Attaining inter-subgeneric hybrids in fragrant azalea breeding and the inheritance of organelle DNA

Inter-subgeneric hybrids were successfully obtained in reciprocal cross combinations between evergreen azaleas (Rhododendron nakaharae and its hybrids) and fragrant deciduous azaleas (R. arborescens and R. viscosum) for the purpose of fragrant evergreen azalea breeding. Nuclear and organelle DNA of these hybrids was investigated using PCR-RFLP markers. Viable hybrid seedlings have nuclear ribosomal DNA (nrDNA) inherited biparentally, mitochondrial DNA (mtDNA) from the seed parent, and chloroplast DNA (cpDNA) from the deciduous azalea, regardless of cross combination. These results suggest that the chloroplast genome from deciduous azaleas and the nuclear genome from evergreen azaleas are compatible in viable hybrid progenies.     

Chimeric inheritance of organelle DNA in variegated leaf seedlings from inter-subgeneric crossing of azalea

The inheritance of organelle DNA was investigated using PCR–RFLP markers in reciprocal cross combinations of inter-subgeneric azalea hybrids between evergreen azaleas (Rhododendron nakaharai and its hybrids) and fragrant deciduous azaleas (R. arborescens and R. viscosum) for the purpose of breeding fragrant evergreen azaleas. The hybrid progenies included green leaf, pale green leaf, variegated leaf and albino seedlings. Most viable green leaf seedlings had inherited ptDNA from the deciduous parent and non-viable albino and pale green leaf seedlings had inherited ptDNA from the evergreen parent. On the other hand, variegated leaf seedlings had chimerically inherited ptDNA from both parents. Their green leaf segments had ptDNA from the deciduous parent, and the pale green and white segments had biparental or maternal ptDNA depending on the progeny. In this study, we obtained interesting inter-subgeneric azalea hybrid progenies that had chimerically inherited organelle DNA and had different colored leaf segments corresponding to the composition of ptDNA from each parent. These results suggest that variegated leaf progenies with chimeric ptDNA from both parents can be subsistent, whereas albino seedlings resulting from plastome–genome incompatibility between the plastid genome from evergreen azalea and the nuclear genome from deciduous azalea are non-viable.     

Inheritance of albinism in certain interspecific and intersubspecific crosses in groundnut (Arachis hypogaea L.)

Summary A study of the several interspecific and inter-subspecific crosses indicated that albinism in groundnut is recessive and in these crosses is controlled by triplicate recessive loci.ICRISAT Paper No. JA-336     

The influence of cytokinins and ionic strength of Anderson's medium on shoot establishment and proliferation of evergreen azalea

Shoot-tip explants of evergreen azalea cv. Fuchsia grown on Anderson's medium and containing different cytokinins produced the highest proliferation rate on a medium containing thidiazuron (TDZ). TDZ concentrations ranging from 0.23 to 2.3 M resulted in both good bud-break rate (4 to 5) and shoot quality (> 0.5 cm in length). Adding 2.3 M zeatin to Anderson's medium containing 0.23 or 2.3 M TDZ increased the number of axillary shoots/explant. However, increasing the zeatin concentration to 4.6 M resulted in a reduced shoot proliferation rate. A medium containing 1.15 M TDZ and 2.3 M zeatin resulted in an 18-fold increase for 'Fuchsia' and a 9-fold increase for 'Hino Crimson' after 6 weeks of culture. It was found that explants grown on a half-strength Anderson's medium with 87.6 mM sucrose generally had better shoot proliferation rate and shoot quality than at higher ionic strength.     

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.     

New interspecific hybrids in the genus Agropyron

Summary Fifteen species from the genus Agropyron were crossed together. Fourteen of the crosses did not produce fruits. Twenty crosses produced varying numbers of caryopses. Of the fruits produced, 22% contained no embryos. For a variety of reasons it was possible to obtain only 2 plants from the remaining 140 embryos. The crosses which yielded viable plants were between A. trachycaulum cv. Primar (2n=28) and A. intermedium cv. Chief (2n=42) as well as between A. trachycaulum from Lethbridge (2n=28) and A. desertorum cv. Nordan (2n=28). The somatic chromosome numbers for the hybrids are 37 and 28. respectively. Studies of vegetative plant character are presented.     

