Development of thermo-sensitive cytoplasmic male sterile (TCMS) lines of wheat characterized by complete male sterility at lower-temperatures and partially restored fertility at higher-temperatures

A ‘‘two-line hybrid system’’ was developed using thermo-sensitive cytoplasmic male sterility (TCMS) from Aegilops kotschyi and is proposed as a means for seed multiplication and production of hybrid wheat. The TCMS line is crossed to a pollinator line during the normal wheat-growing season for production of hybrid wheat seeds, whereas the TCMS line is maintained through self-pollination in an environment with high temperatures. The male sterile wheat line KTP3314A, containing Ae. kotschyi cytoplasm and the short arm of chromosome 1B from Triticum spelta, was studied in different temperature regimes. KTP3314A was completely male sterile under normal wheat growing temperatures (<18 °C). Sterility of KTP3314A was partially restored when plants were grown at temperatures above 20 °C during growth stages 45–52. Male fertility was not correlated with day-length. A series of TCMS lines derived from KTP3314A showed similar environmental responses and performed well in hybrid wheat production at lower temperatures. Fertility of these TCMS lines at higher temperatures makes it possible to produce large quantities of male sterile parental seeds. Based on these findings, application of the TCMS-dependent two-line system in hybrid wheat breeding appears to have a promising future for hybrid wheat production.     

洋葱细胞质雄性不育系及保持系小孢子发生的细胞学观察

为揭示洋葱细胞质雄性不育系小孢子的败育时期,利用石蜡切片技术对洋葱细胞质雄性不育系8A及其保持系8B的小孢子发育过程进行观察和比较。结果表明：石蜡切片洋葱细胞质雄性不育系8A和保持系8B造孢组织时期小孢子的生长发育没有明显的差异;洋葱不育系8A花药绒毡层在花粉母细胞时期就与中层完全分离,并逐渐膨大挤压花粉母细胞,最后逐渐降解。其降解后的残余物侵入药室,与小孢子混合粘连在一起,占据药室的一部分空间,并且不能供给小孢子生长发育所需要的营养物质,最终小孢子降解,花药败育。试验明确了洋葱细胞质雄性不育系8A的小孢子败育时期为花粉母细胞时期。因此,在洋葱雄性不育系8A的不育机理研究中,应重点在花粉母细胞时期进行深入研究。     

玉米细胞质雄性不育性与乙烯的关系

研究了玉米Ｍｏ１７核背景三种雄性不育细胞质（Ｔ、Ｃ、Ｓ）系及正常可育细胞质（Ｎ）系小孢子发育不同阶段花药组织的乙烯释放量，发现不育系小孢子败育前的一、二个时期内花药组织乙烯释放量显著增加。同时设计的乙烯利及其合成抑制剂外源喷施实验也表明了其对雄性育性的逆转效应。讨论了植物激素对玉米细胞质雄性不育性的调控作用     

大白菜CMS96细胞质雄性不育系的特点分析

为了筛选和利用大白菜细胞质雄性不育系,对大白菜的3种细胞质雄性不育系：CMS96细胞质雄性不育系、波里马细胞质雄性不育系(Pol-CMS)和萝卜细胞质雄性不育系(Ogu-CMS)及其相应的保持系进行了主要植物学性状的比较研究。结果表明：Ogu-CMS存在多代转育后植株长势退化、杂交优势差,Pol-CMS存在育性受环境条件影响等缺点,而CMS96细胞质雄性不育系则克服了二者的缺点。另外,CMS96细胞质雄性不育系早期的晚抽薹性比保持系强、杂交种子的产量比保持系杂交种子产量高。CMS96细胞质雄性不育系不仅不育性稳定,多代回交不退化,而且杂交优势强,繁种产量高,表现出了广阔的育种应用前景。     

Glucosinolates Determined by HPLC in the Seeds of Microspore-Derived Homozygous Lines of Rapeseed (Brassica napus L.)*

