Induction of an alloplasmic male-sterile Brassica oleracea by substituting cytoplasm from ‘Early Scarlet Globe’ radish (Raphanus sativus)

Summary Alloplasmic male-sterile Brassica oleracea L. was synthesized in a backcrossing program through amphidiploid Raphanobrassica by using Early Scarlet Globe radish (Raphanus sativus L.) as the donor of cytoplasm and B. oleracea broccoli and cabbage as recurrent pollen parents. Persistence of radish chromosomes and high female sterility were encountered in the first four backcrosses. Following use of colchiploid 4x broccoli as pollen parent, a BC₅ plant was obtained that had 2n=3x+1=28 chromosomes, improved seed set, and no radish traits. The BC₆ with recurrent 2x broccoli contained male-sterile plants with 2n=18 or 19 chromosomes, increased seed set, and broccoli morphology. Subsequent generations segregated for male-sterile and restored male-fertile plants, some with variable development of stamens and pollen. Leaf color of the alloplasmic plants, especially seedlings, was lighter green than normal.     

Effect of Ogura male-sterile cytoplasm on the performance of cabbage hybrid variety. II. Commercial characteristics

Summary The main constraint of using Ogura male-sterile cytoplasm in Brassica is the induction of leaf yellowing at low temperature and the low seed set. But whether or not the cytoplasm can disturb the general commerical performance of the plants is not well-known. Therefore, this work was carried out with the purpose of evaluating Ogura cytoplasmic male-sterile cabbage hybrids and compare them to genomic similar male-fertile ones and to the most popular cabbage hybrid cultivated in Brazil. Harvest data showed an effect of the cytoplasm on reducing plant and head weight, core length and head length and width, although not altering heading index, shape and core indexes, nor head compactness. On the other hand, it was observed also that a careful selection of the parental lineages can produce male-sterile hybrids as good as their male-fertile analogues. All the evaluated hybrids were similar or better than the check for the characteristics under study.Abbreviations dat - days after transplanting - CNPH - National Center for Vegetable Crops Research     

A novel environment-induced genic male sterile (EGMS) mutant in indica rice

Summary Male-sterile mutants were isolated from M₂ and M₃ generations of indica rice variety 26 Zhaizao, dry seeds of which had been exposed to ⁶⁰Co- rays at a dose of 290 Gy. The mutants were planted in early season and ratooned in late season for two successive years for identification of fertility conversion in different growing seasons. One of the mutant lines was further observed in a growth chamber and in the field. Results showed that daily average temperature might be the major factor conditioning the male fertility conversion at a moderate daylength. The critical temperature for the male fertility conversion of the mutant grown under 12.5 h and 14.0 h daylength is about 23°C, below which the plant becomes completely male sterile. Its male fertility conversion character differs from other EGMS lines so far developed. The performance of the hybrids between the mutant and some other indica varieties demonstrated its good combining ability and its potential value in hybrid rice production. The obtained mutant line still sheds KI-stainable pollen grains under male sterilizing conditions. Nevertheless, pollen grains shed from the male sterile plants were much more vulnerable than from normal plants. At sucrose concentration below 1.5 M, the pollen grains from the mutant grown under male sterilizing conditions almost completely broke down, while above 1.5 M they became plasmolysed and shrunken. This is indicative of poor development of the membrane and walls of the pollen grains from the male sterile mutant, causing the pollen grains to be unfunctional. NBT test also clarified the abortion of the pollen grains from the mutant, which were formed in the male sterilizing environment.     

Identification and Inheritance of Fertility Restorer Genes for 'tour' CMS in Rapeseed (Brassica napus L.)

Eighteen genotypes of Brassica napus were crossed to a cytoplasmic male sterile (CMS) line of B. napus BO 15 carrying B. tournefortii cytoplasm (‘tour’ cytoplasm). Fourteen genotypes were found to be stable maintainers of the ‘tour’ CMS. Of the remaining four genotypes, GSL-1 and ‘Asahi-natane’ were found to be heterozygous and ‘Mangun’ and ‘Yudal’ were homozygous for the restorer gene. Analysis of the F₁ and F₂ progenies of (CMS) BO 15 בMangun’ and (CMS) BO 15 בYudal’ showed that fertility restoration is controlled by a single dominant gene. The availability of a number of stable maintainer lines and the simple inheritance pattern of fertility restorer gene makes ‘tour’ CMS a useful system for hybrid seed production in rapeseed.     

Establishing Cytoplasmic Male Sterility in Brassica napus by Mitochondrial Recombination with B. tournefortii

Fusion experiments between B. napus and X-ray-treated B. tournefortii protoplasts were carried out to develop cytoplasmic male sterility (ems) in B. napus. From the regenerants, six lines containing male sterile plants were selected; five lines segregated for male sterility, but one line (25–143) was completely male-sterile from the beginning. Molecular analyses of mitochondrial (mt) and chloroplast (cp) DNA of B. napus, B. tournefortii, B. juncea and cms juncea indicated that the original cytoplasmic donor of the cms juncea-system in B. napus was a B. tournefortii form, while the B. napus genotype used for the fusion experiments had a B. campestris cytoplasm. By analysis (it regenerated plants, line 25–143 was identified as possessing mt-DNA recombined between B. campestris and B. tournefortii. with the major part derived from B. campestris. No differences were detected between epDNAs from H. campestris and from line 25—143. The other five lines were similar to B. campestris with all the probes used. The low frequency of sterile lines from the fusion experiments and the inheritance of the cms in segregating progenies are both discussed.     

