38–48 kDa subunits of buckwheat 13S globulins are controlled by a single locus

The inheritance of buckwheat (Fagopyrum esculentum Moench) seed storage proteins was investigated by control crosses between accessions containing different SDS‐PAGE protein patterns in the range of 38–48 kDa. Analysis of segregation of individual F₁ and F₂ seeds obtained by 17 crosses showed segregation of eight electrophoretic bands, from which 12‐banding variants were inferred. Two variants contained one band, nine contained two bands and one variant contained three bands and behaved as a single co‐dominant unit. The segregation of banding variants fits the expected ratios and thus supports the hypothesis of single Mendelian gene inheritance and that alleles are co‐dominant. The results therefore suggest that the genes controlling globulin subunits are tightly linked and inherited as a single locus. The locus was designated Glob‐1 and the twelve segregated protein variants as alleles a‐l. This is the first buckwheat globulin locus identified with multiple alleles, phenotypically expressed as groups of protein bands and thus applicable as a functional marker in buckwheat genomic studies.     

Cytological analyses of hybrids and derivatives of hybrids between durum wheat and Thinopyrum bessarabicum,using multicolour fluorescent GISH*

The wheat progenitors and other wild relatives continue to be important sources of genes for agronomically desirable traits, which can be transferred into durum wheat (Triticum turgidum; 2n = 4x = 28; AABB genomes) cultivars via hybridization. Chromosome pairing in durum × alien species hybrids provides an understanding of genomic relationships, which is useful in planning alien gene introgression strategies. Two durum cultivars, ‘Lloyd’ and ‘Langdon’, were crossed with diploid wheatgrass, Thinopyrum bessarabicum (2n = 2x = 14; JJ), to synthesize F₁ hybrids (2n = 3x = 21; ABJ) with Ph1. ‘Langdon’ disomic substitution 5D(5B) was used as a female parent to produce F₁ hybrids without Ph1, which resulted in elevation of pairing between durum and grass chromosomes – an important feature from the breeding standpoint. The F₁ hybrids were backcrossed to respective parental cultivars and BC₁ progenies were raised. ‘Langdon’ 5D(5B) substitution × Th. bessarabicum F₁ hybrids were crossed with normal ‘Langdon’ to obtain BC₁ progeny. Chromosome pairing relationships were studied in F₁ hybrids and BC₁ progenies using both conventional staining and fluorescent genomic in situ hybridization (fl‐GISH) techniques. Multicolour fl‐GISH was standardized for characterizing the nature and specificity of chromosome pairing: A–B, A–J and B–J pairing. The A–J and B–J pairing will facilitate gene introgression in durum wheat. Multicolour fl‐GISH will help in characterizing alien chromosome segments captured in the durum complement and in their location in the A and/or B genome, thereby accelerating chromosome engineering research.     

Investigations into the causes of segregation of Ogura male sterility in Bangladeshi cultivars of radish

In a series of three experiments during 1998‐99 and 1999‐2000 at Gazipur, Bangladesh, the causes of segregation of Ogura cytoplasmic genetic male sterility in local cultivars of radish were studied. Male‐sterile populations at the BC₅ and BC₆ generations were grown under a range of field temperatures for 2 years and the results on pollen fertility tests revealed that the expression of male sterility was not affected by temperature. Neither was a genotype‐year interaction found. The unexpected segregation observed in the male‐sterile backcross generations might be due to the presence of restorer alleles in the maintainer parents.     

Cytology and pollen grain fertility in creeping bentgrass interspecific and intergeneric hybrids

Creeping bentgrass (Agrostis stolonifera L. or A. palustris Huds.) is a highly outcrossing allotetraploid species. It can form hybrids with a number of other Agrostis species and Polypogon genus. However, cytology and pollen grain fertility of the creeping bentgrass interspecific and intergeneric hybrids are not well known. In this research, chromosome pairing behaviors during meiosis I in F₁ and pollen viability of F₁ hybrids, as well as seed set rate and seed germination rate of backcrosses were studied in hybrids between creeping bentgrass, and other bentgrass species and three species of Polypogon genus. Abnormal chromosome pairing, laggard chromosomes, and premature segregation in F₁ hybrids were found. Pollen viability ranged from 1.6 to 48.5% amongst F₁ hybrids, significantly lower than that of the parents (85.5–94.1%). Some hybrids produced pollens of different sizes within the same anther. Seed set following backcrosses using F₁ hybrids as the male parent and creeping bentgrass as the recurrent parent was significantly lower than their parents. The study of chromosome paring behaviors and progeny fecundity are important in utilizing the alien genes to improve bio-stress and abio-stress resistance, and in assessing the potential transgene risks of creeping bentgrass.     

