Development of dominant nuclear male-sterile lines with a blue seed marker in durum and common wheat

In order to develop genie male‐sterile lines with a blue seed marker, male‐sterile plants, controlled by a dominant nuclear gene Ms2, were used as female parents against a 4E disomic addition line ‘Xiaoyan Lanli’(2n= 44, AABBDD+4EII) as the male parent to produce monosomic addition lines with blue seed. Male‐sterile plants from the monosomic addition lines were pollinated with durum wheat for several generations and in 1989 a male‐sterile line with the blue grain gene and the male‐sterile gene Ms2 on the same additional chromosome was detected and named line 89‐2343. Using this line, the blue seed marker was successfully added to a short male‐sterile line containing Ms2 and Rht10. The segregation ratios of male sterility and seed colour as well as the chromosome figurations of different plants indicated that the blue grain genes, Ms2 and Rht10 were located on the same additional chromosome. Cytological analysis showed that the blue marker male‐sterile lines in durum wheat and common wheat were monosomic with an additional chromosome 4E. The inheritance ratio for blue seed male‐sterile plants and white seed male‐fertile plants was 19.7% and 80.3%, respectively, in common wheat. The potential for using blue marker sterile lines in population improvement and hybrid production is discussed.     

Chromosomal location of fertility restoring genes for ‘wild abortive’ cytoplasmic male sterility using primary trisomics in rice

Summary Identification and location of fertility restoring genes facilitates their deployment in a hybrid breeding program involving cytoplasmic male sterility (CMS) system. The study aimed to locate fertility restorer genes of CMSWA system on specific chromosomes of rice using primary trisomics of IR36 (restorer), CMS (IR58025A) and maintainer (IR58025B) lines. Primary trisomic series (Triplo 1 to 12) was crossed as maternal parent with the maintainer line IR58025B. The selected trisomic and disomic F₁ plants were testcrossed as male parents with the CMS line IR58025A. Plants in testcross families derived from disomic F₁ plants (Group I crosses) were all diploid; however, in the testcross families derived from trisomic F₁ plants (Group II crosses), some trisomic plants were observed. Diploid plants in all testcross families were analyzed for pollen fertility using 1% IKI stain. All testeross families from Group I crosses segregated in the ratio of 2 fertile: 1 partially fertile+partially sterile: 1 sterile plants indicating that fertility restoration was controlled by two independent dominant genes: one of the genes was stronger than the other. Testcross families from Group II crosses segregated in 2 fertile: 1 partially fertile+ partially sterile: 1 sterile plants in crosses involving Triplo 1, 4, 5, 6, 8, 9, 11 and 12, but families involving triplo 7 and triplo 10 showed significantly higher X² values, indicating that the two fertility restorer genes were located on chromosome 7 and 10. Stronger restorer gene (Rf-WA-1) was located on chromosome 7 and weaker restorer gene (Rf-WA-2) was located on chromosome 10. These findings should facilitate tagging of these genes with molecular markers with the ultimate aim to practice marker-aided selection for fertility restoration ability.     

Isolation of a Spontaneous Chromosome Translocation in Common Wheat

The genes Ms2 for male sterility and Rht10 for dominant dwarfing located on the short arm of chromosome 4D in common wheat arc closely linked. Male sterile, dwarf F₁ plants from the cross of male sterile‘Chinese Spring’× dwarf‘Ai-bian’were backcrossed with the variety‘Chinese Spring, From this offspring a spontaneous chromosome translocation was isolated resulting in a recombinant male sterile and dwarf genotype.     

Development of new cytoplasmic genetic male sterile lines through Indica × Japonica hybridization in rice

Summary From 28 Indica-Japonica crosses, two Indica cultivars, V.20B and Sattari were identified to possess male sterile cytoplasm with fertility restoring genes. It was possible to develop a new Japonica cytoplasmic genetic male sterile line (Zhunghua-1) on Indica male sterile cytoplasm (V 20B) by repeated backcrossing the complete pollen sterile plants of V 20B x Zhunghua-1 to the recurring male parent, Zhunghua-1. The study indicated that it would be possible to develop male sterile lines rom indica-japonica crosses only when there is sufficient amount of reciprocal differences with respect to pollen sterility. Further, it was inferred that it would be easier to develop Japonica male sterile lines on Indica cytoplasm than developing Indica male sterile line with japonica cytoplasm.     

