A novel intergeneric hybrid in the Triticeae: Triticum aestivum x Psathyrost achys juncea

Summary Two hybrid embryos of intergeneric origin between Triticum aestivum cv Fukuho (2n=6x=42, AABBDD) and Psathyrostachys juncea (2n=2x=14, NN) were successfully rescued. One hybrid plant had the expected chromosome number of 28 (ABDN), whereas the second plant had 35 chromosomes. The average meiotic chromosome pairing in the 35-chromosome hybrid was 21.87 univalents + 6.38 bivalents + 0.11 trivalents + 0.009 quadrivalents, which indicates that two copies of the N genome were present. Chromosome pairing in the 28-chromosome hybrid was low (1.35 bivalents), and pointed out the lack of homology between the wheat genomes and the P. juncea genome. These new hybrids showed some necrosis and chlorosis, which caused severe floral abortion in the plant that had 35 chromosomes. These problems became gradually less severe after 18 months.Contrib. no. 372     

Male and female meiosis in diploid and colchitetraploid Phlox drummondii Hook. (Polemoniaceae)

Comparative studies on male and female meiosis in diploid (2n = 2x = 14) and colchitetraploid (2N = 4x = 28) ornamental Phlox drummondii reveal higher chiasma frequency in embryo-sac mother cells as compared with pollen mother cells in the diploid, and significant differences in the pairing properties of chromosomes in the pollen mother cells and embryo-sac mother cells of the colchitetraploid. Male meiosis in the colchicine-induced autotetraploid was abnormal with 51.70% of chromosomes associated in quadrivalents and tridents in 96% of the cells. This was followed by discordant anaphase I disjunction. In the female meiocytes the chromosomes formed 14 bivalents in 86.66% of the cells. Quadrivalents (1–3) appeared only in 13.34% of the cells. It is concluded that meiosis in male and female cells of the colchitetraploid is governed and regulated by different controlling systems of gene(s).     

Meiotic studies of wheat,rye and 42-chromoaome-Triticales

In 42-chromosome — triticales, chromosome pairing was extremely erratic with very low frequency (60–63 %) of regular bivalents per pollen mother cell. The frequency of univalents was very low or nil in the wheat and rye parents.     

Cytological investigations of the interspecific hybrids of Nicotiana tabacum L. x N. glauca Grah

Interspecific amphihaploid and amphidiploid hybrids between Nicotiana glauca Grah. (2n = 24) and N. tabacum L. (2n = 48) cultivars BY 103 and K 326 were analysed. F1 amphihaploids (2n = 36) were viable and completely self- and cross-sterile, and mostly univalents were present during meiosis (with pairing range from 0 to 5). In some meiocytes, meiotic irregularities were observed, such as sporadic chromatin bridges and formation of restitution nuclei. The resultant F1 hybrids were easily converted to amphidiploids (2n = 72) via colchicine treatment of seedlings. The number of univalents and the frequency of PMCs containing unpaired chromosomes indicated that amphidiploids N. tabacum cv. BY 103 or K 326 x N. glauca represented quite a high pairing category. However, they were male sterile because pollen mother cells were arrested at the tetrad stage. The termination of development of PMCs, and consequently male sterility, are very rare in this kind of tobacco hybrids.     

The fate of recombinant chromosomes and genome interaction in Nicotiana asymmetric somatic hybrids and their sexual progeny

Genomic in-situ hybridization (GISH) was used to monitor the behaviour of parental genomes, and the fate of intergenomic chromosome translocations, through meiosis of plants regenerated from asymmetric somatic hybrids between Nicotiana sylvestris and N. plumbaginifolia. Meiotic pairing in the regenerants was exclusively between chromosomes or chromosome segments derived from the same species. Translocation (recombinant) chromosomes contained chromosome segments from both parental species, and were detected at all stages of meiosis. They occasionally paired with respectively homologous segments of N. sylvestris or N. plumbaginifolia chromosomes. Within hybrid nuclei, the meiotic division of N. plumbaginifolia lagged behind that of N. sylvestris. However, normal and recombinant chromosomes were eventually incorporated into dyads and tetrads, and the regenerants were partially pollen fertile. Recombinant chromosomes were transmitted through either male or female gametes, and were detected by GISH in sexual progeny obtained on selfing or backcrossing the regenerants to N. sylvestris. A new recombinant chromosome in one plant of the first backcross generation provided evidence of further chromosome rearrangements occurring at, or following, meiosis in the original regenerants. This study demonstrates the stable incorporation of chromosome segments from one parental genome of an asymmetric somatic hybrid into another, via intergenomic translocation, and reveals their transmission to subsequent sexual progeny.     