Inheritance of petal colour and its independent segregation from seed colour in Brassica rapa

The inheritance of petal (flower) colour and seed colour in Brassica rapa was investigated using two creamy‐white flowered, yellow‐seeded yellow sarson (an ecotype from Indian subcontinent) lines, two yellow‐flowered, partially yellow‐seeded Canadian cultivars and one yellow‐flowered, brown‐seeded rapid cycling accession, and their F₁, F₂, F₃ and backcross populations. A joint segregation of these two characters was examined in the F₂ population. Petal colour was found to be under monogenic control, where the yellow petal colour gene is dominant over the creamy‐white petal colour gene. The seed colour was found to be under digenic control and the yellow seed colour (due to a transparent coat) genes of yellow sarson are recessive to the brown/partially yellow seed colour genes of the Canadian B. rapa cvs.‘Candle’ and ‘Tobin’. The genes governing the petal colour and seed colour are inherited independently. A distorted segregation for petal colour was found in the backcross populations of yellow sarson × F₁ crosses, but not in the reciprocal backcrosses, i.e. F₁× yellow sarson. The possible reason is discussed in the light of genetic diversity of the parental genotypes.     

Blue aleurone trait diagnoses developmental origin of three pistils in a floret in common wheat

The common ‘three‐pistil’ (TP) wheat mutation line expresses TPs in a floret normally containing TPs forming three grains set close back‐to‐back. The developmental origin of the TP trait in common wheat had been diagnosed non‐destructively using the blue aleurone trait. The aleurone colour of F₂ seeds grown in F₁ plants of cross TP/UC66049 was evaluated. Due to xenia, the hue of blue grain colour depended on dose of the Ba1 gene for blue aleurone in the triploid endosperm. The TP trait produced four types of segregation in three‐seed clusters: (i) white grain only, (ii) two white grains and one blue, (iii) one white grain and two blue, and (iv) three blue grains only. The observed frequency of blue–white seed within clusters followed the binominal distribution ₃C_r (0.75)^r·(0.25)^3–r, where r is the number of colour variants in three‐seed clusters (r = 0–3). Intrafloret segregation of seed colour and F₂ segregation derived from aleurone colour of F₃ seeds indicated an independent origin of the TP trait.     

Genetic analysis of presence and absence of lint and fuzz in cotton

Cotton fibre mutants that were fuzzless and/or lintless were crossed with each other and a normal genotype (fuzzy, linted) to produce F₂ and BC₁ generations. F₂ segregation ratios from the cross of fuzzless‐lintless × fuzzy‐linted, for fuzzy‐linted, fuzzless‐linted and fuzzless‐lintless were 45 : 15 : 4. From the cross of fuzzless‐lintless × fuzzy‐linted, the F₂ segregation ratios were 9 : 39 : 16 whereas the BC₁F₁ segregation ratios from the F₁ backcrossed to fuzzless‐lintless were 1 : 3 : 4. These data suggest that the presence or absence of lint and fuzz are controlled by the interaction of four gene loci on non‐homologous chromosomes. We designate these loci as N₁, N₂, Li₃ and Li₄, where N₁ N₁ confers the presence of fuzzy, N₂N₂ confers inhibition of fuzzy initiation and development, and duplicate gene pairs, Li₃Li₃ and Li₄Li₄, determine the presence of lint. Homozygosity for li₃li₃ and li₄li₄ might also inhibit fuzz from development. In other words, they were recessive epistatic to fuzz genes.     

Aluminium tolerance in lentil (Lens culinaris Medik.) with monogenic inheritance pattern

The objectives of this study were to determine genetics of Al tolerance and whether the Al tolerance observed is governed by the same gene. The lines ‘L‐7903’ and ‘L‐4602’ have been developed through breeding programme as Al‐tolerant lines. These lines showed maximum root regrowth and minimum accumulation of Al and callose as compared to sensitive genotypes (‘BM‐4’ and ‘L‐4147’). Al tolerance in the parents, F₁, F₂ and backcross generations was estimated using the regrowth of the primary root after staining and scoring of fluorescent signals. The F₁ hybrids responded similarly to the tolerant parents, indicating dominance of Al tolerance over sensitivity. The segregation ratios obtained for Al tolerance and sensitivity in the F₂ and backcross generations were 3 : 1 and 1 : 1, respectively. Test of allelism confirmed the same gene was conferring Al tolerance in both genotypes (‘L‐7903’ and ‘L‐4602’) as the F₁ was also tolerant and no segregation of tolerant : sensitive was recorded. These results indicated that Al tolerance is a monogenic dominant trait that can be easily transferred to agronomic bases through backcross breeding technique.     