Development of black rot resistant interspecific hybrids between Brassica oleracea L. cultivars and Brassica accession A 19182, using embryo rescue

Black rot is a bacterial disease of Brassica oleracea caused by Xanthomonas campestris pv. campestris. Resistance to the major black rot races 1 or 4 has been identified in related Brassica species including B. carinata and B. napus. In this study, two B. juncea accessions (A 19182 and A 19183) that are resistant to races 1 and 4 of Xcc were used as maternal and paternal parents to generate interspecific hybrids with B. oleracea cultivars. Interspecific hybrids were recovered using the embryo rescue technique and confirmed through inheritance of paternal molecular markers. Twenty-six interspecific hybrid plants were obtained between A 19182 and B. oleracea cultivars, but no interspecific hybrids were obtained using A 19183. Although interspecific hybrid plants were male sterile, they were used successfully as maternal parents to generate backcross plants using embryo rescue. All hybrid and BC₁ plants were resistant to black rot races 1 and 4.     

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.     

Induction of cytoplasmic male sterility in the seed progeny derived from artificially-synthesized interspecific chimera in Brassica

We produced artificial interspecific chimeras by in vitro grafting, and obtained cytoplasmic male sterile (CMS) variants in the seed progenies derived from backcrossing the chimera with one of the mother plants, B. campestris cv. Komatsuna. The induced CMS has been stably inherited by crossing it with `Komatsuna', not with `Ruby Ball' cabbage. The nuclear component of CMS is complete `Komatsuna' type in morphology, chromosome number (2n = 20) and Southern blot using ribosomal 17S RNA gene as a probe. PCR analysis by using mitochondrial atpA primer showed the complete `Ruby Ball' type, suggesting nuclear-cytoplasmic exchange. However, Southern blot patterns were different among those of the CMS and both parents by using atpA. Recombination or some unknown change is supposed in the mitochondrial genome via the processes of synthesis and propagation of the chimeras.     

Analysis of genic male sterility in Brassica oleracea

Summary The male sterility system MS-1 of Brassica oleracea was studied in order to elucidate if nucleo-cytoplasmic interactions determine this system. Crosses of male sterile MS-1 genotypes with heterozygous MS-5 genotypes gave fully fertile F₁ progenies. Selfing of seven F₁ plants resulted in five F₂ populations showing a 9:7 segregation ratio and two a 3:1 ratio for fertile and male sterile plants. Two F₂ progenies deviated from the expected 9:7 or 3:1 segregation ratios for fertile and male sterile plants. Thermosensitivity and distortion of the meiosis are suggested as the causal factors underlying the deviation of the segregation ratios. It was concluded that nuclear factors determine the male sterility in the MS-1 system, because the presence of a nucleocytoplasmic interaction in this system should have given only a 3:1 segregation ratio for fertile and male sterile plants in the F₂ generation.     

Glandular hair densities in three perennial Medicago species

Summary Eight triazine resistant (Brassica napus x B. oleracea) x B. oleracea interspecific hybrids with chromosome numbers ranging from 25 to 27 were backcrossed a second time to B. oleracea but no seed was formed. However, in vitro embryo rescue on 77 developing ovules yielded nine BC₂ plants with chromosome numbers between 19 and 25 and in which the herbicide resistance was still strongly expressed. Three of these plants (NOH-8B2B1, 2n=20; NOH-8B2B3 and NOH-8B2B4, 2n=19) were backcrossed again to B. oleracea. Two of the three plants produced seed which germinated to produce triazine resistan BBC₃s with 18, 19 or 20 chromosomes. The triazine resistant B. campestris cytoplasm has now been stabilized in B. oleracea.     

Identification and inheritance of male sterility in groundnut (Arachis hypogaea L.)

Summary Some plants without pods but with gynophores were observed in two F₄ progenies of two crosses of goundnut (Arachis hypogaea L.). The flowers on these plants had translucent white anthers with no or a few sterile pollen grains. Three such plants in the succeeding generation were hand pollinated with pollen from a short-duration Indian cv. JL 24. The resulting F₁ hybrid plants (male sterile x JL 24) were normal. Chi-square tests for segregation for male fertile and male sterile plants in F₂ and F₃ generations indicated that the male sterility in these crosses of groundnut is governed by two recessive genes. We designate these genes as ms₁ and ms₂ with ms₁ms₁ms₂ms₂ being a male sterile genotype.Submitted as ICRISAT J. A. No. 1812.     

