Transfer of Ph I genes promoting homoeologous pairing from Triticum speltoides to common wheat

Diploid-like chromosome pairing in polyploid wheat is controlled by several Ph (pairing homoeologous) genes with major and minor effects. Homoeologous pairing occurs in either the absence of these genes or their inhibition by genes from other species (Ph ^I genes). We transferred Ph ^I genes from Triticum speltoides (syn Aegilops speltoides) to T. aestivum, and on the basis of further analysis it appears that two duplicate and independent Ph ^I genes were transferred. Since Ph ^I genes are epistatic to the Ph genes of wheat, homoeologous pairing between the wheat and alien chromosomes occurs in the F₁ hybrids. Using the Ph ^I gene stock, we could demonstrate homoeologous pairing between the wheat and Haynaldia villosa chromosomes. Since homoeologous pairing occurs in F₁ hybrids and no cytogenetic manipulation is needed, the Ph ^I gene stock may be a versatile tool for effecting rapid and efficient alien genetic transfers to wheat.Contribution no. 93-435-J from the Kansas Agricultural Experiment Station, Kansas State University, Manhattan, KS 66506-5502, USA     

Cytogenetic processes in the course of Triticum aestivum and Haynatricum hybridization

Meiosis in hybrids obtained in direct and return crossings between Haynatricum and wheat was studied. In F1 hybrids the possibility of stimulation of homoeologous pairing between the chromosomes of T. aestivum and T. dicoccum and, probably, H. villosum, if Haynatricum was used as pollinator, has been shown. This process is considerably intensified when the genes regulating chromosome pairing, in particular ph1b mutation, are used. In reciprocal crosses it was shown that wheat genotypes can differently influence on homoeologous chromosome pairing. In BC1 and F2 generations the chromosome compositions are determining which arise as a result of stochastic processes in premeiotic mitoses and in meiosis in F1 hybrids.     

Comparison of homoeologous chromosome pairing between hybrids of wheat genotypes Chinese Spring ph1b and Kaixian-luohanmai with rye

The ph-like genes in the Chinese common wheat landrace Kaixian-luohanmai (KL) induce homoeologous pairing in hybrids with alien species. In the present study, meiotic phenotypic differences on homoeologous chromosome pairing at metaphase I between hybrids of wheat genotypes Chinese Spring ph1b (CSph1b) and KL with rye were studied by genomic in situ hybridization (GISH). The frequency of wheat-wheat associations was higher in CSph1b×rye than in KL×rye. However, frequencies of wheat-rye and rye-rye associations were higher in KL×rye than in CSph1b×rye. These differences may be the result of different mechanisms of control between the ph-like gene(s) controlling homoeologous chromosome pairing in KL and CSph1b. Wheat-wheat associations were much more frequent than wheat-rye pairing in both hybriods. This may be caused by lower overall affinity, or homoeology, between wheat and rye chromosomes than between wheat chromosomes.     

Comparative analysis of the meiotic effects of wheat ph1b and ph2b mutations in wheat×rye hybrids

 Wheat-wheat and wheat-rye homoeologous pairing at metaphase I and wheat-rye recombination at anaphase I were examined by genomic in situ hybridization (GISH) in wild-type (Ph1Ph2) and mutant ph1b and ph2b wheat×rye hybrids. The metaphase-I analysis revealed that the relative contribution of wheat-rye chromosome associations in ph2b wheat×rye was similar to that of the wild-type hybrid genotype but differed from the effect of the ph1b mutation. The greater pairing promotion effect of the ph1b mutation appears to be relatively more on distant homoeologous partner metaphase-I associations, whereas the lower promoting effect of ph2b is evenly distributed among all types of homoeologous associations. This finding reveals that distinct mechanisms are involved in the control of wheat homoeologous pairing by the two Ph genes. The frequency of wheat-rye recombination calculated from anaphase-I analysis was lower than expected from the metaphase-I data. A greater discrepancy was found in ph2b than in ph1b wheat×rye hybrids, which may suggest a more distal chiasma localization in the former hybrid genotype. Received: 20 June 1997 / Accepted: 9 December 1997     

