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 marker Ms2 gene with a blue seed in durum and common wheat

In order to marker dominant nuclear gene Ms2 with a blue grain, a 4E disomic addition line ＇xiaoyanlanli＇（2n=44, AABBDD＋4EII） as the male parent to pollinate with male-sterile plants of durum wheat, controlled by a dominant nuclear gene Ms2, and a durum wheat line 89-2343 with Ms2 and blue seed marker on the same addition chromosome was developed. The genotype 89-2343 was crossed and backcrossed with a common wheat genotype 7739-3 to produce male fertile plants with blue seeds （MFP-BS）. To combine the blue seed marker, dwarf male-sterile plants carrying RhtlO and Ms2 were fertilized by pollen from selected MFP-BS. At last, the combination of blue seed marker, Ms2 and RhtlO was successfully produced. The segregation ratio of male sterility, seed color as well as chromosome configurations of the combinations suggested that the blue seed marker, Ms2 and RhtlO were located on the same chromosome. Cytological analysis indicated that the male sterile wheat line with a blue seed marker was 43 in chromosome number, with an additional chromosome. The transmission rate for blue seed male-sterile plants was 22.1% in common. In addition, the potential value for blue marker sterile lines in wheat breeding and hybrid production is discussed.     

Male Sterile Lines of Zinnia elegans and Their Cytological Observations

In order to find out a new pathway for utilizing heterosis of Zinnia elegans and accelerate breeding process, the mechanism of anther development of a male sterile line was explored. Backcross, sibmating, selling of fertile plants and testcross with inbred lines were analyzed and identified in the field, and cytology was observed. Recessive nucleus male sterile line AH209AB capable of being a maintainer was obtained by successive backcrosses with male sterile plants and fertile F1 plants as male parents. Cytological and anatomical studies indicated that： （1） The wall of normal anther was constituted of four layers of cells such as epidermis, powder chamber wall, middle level and tapetum cells. The process in meiosis of pollen mother cell in Zinnia elegans was normal and cytoplasm divided simultanously. Mature pollen grain was tricellular type. （2） The petal of male sterile plant degraded as a thread-like structure, the stamens were villiform in appearance and no pollens were formed. The result showed that the anther of male sterile plant no longer proceed to differentiate spore mother cell and the pollen sac after the formation of the tissue of sporogenous cells, there was no evident boundary between tapetum cell, middle lamella and inner wall of PMC, tapetal cells did not develop from the very beginning. So the abortion type was completely structural male sterility. The male sterile line belongs to non-sporange male sterile type and is of great use in F1 seeds production.     

Difference and Inheritance of the Sterility Maintenance of CMS in Indica Rice

The inheritance of the sterility maintenance ability was studied for three types of CMS lines (Zhenshan97A, XieqingzaoA and Ⅱ-32A) and several lines in indica rice. The result showed that the abilities to keep the male sterility of the maintainer lines strengthened obviously with the progressive backcrossing. The critical stage to maintain the sterility of CMS lines happened usually in BC5-6 generation. The selection of the recurrent parent gave an obvious influence on the pollen fertility distribution of the later generations. The author has discussed ways of improving of the maintainer lines.     

Development of Insect-Resistant Hybrid Rice by Introgressing the Bt Gene from Bt Rice Huahui 1 into 11-32A/B,a Widely Used Cytogenic Male Sterile System

Huahui 1 is an elite transgenic male sterile restorer line of wild rice abortive-type that expresses a Bacillus thuringiensis （Bt） δ-endotoxin and provides effective and economic control oflepidopteran insects. To exploit Huahui 1 to develop a new Bt rice, the insertion site of the Bt gene was determined by thermal asymmetric interlaced PCR （TAIL-PCR）. Bt was located in the promoter region ofLOC. Os10g10360, approximately 5.35 Mb from the telomere of the short arm of chromosome 10. For the first time, a Bt cytoplasmic male sterile （CMS） system was developed by introgressing Bt from Huahui 1. The recipient CMS system used consisted of Indonesia paddy rice-type II-32B （maintainer line） and II-32A （male sterile line）. Marker-assisted selection was used to increase selection efficiency in the backcrossing program. In BC5F1, the Bt plant 85015-8 was selected for further analyses, as it had the highest SSR marker homozygosity. In addition, the linkage drag of the foreign Bt gene in 85015-8 was minimized to 8.01-11.46 Mb. The foreign Bt gene was then delivered from 85015-8 into II-32A. The resultant Bt II-32A and Bt II-32B lines were both resistant to lepidopteran in field trials, and agronomic traits were not disturbed. The maintainability of II-32B, and the male sterility and general combining ability of II-32A, were not affected by the Bt introgression. This study demonstrates a simple and fast approach to develop Bt hybrid rice.     