Genetic and histological characterization of a novel recessive genic male sterile line of <Emphasis Type="Italic">Brassica napus</Emphasis> derived from a cross with <Emphasis Type="Italic">Capsella bursa</Emphasis>-<Emphasis Type="Italic">pastoris</Emphasis>

In a previously made cross Brassica napus cv. Oro (2n = 38) × Capsella bursa-pastoris (2n = 4x = 32), one F₁ hybrid with 2n = 38 was totally male sterile. The hybrid contained no complete chromosomes from C. bursa-pastoris, but some specific AFLP (amplified fragment length polymorphism) bands of C. bursa-pastoris were detected. The hybrid was morphologically quite similar to ‘Oro’ except for smaller flowers with rudimentary stamens but normal pistils, and showed good seed-set after pollination by ‘Oro’ and other B. napus cultivars. The fertility segregation ratios (3:1, 1:1) in its progenies indicated that the male sterility was controlled by a single recessive gene. In the pollen mother cells of the male sterile hybrid, chromosome pairing and segregation were normal. Histological sectioning of its anthers showed that the tapetum was multiple layers and was hypertrophic from the stage of sporogenic cells, and that the tetrads were compressed by the vacuolated and disaggregated tapetum and no mature pollen grains were formed in anther sacs, thus resulting in male sterility. The possible mechanisms for the production of the male sterile hybrid and its potential in breeding are discussed.     

Effects of low temperature storage on the pollen of Brassica campestris,B. oleracea and B. napus

Summary Four indica cultivars viz. Kalinga-I, Ptb. 10, IR 27280-13-3-3-3 and Co. 41 were found to possess male sterile cytoplasm with fertility restoring genes while the cultivar Krishna was found to maintain the male sterility in all the cases. All the plants in the F₁ of Kalinga-I × Krishna were observed to be completely male sterile and continued to show complete pollen sterility in subsequent backcross generations when backcrossed with recurring pollen parent, Krishna. Thus, it was posible to develop a new cytoplasmic-genetic male sterile line in indica rice (Krishna A) with Kalinga-I male sterile cytoplasm and this male sterile cytoplasm was found to be genetically different from others. Further, the newly developed male sterile line (Krishna A) was observed to be tolerant for low temperature at seedling stage.     

An assessment of the potential of 4DS.4DL-4s l L translocation lines as a means of eliminating tall off types in semi-dwarf wheat varieties

Summary Wheat varieties tend to be chromosomally unstable producing on average 2–3% of plants with abnormal chromosome numbers. A number of semi dwarf wheat varieties, carrying the gibberellic acid insensitive dwarfing genes Rht1 or Rht2, have been seen to produce distinct tall off types due to reduction in dosage of the chromosome carrying the dwarfing gene. The UK variety Brigand, carrying Rht2 on chromosome 4D, produced very distinct tall off types when this chromosome was reduced in dosage. The frequency of tall off types was sufficiently high to cause the variety to fail United Kingdom statutory uniformity tests. An attempt to prevent the loss of chromosome 4D was made by constructing translocation chromosomes involving the short arm of chromosome 4D, which carries Rht2, and the long arm of chromosome 4S ^l from Aegilops sharonensis, which carries a gene(s) conferring preferential transmission. The work in this paper describes the field evaluation of two lines carrying 4DS.4DL-4S ^l L translocations, and demonstrates their success in preventing spontaneously occurring monosomy of chromosome 4D in semi-dwarf wheats.     

