Cytogenetic studies on the interspecific hybrid Capsicum baccatum L.× C. frutescens L. and its progeny

Summary Morphological and cytogenetical studies were carried out on the F₁ and F₂ hybrids and backcross derivatives of the interspecific cross Capsicum baccatum L. x C. frutescens L. The F₁ and F₂ hybrids displayed irregular meiosis with a maximum association of eight chromosomes in the former and one quadrivalent in the latter with the appropriate number of bivalents and univalents. It is inferred that C. baccatum differs from C. frutescens (yellow) by at least tow or three interchanges and from the white cultivar by a single interchange. Structural repatterning of chromosomes, erratic meiotic behaviour, genes for pollen sterility, segregational imbalances following intergenomic recombination are believed to be major factors causing sterility in the hybrids. The two species are sympatric and natural hybrids have not been realised.     

Pollination systems, hybridization barriers and meiotic chromosome behaviour in Nemesia hybrids

The ability of 13 Nemesia species (six annual and seven perennial) to sexually hybridize was investigated. Six of the perennial Nemesia species investigated were inter-fertile with one another. Two of the annual species, N. macroceras and N. strumosa, were inter-fertile. Thirty three crosses were successful and resulted in viable seeds. The analysis of meiotic chromosome behaviour in interspecific hybrids indicated that Nemesia chromosomes in different parental species were homeologous. No evidence of chromosome inversions or chromosome translocations was observed during meiosis in interspecific hybrids between the six perennial Nemesia species. In the hybrids produced between N. macroceras and N. strumosa, a quadrivalent was observed during meiotic metaphase I, indicating that these two species differ by a reciprocal translocation. A successful hybridization was made between N. anisocarpa (annual) and N. foetans (perennial), producing two triploid hybrids. In the unsuccessful crosses, pollen tubes were observed entering ovaries and ovules, suggesting that post-fertilization barriers were preventing sexual hybridization. Many of these crosses produced nonviable, shrunken, empty seeds, suggesting that endosperm breakdown and embryo abortion prevent interspecific hybridization in unsuccessful crosses. The manipulation of ploidy levels in N. fruticans and N. strumosa and tissue culture of N. strumosa × N. fruticans ovules failed to overcome post-fertilization barriers between these species.     

Cytogenetics of interspecific hybrids in the genus Capsicum L.

Summary Crosses between a wild species C. chacoense and three cultivated species of chili pepper viz. C. annuum, C. frutescens and C. chinense yielded hybrids when C. chacoense was the seed parent but the reciprocal crosses were unsuccessful. C. chacoense × C. annuum F₁ hybrids were partly fertile and therefore an F₂ population could be raised; the other two F₁ hybrids were totally sterile. Chromosome pairing in the F₁ plants resulted largely in bivalents and a few multivalents and univalents. The genomes of the four species share large homologies and the role of chromosome structural changes in genome differentiation is suggested. Hybrid sterility is the major reproductive isolation mechanism.     

Micropropagation of intra and interspecific Lens hybrids

An intraspecific (Tetir × ILL 323) and an interspecific (Alpo × L. odemensis) lentil hybrid were multiplied in vitro in three consecutive micropropagation cycles to increase the production of F₂ seeds. Cloning efficiencies were slightly higher for Tetir × ILL323 (83%) compared to Alpo × L. odemensis (67%). A total of 982 F₂ seeds were produced in the experiment with Alpo × L. odemensis, consisting of the 334 F₂ seeds of the original hybrid and 648 F₂ seeds produced by the 12 plants cloned; consequently, F₂ seed production was increased three-fold over the original hybrid (194%). A total of 6050 F₂ seeds were produced in the experiment with Tetir × ILL 323, made up of the 483 F₂ seeds of the original hybrid and 5567 F₂ seeds produced by the 15 plants cloned; therefore, F₂ seed production was increased by more than twelve-fold over the original hybrid (1153%). In both hybrids the F₂ seed production of cloned plants diminished in the three consecutive multiplication cycles, with the plants having experienced less vegetative development. In lentil, F₂ seed production of cloned plants is related for the first time to the length of their period of vegetative development. In conclusion, micropropagation of hybrids is an interesting tool to construct from a single individual large F₁ populations that enable to increase by a manifold the production of F₂ seeds useful for genetic studies and breeding.     