利用荧光原位杂交技术分析新合成异源四倍体拟南芥

在植物异源倍化育种中或者通过异源多倍化导入有利基因时,初级杂交后代异源多倍体减数分裂期间,避免部分同源染色体干扰,使同源染色体正常联会和配对以及正确分离是产生功能性雌雄配子的细胞学基础。本研究通过种间杂交技术,获得初级杂交后代异源四倍体拟南芥A. suecica,并重点分析其花粉母细胞减数分裂期同源染色体联会和配对情况。利用DAPI技术染色证明了花粉母细胞减数分裂期间核内染色体数目的正确性和均等分裂;而用荧光原位杂交技术进一步证明了核内染色体组的来源的正确性和同源染色体联会和配对的精准性。该结果证实新合成异源多倍体能进行正常的减数分裂和产生功能性雌雄配子,为种属间杂交和异源多倍体育种以及植物杂种优势利用提供了有利的细胞遗传学证据。     

Cytogenetic analysis of a spontaneous 5B/6B translocation in tetraploid wheat landraces from Ethiopia, and implications for breeding

Three tetraploid (2n= 4x= 28) wheat Triticum turgidum L. landrace morphotypes (= genotypes) from Ethiopia were found to carry a variant karyotype directly discernible under the microscope. This was possible because the rearrangement involved one of the satellited chromosomes. Giemsa C-banding revealed that the rearrangement resulted from a 5BS.6BS(5BL.6BL) centric reciprocal translation. The banding pattern on 5BL was polymorphic, suggesting that this translocation might have occurred more than once. There was little C-band polymorphism for the remaining chromosomes, except for 2A. As pure lines, all three morphotypes showed normal chromosome pairing at metaphase I (MI) in pollen mother cells (PMCs). indicating that they are genomically stable. Meiotic analyses of F₁ hybrids and F₂ segregates derived from crosses with tester varieties clearly indicated that one of them (B-l–9) carried another translocation. However, we were not successful in delecting the chromosomes involved, presumably the interchanged segments did nol include C-banding regions. By using T5BS.6BS, direct evidence for segregation distortion against translocation homozygotes in intervarietal hybrids was obtained. The distorted segregation was attributed lo zygotic selection. No aneuploid plants were obtained from the F₂ segregates. However, translocation heterozygotes resulting in unstable meiosis were abundant in the F₂ generation. The implications of the results in using the indigenous landraces in hybridization breeding are discussed.     

Recessive genes controlling resistance to Helminthosporium leaf blight in synthetic hexaploid wheat

A study was conducted under controlled environment conditions in a phytotron to determine the nature of the inheritance of resistance Helminthosporium leaf blight (HLB) in a synthetic hexaploid wheat line, ‘Chirya‐3’, against the isolate KL‐8 of Bipolaris sorokiniana from the major wheat growing region of India. Crosses were made between two susceptible lines ‘WH 147’ and ‘Chinese Spring’. Analyses of F₁ and F₂ populations of these two crosses (‘WH 147’בChirya‐3’ and ‘Chinese Spring’בChirya‐3’) showed that resistance against the isolate in ‘Chirya‐3’ was governed by two recessive genes functioning in a complementary interaction giving an F₂ segregation pattern of 1 : 15 (resistant : susceptible). The segregation pattern of the resistant F₂ progenies in F₃ families from both crosses confirmed that two homozygous recessive genes were responsible for resistance to the isolate of Bipolaris sorokiniana in the synthetic line ‘Chirya‐3’. It is proposed that the genes be designated as hlbr1 and hlbr2.     