Development of molecular markers linked to the Leptosphaeria maculans resistance gene Rlm6 and inheritance of SCAR and CAPS markers in Brassica napus × Brassica juncea interspecific hybrids

Leptosphaeria maculans causes blackleg disease on Brassica napus, an economically important oilseed crop. Brassica juncea has high resistance to blackleg and is a source for the development of resistant B. napus varieties. To transfer the Rlm6 resistance gene from B. juncea into B. napus, an interspecific cross between B. napus “Topas DH16516” and B. juncea “Forge” was produced, followed by the development of F₂ and F₃ generations. Sequence characterized amplified region (SCAR) and cleaved amplified polymorphic sequence (CAPS) markers linked to the L. maculans resistance gene Rlm6 were developed. Segregation of SCAR and CAPS markers linked to Rlm6 were confirmed by genotyping of F₂ and F₃ progeny. Segregation of CAPS markers and phenotypes for blackleg disease severity in F₂ plants had a Mendelian ratio of 3:1 in resistant vs. susceptible plants, respectively, supporting the assumption that genetic control of resistance was by a single dominant gene. The molecular markers developed in this study, which show linkage with the L. maculans resistance gene Rlm6, would facilitate marker‐assisted backcross breeding in a variety development programme.     

The inheritance of resistance to powdery mildew in interspecific hybrids and induced amphiploids of Zinnia elegans Jacq. and Z. angustifolia HBK

Summary Sterile interspecific hybrids and colchicine-induced amphiploids of Zinnia elegans Jacq. and Z. angustifolia HBK were examined to determine the mode of inheritance of resistance to Erysiphe cichoracearum DC ex Merat. Fertility was restored through colchicine treatment of two sterile hybrids of species reciprocal parentage which differed in ray petal response to the pathogen. Derived amphiploids were subsequently intercrossed to overcome the lack of segregation for this trait due to genetic control of pairing upon chromosome doubling. Resistance to E. cichoracearum appears to be complexly inherited in both leaves and ray florets of sterile hybrids and induced amphiploids. Two major dominant genes have been implicated in conferring resistance in ray petal tissue of derived amphiploids. Data obtained from the F₁ hybrid progeny of the intercrossed amphiploids indicate that this trait is not cytoplasmically inherited. It is speculated that the genes conferring resistance in the ray florets are acting independently from those controlling leaf resistance and that most, if not all, of the resistance genes are inherited from Z. angustifolia.Scientific Article No. A-3825, Contribution No. 6804 of the Maryland Agricultural Experiment Station.     

Reproductive behaviour and broomrape resistance in interspecific hybrids of sunflower

Interspecific hybrids and backcross generations between the wild perennial species Helianthus resinosus, Helianthus paucifiorus, Helianthus laevigatus, Helianthus nuttallii ssp. nuttallii T. & G. and Helianthus giganteus, resistant to broomrape (Orobanche cernua) and susceptible inbred lines were obtained to study crossability to cultivated sunflower and the transmission and expression of resistance to this parasitic weed. Conventional crosses with all the species tested were successful except for the crosses with diploid H. giganteus, for which embryo rescue techniques were needed to overcome hybrid incompatibility. Pollen viability and seed set were highest for F₁ hybrids with hexaploid species and lowest for those with the diploid H. giganteus. We evaluated F₁, BC₁F₁, some BC₂F₁ plants and the wild and cultivated parents. The wild species and interspecific hybrids were resistant to broomrape infection except for H. nuttallii, which showed segregation, indicating that the resistance is dominant. The crossability and resistance of F₁, and back-cross generations of species with different ploidy levels indicate that the transfer of broomrape resistance to cultivated sunflower is feasible.     