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.     

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.     

Cloning and genetic transformation of a novel rice gene OsAPT2

A novel rice gene OsAPT2,which encodes a putative adenine phosphoribosyl transferase(APRT),was cloned.Its full-length cDNA is 1125bp,composing an ORF encoding 212 amino acid residues and a stop cordon,a 5' UTR of 123 bp and a 3' UTR of 363 bp.The sequence data have been submitted to the DDBJ/EMBL/GenBank databases(accession number:AY238894).The deduced amino acid sequence of OsAPT2 is highly homologous to those of previously reported APRTs.The genomic OsAPT2 gene contains 7 exons and 6 introns.Its total length is 4758 bp.Then,an antisense expression vector of the full-length OsAPT2 cDNA was constructed and transformed into rice variety Taibei309 by Agrobacterium tumefaciens mediated transformation method.In total,650 T0 transgenic plants were obtained based on both antibiotic screening and specific PCR identification.One hundred individuals of them were selected and planted in Hainan Island.From those 11 male sterile lines with seed-setting rate lower than 3% in bagged spike were obtained.Results suggest that OsAPT2 is involved in male sterility.Nine of the 11 male sterile lines were constitutive sterile lines;two of the 11 male sterile lines were thermo-sensitive genic male sterile lines,which may be useful in hybride rice breeding.     

新型甘蓝型油菜核不育材料绵7AB-4-2农艺性状鉴定及分子检测(英文)

[Objective] The study was to investigate the agronomic traits and breeding characteristics of genic male sterile material Mian 7AB-4-2 in Brassica napus. [Method] The differences in agronomic traits and polymorphisms in SSR markers, between the genic male sterile material Mian 7AB-4-2 in Brassica napus and its sisterly line Mian 7AB-4-1 were investigated by hybridization and molecular identification; and the percentage of sterile individuals of Mian 7AB-4-2 and of the hybrids with its sisterly line Mian 7AB-4-1 from test cross and back cross were also studied. [Result] Mian 7AB-4-2 was not significantly different in agronomic traits from its sisterly line Mian 7AB-4-1 at 0.05 probability level. The percentages of sterile individuals in the pollinated fertile Mian 7AB-4-2 plants were over 60%, and that in its sisterly line Mian 7AB-4-1 was about 25%. In test crosses with other nine sterile lines, Mian 7AB-4-1 kept the percentage of sterile individuals of sterile lines over 90%, and the percentage of sterile individuals from back cross over 80%. With regard to molecular markers, Mian 7AB-4-2 and its sisterly line Mian 7AB-4-1 were different in the band number from SSR primers a2 and E10. [Conclusion] The results indicate that Mian 7AB-4-2 is helpful for rapeseed breeding, quickening the application of new materials in field breeding.     

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.     

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）]。     

洋葱CMS-T型育性分子标记的筛选与鉴定

为筛选便于鉴定洋葱细胞质雄性不育类型及育性相关基因型的分子标记,采用cob(s)、cob(n)、orfA501、Cpp1和orf725共5个与CMS相关分子标记,jnurf13、AcSKP1、DNF-566、RNS-357、OPT和PsaO共6个与育性Ms位点相关分子标记分别对洋葱CMS类型和育性基因型筛选与鉴定。在洋葱不育类型分子标记鉴定中,Cpp1标记不适合本材料CMS分类,cob(s)、cob(n)和orfA501 3个标记联合鉴定洋葱CMS-T类型,筛选出orf725标记适合区分不育株及其不育类型;在34份洋葱育性相关基因型鉴定中,DNF-566、RNS-357、OPT和PsaO4个标记鉴定结果与表型不一致,筛选出jnurf13和AcSKP1 2个分子标记能够鉴定洋葱保持系基因型msms、不育系基因型msms、育性恢复基因型MsMs及杂合子(Msms)。结果表明洋葱CMS-T材料由单核基因Ms位点控制,并且首次报道AcSKP1标记用于CMS-T材料验证,但AcSKP1标记采用混合PCR,鉴定成本相对较高。所以,orf725和jnurf13标记适合田间筛选洋葱不育株类型和育性相关基因型。     