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.     

Novel interspecific hybridization between sweetpotato (<Emphasis Type="Italic">Ipomoea batatas</Emphasis> (L.) Lam.) and its two diploid wild relatives

Interspecific hybridization is an important approach to broaden the genetic base and create novel plant forms in breeding programs. However, interspecific hybridization in Ipomoea is very difficult due to the cross incompatibility. Here we report two novel interspecific F₁ hybrids between I. batatas (L.) Lam. (2n = 6x = 90) and two wild species, I. grandifolia (2n = 2x = 30) and I. purpurea (2n = 2x = 30). Hybridization was stimulated by applying plant growth hormones. Morphological, molecular and cytological tests were conducted to confirm their hybridity. We found that the two hybrids were quite distinctive in leaf color and morphology, and yielded intermediate sizes of storage roots compared to their respective parents. Inter-simple sequence repeat analysis showed that the unique DNA bands from the wild parents could be detected in these two hybrids. The cluster analysis also showed that the two F₁ hybrids were closer to I. batatas in phylogeny relationship. The number of chromosomes of the two hybrids was both 60, indicating that the hybrids were tetraploid. The meiotic configuration analysis of the H₁ of I. batatas × I. grandifolia revealed the occurrence of 17.58I + 14.28II + 1.36III + 2.48IV at metaphase I in average chromosome association per pollen mother cells (PMCs), 4.26I + 18.32II + 2.56III + 3.12IV was average meiotic configuration in the H₂ of I. batatas × I. purpurea. Both hybrids appeared to be polyads and multi-microcytes at tetrad phase and differed in their pollen fertility.     

Fertility restoration of hybrids between Solanum melongena L. and S. aethiopicum L. Gilo Group by chromosome doubling and cytoplasmic effect on pollen fertility

Reproductive fertility traits were studied in the reciprocal hybrids of the eggplant(Solanum melongena L.) and S. aethiopicum L. Gilo Group, and in synthetic amphidiploids to discover whether fertility in these reciprocal hybrids was restored by chromosome doubling. Isozyme and RAPD analyses confirmed hybridity of the hybrids and amphidiploids. Analyses of chloroplast and mitochondrial DNAs confirmed that the cytoplasm of each of the hybrids and amphidiploids was from the maternal parent. Pollen sterility of S. melongena × S. aethiopicum Gilo Group [F₁ (Mel × Aet)] was restored by chromosome doubling, while the reciprocal hybrid S. aethiopicum Gilo Group ×S. melongena [F₁ (Aet × Mel)]and its amphidiploid did not produce any pollen grains; their microspores degenerated without being released from tetrads. Hence the cytoplasm of S. aethiopicum Gilo Group seems to beresponsible for their pollen-non-formation type sterility of the hybrid. Both the F₁ hybrids did not set any fruits by either selfing or backcrossing, while their amphidiploids set fruits after pollinating with pollen from the amphidiploid of F₁ (Mel × Aet). Seeds obtained from both the amphidiploids germinated normally. Chromosome doubling has been effective in restoring fertility of the hybrids. This revised version was published online in July 2006 with corrections to the Cover Date.     