Studies on male and female meiosis in Indian Allium

Male meiosis in autotetraploid Allium tuberosum (4×=32) is fairly regular, keeping in view its cytological status, with 81 percent of the chromosomes associated in quadrivalents and trivalents. About 5% of the cells have 32 univalents. Anaphase segregation is slightly irregular. While 48% of the pollen mitoses show 16 chromosomes, 87% of the mature pollen is viable as indicated by carmine or iodine staining. — Megaspore mother cells have 64 chromosomes associated in 32 bivalents at metaphase I. Anaphase segregation is normal. In three out of 56 cells studied multivalents, bivalents and univalents are observed as in male meiosis. — It is concluded that the species reproduces by pseudogamous parthenogenesis made possible by meiotic modification. This modification is almost perfect and almost completely specific for female meiosis. Slight effects are observed in male meiosis.     

S RNase and Interspecific Pollen Rejection in the Genus Nicotiana: Multiple Pollen-Rejection Pathways Contribute to Unilateral Incompatibility between Self-Incompatible and Self-Compatible Species

In self-incompatible (SI) plants, the S locus acts to prevent growth of self-pollen and thus promotes outcrossing within the species. Interspecific crosses between SI and self-compatible (SC) species often show unilateral incompatibility that follows the SI x SC rule: SI species reject pollen from SC species, but the reciprocal crosses are usually compatible. The general validity of the SI x SC rule suggests a link between SI and interspecific pollen rejection; however, this link has been questioned because of a number of exceptions to the rule. To clarify the role of the S locus in interspecific pollen rejection, we transformed several Nicotiana species and hybrids with genes encoding SA2 or SC10 RNase from SI N. alata. Compatibility phenotypes in the transgenic plants were tested using pollen from three SC species showing unilateral incompatibility with N. alata. S RNase was implicated in rejecting pollen from all three species. Rejection of N. plumbaginifolia pollen was similar to S allele-specific pollen rejection, showing a requirement for both S RNase and other genetic factors from N. alata. In contrast, S RNase-dependent rejection of N. glutinosa and N. tabacum pollen proceeded without these additional factors. N. alata also rejects pollen from the latter two species through an S RNase-independent mechanism. Our results implicate the S locus in all three systems, but it is clear that multiple mechanisms contribute to interspecific pollen rejection.     

羽衣甘蓝花粉母细胞减数分裂及雄配子体发育观察

采用常规压片法对羽衣甘蓝花粉母细胞减数分裂及雄配子体发育进行了细胞学观察,结果显示:羽衣甘蓝减数分裂类似甘蓝种,细胞质分裂为同时型,四分体以正四面体型或十字交叉型为主;终变期有9个二价体,此时可进行染色体计数;中期Ⅰ和Ⅱ少数细胞中可见赤道板外染色体,后期Ⅰ和Ⅱ存在落后染色体,四分体时期可观察到少量含微核的异常四分体;单核靠边期时花蕾长度约为2.0~2.2 mm,小孢子经过发育最终成为3-细胞型花粉并具3个萌发孔,成熟花粉中败育花粉比率为1.3%.     

Cytogenetics of a new type of barley with 16 chromosomes

In the progeny of a trisomic type for chromosome 6, Purple, a 16-chromosome type was obtained, which had a pair of new metacentric chromosome 6 in excess. The new metacentric chromosome 6 was shorter than any of the 14 chromosomes of normal barley complement and showed a heteropycnotic nature at late prophase in somatic mitosis. At metaphase I in the plants with 14+one metacentric chromosome 6 (2n=15) the chromosome configuration was exclusively 7_II+1_I indicating that the extra metacentric chromosome 6 could not associate with the normal chromosome 6. At diakinesis and metaphase I in the new 16-chromosome plants most of the sporocytes showed 8_IIor 7_II+2_I. Neither tetravalents nor trivalents were observed at meiosis. The chromosome behaviour at anaphase I and later stages of meiosis was regular in general, resulted in a fairly high pollen fertility of about 61 per cent. Seed fertility however, was very low. The transmission rate of the new metacentric chromosome 6 through the pollen was extremely low in 16-chromosome plants. Possible origin of new basic number and B-chromosome in diploid level through trisomic condition was suggested (Summary see p. 138).Contribution No. 141 of the Department of Plant Science, University of Manitoba.     