Microspore culture was employed to measure the relative efficiencies of anther culture and isolated microspore culture for the regeneration of embryoids and plants of Brassica napus. The yield of embryoids and plants was at least 10-fold greater from isolated microspores than from anther cultures. Approximately 1400 microspore-derived homozygous line's, the parental varieties and the corresponding F₂ plants were grown in a field trial. Important agricultural characteristics, such as morphological homogeneity, growth rate, onset of flowering and seed setting were evaluated subjectively and seed yield and glucosinolate content of individual plants were determined. The relative concentrations of up to S different glucosinolates in these seeds were measured via an automated high performance liquid chromatography (HPLC) system. The alkenyl and indole glucosinolates, the two most important categories of glucosinolates, were found in varying proportions and were independently determined in these line's. Our results do not support the previously suggested connection between low concentrations of glucosinolates and weak growth and/or poor seed yield. Additionally, no evidence was found that the lines derived from isolated microspore culture were subjected to unexpected selection pressures that might adversely affect the diversity of the lines obtained. These results demonstrate that microspore culture is a powerful tool not only for genetic analysis bur also for practical plant breeding.     

普通小麦T型和K型胞质雄性不育系败育过程细胞学观察和同工酶检测

利用细胞学观察和同工酶电泳测试，分析了普通小麦Ｔ型和Ｋ型胞质雄性不育系、保持系和恢复系花药的发育过程，结果表明：（１）用同工酶电泳检测小麦Ｔ型和Ｋ型雄性不育胞质效应的区别有效。（２）酯酶酶谱较过氧化物酶酶谱更能清楚地检测两类不同胞质不育系败育发生的时期和程度。（３）细胞学观察表明，Ｔ型不育系小孢子在双核期其细胞核绝大部分不存在，成为空壳，败育较早，较迅速，这可能是其恢复源较狭窄的原因之一；而Ｋ型不     

萝卜胞质紫菜薹雄性不育系花药发育的细胞形态学研究

通过2种细胞质紫菜薹雄性不育系小孢子发生的细胞形态学观察表明,供试不育系的雄蕊都深度退化,花药内无花粉。改良不育系小孢子在四分体前发育正常,在单核小孢子期,绒毡层异常膨大挤压小孢子,造成小孢子发育营养不良,引起小孢子败育。Ogura不育系的小孢子发育有所不同,它在孢原细胞分化期之前就已有53%败育,不形成花粉囊;形成花粉囊的孢原细胞大多只形成2个体积很小的的花粉囊,其中的小孢子发育与改良不育系的小孢子发育相似:单核早期绒毡层膨大挤压小孢子,使小孢子败育。     

Temperature related instability of a naturally induced cytoplasmic male sterility in industrial chicory (Cichorium intybus L.)

Natural alloplasmic cytoplasmic male sterile (CMS) clones of industrial chicory were obtained after crossing wild chicory with selected breeding lines. We investigated the CMS stability of 10 clones in various environmental growing conditions. CMS was stable under cool growing conditions in most of them. Fertility restoration, based on pollen production scores, was observed in all clones after a period of hot temperatures. The early flower bud stage was sensitive, resulting in fertile flowers 12–17 days after exposure to high temperatures. Experiments under controlled growing conditions at 15°C demonstrated that a heat shock of 2 days at 25 or 30°C was sufficient to restore fertility. Sterile flowers were formed when plants were again grown at lower temperatures. Significant differences between individual clones were observed, indicating the potential of genetic selection to obtain stable CMS parent lines.     

甜椒雄性不育两用系小孢子发育的显微观察

以甜椒雄性不育两用系为试材,对不育株与可育株花粉母细胞减数分裂过程中染色体行为、花药和小孢子发育过程进行了研究。结果表明,不育株花粉母细胞减数分裂染色体行为未见异常,败育发生在四分小孢子形成之后。导致小孢子败育的原因与四分体胼胝质壁不适时解体和绒毡层细胞发育异常、延迟解体有关。     

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.     

玉米小孢子高频再生培养技术程序研究

以大田种植的玉米供体植株为实验材料,以提高游离小孢子存活率及成胚率为重点进行了技术优化研究。摸索出了一些行之有效的技术措施,提高了游离小孢子的离体培养反应,获得了一批再生植株。通过技术优化,建立玉米小孢子离体培养高频再生技术程序,在此基础上构建了玉米DH育种技术平台。应用这一技术平台,可以在当年获得玉米DH纯系的种子,第二年即可安排大田纯系鉴定。     

Identification of proteins related to microspore embryogenesis responsiveness in anther cultures of winter triticale (×<Emphasis Type="Italic">Triticosecale</Emphasis> Wittm.)