Inheritance of male sterility mutations induced in haploid sorghum tissue culture

Summary A high frequency of male sterile mutants regeneration was shown in callus cultures derived from leaves and panicles of haploid sorghum (Ms_c1, A1 cytoplasm) and a spontaneous autodiploid obtained from this haploid. The cultures derived from the embryos of this autodiploid yielded significantly fewer mutants. Absolutely or partially male sterile mutants appeared among the regenerants or in the progeny of fertile regenerants. In the self-fertilized progenies of partially male sterile mutants and in the hybrids of sterile mutants with autodiploid line (i.e. under one and the same nuclear genome) male sterility mutations were inherited as cytoplasmic. Non-Mendelian segregation of sterile, partially male sterile and fertile plants was observed in these progenies. Partially male sterile plants were characterized by somatic segregation of male sterility genetic factors. In test-crosses with some CMS A1 fertility restorers, mutations were manifested as nuclear recessive while with others as nuclear dominant. These differences are supposed to be the result of interaction of fertility restorer genes of these testers with the novel cytoplasm. Male sterility mutations accompanied with female sterility were inherited as nuclear recessives.Abbreviations f fertile - ps partially male sterile - s male sterile plants     

Crosses between Beta vulgaris L. and Beta lomatogona F. et M.

Summary A population of 2x^ms sugar beets was crossed with 4x Beta lomatogona F. et M. The 3x F₁-plants were male sterile and were backcrossed with 2x and 4x sugar beets and multiplied without pollination as well. After the 1st backcross mainly 3x apomict types arose again and, among others, a small number of successful 4x backcrosses. After pollination by 4x sugar beets this 4x F₁ B₁ produced. besides predominatly apomictically multiplied 4x plants, also about 7% haploid 2x hybrids. The latter probably possess 1 genome from B. vulgaris and 1 genome from B. lomatogona. In the meiosis of the PMC's a certain amount of homeology between a number of chromosomes of both species could be established. The amphihaploid hybrids can be used as breeding parents for the creation of types in which introgession can occur. During hybridization in addition to 2x and 4x B. vulgaris types a number of 2x-, 3x-, 5x- and 6x-hybrids arose. This is presumably caused by the presence of gametes with the somatic number of chromosomes and the occurrence sometimes of haploid apomictic multiplication.The presence of large numbers of bolters in the F₁ and F₁ B₁ suggests that the bolting tendency of both species is based on different genes.     

Cytological identification of 1B/1R wheat-rye translocations in winter wheat breeding lines

Summary The incorporation of rye (S. cereale L.) chromatin into winter wheat (T. aestivum L.) cultivars is often achieved via hybridization of unadapted wheat-rye translocation lines with adapted wheat germplasm. Identification of progenies possessing the translocated chromosome has traditionally involved phenotypic screening for the desired rye characteristics. In this study, the Giemsa N-banding technique was evaluated as a potential screening tool for detection of 1B/1R wheat-rye translocations. Five breeding lines were examined from the pedigree Aurora/2^*TAM W-101. The differential banding patterns of chromosome 1B contributed by TAM W-101 and chromosome 1B/1R contributed by Aurora allowed unequivocal identification of translocation genotypes. Three of the lines were found to be heterogeneous, whereby plants were homozygous for either the normal 1B or the translocated 1B/1R chromosome. The remaining two lines were observed to be homozygous and homogeneous for the translocated 1B/1R chromosome. The implication of N-banding chromosomal analyses to wheat breeding is presented.Contribution No. J-5172, Department of Agronomy, Oklahoma Agriculture Experiment Station, Oklahoma State University, Stillwater, OK74078.     

Molecular markers linked to <Emphasis Type="Italic">Bn</Emphasis>;<Emphasis Type="Italic">rf</Emphasis>: a recessive epistatic inhibitor gene of recessive genic male sterility in <Emphasis Type="Italic">Brassica napus </Emphasis>L.