水稻短根白化突变体sra1生理生化分析及基因定位

叶色突变体是研究光合作用及叶绿素合成与降解途径的理想材料,有助于了解高等植物叶绿体发育和光合作用的调控机制。利用甲基磺酸乙酯(EMS)处理西农1B,获得一个短根白化突变体sra1 (short radicle and albino 1),从出芽至第三叶期叶片始终为白色,胚根较同时期野生型明显变短。对相同位置的叶片观察发现,西农1B叶肉细胞叶绿体含量丰富且膜系统发育完整,而sra1叶肉细胞液泡化严重,叶绿体数目明显减少或没有,基粒类囊体垛叠松散且稀少。生理生化分析发现sra1叶绿素a、叶绿素b和类胡萝卜素含量接近于零,净光合速率为负值。遗传分析表明该短根白化表型由单个隐性核基因控制,最终将SRA1定位于水稻第3染色体长臂InDel标记Z-20和Z-42间,物理距离约657 kb,是一个未报道的新基因。     

Genetic diversity in advanced derivatives of Brassica interspecific hybrids

Genetic diversity among the 88 entries including eighty F₄ derivatives i.e., 20 each selected from Brassica crosses viz., B. juncea × B. napus, B. juncea × B. rapa var. toria, B. juncea ×B. rapa var. yellowsarson and B. tournefortii × B. juncea, and eight parent genotypes was assessed through multivariate analysis (D² statistic). Significant differences among the family groupsas well as within the family were recorded for all the 14 characters studied. The D² analysis revealed enormous diversity among the interspecific cross derivatives. The genetic distances calculated among different Brassica species revealed that B. tournefortii had maximumdiversity with B. juncea followed by B. napus, B.rapa var. toria and B. rapa var. yellow sarson.Amongst interspecific crosses, maximum diversity was noticed indescendants of cross B. tournefortii × B. juncea followed byB. juncea × B. napus, B. juncea × B.rapa var. toria and the least in the cross B. juncea ×B. rapa var. yellow sarson. These results indicated that the derivatives selected from cross of diverse parents revealed greater diversity. The clustering pattern showed that many derivatives of the cross fell into the same cluster but in many cases in spite of common ancestry many descendants of the cross spread over different clusters. The characters, namely, plant height, secondary branches per plant, days to flowering and1000-seed weight were contributed maximum towards genetic divergence. This revised version was published online in August 2006 with corrections to the Cover Date.     

Chromosome association and pollen fertility in some interspecific hybrids of Lilium

Summary Meiotic observations in PMCs were made in eight kinds of interspecific hybrids in Lilium. Three hybrids, 6134-S × L. cernuum. L. regale × L. leichtlinii maximowiczii and L. auratum platyphyllum × L. henryi showed respectively the mean chromosome association frequencies of 1.4_II, 0.8_II and 1.5_II per cell at MI which were the lowest values hitherto reported in Lilium hybrids. In L. longiflorum × L. henryi, 33.3° of the cells had 1 or 2 multivalents of three to five chromosomes in addition to uni- and bivalents. 12° bivalents and 12 univalents were invariably observed in the triploid hybrid L. longiflorum × L. cernuum. The hybrids between the species belonging to the different sections of the genus generally showed high pollen sterility, with some exceptions. A more or less remote genomic homology was found between the different sections in Lilium.     

Maternal inheritance of cytoplasmic organelles in intergeneric hybrids of Carica papaya L. and Vasconcellea spp. (Caricaceae Dumort., Brassicales)

The mode of inheritance of chloroplast and mitochondrial DNA has been determined in intergeneric hybrids between C. papaya and four different Vasconcellea species by employing a restriction fragment length polymorphism analysis of a PCR-amplified chloroplast and mitochondrial DNA region. Artificial F₁ hybrids were produced between a female specimen of C. papaya and male specimens of either V. parviflora, V. goudotiana, V. cundinamarcensis or V. quercifolia. The hybridization patterns of all hybrids correspond in all cases with that of the C. papaya mother, and are different from that of the paternal Vasconcellea species, thus indicating the maternal inheritance of cpDNA and mtDNA in intergeneric hybrids between C. papaya and wild relatives of the Vasconcellea genus.     