鹅观草属——人工合成双二倍体(2n=10x=70)的细胞学及育性的研究

本文对缘毛鹅观草(Roegneria pendulina)、鹅观草(R.tsukushiensis var.transiens)及其人工合成杂种F_1、双二倍体进行了细胞学,育性等的分析和研究。结果表明双亲的减数分裂,花粉育性和结实性均正常,杂种F_1的减数分裂不规则且完全不育;当代双二倍体的染色体数目为70,其减数分裂构型为:6.04 Ⅰ+26.21 Ⅱ+1.52 Ⅲ+1.59 Ⅳ+0.02 Ⅴ:第二代双二倍体的染色体数目为70,个别植株为69,减数分裂构型分别为:4.16 Ⅰ+27.33 Ⅱ+0.50 Ⅲ+2.16 Ⅳ和4.79 Ⅰ26.26 Ⅱ+1.13 Ⅲ+2.13 Ⅳ。与期望染色体配对模式相比,双二倍体中二价体出现的频率有明显增大的趋势。在减数分裂AⅠ和AⅡ分别观察到数目不定的落后单价体,大部份的四分体中出现了微核。双二倍体的育性得到了很大程度的恢复,其花粉可染色性及结实率分别为54.4％和64.0％。     

黄芩的花粉母细胞减数分裂及核型分析

采用压片法,对黄芩花粉母细胞减数分裂及核型进行了研究。结果表明:黄芩的大多数花粉母细胞减数分裂中染色体的行为正常,在终变期同源染色体配对后可形成9个二价体,后期Ⅰ染色体以9∶9的方式向细胞两极分离,其减数分裂为同时型;在少数花粉母细胞减数分裂中观察到落后染色体、染色体桥等异常行为;其花粉粒育性为76.49%。黄芩的染色体数目为2n=2X=18,核型公式为K(2n)=2X=18=16m+2 sm,染色体相对长度组成为2n=1 s+4M1+3M2+1L,其核型为"1A"型。     

Pollination in Nicotiana alata stimulates synthesis and transfer to the stigmatic surface of NaStEP, a vacuolar Kunitz proteinase inhibitor homologue

After landing on a wet stigma, pollen grains hydrate and germination generally occurs. However, there is no certainty of the pollen tube growth through the style to reach the ovary. The pistil is a gatekeeper that evolved in many species to recognize and reject the self-pollen, avoiding endogamy and encouraging cross-pollination. However, recognition is a complex process, and specific factors are needed. Here the isolation and characterization of a stigma-specific protein from N. alata, NaStEP (N. alata Stigma Expressed Protein), that is homologous to Kunitz-type proteinase inhibitors, are reported. Activity gel assays showed that NaStEP is not a functional serine proteinase inhibitor. Immunohistochemical and protein blot analyses revealed that NaStEP is detectable in stigmas of self-incompatible (SI) species N. alata, N. forgetiana, and N. bonariensis, but not in self-compatible (SC) species N. tabacum, N. plumbaginifolia, N. benthamiana, N. longiflora, and N. glauca. NaStEP contains the vacuolar targeting sequence NPIVL, and immunocytochemistry experiments showed vacuolar localization in unpollinated stigmas. After self-pollination or pollination with pollen from the SC species N. tabacum or N. plumbaginifolia, NaStEP was also found in the stigmatic exudate. The synthesis and presence in the stigmatic exudate of this protein was strongly induced in N. alata following incompatible pollination with N. tabacum pollen. The transfer of NaStEP to the stigmatic exudate was accompanied by perforation of the stigmatic cell wall, which appeared to release the vacuolar contents to the apoplastic space. The increase in NaStEP synthesis after pollination and its presence in the stigmatic exudates suggest that this protein may play a role in the early pollen-stigma interactions that regulate pollen tube growth in Nicotiana.     

In vitro propagation and chromosome doubling of a Triticum crassum x Hordeum vulgare intergeneric hybrid

Summary In vitro culture of inflorescence tissue of a Triticum crassum (6x) x Hordeum vulgare cv. Bomi (2x) intergeneric hybrid resulted in the proliferation of totipotent callus from which plants were regenerated. Regeneration was also achieved from immature inflorescence callus of T. crassum but not from H. vulgare. T. crassum x H. vulgare regenerates had a somatic chromosome number of 28, identical to that of the original hybrid. Four chimeric plants with a partially doubled chromosome number were obtained by in vitro colchicine treatment of hybrid callus prior to induction of plant regeneration. All T. crassum regenerates had 35 chromosomes rather than the expected number of 42. Meiotic analysis of a 35-chromosome plant revealed an extremely abnormal meiosis which might be attributable to a complete disturbance in meiotic control system(s) including that of meiotic pairing.Dedicated to Professor Georg Melchers on the occasion of his 75th birthday.Contribution No. 624 Ottawa Research Station, Research Branch, Agriculture Canada, Ottawa, Ont. K1A OC6 (Canada)     