Formation of unreduced gametes is impeded by homologous chromosome pairing in tetraploid <Emphasis Type="Italic">Triticum turgidum</Emphasis> × <Emphasis Type="Italic">Aegilops tauschii</Emphasis> hybrids

It is believed that unreduced gametes with somatic chromosome numbers play a predominant role in natural polyploidization. Allohexaploid bread wheat originated from spontaneous hybridization of Triticum turgidum L. with Aegilops tauschii Coss. Unreduced gametes originating via meiotic restitution, including first-division restitution (FDR) and single-division meiosis (SDM), are well documented in triploid F₁ hybrids of T. turgidum with diploid Ae. tauschii (genomic constitution ABD, usually with 21 univalents in meiotic metaphase I). In this study, two T. turgidum lines known to carry genes for meiotic restitution were crossed to tetraploid Ae. tauschii. The resulting F₁ hybrids (genomes ABDD), had seven pairs of homologous chromosomes and regularly formed 14 univalents and seven bivalents at metaphase I. Neither FDR nor SDM were observed. The distribution of chromosome numbers among progeny obtained by self pollination and a backcross to T. turgidum showed the absence of unreduced gametes. These results suggest that high homologous pairing interfered with meiotic restitution and the formation of unreduced gametes. This may be related to asynchronous movement during meiosis between paired and unpaired chromosomes or to uneven distribution of chromosomes in anaphases, resulting in nonviable gametes.     

A diallel study of yield components and fruit quality in chilli pepper (<Emphasis Type="Italic">Capsicum baccatum</Emphasis>)

The aim of this work was to estimate the general and specific combining ability of peppers by measuring fruit quality and yield traits. This experiment was carried out on the garden field from Universidade Federal de Viçosa (UFV), Minas Gerais State—Brazil. Eight lines of Capsicum baccatum belonging to the UFV Horticultural Germplasm Bank were chosen based on their broad genetic and phenotypical background variation and then they were crossed in a complete diallel way. The F₁ seeds of the 56 hybrids and eight parents were planted in the field in a randomized complete block design. The data were submitted to ANOVA and the means were grouped by Scott–Knott test (P ≤ 0.01). Significant variation for fruit quality and yield components was observed among parents and F₁ generation. Analysis of variance for the combining ability showed that GCA effects exhibited significant difference and SCA effects of the crosses were significant, except for the height of first bifurcation. For almost all characters both additive and non-additive effects influenced the performance of hybrids.     

Relative Efficiency of Anther Culture and Chromosome Elimination Techniques for Haploid Induction in Triticale × Wheat and Triticale × Triticale Hybrids

Summary The study was undertaken to evaluate the relative efficiency of anther culture and chromosome elimination (by crosses with maize) techniques of haploid induction in intergenotypic triticale and triticale × wheat hybrids. For this, 15 triticale × wheat and 8 triticale × triticale F₁ hybrids were subjected to anther culture and were also simultaneously crossed with the `Madgran Local' genotype of maize (Zea mays L.) to induce haploids through the chromosome elimination technique. The haploid embryo formation frequency through the chromosome elimination technique was significantly higher in both, triticale × wheat (20.4%) and triticale × triticale (17.0%) F₁ genotypes, as compared to the calli induction frequencies through anther culture (1.6 and 1.4%, respectively). Further, four triticale × wheat and three triticale × triticale F₁ genotypes failed to respond to anther culture, whereas, all the F₁ genotypes formed sufficient number of haploid embryos through the chromosome elimination technique with no recovery of albino plantlets. The haploid plantlet regeneration frequencies were also significantly higher through the latter technique in both triticale × wheat (42.7%) and triticale × triticale (49.4%) F₁s as compared to anther culture (8.2 and 4.0%, respectively), where the efficiency was drastically reduced by several constraints like, high genotypic specificity, low regeneration frequency and albinism. The overall success rates of obtaining doubled haploids per 100 pollinated florets/anthers cultured were also significantly higher through the chromosome elimination technique (1.1% in triticale × wheat and 1.5% in triticale × triticale hybrids), proving it to be a highly efficient and economically more viable technique of haploid induction as compared to anther culture, where the success rates were only 0.2% and 0.1%, respectively.     