Single nucleotide polymorphism genotyping of the barley waxy gene by polymerase chain reaction with confronting two-pair primers

A high‐throughput single nucleotide polymorphism (SNP) genotyping procedure was developed to select amylose‐free barley mutants whose waxy genes had a C‐ to T‐base substitution in exon 5, which converted Gln‐89 of the wild‐type gene into a termination codon. An F₂ population carrying an amylose‐free waxy gene was checked for segregation. Polymerase chain reaction with confronting two‐pair primers (PCR‐CTPP) produced allele‐specific PCR products that have different sizes and are inherited in a co‐dominant manner. Two alleles of the barley waxy gene with SNP were correctly identified in parental strains using the PCR‐CTPP procedure. Segregation of the SNP as detected by PCR‐CTPP in an F₂ population fitted the expected 1:2:1 ratio. The PCR‐CTPP procedure can provide a time saving and cost‐effective alternative to derived cleaved amplified polymorphic sequence in marker‐assisted selection.     

Molecular cytogenetic characterization of Thinopyrum genomes conferring perennial growth habit in wheat- Thinopyrum amphiploids

Seven wheat‐Thinopyrum amphiploids, AT 3425, AgCs, PI 550710, PI 550711, PI 550712, PI 550713 and PI 550714, were evaluated for perennial growth habit in the field. Three of them, AgCs, AT 3425, and PI 550713, were identified as perennials. Fluorescent genomic in situ hybridization (FGISH) patterns of mitotic chromosomes indicated that AgCs had seven pairs of Thinopyrum chromosomes and 21 pairs of wheat chromosomes. PI 550713 and AT 3425 showed similar FGISH patterns of mitotic chromosomes with three pairs of wheat‐Thinopyrum translocated chromosomes, seven pairs of Thinopyrum chromosomes, and 18 pairs of wheat chromosomes. Thinopyrum chromosome pairing in the Fi hybrid of AT 3425 with AgCs demonstrated differences between Thinopyrum genomes in these two amphiploids. Based on chromosome constitutions, pairing and reported pedigrees, AgCs and AT 3425 were identified as a wheat‐Thinopyrum elongatum amphiploid and partial wheat‐Thinopyrum ponticum amphiploid, respectively. Chromosome pairing in the F₁ hybrid between AT 3425 and PI 550713 revealed that these two amphiploids contained the same Thinopyrum genome. Two different Thinopyrum genomes conferring perennial growth habit were identified from the perennial amphiploids and characterized cytogenetically.     

Inheritance of siliqua locule number and seed coat colour in Brassica juncea

The inheritance of siliqua locule number and seed coat colour in Brassica juncea was investigated, using three lines each of tetralocular brown seeded and bilocular yellow seeded. Three crosses of tetralocular brown seeded × bilocular yellow seeded lines were attempted and their F₁, F₂ and backcross generations were examined for segregation of these two traits. Brown seed colour and bilocular siliqua characters were found to be dominant over yellow seed and tetralocular siliqua, respectively. Chi‐square tests indicated that each trait is controlled by different sets of duplicate pairs of genes. Bilocular siliquae or brown seeds can result from the presence of either of two dominant alleles, whereas tetralocular siliquae or yellow seeds are produced when alleles at both loci are recessive. A joint segregation analysis of F₂ data indicated that the genes governing siliqua locule number and seed colour were inherited independently.     

Inheritance of tolerance to zinc deficiency in barley

Barley (Hordeum vulgare L.) is often grown on alkaline zinc (Zn)‐deficient soils where reductions in yield and grain quality are frequently reported. Currently, the use of Zn‐based fertilizer along with Zn‐deficiency‐tolerant genotypes is considered the most thorough approach for cropping the Zn‐deficient soils; however, developing or breeding genotypes with higher Zn efficiency requires a good understanding of the inheritance of tolerance to Zn deficiency. This study was conducted to determine genetic control of this trait in barley. Two parental cultivars ('Skiff, moderately tolerant; and ‘Forrest’, sensitive), 185 F₂ plants, and 48 F₂‐derived F₃ families from this cross were screened to determine inheritance of tolerance to Zn deficiency using a visual score of deficiency symptoms. The segregation ratios observed indicated that greater tolerance to Zn deficiency in ‘Skiff compared with ‘Forrest’ at the seedling stage is controlled by a single gene with no dominance. The results also indicate that visual scores are useful for genetic analysis of tolerance to Zn deficiency.     

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.     