麦蓝菜的花粉母细胞减数分裂及核型分析

采用压片法,对麦蓝菜的花粉母细胞减数分裂及体细胞核型进行了研究。结果表明,麦蓝菜的绝大多数花粉母细胞减数分裂中染色体的行为正常,在终变期同源染色体配对后可形成15个二价体;在少数花粉母细胞减数分裂中观察到落后染色体、染色体桥等异常行为;其减数分裂为同时型,其花粉粒育性为88.04%。麦蓝菜的染色体数目为2n=2X=30,核型公式为K(2n)=2X=30=22m(2SAT)+8 sm,染色体相对长度组成为2n=16M2+14M1,其核型为"1A"型。     

Syncytes during male meiosis resulting in 2n pollen grain formation in Lindelofia longiflora var. falconeri

Abstract Lindelofia longiflora (Royle ex Benth.) Baill. var. falconeri (Cl.) Brand (Family: Boraginaceae) is investigated cytologically (n= 12) for the first time from the cold deserts of Pangi Valley, Chamba District (Himachal Pradesh) in India. We report the formation of syncytes and 2n pollen grains in the species. During meiosis, the majority of the pollen mother cells (PMCs) exhibited 12 bivalents, equal segregation of chromosomes during anaphases, regular tetrads, and normal‐sized pollen grain formation. Occasionally, two proximate PMCs fused during the early stages of prophase‐I and resulted in the formation of syncytes. The frequency of syncytes in the accession is rather low, at 25 out of 1866 (1.33%). Such syncyte PMCs are detectable during meiosis due to their larger size compared to typical PMCs. The syncytes or polyploid cells showed normal 24 bivalents and depicted perfectly regular meiotic course. But the products of such PMCs yield 2n or larger sized pollen grains that are almost double the size of typical normal or n pollen grains. The origin of syncytes as a consequence of the fusion of meiocytes during the early stages of meiosis‐I could be attributed to low temperature stress conditions prevailing in the Pangi Valley, where temperature during May and June dip to below freezing, the time the plants enters the reproductive/flowering bud stage. It is possible that such apparently fertile 2n pollen grains originating from syncytes might play a role in the origin of intraspecific polyploids in the species.     

Syncytes during male meiosis resulting in 2n pollen grain formation in Lindelofia longiflora var.falconeri

Lindelofia longiflora (Royle ex Benth.) Baill.var.falconeri (Cl.) Brand (Family:Boraginaceae) is investigated cytologically (n =12) for the first time from the cold deserts of Pangi Valley,Chamba District (Himachal Pradesh) in India.We report the formation of syncytes and 2n pollen grains in the species.During meiosis,the majority of the pollen mother cells (PMCs) exhibited 12 bivalents,equal segregation of chromosomes during anaphases,regular tetrads,and normal-sized pollen grain formation.Occasionally,two proximate PMCs fused during the early stages ofprophase-I and resulted in the formation of syncytes.The frequency of syncytes in the accession is rather low,at 25 out of 1866 (1.33％).Such syncyte PMCs are detectable during meiosis due to their larger size compared to typical PMCs.The syncytes or polyploid cells showed normal 24 bivalents and depicted perfectly regular meiotic course.But the products of such PMCs yield 2n or larger sized pollen grains that are almost double the size of typical normal or n pollen grains.The origin of syncytes as a consequence of the fusion of meiocytes during the early stages of meiosis-I could be attributed to low temperature stress conditions prevailing in the Pangi Valley,where temperature during May and June dip to below freezing,the time the plants enters the reproductive/flowering bud stage.It is possible that such apparently fertile 2n pollen grains originating from syncytes might play a role in the origin of intraspecific polyploids in the species.     

Evolutionary implications of permanent odd polyploidy in the stable sexual, pentaploid of Rosa canina L

In Rosa canina (2n = 5x = 35), the pollen and ovular parents contribute, respectively, seven and 28 chromosomes to the zygote. At meiosis I, 14 chromosomes form seven bivalents and 21 chromosomes remain as univalents. Fluorescent in situ hybridization to mitotic and pollen mother cells (PMC) of R. canina showed that 10 chromosomes (two per genome) carry ribosomal DNA (rDNA) loci. Five chromosomes carry terminal 18S-5.8S-26S rDNA loci; three of these also carry paracentric 5S rDNA loci and were designated as marker chromosomes 1. Five chromosomes carry only 5S rDNA loci and three of these were designated as marker chromosomes 2. The remaining four of the 10 chromosomes with rDNA loci were individually identifiable by the type and relative sizes of their rDNA loci and were numbered separately. At PMC meiosis, two marker chromosomes 1 and two marker chromosomes 2 formed bivalents, whereas the others were unpaired. In a gynogenetic haploid of R. canina (n = 4x = 28), obtained after pollination with gamma-irradiated pollen, chromosomes at meiosis I in PMC remained predominantly unpaired. The data indicate only one pair of truly homologous genomes in R. canina. The 21 unpaired chromosomes probably remain as univalents through multiple generations and do not recombine. The long-term evolutionary consequence for the univalents is likely to be genetic degradation through accumulated mutational change as in the mammalian Y chromosome and chromosomes of asexual species. But there is no indication that univalents carry degenerate 5S rDNA families. This may point to a recent evolution of the R. canina meiotic system.     