Plant Regeneration and Chromosomal Stability in Tissue Cultures of the Hybrids of Elymus canadensis with Psathyrostachys juncea and Secale cereale

Callus was induced from segments of immature inflorescences of Elymus canadensis×Psathyrostachys juncea and Elymus canadensis×Secale cereale F₁ hybrids on MS medium supplemented with 2 mg/l of 2,4-D and plants were regenerated from the calluses on a hormone free MS medium. The plants regenerated from both the hybrids exhibited a high degree of stability in morphology, chromosome number and chromosome pairing. The reasons for this are discussed.     

辣椒过氧化物酶同工酶研究及模糊聚类分析

以辣椒属(Capsicum)的7个种(包括4个变种、3个类型)计172个品种为试材,于成株期取功能叶片进行制样分析,得出过氧化物酶(POD)同工酶的基本酶谱为12条酶带,并将172份材料中各个种、类型出现的过氧化物酶同工酶表型频率用模糊聚类分析(fuzzy clusber analysis)方法进行动态聚类.结果表明,C.annuum的两个变种关系极近;C.baccatum的两个变种关系也很近; C.chi-nense与C.frutescens关系较近,这两个种及C.praetermissum与C.annuum关系亦较近,而C.annuum与C.baccatum、eximium C.chacoense关系则较远.从而得出,普通栽培辣椒,首先应该利用其野生种C.annuum var.annuum的有用基因,其次是C.frutescens、C.chinense及C.praetermissum,最后才应当考虑利用C.eximium、C.baccatum及C.chacoense的有用基因.据此对辣椒属的起源、分类及上述种质材料在育种上的应用问题进行了讨论.     

Rapid introduction of disease resistance from rye into common wheat by anther culture of a 6x triticale × nulli-tetrasomic wheat

Powdery mildew (caused by Erysiphe graminis) and yellow rust (caused by Puccinia striiformis) are the two most serious wheat diseases found in China. Rye chromosomes, carrying genes for resistance to these diseases, were introduced into common wheat in two generations using chromosome engineering and anther culture. The F₁ hybrids from a cross involving a hexaploid triticale (×Triticosecale Wittmack) בChinese Spring’ nulli‐tetrasomic N6DT6A wheat aneuploid line were anther cultured and doubled‐haploid plants were regenerated. Using genomic in situ hybridization, C‐banding and biochemical marker analyses, one of the anther‐cultured lines (ZH‐1)studied in detail, proved to be a doubled‐haploid with one rye chromosome pair added (1R) and a homozygous 6R/6D substitution (2n= 44). The line was tested for expression of disease resistance and found to be highly resistant to powdery mildew and moderately resistant to yellow rust.     

A New Intergeneric Hybrid between Triticum aestivum L. and Agropyron fragile (Roth) Candargy: Variation in A. fragile for Suppression of the Wheat Ph-Locus Activity

New intergeneric hybrids were obtained between Triticum aestivum L. cv. Tukuho’ (2n = 6x = 42, AABBDD) and Agropyron fragile (Roth) Candargy PGR 8097 (2n = 4x = 28, PPPP) at a frequency of 1.06 %, through the use of direct embryo culture and in ovulo embryo culture. Such hybrids could be used to transfer barley yellow dwarf virus (BYDV) resistance and winterhardiness into bread wheat. The somatic chromosome number in all the hybrid plants was 2n = 5x = 35, as expected. Considerable variation in chromosome pairing was observed among the different hybrid plants. Average meiotic chromosome configuration at metaphase I was 17.29 Is + 6.57 rod Us + 1.97 ring Us + 0.18 III + 0.03 IV + 0.002 VI. The high level of chromosome pairing in some F₁ hybrids was attributed to Ph-suppressor gene(s) present in A. fragile. The hybrids could not be backcrossed to wheat, but amphiploid seeds have been obtained by colchicine treatment.     