Homoeologous relationships of Aegilops speltoides chromosomes to bread wheat

 Homoeologous pairing at metaphase I was analysed in the standard-type, ph2b and ph1b hybrids of Triticum aestivum (AABBDD) and Aegilops speltoides (SS). Data from relative pairing affinities were used to predict homoeologous relationships of Ae. speltoides chromosomes to wheat. Chromosomes of both species, and their arms, were identified by C-banding. The Ae. speltoides genotype carried genes that induced a high level of homoeologous pairing in the three types of hybrids analyzed. All arms of the seven chromosomes of the S genome showed normal homoeologous pairing, which implies that no apparent chromosome rearrangements occurred in the evolution of Ae. speltoides relative to wheat. A pattern of preferential pairing of two types, A-D and B-S, confirmed that the S genome is very closely related to the B genome of wheat. Although this pairing pattern was also reported in hybrids of wheat with Ae. longissima and Ae. sharonensis, a different behaviour was found in group 5 chromosomes. In the hybrids of Ae. speltoides, chromosome 5B-5S pairing was much more frequent than 5D-5S, while these chromosome associations reached similar frequencies in the hybrids of Ae. longissima and Ae. sharonensis. These results are in agreement with the hypothesis that the B genome of wheat is derived from Ae. speltoides. Received: 8 January 1998 / Accepted: 4 February 1998     

Chromosome pairing in meiosis of partially fertile wheat/rye hybrids

The normal course of meiosis depends on regular pairing of homologous chromosomes. In intergeneric hybrids, including those of wheat, there is no chromosome pairing because there are no homologs. In F₁ wheat/rye hybrids, pairing is largely prevented by the pairing homoeologous1 (Ph1) gene. In its presence, there are only rare instances of pairing; most chromosomes are univalent, and their orientation at metaphase I initiates different pathways of the meiotic cycle. The meiotic-like pathway includes a combination of the reductional and the equational + reductional steps at AI followed by the second division. The resulting gametes are mostly non-functional. The mitotic-like pathway involves equational division of univalents at AI and the absence of the second division. Any fertility of wheat/rye hybrids depends on the production of unreduced gametes arising from meiotic restitution (mitotic-like division). We examined the meiotic pairing in wheat/rye hybrids created from wheat lines with single rye chromosome substitutions and Ph1 present. This guaranteed F₁ meiosis with one pair of rye homologs. All hybrids formed bivalents, but proportions of meiocytes with bivalents varied. In the meiocytes where bivalents were present, there was a higher tendency for the meiotic-like pathway, while in meiocytes where bivalent pairing failed, the tendency was stronger for the mitotic-like pathway. Among the equationally dividing cells, we observed more than 90 % of meiocytes without bivalents, where rye homologs did not form bivalents, too. The data indicate a potential application of wheat/rye lines in producing genetic stocks of amphidiploids with designated genomic constitutions.     

Direct evidence for high level of autosyndetic pairing in hybrids of Thinopyrum intermedium and Th. ponticum with Triticum aestivum

 Genomic in situ hybridization (GISH) was used to distinguish autosyndetic from allosyndetic pairing in the hybrids of Thinopyrum intermedium and Th. ponticum with Triticum aestivum cv ‘Chinese Spring’ (CS). All hybrids showed high autosyndetic pairing frequencies among wheat chromosomes and among Thinopyrum chromosomes. The high autosyndetic pairing frequencies among wheat chromosomes in both hybrids suggested that Th. intermedium and Th. ponticum carry promoters for homoeologous chromosome pairing. The higher frequencies of autosyndetic pairing among Thinopyrum chromosomes than among wheat chromosomes in both hybrids indicated that the relationships among the three genomes of Th. intermedium and among the five genomes of Th. ponticum are closer than those among the three genomes of T. aestivum. Received: 19 September 1996 / Accepted: 18 April 1997     

应用原位杂交技术对小麦—黑麦代换系间杂交的细胞遗传研究

应用原位杂交技术结合染色体组型分析方法,对两个小麦-黑麦异源双代换系5R/5A和6R/6A杂交后代的遗传进行了研究,探讨同祖染色体配对的可能性并获得小麦-黑麦易位系。实验中对杂种F1代植株减数分裂各时期的花粉母细胞染色体行为进行分析,结果发现有22.91%的花粉母细胞中黑麦染色体与小麦染色体发生同祖配对。F2代通过C-分带、原位杂交鉴定,在45株中检测到9株易位,易位频率为20%,是目前报道易位频率最高的。染色体易位有的来源于同祖配对交换,有的来源于单价体错分裂或断裂的重建。     

Genotypic variation in tetraploid wheat affecting homoeologous pairing in hybrids with Aegilops peregrina.