水稻雄性不育性及其分子标记辅助选择

综述了水稻质核互作雄性不育性及光温敏雄性不育性的遗传规律及分子标记辅助选择。质核互作雄育性由细胞质不育（Ｓ）和两对隐性细胞核基因控制,为孢子体不育的典败类型。光温敏核不育系,长日高温雄性不育,短日低温雄性可育,其不育性受两对隐怀基因控制,与细胞质无关,为孢子体不育的典败类型。ｐｍ１位于第７染色体,上与分子标记ＲＧ４７７的遗传距离３．５ｃＭ,下与ＲＧ５１１和ＲＺ２７２相距１５．０ｃＭ;ｐｍ２display     

水稻品种对细菌性条斑病的抗性研究

选用3套同核异质水稻不育系及其相应的保持系,以及生产上推广应用的13个水稻主栽品种,于水稻孕穗接种具有不同致病力的4个水稻细菌性条斑病菌菌株。不育胞质的抗病性鉴定结果表明,不同胞质类型的不育系之间抗病性差异明显,均比其同名保持系感病,其中野败型不育胞质（W汕A、W协A）高感细条病,T型胞质（T新A、T协A）对细条病的抗性较好。但在G型（如G新A等）少数不育胞质与条斑病菌如Syt等少数菌株的互作中,表现出明显的胞质效应,不育胞质较之正常胞质更感病。水稻品种抗性鉴定结果表明,江苏省水稻细条病发生地区目前推广的13个主栽品种中,武育粳3号、镇稻88等6个粳稻品种表现为高抗,5个籼型品种和1个粳品种高感条斑病。根据调查,在病区种植高抗品种完全可以控制水稻细条病的危害。     

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     

洋葱雄性不育性及其应用研究进展*

 概述了洋葱雄性不育系发现及利用的演化过程,介绍了S型和T型不育细胞质的特点及在全世界的利用情况;在质核互作不育类型利用上,介绍了洋葱和细香葱的最新研究进展; 并对利用不育系进行大面积制种生产过程中可能出现的风险进行了分析。最后提出了我国南方各省洋葱育种的方向。     

红麻雄性不育细胞质相关的cSNP位点发掘

在前期已获得红麻atp9基因编码区的基础上,以红麻细胞质雄性不育系P3A、保持系和F1代为材料,随机选取不同材料的3个atp9独立gDNA克隆子进行测序和序列比对。结果显示:具有不育细胞质的不育系和F1代与具有可育细胞质的保持系在第290位点处存在碱基差异,具有不育细胞质的材料均为碱基T,而具有可育细胞质的材料在相应位点为碱基A;atp9基因第290位点处T/A碱基转换的cSNP位点经已知细胞质的红麻种质资源的进一步检测,同时结合这些资源的田间育性进行分析发现,红麻种质资源UG93-2MS-1、UG93-2MS-2、UG93-2MS-3、UG93-2-22、KN250、KN142、ZB90在第290位点处为碱基T,能扩增出319bp条带,在田间对应的育性除品种UG93-2-22为可育外,其余均为不育或半不育;而福红992在第290位点处为碱基A,未能扩增出319bp条带,在田间对应的育性为可育。因此,位于atp9基因的第290位点处的T/A碱基转换的cSNP位点与红麻不育细胞质相关或连锁,为红麻不育细胞质鉴定提供了新的检测手段。     

大葱和洋葱种间杂种的获得及其特性

【目的】构建大葱和洋葱的种间杂种。【方法】以章丘大葱和洋葱自交系06-38为材料,进行远缘杂交,对杂种子房进行培养;利用CAPS技术对所获得植株进行鉴定;利用花粉染色法、离体萌发法和自交对其进行育性鉴定。【结果】大葱和洋葱正反交,自然状态下胚败育。在未添加激素的MS培养基上进行大葱与洋葱种间正反交后的子房培养,均获得种间杂种F1,授粉后第7天为最佳培养时期;通过子房培养分别获得了大葱×洋葱、洋葱×大葱杂种植株26株和22株;经CAPS检测,杂交后代全部为真杂种。F1杂种植株具有明显的杂种优势,杂种雄性不育。【结论】利用子房培养技术,克服了大葱和洋葱远缘杂种在自然状态下胚败育的情况发生,获得了种间杂种F1;所获得的杂种植株表现出明显的杂种优势,且雄性不育。     

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。