植株花蕾中得到表达, 但是表达水平在不同转基因植株间存在差别; 转基因植株开花后, 均表现雄性不育, 不能产生花粉或产生没有活力的少量花粉, 自交不能结实; 用未转化植株正常花粉对雄性不育植株进行授粉, 能够正常结实; 保持系(未转化植株)与不育株杂交后代中雄性不育株与可育株的比例为1:1, 在杂交后代植株子叶期, 喷洒10 mg/L的PPT可以完全杀死可育株; 利用其他菜心品种为父本与不育株进行杂交, 获得的F1植株在生长势和产量方面表现优势, 表明开展菜心优势育种具有一定的潜力。     

TA29-barnase基因转化菜心

利用根癌农杆菌导入法, 以菜心带柄子叶为外植体, 对TA29-barnase基因转化菜心进行研究。获得转化植株,进行PCR、Southern blotting杂交和半定量RT-PCR检测, 表明目的基因已经整合到转化植株中, 并且目的基因在转基因植株花蕾中得到表达, 但是表达水平在不同转基因植株间存在差别; 转基因植株开花后, 均表现雄性不育, 不能产生花粉或产生没有活力的少量花粉, 自交不能结实; 用未转化植株正常花粉对雄性不育植株进行授粉, 能够正常结实; 保持系(未转化植株)与不育株杂交后代中雄性不育株与可育株的比例为1:1, 在杂交后代植株子叶期, 喷洒10 mg/L的PPT可以完全杀死可育株; 利用其他菜心品种为父本与不育株进行杂交, 获得的F1植株在生长势和产量方面表现优势, 表明开展菜心优势育种具有一定的潜力。     

水稻不育系安农S-1育性转换及相关基因的表达分析

通过在自然环境和高温温室内对安农S-1的不同部位进行高温、低温诱导处理,对水稻温敏核不育系安农S-1的温度敏感时期和诱导部位进行了研究。总共进行了8种处理,结果表明：安农S-1的育性转换时期是从花粉母细胞形成到减数分裂的四分体时期之前。在育性转换时期,处于高温的条件下,根部低温处理不能诱导安农S-1可育,穗部低温处理可以使安农S-1保持可育,可见安农S-1的温度敏感部位在幼穗。aprz基因和育性相关,用RT-PCR方法研究了aprt基因在安农S-1不同部位和不同温度环境的表达变化,结果显示,在幼穗中aprt基因的表达在高温环境中被大幅度下调,而在叶中和根中的变化比较小,这说明幼穗对温度最敏感,从侧面验证了引起安农S-1育性转换的温度敏感部位是在幼穗。     

Targeted mutagenesis of a conserved anther‐expressed P450 gene confers male sterility in monocots

Targeted mutagenesis using programmable DNA endonucleases has broad applications for studying gene function in planta and developing approaches to improve crop yields. Recently, a genetic method that eliminates the need to emasculate the female inbred during hybrid seed production, referred to as Seed Production Technology, has been described. The foundation of this genetic system relied on classical methods to identify genes critical to anther and pollen development. One of these genes is a P450 gene which is expressed in the tapetum of anthers. Homozygous recessive mutants in this gene render maize and rice plants male sterile. While this P450 in maize corresponds to the male fertility gene Ms26, male fertility mutants have not been isolated in other monocots such as sorghum and wheat. In this report, a custom designed homing endonuclease, Ems26+, was used to generate in planta mutations in the rice, sorghum and wheat orthologs of maize Ms26. Similar to maize, homozygous mutations in this P450 gene in rice and sorghum prevent pollen formation resulting in male sterile plants and fertility was restored in sorghum using a transformed copy of maize Ms26. In contrast, allohexaploid wheat plants that carry similar homozygous nuclear mutations in only one, but not all three, of their single genomes were male fertile. Targeted mutagenesis and subsequent characterization of male fertility genes in sorghum and wheat is an important step for capturing heterosis and improving crop yields through hybrid seed.     

Aanlysis of short photo-periodic sensitive genic male sterility and molecular mapping of rpms3(t) gene in rice (Oryza sativa L.) using SSR markers

A research was conducted on the pollen fertility of rice sterile lines D52S and D38S responsive to photoperiod during the sensitive stage under natural and controlled conditions. Bulk segregant analysis (BSA) and recessive class approach were applied to identify DNA markers that co-segregate with gene conferring male-sterility in D52S mutant rice. The results showed that in day-light higher or equal to 14.00 h, D52S and D38S rice pollen were fertile; however, they were sterile when day-length was less than 14.00 h. They were therefore considered to be short photo-periodic sensitive genic male sterile lines(Short PGMS lines). Under short day-light conditions, the pollen fertility segregation of F₂ populations from crosses between D52S/Shuhui527 and D52S/Gui99showed 3:1 ratio of fertile to sterile plants suggestingthat male sterility in D52S was controlled by one recessive gene. Two markers RM244 and RM216 located on chromosome number 10 co-segregated completely with the rpms locus. The locus was mapped to the interval between SSR markers RM2571 (6.6 cM) and RM244 (4.6 cM).     

桔梗雄性不育花药的发育解剖学研究

通过田间观察和石蜡切片法对桔梗不育和可育花药的形态及其解剖结构进行了比较研究,结果表明:桔梗可育和不育花器形态、花药长度无明显差异,而开花后,可育花药饱满、有光泽、有花粉粒散出,不育花药皱瘪、无光泽、无花粉粒散出;不育花药绒毡层提前退化解体,导致小孢子发育异常。     

光温条件对短光低温不育水稻育性转换的影响

短光低温不育水稻是一类与长光高温不育水稻育性转换特性相反的新种质,通过对分期播种材料自然条件下的育性及人工短光处理后的育性比较分析表明,宜DIS的光敏期为第一苞分化期至第二次枝梗原基和小穗原基分化期（幼穗分化I－III期）,在常温下光敏期的短日处理可使育性趋于不育,但在高温下光敏期短日不育效应被削弱,宜DIS的花粉育性在高,低温作用下育性的均出现下降,根据高,低温影响育性的资料分析得到的敏感期基本一致,即花粉母细胞形成期至花粉内容物充实期（幼穗分化V－Ⅶ期）。