The Ph1 gene has long been considered the main factor responsible for the diploid-like meiotic behavior of polyploid wheat. This dominant gene, located on the long arm of chromosome 5B (5BL), suppresses pairing of homoeologous chromosomes in polyploid wheat and in their hybrids with related species. Here we report on the discovery of genotypic variation among tetraploid wheats in the control of homoeologous pairing. Compared with the level of homoeologous pairing in hybrids between Aegilops peregrina and the bread wheat cultivar Chinese Spring (CS), significantly higher levels of homoeologous pairing were obtained in hybrids between Ae. peregrina and CS substitution lines in which chromosome 5B of CS was replaced by either 5B of Triticum turgidum ssp. dicoccoides line 09 (TTD09) or 5G of Triticum timopheevii ssp. timopheevii line 01 (TIMO1). Similarly, a higher level of homoeologous pairing was found in the hybrid between Ae. peregrina and a substitution line of CS in which chromosome arm 5BL of line TTD140 substituted for 5BL of CS. It appears that the observed effect on the level of pairing is exerted by chromosome arm 5BL of T turgidum ssp. dicoccoides, most probably by an allele of Ph1. Searching for variation in the control of homoeologous pairing among lines of wild tetraploid wheat, either T turgidum ssp. dicoccoides or T timopheevii ssp. armeniacum, showed that hybrids between Ae. peregrina and lines of these two wild wheats exhibited three different levels of homoeologous pairing: low, low intermediate, and high intermediate. The low-intermediate and high-intermediate genotypes may possess weak alleles of Ph1. The three different T turgidum ssp. dicoccoides pairing genotypes were collected from different geographical regions in Israel, indicating that this trait may have an adaptive value. The availability of allelic variation at the Ph1 locus may facilitate the mapping, tagging, and eventually the isolation of this important gene.     

Mathematics of chromosome pairing

Analysis of frequencies of chromosome configurations in wheat-related species hybrids is extended to include cases involving homoeologous groups of size six. Further, the expected frequencies of the various configurations emanating from homologous and homoeologous pairing events have been determined for groups consisting of two and three homoeologous pairs of homologous chromosomes. Analysis of configuration frequencies of nullisomic 5B of wheat, in which both homologous and homoeologous pairing occurs, led to estimation of the relative frequencies of homologous and homoeologous pairing events as 17.9:1. In the samples analyzed, slightly less than one homoeologous exchange occurred per gamete.     

不同细胞质的普通小麦"中国春"(Triticum aestivum var.Chinese Spring)与小偃麦杂交F1代的育性与减数分裂行为的研究

15个不同细胞质“中国春”小麦与八倍体小偃麦杂交 ,杂种F1减数分裂的染色体行为表明 :普通小麦与天蓝偃麦草的F或E组染色体之间存在着部分同源关系 ;D2 型细胞质促进部分同源染色体配对、但却抑制同源染色体配对 ;Sv 型细胞质对同源染色体或部分同源染色体的配对均有抑制作用 ;G型细胞质促进同源染色体配对。1 5个不同细胞质“中国春”小麦与六倍体小偃麦杂交 ,F1结实率很低 ,减数分裂中期的染色体行为混乱 ,单价体过多 ,或许意味着在天蓝偃麦草 (Elytrigiain termedium)与长穗偃麦草 (E .elongatum)的E组染色体之间存在着很大差别。随着回交代数的增加 ,选出G型、D2 型、Mt 型、Mu 型等细胞质雄性不育的八倍体小偃麦品系 ,其中D2 型细胞质八倍体小偃麦具有光周期敏感性雄性不育的特征 ;G型细胞质“远中 3”育性正常 ,表明八倍体小偃麦“远中 3”的E组染色体中存在G型胞质的育性恢复基因。     

Effect of a rye B chromosome and its segments on homoeologous pairing in hybrids between common wheat and Aegilops variabilis

Rye B chromosomes, which are supernumerary chromosomes dispensable for the host but increase in number by non-disjunction after meiosis, have been reported to affect meiotic homoeologous pairing in wheat-rye hybrids. The effect of a rye B chromosome (B) and its segments (B-9 and B-10) on homoeologous pairing was studied in hybrids between common wheat (2n=42) and Aegilops variabilis (2n=28), with reference to the Ph1 gene located on wheat chromosome 5B. The B-9 and B-10 chromosomes are derived from reciprocal translocations between a wheat and the B chromosomes, and the former had the B pericentromeric segment and the latter had the B distal segment. Both the B and B-9 chromosomes suppressed homoeologous pairing when chromosome 5B was absent. On the other hand, the B-9 and B-10 chromosomes promoted homoeologous pairing when 5B was present. On pairing suppression, B-9 had a greater effect in one dose than in two doses, and B-9 had a greater effect than B-10 had in one dose. These results suggested that the effect of the B chromosomes on homoeologous pairing was not confined to a specific region and that the intensity of the effect varied depending on the presence or absence of 5B and also on the segment and dose of the B chromosome. The mean chiasma frequency (10.23) in a hybrid (2n=36) possessing 5B and one B-9 was considerably higher than that (2.78) of a hybrid (2n=35) possessing 5B alone, and was comparable with that (14.09) of a hybrid (2n=34) lacking 5B. This fact suggested that the B chromosome or its segment can be used in introducing alien genes into wheat by inducing homoeologous pairing between wheat and alien chromosome.     