For a better understanding of the physiological background of microspore embryogenesis (ME), the protein profile was analyzed in four winter triticale DH lines, which show extremely different embryogenic potential. The analysis were conducted with anthers at the phase of development optimal for ME induction and then after low temperature (LT, 3 weeks at 4 °C) ME-inducing tillers treatment. The sub-proteome of anthers was mapped by two-dimensional gel electrophoresis (2-DE). The protein species significantly more abundant (at least 2-fold) in responsive DH lines after LT treatment were chosen for identification by MALDI-TOF/TOF analysis. In total, 31 protein species were successfully identified as involved in the determination of microspore competence, stress response and in the regulation of ME induction. Microspore competence required sufficient energy supply and efficient system of cell protection that determine survival under prolonged LT stress treatment. LT stress was associated with increased accumulation of proteins typical for cell defence against oxidative stress (e.g., l-ascorbate peroxidase), chaperons (e.g., HSP70) and other enzymes/factors ensuring protein biosynthesis, stability and active cell divisions. Also here, effective cell defence required undisturbed energy supply. Among proteins that accumulated differentially in accordance with microspore embryogenic potential again the most important role seems to be played by the enzymes ensuring energy production and determining ability of plant stress adaptation. Two protein species (enolase, 12S storage protein), proposed earlier as candidates for markers of embryogenesis in other in vitro plant culture systems confirmed their utility for triticale anther cultures.     

玉米小孢子培养高频再生技术程序的研究

以大田种植的玉米供体植株为实验材料，以提高游离小孢子存活率及成胚率为重点进行了技术优化研究。摸索出了一些行之有效的技术措施，提高了游离小孢子的离体培养反应，获得了一批再生植株。通过技术优化，建立玉米小孢子离体培养高频再生技术程序，在此基础上构建了玉米DH育种技术平台。应用这一技术平台，可以在当年获得玉米DH纯系的种子，第二年即可安排大田纯系鉴定。     

野生稻（O.rufipogon）新胞质改良不育系稻米品质的研究

通过核置换回交，获得一种普通野生稻（O. rufipogon）雄性不育细胞质，称为FA细胞质，育成新质源不育系金农1A。研究表明，FA细胞质不育系与WA和HL不育系的恢保关系不同，野败型的保持系和恢复系、红莲型的保持系和恢复系都可以作为FA细胞质的保持系，而不能成为FA细胞质的恢复系。这是一种新发掘的水稻雄性不育细胞质源。FA细胞质突破了WA和HL细胞质恢保关系的遗传局限，扩大了保持系育种范围，可以用品质优良的栽培品种培育成优质稻米不育系。新质源不育系金农1A不育度高，不育性稳定，农艺性状优良，抗稻瘟病和白叶枯病，开花时间早，稻米12项品质指标全部达到农业部颁发的优质米一级或二级标准，实质性地提高了不育系的稻米品质和综合水平，为培育优质米杂交稻奠定良好的遗传基础。     

不结球白菜Ogura雄性不育花器官形态及败育细胞学的研究

对不结球白菜细胞质雄性不育系S2108A和Y3611A及其相应的保持系S2108B和Y3611B进行了花器官形态比较和花药发育细胞学观察,结果表明:两套不育系和保持系之间在花器官的多个性状上差异极显著,而两个不育系S2108A和Y3611A之间除花蕾大小、雌蕊长和花瓣长有差异外,其他均不显著,说明从不同不育源转育来的两个不育系在花器官外形上差异不大;两个不育系花药败育表现均起始于四分体时期,且由于绒毡层的液泡化和径向膨大,挤压四分体小孢子导致败育,表明败育与绒毡层的不正常发育有关.此外,与Y3611A不同,在不育系S2108A中还存在另一种败育形式,绒毡层细胞壁发生融合,成为紧贴药室壁的类似于变形型绒毡层的周原质团,部分细胞质流人药室腔,包裹粘连四分体小孢子,成为染色很深的不连续团块状物,花药败育;两不育系花药发育存在花粉囊数目的变异,且花粉囊发育时期不同步,个别花粉囊较正常花粉囊发育推迟,在不育系Y3611A中还有巨型花粉囊的现象.     

