Conversion of the genic male sterility (GMS) system of bell pepper (Capsicum annuum L.) to cytoplasmic male sterility (CMS)

Non‐pungent bell pepper (Capsicum annuum L.) lacks the cytoplasmic male sterility (CMS) nuclear restorer allele, Rf, and CMS cannot be employed in its F₁ hybrid seed production. To demonstrate that the genic male sterility (GMS) system in non‐pungent bell pepper can be converted to the CMS male sterility system, the conversion of GMS to CMS for non‐pungent bell pepper line GC3 was conducted by introgression of S‐type cytoplasm and the Rf allele from tropical pungent donors. After morphological traits were evaluated, two lines from BC₁F₁ containing S‐type cytoplasm and four lines from BC₂F₂ containing Rf allele, phenotypically similar to GC3, were obtained and could be employed as CMS male sterile lines and restorer lines for non‐pungent bell pepper. Four molecular markers potentially linked to traits of interest were also evaluated in BC₁F₁ and BC₁F₂ populations. This is the first time that GMS has been successfully converted to CMS in bell pepper, a significant contribution for bell pepper hybrid seed production.     

Genetic linkage between male sterility and non‐spiny trait in safflower (Carthamus tinctorius L.)

Genetic male sterility (GMS) exists naturally in safflower (Carthamus tinctorius L.). In the existing safflower GMS lines, sterile and fertile plants are distinguishable at flowering. This causes delay in fertile plants rouging and reduction in hybrid purity. In this investigation, a cross between a spiny GMS parent 13‐137 and a spiny non‐GMS parent ‘A1’ was effected. One sib cross, SC‐67, producing non‐parental‐type non‐spiny sterile and spiny fertile plants in F₃ was advanced to F₉ through sib crossing between non‐spiny sterile and spiny fertile plants. Mendelian digenic segregation was not observed for non‐spiny trait and male sterility. The results revealed strong linkage between these traits. The linkage was confirmed in F₂ generations of crosses between a non‐spiny marker‐linked GMS line (MGMS) and five elite lines. Male sterility–linked non‐spiny trait could distinguish sterile and fertile plants at elongation stage. The MGMS would be useful in production of pure F₁ hybrid seed and development of elite populations.     

Chloroplast-DNA variation in the wild and cultivated olives (Olea europaea L.) of Morocco

The male sterile plants that segregated in a BC₅F₂ of `C. sericeus × C. cajan var. TT-5' population were maintained by sib mating. The male sterile plants were crossed with ICPL-85012.Approximately 50% of the F₁ plants were sterile. F₂ plants derived from the fertile F₁ plants did not segregate for male sterility. The reciprocal hybrid i.e. ICPL-85012 × Fertile derivatives from C. sericeus × TT-5, did not express male sterility. However, among the 12 F₂ plant to row progenies, two segregated 25% male sterile plants and remaining 10 did not segregate. The segregation pattern in subsequent progenies revealed that the sterility was under control of a single recessive allele. Studies on the backcross and their BC₁F₂ and BC₁F₃progenies revealed another sterility gene which was found to be dominant in inheritance. This paper shows that what was thought to be cytoplasmic male sterility from C. sericeus cytoplasm is actually a single dominant gene possibly acting in concert with a single recessive gene to mimic cytoplasmic male sterility. This revised version was published online in July 2006 with corrections to the Cover Date.     

小麦花粉致死基因Ki的SSR标记分析

小麦雄性不育主要是通过花粉的败育表现,其不育材料对小麦杂种优势的利用研究具有重要意义和价值,国外研究表明,某些特定普通小麦品种间杂交F₁表现的花粉部分不育现象,受控于核基因组花粉致死基因Ki,为了筛选小麦花粉致死基因Ki的连锁标记,利用现代分子生物学技术通过定位该基因,克隆出花粉致死基因连锁标记片段,为小麦雄性不育种质材料的转育提供有效的选择标记。对小麦花粉致死基因Ki进行了分子标记定位,以‘中国春’和澳大利亚春小麦品种的BC₁F₁代作为定位群体,利用分离群体分组分析法(BSA)对位于小麦6B染色体上85对SSR引物进行多态性筛选,具有多态性的引物再通过BC₁F₁定位群体进行验证,从中筛选出与目的基因连锁的2个SSR标记Xgwm626和Xgpw4138。运用Mapmaker 3.0软件进行连锁分析。结果表明,Xgwm626和Xgpw4138与Ki基因的遗传距离分别为9.2 cM和6.9 cM,且2个SSR标记位于目的基因两侧,并将Ki定位于小麦6BL染色体上。研究结果为Ki基因的分子标记辅助选择和进一步精细定位奠定了基础。     