Patterns of meiosis in ABDR amphihaploids depend on the specific type of univalent chromosome division

Meiosis in wide F₁ hybrids in the tribe Triticeae is complex and irregular. Despite extensive research, the meiotic pathways in such hybrids remain largely unknown. In this study, the effect of the genotypes of disomic wheat–rye substitution lines on the regulation of meiosis in ABDR amphihaploids was analysed by comparing microsporogenesis and seed set under self pollination in hybrids of normal bread wheat (Triticum aestivum L.) and wheats with single chromosome substitutions 1R(1A), 1Rv(1A), 2R(2D)₁, 2R(2D)₂, 2R(2D)₃, 5R(5D), 5R(5A), 6R(6A) with rye (Secale cereale L.). Three types of meiotic pathways were identified in the hybrids: reductional, equational, and equational+reductional. During the reductional type division, chromosomes randomly moved toward the poles at Anaphase I (AI) and separated sister chromatids at AII. Meiosis ended with tetrad formation, and the resulting plants were sterile ((2R(2D)₁ × R, 2R(2D)₂ × R). In the equational type division, the chromosomes aligned along the equator at Metaphase I and at AI separated into chromatids that moved toward the poles. This meiotic pathway led to dyad formation and partial fertility (especially in hybrids of 1Rv(1A) and 6R(6A)). In the equational+reductional pathway, some chromosomes divided in an equational manner while others were segregated reductionally within the same microsporocyte and this led to formation of tetrads with groups of uncoiled chromosomes. Comparison of the frequencies of each of the three meiotic pathways in different hybrid combinations suggests differences in contributions of individual chromosomes to the control of the entire meiotic process.     

Detection of wheat-barley translocations by genomic in situ hybridization in derivatives of hybrids multiplied in vitro

Wheat-barley translocations were identified by genomicin situ hybridization (GISH) in backcross progenies originating from in vitro regenerated wheat (Triticum aestivum L. cv. Chinese Spring) × barley (Hordeum vulgare L. cv. Betzes) hybrids. The regenerated hybrids were pollinated with the wheat line Martonvásári 9 kr1. Five translocated wheat-barley chromosomes were recovered among 51 BC₂F₂ progeny from the in vitro regenerated wheat × barley hybrids. All were single breakpoint translocations with the relative positions of the breakpoints ranging from the centromere to about 0.8 of the relative arm length. Of the four translocations with intercalary breakpoints, three were transfers of terminal barley segments to wheat chromosomes; one was a transfer of a terminal wheat segment to a barley chromosome. Because of the absence of diagnostic N-bands, the identity of three barley segments could not be determined; in one translocation the barley chromosome involved had a NOR so it must have been 5H or 6H, and the centric translocation was 4HS.2BL. Following selfing, homozygotes of four translocations were selected. The experiment suggests that in vitro culture conditions are conducive for major genome rearrangements in wheat-barley hybrids. This revised version was published online in July 2006 with corrections to the Cover Date.     

Attempts to overcome postfertilization barrier in interspecific crosses of the genus Lens

Wild relatives are a potential source of genetic diversity to lentil (Lens culinaris Medik). The objective of this research was to obtain viable interspecific hybrids between the domesticated lentil and its wild relatives. The paper details the results of a number of interspecific crosses among L. culinaris, L. orientalis, L. odemensis, L. ervoides and L. nigricans. Viable hybrids were produced between L. culinaris × L. orientalis, L. culinaris × L. nigricans, L. culinaris × L. ervoides and between L. culinaris × L. odemensis. Further viable hybrids were obtained between L. culinaris and L. ervoides, which have the potential to be a ‘bridge’ in hybridization to L. culinaris for specific L. nicrigans lines which proved recalcitrant in L. culinaris × L. nigricans crosses. This is the first time that four wild species of lentils have been used successfully in hybridization with cultivated lentils, and viable hybrids produced. This paper also suggests that the artificial supplement of GA₃, hormone is needed after fertilization for the normal growth of the hybrid embryo, possibly as the natural GA₃ production is restricted with alien pollinations in cultivated lentils in both F₁ and backcross hybrids.     