Genetic studies in four interspecific hybrids of the genus oryza

The paper reports the results of genetic studies made in four interspecific hybrids of the genus Oryza, and the data are employed to discuss the inter-relationships between the different species. The characters of wild parents were found to be dominant in the F₁ hybrids.In O. sativa (variety Fukoku) × O. perennis subsp. balunga, the inheritance of the eight characters studied was found to be of the same nature as that of complementary, modifying, duplicating and polygenic types involving two or three genes. In O. perennis (Cuban type) × O. perennis subsp. balunga, the interesting feature was the occurrence of recessive characters (typical of cultivated rices) in the F₂ population, even though the two parents carried the dominant wild characters. This should be possible only by the operation of duplicate genes and such a mode of inheritance was observed for three characters. No definite Mendelian ratios could be got for the remaining five characters studied. The appearance of characters of cultivated rices has been suggested to point to the ability of O. perennis to give rise to O. sativa through natural crossing and selection. In O. glaberrima × O. perennis subsp. balunga, the ten characters studied segregated in monogenic or digenic fashion. When digenic, these characters exhibited complementary or inhibitory factor interactions. In O. glaberrima × O. breviligulata, the simple mode of inheritance in the F₂ generation has been suggested to indicate the close relationship of the two species.Some of the general features of the hybrids and segregating populations studied were the vigorous and fertile F₁s followed by the appearance of weak and unthrifty plants in the F₂ generation and the tendency for the parental characters to inherit together. The appearance of plants looking similar to O. sativa var. fatua in the F₂ populations of O. sativa × O. perennis, and of O. stapfii in the F₂s of O. glaberrima × O. perennis has been supposed to provide further evidence on the hybrid origin of these two forms.     

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.     

The mechanism and inheritance of intraflower self-pollination in self-pollinating variant strains of periwinkle

The mechanism and inheritance of intraflower self‐pollination in self‐pollinating variant strains of periwinkle was studied using three self‐pollinating and two non‐self‐pollinating but fully self‐fertile strains. Intraflower self‐pollination in self‐pollinating strains occurred due to the continued growth of the gynoecium beyond the base of the anthers, even after anthesis, resulting in intraflower self‐pollination. In non‐self‐pollinating strains, the stigma remained below the base of the anthers and did not come into contact with the pollen up to flower drop. Intraflower self‐pollination was found to be governed by duplicate recessive genes. The utility of the intraflower self‐pollination trait in the breeding of periwinkle cultivars is discussed.     

Development of multiplex PCR markers to distinguish evergreen and deciduous azaleas

We successfully developed new PCR‐based markers for analysing the DNA inheritance patterns of vigorous inter‐subgeneric hybrids of azalea. The hybrids were between evergreen azaleas (Rhododendron nakaharae and its hybrids) and fragrant deciduous azaleas (Rhododendron arborescens and Rhododendron viscosum). We determined the sequence of the nuclear internal transcribed spacer (ITS) region in evergreen and deciduous azaleas, and found a deletion of five base pairs (bp) in evergreen azaleas compared with deciduous azaleas. Sequences for the chloroplast matK‐trnK region were obtained from GenBank. Primers for the multiplex PCRs were designed using these sequence data. In the multiplex PCR of the ITS region, evergreen azaleas showed a specific band of 140 bp and deciduous azaleas showed a specific band of 800 bp. In the multiplex PCR of the matK‐trnK region, evergreen azaleas showed a specific band of 560 bp and deciduous azaleas showed a specific band of 300 bp. These results indicate that multiplex PCR methods are useful to distinguish evergreen and deciduous azaleas easily and rapidly.     

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.     

Fertilisation assessment and postzygotic development in several intra- and interspecific Prunus hybrids

Summary the clearing technique was applied to assess fertilisation and to follow the postzygotic development in ten intra- and interspecific crosses among Prunus species used as rootstocks. The technique allowed to distinguish fertilised ovules from unfertilised ones, as well as to calculate the fertilisation percentage in intra-and interspecific compatible crosses. All the stages from unfertilised to fertilised ovules were analysed in order to determine the best time for embryo rescue. Differences in postzygotic development among the crosses are discussed in terms of the genotype of the seed parent in the pollen-pistil interactions.Abbreviations DAP Days After Pollination