Development of cytoplasmic-genic male sterility in safflower

An interspecific cross was made between Carthamaus oxyacantha and the cultivated species C. tinctorius to develop a cytoplasmic‐genic male sterility (CMS) system in safflower. C. oxyacantha was the donor of sterile cytoplasm. The 3: 1 segregation pattern observed in BC₁F₂ suggested single gene control with dominance of male‐fertility over male‐sterility. The information obtained from crossing male sterile X male fertile plants in BC₁F₃ and BC₁F₄ generations showed statistically significant single gene (1: 1) segregation for male sterility vs. male fertility. The results demonstrated that C. tinctorius possesses a nuclear fertility restorer gene and that a single dominant allele restored fertility (Rf) in progeny carrying CMS cytoplasm of C. oxyacantha. Male sterility occurred with the homozygous recessive condition (rfrf) in a sterile C. oxyacantha cytoplasm background and not in the normal cytoplasm of C. tinctorius. The genetic background of different restorer lines of C. tinctorius having normal cytoplasm did not effect fertility restoration. The absence of male sterile plants in C. tinctorius populations ruled out the possibility of genetic male sterility. Normal meiosis in F₁ and BC₁F₂ ruled out a cytogenetic basis for the occurrence of male sterility.     

Identification of AFLP markers linked to one recessive genic male sterility gene in oilseed rape, Brassica napus

The commercial utilization of heterosis in seed yield by means of hybrid varieties is of great importance for increasing oilseed rape production in China. This requires a functional system for the production of hybrid seed. The Brassica napus oilseed rape line 9012AB is a recessive epistatic genic male sterility (GMS) two‐type line, in which the sterility is controlled by two pairs of recessive duplicate sterile genes (ms1 and ms2) interacting with one pair of a recessive epistatic inhibitor gene (rf). Homozygosity at the rf locus (rfrf) inhibits the expression of the recessive male sterility trait in homozygous ms1ms1ms2ms2 plants. This study was conducted to identify molecular markers for one of the male fertility/sterility loci in the B. napus male sterility line 9012AB. Sterile bulk (BS) and fertile bulk (BF) DNA samples prepared from male sterile and male fertile plants of the homozygous two‐type line 9012AB were subjected to amplified fragment length polymorphic (AFLP) analysis. A total of 256 primer combinations were used and seven markers tightly linked to one recessive genic male sterile gene (ms) were identified. Among them, six fragments co‐segregated with the target gene in the tested population, and the other one had a genetic distance of 4.3 cM. The markers identified in this study will greatly enhance the utilization of recessive GMS for the production of hybrid seed in B. napus oilseed rape in China.     

Induction of a mutation in the male fertility gene of the preferentially transmitted Aegilops sharonensis chromosome 4S1 and its application for hybrid wheat production

Summary Wheat plants nullisomic for chromosome 4B are male sterile due to the absence of the male fertility gene Ms1. However, plants in which chromosome 4B has been substituted by the preferentially transmitted chromosome 4S¹ of Ae. sharonensis are male fertile due to the compensating effect of Ms4 on the alien chromosome. This substitution line has been mutated and three recessive mutation of Ms4 have been selected. Plants homozygous for these mutations are male sterile. The implication of these mutations for hybrid wheat production is discussed.     

Mapping three new interspecific hybrid sterile loci between Oryza sativa and O. glaberrima

Hybrid sterility hinders the transfer of useful traits between Oryza sativa and O. glaberrima. In order to further understand the nature of interspecific hybrid sterility between these two species, a strategy of multi-donors was used to elucidate the range of interspecific hybrid sterility in this study. Fifty-nine accessions of O. glaberrima were used as female parents for hybridization with japonica cultivar Dianjingyou 1, after several backcrossings using Dianjingyou 1 as the recurrent parent and 135 BC₆F₁ sterile plants were selected for genotyping and deducing hybrid sterility QTLs. BC₆F₁ plants containing heterozygous target markers were selected and used to raise BC₇F₁ mapping populations for QTL confirmation and as a result, one locus for gamete elimination on chromosome 1 and two loci for pollen sterility on chromosome 4 and 12, which were distinguished from previous reports, were confirmed and designated as S37(t), S38(t) and S39(t), respectively. These results will be valuable for understanding the range of interspecific hybrid sterility, cloning these genes and improving rice breeding through gene introgression.     