Synthesis and cytological characterization of trigeneric hybrids of durum wheat with and without Ph1.

Wild grasses in the tribe Triticeae, some in the primary or secondary gene pool of wheat, are excellent reservoirs of genes for superior agronomic traits, including resistance to various diseases. Thus, the diploid wheatgrasses Thinopyrum bessarabicum (Savul. and Rayss) A. Love (2n = 2x = 14; JJ genome) and Lophopyrum elongatum (Host) A. Love (2n = 2x = 14; EE genome) are important sources of genes for disease resistance, e.g., Fusarium head blight resistance that may be transferred to wheat. By crossing fertile amphidiploids (2n = 4x = 28; JJEE) developed from F1 hybrids of the 2 diploid species with appropriate genetic stocks of durum wheat, we synthesized trigeneric hybrids (2n = 4x = 28; ABJE) incorporating both the J and E genomes of the grass species with the durum genomes A and B. Trigeneric hybrids with and without the homoeologous-pairing suppressor gene, Ph1, were produced. In the absence of Ph1, the chances of genetic recombination between chromosomes of the 2 useful grass genomes (JE) and those of the durum genomes (AB) would be enhanced. Meiotic chromosome pairing was studied using both conventional staining and fluorescent genomic in situ hybridization (fl-GISH). As expected, the Ph1-intergeneric hybrids showed low chromosome pairing (23.86% of the complement), whereas the trigenerics with ph1b (49.49%) and those with their chromosome 5B replaced by 5D (49.09%) showed much higher pairing. The absence of Ph1 allowed pairing and, hence, genetic recombination between homoeologous chromosomes. Fl-GISH analysis afforded an excellent tool for studying the specificity of chromosome pairing: wheat with grass, wheat with wheat, or grass with grass. In the trigeneric hybrids that lacked chromosome 5B, and hence lacked the Ph1 gene, the wheat-grass pairing was elevated, i.e., 2.6 chiasmata per cell, a welcome feature from the breeding standpoint. Using Langdon 5D(5B) disomic substitution for making trigeneric hybrids should promote homoeologous pairing between durum and grass chromosomes and hence accelerate alien gene transfer into the durum genomes.     

小麦-黑麦代换系5A/5R与6A/6R杂交诱导同祖染色体配对与易位的研究

利用两个小麦-黑麦异源双代换系DS 5A/5R与DS 6A/6R杂交,探讨同祖染色体配对的可能性与创制小麦黑麦异源易位系.在方法上对杂种F1的减数分裂行为进行研究,观察5R与5A、6R与6A配对频率,探讨同祖染色体配对规律.实验结果看到杂交F1减数分裂中有22.91%的花粉母细胞有小麦染色体(ABD组)与黑麦染色体(R组)发生同祖配对.在F2及以后世代,通过染色体C分带、原位杂交检测,选择小麦-黑麦易位系.在F2代的45株中检测到9株有易位,易位频率为20%,是目前小麦-黑麦染色体易位频率最高的.染色体易位有的来源于同祖配对的交换,有的来源于单价体错分裂或断裂的重建.     

Unique chromosome behavior and genetic control in Brassica×Orychophragmus wide hybrids: a review

Researchers recognized early that chromosome behavior, as other morphological characters, is under genetic control and gave some cytogenetical examples such as the homoeologous chromosome pairing in wheat. In the intergeneric sexual hybrids between cultivated Brassica species and another crucifer Orychophragmus violaceus, the phenomenon of parental genome separation was found under genetic control during mitosis and meiosis. The cytogenetics of these hybrids was species-specific for Brassica parents. The different chromosome behavior of hybrids with three Brassica diploids (B. rapa, B. nigra and B. oleracea) might contribute to the different cytology of hybrids with three tetraploids (B. napus, B. juncea and B. carinata). The finding that genome-specific retention or loss of chromosomes in hybrids of O. violaceus with B. carinata and synthetic Brassica hexaploids (2n=54, AABBCC) is likely related to nucleolar dominance gives new insight into the molecular mechanisms regarding the cytology in these hybrids. It is proposed that the preferential expressions of genes for centromeric proteins from one parent (such as the well presented centromeric histone H3) are related with chromosome stability in wide hybrids and nucleolar dominance is beneficial to the production of centromere-specific proteins of the rRNAs-donor parent and to the stability of its chromosomes.     