7–7365AB is a recessive genic male sterile (RGMS) two-type line, which can be applied in a three-line system with the interim-maintainer, 7–7365C. Fertility of this system is controlled by two duplicate dominant epistatic genes (Bn;Ms3 and Bn;Ms4) and one recessive epistatic inhibitor gene (Bn;rf). Therefore an individual with the genotype of Bn;ms3ms3ms4ms4Rf_ exhibits male sterility, whereas, plant with Bn;ms3ms3ms4ms4rfrf shows fertility because homozygosity at the Bn;rf locus (Bn;rfrf) can inhibit the expression of two recessive male sterile genes in homozygous Bn;ms3ms3ms4ms4 plant. A cross of 7–7365A (Bn;ms3ms3ms4ms4RfRf) and 7–7365C (Bn;ms3ms3ms4ms4rfrf) can generate a complete male sterile population served as a mother line with restorer in alternative strips for the multiplication of hybrid seeds. In the present study, molecular mapping of the Bn;Rf gene was performed in a BC₁ population from the cross between 7–7365A and 7–7365C. Bulked segregant analysis (BSA) and amplified fragment length polymorphism (AFLP) technique was used to identify molecular markers linked to the gene of interest. From a survey of 768 primer combinations, seven AFLP markers were identified. The closest marker, XM5, was co-segregated with the Bn;Rf locus and successfully converted into a sequence characterized amplified region (SCAR) marker, designated as XSC5. Two flanking markers, XM3 and XM2, were 0.6 cM and 2.6 cM away from the target gene, respectively. XM1 was subsequently mapped on linkage group N7 using a doubled-haploid (DH) mapping population derived from the cross Tapidor × Ningyou7, available at IMSORB, UK. To further confirm the location of the Bn;Rf gene, additional simple sequence repeat (SSR) markers in linkage group N7 from the reference maps were screened in the BC₁ population. Two SSR markers, CB10594 and BRMS018, showed polymorphisms in our mapping population. The molecular markers found in the present study will facilitate the selection of interim-maintainer.     

Characterization of a male sterile mutant from progeny of a transgenic plant containing a leaf senescence-inhibition gene in wheat

A male sterile plant of wheat (Triticum aestivum L.) segregated from progenies of a transgenic family containing the leaf senescence-inhibition gene P _SAG12 -IPT in the genetic background of ??Xinong 1376??, a well adapted winter wheat cultivar. The male sterile plant (named TR1376A) showed no phenotypic changes, except for florets and male organs, compared to its male fertile sibling plants (named TR1376B). The glumes and florets of male sterile TR1376A plants widely opened whereas those of the fertile counterpart TR1376B were closed or opened only briefly at flowing. Anthers of TR1376A were slender and indehiscent, and failed to release pollen. Compared to TR1376B, TR1376A anthers contained greatly reduced amounts of pollen, which was inviable or weakly viable. Ultra-structure studies indicated that cells in the endothecium and middle layers of the anther wall were dissolved or poorly developed in the sterile anthers of TR1376A. Molecular studies showed that the male sterility of TR1376A was caused by a sequence deletion or mutation that occurred in the promoter region of the transgene. F₁ hybrids of TR1376A and TR1376B gave 1:1 segregation of male fertility to sterility, indicating that the male sterility of TR1376A was heritable and controlled by a single dominant gene (named Ms1376). To date, only a few dominant nuclear male sterility genes have been characterized and one of them (Ms2) has been successfully used to improve wheat cultivars through recurrent breeding strategies. The discovery of the Ms1376 gene provides another dominant male sterile source for establishing recurrent breeding systems in wheat.     

Cytoplasmic male sterility in quinoa

Summary The quinoa cultivar Apelawa carries both normal and male sterile cytoplasms. Plants with male sterile cytoplasm produce flowers characterized by the complete absence of anthers and prominent exsertion of stigmas. Intraspecific crosses between male sterile quinoa plants and normal male fertile pollen donors consistently produced male sterile offspring under greenhouse conditions. Interspecific hybridization between male sterile quinoa plants and the related weed species Chenopodium berlandieri resulted in offspring with partial restoration of male fertility. The male sterile cytoplasm found in Apelawa has potential for use in the production of quinoa hybrids, although a more complete restorer system than that identified in C. berlandieri would be desirable.     

Transfer of cytoplasmic male sterility by spontaneous androgenesis in tobacco (Nicotiana tabacum L.)

Summary Five cytoplasmic male sterile lines of Nicotiana tabacum homozygous for a dominant rootless mutation [Rac^-] were used as female parents in crosses with three male fertile varieties. Androgenetic haploids were selected by their ability to form plantlets with a normal root system, whereas hybrid plantlets failed to grow. High frequencies of androgenetic haploid plants were obtained with N. debneyi cytoplasm, and with one paternal genotype. Chromosome doubling was performed using three different methods (pollination, in vitro culture, acenaphthen) and cytoplasmic male sterile doubled haploid plants have been obtained. The results demonstrate the efficiency of this method for a rapid cytoplasm transfer in Nicotiana tabacum; the integrity of the cytoplasm is conserved. For the first time, we possess a counter selectable marker universally usable for Nicotiana tabacum and closely related species.