一个水稻雄性不育突变体的遗传分析和基因定位

ms-np是一个源于自然突变的水稻雄性不育突变体,明显较正常植株矮小,叶色浓绿。小花解剖观察发现,突变体小花花丝细长,花药干瘪,呈白色透明状,但雄性器官的数量和雌性器官正常。碘染证实,突变体的花药壁内没有花粉粒着色,是一个典型的无花粉型雄性不育材料。5个F₂和2个BC₁F₁群体的遗传分析显示,该突变性状受1对隐性基因控制。对组合ms-np/M63衍生F₂不育单株的连锁分析表明,ms-np(t)基因位于水稻第6 染色体微卫星标记RM541和RM343之间,遗传距离分别为15.2 cM和7.9 cM。     

一个水稻雄性不育突变体的遗传分析和基因定位

ms-np是一个源于自然突变的水稻雄性不育突变体,明显较正常植株矮小,叶色浓绿。小花解剖观察发现,突变体小花花丝细长,花药干瘪,呈白色透明状,但雄性器官的数量和雌性器官正常。碘染证实,突变体的花药壁内没有花粉粒着色,是一个典型的无花粉型雄性不育材料。5个F₂和2个BC₁F₁群体的遗传分析显示,该突变性状受1对隐性基因控制。对组合ms-np/M63衍生F₂不育单株的连锁分析表明,ms-np(t)基因位于水稻第6 染色体微卫星标记RM541和RM343之间,遗传距离分别为15.2 cM和7.9 cM。     

The Attempted Transfer of Male Sterility from the Cornerstone Mutant of Tetraploid Wheat to Diploid Wheat

An attempt was made to transfer male-sterility, mslc, from the Cornerstone mutant of tetraploid wheat to diploid wheat. The N-banding technique revealed that chromosome 4 A of tetraploid wheat does not pair with chromosome 4 of diploid wheat in triploid F₁ hybrids: consequtntly the transfer of male-sterility gene(s) from tetraploid wheat to diploid was not successful. The culchiane induced ampliploids possessing AAAABB in mslc background were fully fertile indicating the complete compensation of mslc by the newly introduced A genome of T. monocaccum. The fertility compensating gene(S) presumably located on chromosome 4 of diploid wheat may be used to produce hybrid wheat by the XYZ system.     

Identification of AFLP markers linked to the epistatic suppressor gene of a recessive genic male sterility in rapeseed and conversion to SCAR markers

A recessive epistatic genic male sterility (REGMS) two‐type line, 9012AB, has been used for rapeseed hybrid seed production in China. The male sterility of 9012AB is controlled by two recessive duplicate sterile genes (ms1 and ms2) interacting with one recessive epistatic suppressor gene (esp). Homozygosity at the esp locus (espesp) suppresses the expression of the recessive male sterility trait in homozygous ms1ms1ms2 ms2 plants. In this study, we used a combination of bulked segregant analyses and amplified fragment length polymorphism (AFLP) to identify markers linked to the suppressor gene in a BC₁ population. From the survey of 1024 AFLP primer combinations, eight markers tightly linked to the target gene were identified. The two closest markers flanking both sides of Esp, P9M5₃₇₀ and S16M14₇₈₀, had a genetic distance of 1.4 cM and 2.1 cM, respectively. The AFLP fragment from P4M8₁₉₀, which co‐segregated with the target gene was converted into a sequence characterized amplified region marker. The availability of linked molecular markers will facilitate the utilization of REGMS in hybrid breeding in Brassica napus.     

Intergeneric hybridization of Diplotaxis erucoides with Brassica napus. I. cytogenetic analysis of F1 and BC1 progeny

Summary Intergeneric hybrids (F₁) Diplotaxis erucoides (DeDe) x Brassica napus (AACC) and the first backcross to B. napus (BC₁) have been obtained through in vitro culture of excised ovaries. The chromosome numbers of F₁ and BC₁ plants proved the occurrence of unreduced gametes. The study of metaphase I chromosome pairing showed that autosyndesis in De genome and allosyndesis between De and A/C genomes might exist. The male fertility of the F₁ plants was low. Some male-sterile plants were found in F₁ and BC₁ progeny. The possibilities of creating addition lines B. napus-D. erucoides and of obtaining a new cytoplasmic male sterility in B. napus are discussed.     

Development of SCAR and CAPS markers linked to a recessive male sterility gene in lettuce (<Emphasis Type="Italic">Lactuca sativa</Emphasis> L.)