Production and testing of plants regenerated from protoplasts of photoperiod sensitive genic male sterile rice (Oryza sativa L.)

Plants were regenerated from protoplasts isolated from embryonic suspension cultures of N5047S, a photoperiod sensitive genic male sterile (PGMS) Japonica rice line. Flow cytometric analyses of nuclear DNA content identified some tetraploid regenerates whose agronomic traits could be distinguished from diploid regenerates. Pollen and female fertility of diploid protoplast-derived clones grown under different light and temperature conditions was compared. A promising PGMS protoplast clone, ZAU11S, was developed from these clones. Its male sterility was confirmed as a photoperiod × temperature interaction type. This revised version was published online in July 2006 with corrections to the Cover Date.     

哈克尼西棉细胞质陆地棉雄性不育系orf160的克隆及遗传转化

以哈克尼西棉细胞质陆地棉雄性不育系及其保持系、恢复系为材料,克隆并测序线粒体功能基因的侧翼序列。通过生物信息学分析,在atp4下游发现一个哈克尼西棉细胞质不育系特有的orf160。orf160全长480 bp,N端与atp6序列部分同源,C端序列与核序列部分同源;编码的159个氨基酸序列,与膜蛋白、细胞周期蛋白具有部分同源性。以含有线粒体导肽的GFP瞬时表达载体转化后,在洋葱表皮的细胞膜上观察到绿色荧光,表明线粒体定向载体正常表达。构建酵母和植物表达载体,分别转化酵母、烟草和拟南芥;同对照相比,转基因酵母菌斑畸形且生长缓慢;转基因拟南芥和烟草种子大部分畸形,T₁和T₂代植株的结实率和种子发芽率降低,花粉着色比野生型浅。结果表明,orf160基因的表达影响受体正常发育。     

利用分子标记辅助选育白菜薹复等位基因型雄性不育系

为解决白菜薹杂种优势利用中的杂交制种手段问题,以大白菜复等位基因型雄性不育系为不育源,设计定向转育方案,采用连续回交的方法转育不育性和农艺性状,同时利用分子标记辅助选择目标基因型植株,向白菜薹自交系中转育不育基因,创制白菜薹雄性不育系。通过26对SSR引物的筛选鉴定,获得与显性雄性不育基因Ms紧密连锁、且同样可以标记同一位点恢复基因Msf和可育基因ms的分子标记GSSR1。经过3个世代的回交转育,创制出了具有100%不育度和100%不育株率的白菜薹复等位基因型雄性不育系BGMS-3。以BGMS-3为母本,与6个白菜薹自交系杂交,筛选出1个强优势组合C3。     

PPR‐B‐31: a new maintainer allele in the male‐fertility restorer gene of radish (Raphanus sativus) Ogura cytoplasm

The PPR‐B gene is responsible for male‐fertility restoration of the Ogura‐type male‐sterile radish plants, and it is located in the complex Rfo locus in the vicinity of similar PPR‐A gene and PPR‐C pseudogene. The aim of this study was to identify PPR‐B alleles and understand the structure of the Rfo locus in radish breeding lines. Five lines of radish with normal male‐fertile cytoplasm were tested. The entire PPR‐B gene was amplified, sequenced and allelic PPR‐B sequences were identified. The results indicated that the maintainer lines 7, 15 and 21 contained a non‐restoring form of PPR‐B protein. A unique PPR‐B was found in lines 24/15 and 31 that are restorer and maintainer lines, respectively. The substitutions might be responsible for the loss of a restoring function of the PPR‐B‐31 allele. Amplification of the PPR‐A/PPR‐B and PPR‐B/PPR‐C intergenic regions allowed to identify rearrangements within Rfo locus. Obtained results confirm the wide allelic variation within the Rfo locus, as well as high genetic complexity of the fertility restoration mechanism in radish.     

Development of TGMS lines for developing two-line rice hybrids for the tropics

A breeding program was initiated at the International Rice Research Institute (IRRI) in 1990 to develop thermosensitive genic male sterile (TGMS) rice lines for developing two-line rice hybrids for the tropics. The TGMS trait was transferred from a temperate japonica TGMS mutant, Norin PL 12, to indica and tropical japonica rice varieties using the pedigree selection procedure. Six new TGMS rice lines adapted to tropical conditions were developed which showed complete pollen and spikelet sterility when maximum temperature was higher than 30 ^°C 1–2 week after panicle initiation. However, up to 85.5% spikelet fertility was observed when these lines were exposed to 26–29 ^°C during the critical stage. Using two of these TGMS lines, some heterotic rice hybrids showing 1–1.6 t/ha higher grain yield than the inbred check varieties were identified in unreplicated observational yield trial conducted at IRRI. Two of the six two-line hybrids yielded significantly higher than the check variety in a replicated preliminary yield trial. This revised version was published online in July 2006 with corrections to the Cover Date.