Low glucosinolate Brassica juncea breeding line revealed to be nullisomic.

The low glucosinolate Brassica juncea breeding line 1058 was derived from a BC1F3 plant of an interspecific cross between high glucosinolate Indian B. juncea (genome AABB, 2n = 36) line 60143 and B. rapa (genome AA, 2n = 20) canola strain CZY. Line 60143 had 2n = 36 chromosomes (18 bivalents at metaphase I) and strain CZY had 2n = 20 chromosomes (10 bivalents). Line 1058 was nullisomic, with 2n - 2 = 34 chromosomes, with 17 bivalents formed at metaphase I and an even chromosomal segregation of 17:17 at anaphase I. In F1 hybrid plants of the cross 1058 x CZY, 98.3% of the pollen mother cells had 10 bivalents and seven univalents. This is evidence that plants of line 1058 are nullisomic, missing one pair of B-genome chromosomes.     

Studies on Chromosome Behaviour of F1 and Fertility Restoration in Hybrid of Gossypium hirtutum ×G. bickii

By means of dripping GAs(50 ppm) and NAA(40 ppm) on the hybrid boll-embryo culture in vitro, one F1 plant of G. hirsuturn × G. bickii was obtained in 1982, the Fx plant flowered profusely but failed to produce any seeds when selfed or back crossed. In meiosis of the pollen mother cell (PMC), mean chromosome coniugation was 33.24 Ⅰ +2.67 Ⅱ +0.095 Ⅲ + 0.048 Ⅳ with high frequence (77.6%) of one chiasma attenuated bivalents. Mean number of chiasmta per bivalents was 1.23. The univalents were scattered over the achromatic figure for most PMCs. Although a few of the bivalents were located in' the equatorial region, but they did not form a definite plate. At the second anaphase the distribution of chromosomes was very irregular. In the majority of cases, multipolar distripution of chromosomes was observed. At the completion of meiosis highly abnormal sporads occurred, which contained from 2 ro 13 spores of various sizes. Thus, all of the pollen grains produced were sterile. When the F1 branches were grafted onto the upland cotton and thence they were backcrossed under short day (12 hours) and cold night(15–18℃) exposure, BC1 seeds could be harvested. BC1 and BC2 plants could grow up later. In the BC, generation, the fertility of the hybrid was restored. By 1988, ten pure lines of hybrid with the characers of both male parent (viz.red petal with purple spot and strong fibre) and female parent (white fibre, high yield, earliness ect.) were selected for the first time.     

Synaptonemal complex formation in hybrids of Lolium temulentum x Lolium perenne (L.)

Comparisons were made between two kinds of tetraploids derived from the hybrid Lolium temulentum x L. perenne. One hybrid behaves like an autotetraploid with multivalents at first metaphase of meiosis in pollen mother cells. The other behaves like an allotetraploid, in which pairing at first metaphase is restricted to bivalents comprised of strictly homologous chromosomes. The diploidisation of the latter form is controlled by determinants located on both the normal, A chromosomes and on supernumary B chromosomes. Reconstruction of synaptonemal complexes and their elements, from serial sections through pollen mother cell nuclei examined under the electron microscope, reveals that at zygotene pairing in both forms results in multivalent formation involving non-homologous as well as homologous chromosomes. The mechanism responsible for the diploidisation is, therefore, not based on a restriction of pairing at early meiosis to homologous chromosomes but on a correction or transformation of the multivalent chromosome associations to bivalents subsequent to zygotene. The transformation is not completed until late pachytene. In the multivalent-forming tetraploid a maximum of four chromosomes are associated at first metaphase. Yet configurations of a higher valency are found at zygotene. There is, therefore, a partial transformation of multivalents even in this autotetraploid form which restricts configurations at metaphase I to homologous and homoeologous chromosomes only. In both hybrids some homologous bivalents are not the product of resolution of multivalents but result from two-by-two pairing from the beginning of zygotene.