Genetic male sterility (GMS) has been a useful system for the production of hybrid varieties in self-pollinated plants. We obtained a GMS line developed from a spontaneous mutation in lettuce (Lactuca sativa L.). Genetic analysis in our previous study revealed that the sterility was controlled by a recessive gene which was named ms-S. For simple and quick screening of individuals showing male sterility, we attempted molecular mapping of the ms-S locus using an amplified fragment length polymorphism (AFLP) technique. From the examination of 4,096 AFLP primer combinations, 63 AFLP markers were found to be linked to the gene and nine of them were successfully converted into sequence characterized amplified region (SCAR) markers and cleaved amplified polymorphic sequence (CAPS) markers. Linkage analysis indicated that these nine markers were closely linked to the ms-S gene and all were located on the same side of the gene. The minimum genetic distance between the ms-S gene and a marker was 3.1 cM. These results provide additional information for map-based cloning of the ms-S gene and will be of great help for lettuce breeding using GMS to produce F₁ hybrids.     

Mapping of the <Emphasis Type="Italic">ms8</Emphasis> male sterility gene in sweet pepper (<Emphasis Type="Italic">Capsicum annuum</Emphasis> L.) on the chromosome P4 using PCR-based markers useful for breeding programmes

The nuclear male sterility gene ms8 is expected to facilitate the production of sweet pepper (Capsicum annuum L.) hybrids as it provides means for hybridization without the labor-intensive hand emasculation of female inbred lines. The development of molecular markers linked to ms8 locus will help the breeding practice for the selection of hybrid parental lines. In this study, F₂ population resulting from a cross between the sweet pepper male sterile line 320 and the male fertile variety Elf was used to identify DNA markers linked to the ms8 locus. With the use of RAPD–BSA technique, seven markers linked to the ms8 locus were found. Four of them were converted into SCAR markers. In addition, two COSII/CAPS markers linked to the ms8 locus were identified. Comparative mapping with reference pepper maps indicated that the ms8 locus is located on the lower arm of the pepper chromosome P4. Identified markers are useful for molecular breeding, however, at present markers tightly linked to ms8 locus are still lacking. Identification of molecular markers linked to the ms8 locus and determination of its chromosomal localization are useful for fine mapping and also provide the perspective for ms8 gene cloning.     

Cytogenetic study of Helianthus laevigatus and its F1 and BC1F1 hybrids with cultivated sunflower, Helianthus annuus

Although the wild sunflower species Helianthus laevigatus has not been extensively studied it may be considered for sunflower breeding as a potential source of desirable genes for Sclerotinia stalk rot resistance and high contents of proteins and linoleic acid in the seed. A set of six H. laevigatus populations was crossed to cultivated sun~ower lines and produced nine F₁ (2-14 plants) and 66 BC₁F₁ hybrid combinations (1-13 plants). Male sterility occurred in F₁ and BC₁F₁ hybrid combinations and pollen viability was lower in the progenies than in the parents (51.6-77.2%in F₁ and in F₁ and 4.8-34.0% in BC₁F₁). Meiosis was normal in the H. laevigatus populations It was found that this tetraploid species also occurred in a hexaploid form Numerous irregularities were observed in the meiosis of the F₁ interspecific hybrids During diakinesis, quadrivalents and hexavalents were recorded in addition to bivalents Dislocated chromosomes and chromosome bridges were present in the other phases The chromosome number in F₁ was 68 (tetraploid). Irregularities in chromosome pairing were observed in the interspecific hybrids at BC₁F₁. There were many univalents, and trivalents quadrivalents and hexavalents were also present The chromosome number in the BC₁F₁ generation ranged from 34 to 60. The occurrence of meiotic irregularities in the F₁ and BC₁F₁ interspecific hybrids indicates that H. laevigatus and the cultivated sunflower differ in genome constitution.     