phKL基因诱导小麦远缘杂种部分同源染色体配对的能力界于Ph1和Ph2基因突变体之间

在普通小麦地方品种自然群体中天然存在促进小麦-外源杂种部分同源染色体配对的基因phKL。本研究比较了phKL基因与人工Ph基因突变系诱导小麦-Aegilops variabilis及小麦-黑麦杂种部分同源染色体配对的作用大小。研究结果表明,诱导小麦- Ae. variabilis(或黑麦)部分同源染色体配对作用的顺序是ph1b > phKL > ph2b > ph2a,即phKL基因的作用介于Ph1与Ph2突变体之间。     

小麦VE161雄性不育异代换系的染色体分析

VE161是一个具有一对长穗偃麦草(Agropyron elongatum)染色体的小麦雄性不育异代换系。由于不能用做父本,采用一般单体分析法确定其被代换的染色体比较困难。因此用一套中国春缺-四体和重双端体对其进行了分析,并以F_1花粉母细胞MI单价体的N-显带技术进行了验证。结果表明,VE161代换系所代换的染色体为7B,该方法对确定雄性不育代换系所代换的染色体是一个简单易行的方法。染色体行为观察说明,该代换系的一对长穗偃麦草染色体可能与小麦的染色体7B没有部分同源关系,除了引起雄性不育以外,有时与小麦染色体分裂不同步而表现落后,并有抑制Ph基因的作用,能强烈促进小麦部分同源染色体的联会配对,致使在中期Ⅰ出现很高频率的各种多价体。     

Determination of the frequency of wheat-rye chromosome pairing in wheat x rye hybrids with and without chromosome 5B

Genomic in-situ hybridization (GISH) was used to determine the amount of wheat-rye chromosome pairing in wheat (Triticum aestivum) x rye (Secale cereale) hybrids having chromosome 5B present, absent, or replaced by an extra dose of chromosome 5D. The levels of overall chromosome pairing were similar to those reported earlier but the levels of wheat-rye pairing were higher than earlier determinations using C-banding. Significant differences in chromosome pairing were found between the three genotypes studied. Both of the chromosome-5B-deficient hybrid genotypes showed much higher pairing than the euploid wheat hybrid. However, the 5B-deficient hybrid carrying an extra chromosome 5D had significantly less wheat-rye pairing than the simple 5B-deficient genotype, indicating the presence of a suppressing factor on chromosome 5D. Non-homologous/non-homoeologous chromosome pairing was observed in all three hybrid genotypes. The value of GISH for assessing the level of wheat-alien chromosome pairing in wheat/alien hybrids and the effectiveness of wheat genotypes that affect homoeologous chromosome pairing is demonstrated.     

Metaphase I bound arms frequency and genome analysis in wheat-Aegilops hybrids

Summary Meiotic associations of different wheat-Aegilops variabilis and wheat-Ae. kotschyi hybrid combinations with low and high homoeologous pairing were analyzed at metaphase I. Five types of pairing involving wheat and Aegilops genomes were identified by using C-banding. A genotype that seems to promote homoeologous pairing has been found in Ae. variabilis var. cylindrostachys. Its effect is detectable in the low pairing hybrids but not in the high ones. Pairing affinity has been analyzed on the basis of metaphase I associations in the low and high homoeologous pairing hybrids, and in bivalents and multivalents in the high pairing hybrids. The results indicate that the amount of bound arms of each type of identifiable association relative to the total associations formed (relative contribution) was not maintained, either between the different levels of pairing (low and high) or between different meiotic configurations (bivalents and multivalents). These findings seem to indicate that quantifications of genomic relationships based on the amount of chromosome pairing at metaphase I must be carefully done in this type of hybrid combinations.     

Cytogenetics of the trigeneric hybrid, (Hordeum vulgare ×Triticum aestivum) ×Secale cereale

Summary Chromosome pairing was studied in hybrids of (Hordeum vulgare ×Triticum aestivum) ×Secale cereale. Chiasma frequency per cell varied from 1.94 to 3.16 between the different hybrids. This variation was attributed to genetic variability in rye parents which affected homoeologous pairing. The pairing of rye chromosomes as revealed by Giemsa C-banding was a combination of nonhomologous association between rye chromosomes and associations with chromosomes of wheat and barley. Contribution No. 634 Ottawa Research Station