Production of intergeneric hybrids between Brassica juncea and Diplotaxis virgata through ovary culture, and the cytology and crossability of their progenies

The cytogenetic study was investigated in the intergeneric F₁ hybrid, F₂and backcross progenies (BC₁). The plants used were Brassica juncea(2n=36) and Diplotaxis virgata(2n=18). Three intergeneric F₁ hybrids between two species were produced through ovary culture. They showed 36 chromosomes. It might consist one genome of B. juncea and two genomes of D. virgata. The morphology of the leaves resembled that of B. juncea. The color of the petals was yellow that was like in D. virgata. The size of the petal was similar to that of B. juncea. The mean pollen fertility was15.3% and the chromosome associations in the first meiotic division were(0–1)_IV+(0–2)_III+(8–12)_II+(12–20)_I. Many F₂ and BC₁seeds were harvested after open pollination and backcross of the F₁ hybrids withB. juncea, respectively. The F₂seedlings showed different chromosome constitutions and the range was from 28 to54 chromosomes. Most seedlings had 38chromosomes followed by 36, 40 and 54. The BC₁ seedlings also showed different chromosome constitutions and the range was from 29 to 62. Most seedlings had both 40and 54 chromosomes followed by 36, 46 and52. In the first meiotic division of F₂ and BC₁ plants, a high frequency of bivalent associations was observed in all the various kinds of somatic chromosomes. Many F₃ and BC₂ seeds were obtained by self-pollination and open pollination of both F₂ and BC₁ plants, and by backcrossing both F₂ and BC₁plants with B. juncea, respectively,especially, three type progeny with 36, 40or 54 chromosomes. The somatic chromosomes of the F₃ and BC₂ plants were further investigated. The bridge plants between B. juncea and D. virgata with 36 chromosomes may be utilized for breeding of other Brassica crops as well as B. juncea. This revised version was published online in July 2006 with corrections to the Cover Date.     

甘蓝型油菜与播娘蒿原生质体融合杂种后代的遗传研究

甘蓝型油菜(2n=38)与播娘蒿(2n=28)原生质体融合杂种F₁连续自交3代,获得F₂、F₃和F₄后代。用细胞学和SSR分子标记方法,分析杂种后代的染色体数目变异、减数分裂行为以及播娘蒿遗传成分的保留情况。结果表明在F₂、F₃和F₄代中,根尖细胞染色体平均数分别为38.47±3.17、37.65±3.23和36.66±2.95,随着自交世代增加呈减少趋势;在杂种后代减数分裂中,观察到染色体桥、染色体落后、染色体周期不同步、不均等分离等现象;杂种后代F₂、F₃和F₄代中检测到播娘蒿特征条带的平均频率分别为9.62%、2.99%和0.31%,呈减少趋势。因此要实现播娘蒿种质向油菜渗入应该重视F₂世代的选择。     

Culture of embryos and shoot tips for chromosome doubling in Lolium perenne and sterile hybrids between Lolium and Festuca

An improved method is reported for polyploid induction in Lolium (ryegrass), and in sterile F₁ hybrids between Lolium and Festuca (fescue). Two factors greatly increased the survival rate of colchicine‐treated embryos of Lolium perenne (perennial ryegrass) germinated and cultured in vitro (1) a high concentration of sucrose (100 g/1) in the germination medium and (1), maintenance at a low temperature of 10°C for 2 weeks after treatment. The maximum success rate for chromosome doubling among survivors of perennial ryegrass was 79.1%, and for L. perenne×Festuca arundinacea F₁ hybrid embryos it exceeded 40%. The same doubling treatment also works with shoot tip culture in non‐ flowering genotypes obtained by anther culture of L. multiflorum×F. arundinacea hybrids.