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     

Effect of temperature on the expression of male sterility in Brussels sprouts (Brassica oleracea L. var. Gemmifera DC.)

Summary The effect of temperature on the expression of male sterility was studied in clones of partially male sterile and completely male sterile plants of Brussels sprouts.At a low temperature (10°C) most clones of male sterile plants developed normal fertile flowers, but some clones showed an opposite reaction. The female fertility of the clones of the male sterile plants did not differ much from that of the fertile clones.These results point to a possibility of propagating male sterile lines of cole crops by selfing or sib-mating them at low or high temperatures.     

红麻细胞质雄性不育系小孢子败育的细胞学观察

对同质异核的红麻细胞质雄性不育系L23A和K03A的小孢子发生和败育过程进行了细胞学比较观察。结果发现,2个不育系的败育过程有一定的相似性,败育都发生在四分体及单核小孢子时期;单核小孢子外壁发育都出现异常,小孢子原生质收缩变形或解体;花药壁的中层延迟解体,绒毡层出现细胞融合、液泡化、提前脱离药壁进入药室和过早解体等异常行为。但两者败育的过程又有一定差异,单核早中期,L23A的小孢子发育基本正常,而K03A的小孢子外壁发育不良,壁薄且质脆、易破损,刺稀少细小;L23A的小孢子外壁在单核后期解体成为念珠状,而K03A的小孢子外壁在发育过程中破损、逐渐溶解;小孢子外壁解体过程中,L23A的小孢子原生质收缩为核质不分的一团,而K03A的小孢子原生质逐渐解体。     

Genetic investigation and cytological comparison of two genic male sterile lines 9012A and MSL in <Emphasis Type="Italic">Brassica napus</Emphasis> L.

Genic male sterility (GMS) is one of the effective pollination control systems in hybrid rapeseed (Brassica napus L.) breeding. The recessive GMS lines 9012A and Lembke (MSL) have made a great contribution to the utilization of heterosis in rapeseed. Cytological observation and genetic investigation were employed to reveal anther abortion characters and genetic relationship of 9012A and MSL. Cytological and transmission electron microscopy observation revealed that anther abortion of 9012A and MSL initiated at the microsporocyte stage, both lines showed premature or retarded degradation of tapetum. Some anthers in 9012A developed to the microspore stage since their tapetal cells changed to the secretory type. However, MSL anthers did not progress to the tetrad stage, since the tapetum did not change to the secretory type. Genetic investigation revealed that MSL and 9012A had the same temporary maintainer system, while their cytoplasm types were different. The cytoplasm of MSL was similar to Cam while 9012A was similar to Nap type. The results laid a solid foundation for utilization of the two male sterile lines in rapeseed hybrid breeding.     

红麻GMS与CMS小孢子败育过程的细胞学及组织化学比较

以红麻细胞质雄性不育系(GMS)L23A、保持系L23B及从该保持系中发现的细胞核雄性不育系(CMS)L23BS为材料,采用石蜡显微制片技术,在光学显微镜下比较观察了三者花药发育过程中小孢子的发育过程及组织化学变化。结果表明,L23A的花药发育过程中,小孢子发育的不同阶段均出现败育现象,最早的败育表现为花粉母细胞在减数分裂之前的退化解体,最终形成空的花粉囊; 有的因花粉母细胞减数分裂过程异常而导致不能形成小孢子四分体;有的因小孢子在四分体中不能正常释放而败育,同时绒毡层细胞过度液泡化,并提早解体死亡; 花药发育早期含有少量蛋白质和淀粉,随着花药的发育逐渐变少。而L23BS小孢子败育的时期集中于四分体至单核花粉期间,表现为小孢子发育异常,有些小孢子不能从四分体里释放出来而影响其正常发育。在四分体以前与可育系类似,花药含丰富的蛋白质和淀粉。在败育的过程中,花药中的蛋白质和淀粉含量渐渐减少,但药隔组织中的在颗粒淀粉含量几乎不变。