Inheritance of Karnal bunt-free trait in bread wheat

A Karnal bunt (KB)‐free wheat stock (‘KBRL22’) obtained from a cross of two resistant lines (‘HD29’ and ‘W485’) was used as a donor to introgress the KB‐free trait into ‘PBW343’(an ‘Attila’ sib), the most widely grown wheat cultivar in India. The number of KB‐free and KB‐affected plants in BC ₁, BC₂, BC₃ and BC₄ as well the F₂ was recorded after artificial inoculations. The segregation pattern in these generations clearly indicated two independently segregating, dominant genes which jointly confer the KB‐free attribute. The importance of the KB‐free line generated in this experiment is discussed.     

Inheritance of high palmitic acid content in the sunflower mutant CAS-12 and its relationship with high oleic content

CAS‐12 is a sunflower mutant with increased levels of palmitic (C16: 0 = 30%) and oleic (C18: 1 = 55%) acids in its seed oil, hence it has a reduced linoleic acid content (C18: 2 < 5%). This study was conducted to determine the inheritance of high C16: 0 content and its relationship with high C18: 1 content in CAS‐12. Reciprocal crosses involving CAS‐12, CAS‐5 (high C16: 0 content), HAOL‐9 (high C18: 1 content) and HA‐89 (standard fatty acid profile) were made. The F₁, F₂ and BC₁F₁ generations were obtained. The genetic control of the high C16: 0 trait in CAS‐12 was partially recessive and gametophytic. In all cases, this character segregated in the ratio 19: 38: 7 (low: intermediate: high C16: 0 content) in the F₂ generation. These results, together with the lack of segregation for C16: 0 content in crosses between CAS‐12 and CAS‐5, indicated that the genetic control of the high C16: 0 trait in CAS‐12 was similar to that in CAS‐5 in being controlled by partially recessive alleles (p1, p2, and p3) at three loci. Crosses between HA‐89 and CAS‐12, and HAOL‐9 and CAS‐5 (segregating for C16: 0 and C18: 1) demonstrated that the high C16: 0 and the high C18: 1 traits were independently inherited. However, C18: 1 segregation in these crosses exhibited reversal of dominance. Apparently, the low C18: 1 parental lines carried modifier genes causing the deviation.     

Monosomic analysis of Helminthosporium leaf blight resistance genes in wheat

A set of 21 monosomic (2n ‐ 1) and the disomic (2n) lines of the ‘Chinese Spring’ cultivar were crossed with ‘Chirya‐3′, the CIMMYT synthetic wheat line which has been identified as highly resistant for Helminthosporium leaf blight disease (HLB), in order to locate the genes governing disease resistance. The F₁ and segregating populations were challenged and screened against the most virulent pure mono‐conidial HLB isolate KL‐8 (Karnal, India). The F₁ progenies of the crosses were found to be susceptible because of the recessive nature of resistance. The F₂ progeny of the control cross (‘Chinese Spring’בChirya‐3’), segregated in the ratio of 1: 15 (resistant: susceptible), indicating that resistance to HLB was controlled by a pair of recessive genes. While the F₂ progeny of 19 monosomic crosses segregated in the ratio of 1: 15 (resistant: susceptible), the progeny of the remaining two crosses, 7B and 7D, deviated significantly from the ratio, revealing that 7B and 7D were the critical chromosomes for resistance genes that were located one on each chromosome. Moreover, the critical lines, 7B and 7D, confirmed the digenic complementary recessive nature of gene action by fitting well with the overall pooled F₂ segregation ratio of 13: 51 (resistant: susceptible) as expected for digenic complementary recessive resistance. The F₃ segregation ratios of the critical crosses, based on their pooled F₂ analysis, was estimated as 19: 32: 13 (non‐segregating susceptible: segregating as susceptible and resistant: non‐segregating resistant). F₃ progenies when tested with these ratios showed goodness‐of‐fit, confirming that the two pairs of recessive resistance genes were located on chromosomes 7B and 7D.     

Inheritance of rust resistance genes and molecular markers in microspore-derived populations of flax

The inheritance patterns of rust resistance genes and molecular markers in microspore‐derived populations of flax were investigated. Plants were produced from anther culture of F₁ plants from two crosses. Microspore‐derived plants in anther culture of flax were identified using molecular markers. Two rust resistance genes and three out of six molecular markers were inherited in expected Mendelian ratios in microspore‐derived populations. Distorted segregation for the other three molecular markers was shown to be the result of over‐representation of genomic fragments from the more responsive parent in the F₁ donor plant. The implication of this study in relation to androgenesis and flax breeding using anther culture is discussed.