Microsatellite Marker Identification of a Triticum Aestivum -Aegilops Umbellulata Substitution Line with Powdery Mildew Resistance

Summary Aegilops umbellulata acc. Y39 and Triticum carthlicum acc. PS5, immune to many powdery mildew isolates, were crossed to make an amphidiploid line Am9. The powdery mildew resistance of Am9 was transferred to common wheat cultivar Laizhou953 by crossing and backcrossing. In this study, the origin of powdery mildew resistance in a BC₃F_4:5 population derived from a cross of Am9 and Laizhou953 was identified. Microsatellite markers analysis showed that markers Xgwm257, Xgwm296, and Xgwm319, co-segregated with the powdery mildew resistance, whereas markers Xgwm210, Xgwm388/140, Xgwm388/170 and Xgwm526 were related to susceptibility and linked to resistance in repulsion. Of three markers related to resistance, Xgwm257 and Xgwm319 were codominant, whereas Xgwm296 was dominant. All three markers were Ae. umbellulata-specific indicating that resistance in the test population originated from Ae. umbellulata acc. Y39. The chromosome location and mapping of these linked microsatellite markers, the chromosome numbers of derived BC₃F_4:6 families, and chromosome pairing in F₁ plants from a cross of a homozygous resistant BC₃F_4:5 plant and Laizhou953, showed that wheat chromosome 2B was substituted by Ae. umbellulata chromosome 2U. This is the first gene conferring powdery mildew resistance transferred to wheat from Ae. umbellulata, and it should be a novel resistance gene to powdery mildew. It was temporarily designated PmY39.The first two authors made equal contributions     

Inheritance and chromosome location of photoperiod‐thermo sensitive male sterility in wheat line Xinong 291S

The present research aimed to study the inheritance and chromosomal location of a photoperiod‐thermo sensitive male sterility (PTSMS) gene in Xinong 291S (XN291S), which is a new PTSMS wheat line. The inheritance was studied in F₁ and F₂ populations derived from crosses between XN291S and eight wheat cultivars. All F₁ plants were fertile and the F₂ populations segregated in either 15 : 1 or 3 : 1 fertile : sterile ratios indicating that PTSMS was controlled by one or two recessive major genes. Five wheat cultivars carried homozygous dominant alleles for fertility, whereas the other three, including ‘Chinese Spring’ (CS), carried a single homozygous dominant allele. Chromosomal location of the PTSMS gene was studied by crossing a set of CS nulli‐tetrasomic lines to XN291S. Self‐fertility of F₁ hybrids XN291S/N5BT5A and XN291S/N5BT5D were significantly lower than the others. Therefore, the PTSMS gene in XN291S that differed from CS was located on chromosome 5B. Location of the second PTSMS gene needs further study.     

Mapping of male sterility gene ms10 in chilli pepper (Capsicum annuum L.)

Most of the hybrid seed in chilli are produced manually, but the use of male sterility (MS) can reduce the cost of hybrid seed production. MS‐12, a nuclear male‐sterile (NMS) line developed at Punjab Agricultural University, Ludhiana (India), has been utilized to develop commercial F₁ hybrids. A recessive gene, designated as ms10, governs MS in MS‐12. Due to recessive gene control, development of new NMS lines incorporating ms10 gene is tedious and time‐consuming. We identified SSR markers AVRDC‐PP12 and AVRDC_MD997* linked to the ms10 gene. A total of 558 primer pairs were screened following bulked segregant analysis (BSA). Linkage analysis in 210 F₂ plants indicated that the two SSR markers were linked to the ms10 gene and the marker AVRDC‐PP12 was closest to the gene at 7.2 cM distance. The marker was mapped to chromosome 1 at genome position 175 694 513 to 175 694 644. Until more closely linked markers are developed, the marker AVRDC‐PP12 would facilitate transfer of ms10 gene through marker‐assisted selection (MAS). Fine mapping would lead to cloning of the ms10 gene.     

Identification of male sterility maintainer lines for Ogura radish (Raphanus sativus L.)

‘Ogura radish’, a cytoplasmic genetic male sterile line, was crossed with four local and three Japanese cultivars to identify maintainer lines. Out of seven F₁ families, one cross involving a local cultivar, Aushi, produced 100% male sterile (MS) progeny. The crosses involving the other two local cultivars, Tangail Local and Kuni, produced about 90% MS progeny, indicating the presence of maintainer gene(s) for male sterility. The fourth local cultivar, Tasaki, produced 100% male fertile (MF) progeny. All three exotic cultivars appeared to possess the chromosomal gene(s) for controlling the male sterility. In BC₁, BC₂ and BC₃ generations, segregation of MS plants were more frequent when ‘Aushi’ was used as recurrent parent. The expression of male sterility was not affected by seasonal influences. Thus the local cultivar ‘Aushi’ may be used as maintainer line for ‘Ogura radish’. To produce hybrid seed, ‘Tasaki’ can be used as pollinator line as it exhibit high heterosis with ‘Aushi’. This revised version was published online in July 2006 with corrections to the Cover Date.